KR20210065950A - Methods and compositions for the treatment of senescence-associated disorders using CCR3-inhibitors - Google Patents

Abstract

Methods for ameliorating neurodegenerative diseases with CCR3 modulators are provided. The method comprises administering to a subject a therapeutically effective amount of a CCR3 modulator and involves improvement of cognitive, motor, or other neurodegenerative-affecting function. Cognitive and motor diseases in which the method of the present invention can improve cognition include Alzheimer's disease, Parkinson's disease, frontotemporal dementia, Huntington's disease, amyotrophic lateral sclerosis, multiple sclerosis, glaucoma, muscular dystrophy, vascular dementia, progressive supranuclear palsy.

Description

Methods and compositions for the treatment of senescence-associated disorders using CCR3-inhibitors

I. CROSS-REFERENCE TO RELATED APPLICATIONS

35 U.S.C. Pursuant to § 119(e), this application claims priority to the following filing dates: U.S. Provisional Patent Application Serial No. 62/737,017, filed September 26, 2018; The publications of these applications are incorporated herein by reference.

II. intro

The following is provided for background information only and is not a prior art of the present invention.

Aging is an important risk factor for several human diseases, including cognitive impairment, cancer, arthritis, vision loss, osteoporosis, diabetes, cardiovascular disease and stroke. In addition to normal synaptic loss during natural aging, synaptic loss is an early pathological event common to many neurodegenerative diseases and is most associated with the neurological and cognitive impairments associated with these symptoms. Thus, aging remains the single most dominant risk factor for dementia-related neurodegenerative diseases such as Alzheimer's disease (AD) (Bishop, NA et al., Neural mechanisms of aging and cognitive decline. Nature 464(7288), 529) -535 (2010);Heeden, T. et al., Insights into the aging mind: a view from cognitive neuroscience. Nat. Rev. Neurosci. 5(2), 87-96 (2004); Mattson, MP, et al. ., Aging and neuronal vulnerability. Nat. Rev. Neurosci. 7(4), 278-294(2006)). Aging affects all tissues and functions of the body, including the central nervous system, and the decline in functions such as cognitive and motor activity can severely affect quality of life.

Treatments for cognitive and motor decline associated with neurodegeneration have achieved only limited success in preventing and reversing impairment. Therefore, it is important to identify novel therapies to maintain cognitive integrity by protecting, counteracting, or reversing the effects of aging. Unfortunately, drug treatment for cognitive and motor disorders faces major hurdles. For example, it is thought that treatments such as small molecules must cross the blood-brain barrier (BBB) to be effective in treating cognitive decline and motor activity. Transportation through the BBB is not without exception, 98% of all small molecules do not cross it (Pardridge, William M, The Blood-Brain Barrier: Bottleneck in Brain Drug Development , NeuroRx: The J. of the Am. Society for Experimental NeuroTherapeutics, 2:3-14, 2005). Consequently, treatments for neurodegenerative diseases such as Alzheimer's disease, Huntington's disease, Parkinson's disease and amyotrophic lateral sclerosis (ALS) are limited. Indeed, Alzheimer's disease exhibits one of the highest failure rates in clinical development of any disease domain (99.6% failure in the last 20 years). ( Id.; and Burke, M., Renewed focus on dementia checked by drug challenges , ChemistryWorld, Jul. 3, 2014). BBB-penetrating drugs approved for the treatment of cognitive diseases such as Alzheimer's, such as donepezil, have temporary effects and are not effective for all patients (Banks, WA, Drug Delivery to the Brain in Alzheimer's Disease: Consideration of the Blood). -brain Barrier , Adv. Drug. Deliv. Rev. 64(7):629-39(2012); and Burke, M. supra ). Similarly, levodopa, which crosses the BBB and treats Parkinson's disease symptoms, such as exercise deficiency, eventually loses its efficacy. A further obstacle to treatment across the BBB is that making the compound more lipophilic (which tends to increase BBB permeability more) often increases clearance from the blood, reducing bioavailability. Thus, the increase in lipophilicity offsets the plasma area under the curve (AUC) to minimize brain uptake (Pardridge, William M, supra ).

III. summary

The present invention overcomes these shortcomings. For example, compound 1, a compound of the present invention, exhibits resistance to traversing the BBB at significant concentrations, but unexpectedly results in amelioration of symptoms associated with age-related neurodegeneration and age-related reduced movement. Thus, the present invention can act in a peripheral manner, i.e. without taking into account the requirement of a cognitively acting agent effective to act directly on the central nervous system. Furthermore, while certain embodiments of the present invention are capable of traversing the BBB at significant concentrations, their ability to antagonize the CCL11/CCR3 pathway provides an alternative mechanism of action to current therapies for cognitive and motor deficits.

The compounds of the present invention act as antagonists of the cc motif chemokine receptor 3 (CCR3), a receptor for eotaxin-1. Eotaxin-1 (CCL11) is a protein that has been shown to increase plasma levels with aging and cause a decline in cognitive function. (Villeda et al., The aging systemic milieu negatively regulates neurogenesis and cognitive function , Nature, 477 (7362) ):90-94 (2011)). Eotaxin/CC11 acts on the G-protein coupled receptor CCR3, mainly expressed on peripheral eosinophils and central nervous system neurons and glia. (Xia, M, et al. , Immunohistochemical Study of the *?*-Chemokine Receptors CCR3 and CCR5 and Their Ligands in the Normal and Alzheimer's Disease Brains , Am. J. Pathol. 153(1);31-37(1998)). An increase in CCR3 levels was observed in the CNS in Alzheimer's disease (Id.). The cognitive effect of CCR3 antagonism in Alzheimer's disease and related disorders is expected to be mediated by a centrally located eotaxin-1/CCR3 interaction. However, compound 1 (a compound of the present invention) unexpectedly was unable to cross the BBB at significant concentrations, but enhanced cognitive function and stimulated neurogenesis.

age-related disorders, neurodegenerative diseases, including but not limited to Alzheimer's disease, Parkinson's disease, Lewy body dementia, frontotemporal dementia, Huntington's disease, amyotrophic lateral sclerosis, multiple sclerosis, glaucoma, muscular dystrophy, vascular dementia, progressive supranuclear palsy, and Methods of treating a patient for associated cognitive and motor decline are provided. The method of treatment comprises modulation of CCR3, a major receptor of CCL11/eotaxin-1, through administration of a therapeutically effective amount of a CCR3 antagonist of the present invention. The method comprises administering to a subject or patient an effective therapeutic dose of a CCR3 antagonist and monitoring for specific clinical endpoints. Also included are methods of treating neurodegenerative diseases and associated cognitive and motor declines with agents that act peripherally, ie, do not cross the blood-brain barrier to significant concentrations but exhibit amelioration of the disease, such as improved cognition or motor activity.

Methods of using a diagnostic or companion diagnostic test in connection with the described age-related disorders are also provided. By way of example, and not limitation, such diagnosis or companion diagnosis includes a device capable of determining or detecting the presence of a leukocyte subset in a subject. The diagnostic or companion diagnostic device may also determine or detect a relative or absolute concentration, a relative or absolute number of eosinophils, from the subject's blood or tissue. These diagnostic or companion diagnostic devices may be used together.

IV. inserting a reference

All publications, patents, and patent applications mentioned in this specification are herein incorporated by reference in their entirety to the same extent as if each individual publication or patent application was specifically and individually indicated to be incorporated by reference.

V. BRIEF DESCRIPTION OF DRAWINGS
1A shows the sum of doublecortin (DCX) positive cells in the hippocampus, 2 and 18 month old C57B1/6 mice treated with IgG (n = 18 for both groups); 18-month-old mice treated with Compound 1 for 2 weeks at either high dose (n = 16) or low dose (n = 31); and 18 month old mice treated with compound 1 (“Cmpd 1”) for 4 weeks at a low dose (n = 16). Data shown are mean ± sem, *P<0.05, ***P<0.001.
1B shows the sum of 5-bromo-2'-deoxyuridine (BrdU) positive cells in the hippocampus, 2 and 18 month old C57B1/6 mice treated with IgG (n = 19 for both groups). ) ; 18-month-old mice treated with Compound 1 for 2 weeks at either high dose (n = 16) or low dose (n = 24). Data shown are mean ± sem where ***P < 0.001
2A depicts the effect of Compound 1 on time spent by young and old C57B1/6 mice in new (N) versus familiar/old (“F” or “O”) cancers during the Y-maze test. Two-month-old mice were treated with IgG for two weeks (n = 18). 18 month old mice were treated with IgG for 2 weeks (n = 18); high dose of compound 1 (n = 15) for 2 weeks; low dose of compound 1 (n = 31) for 2 weeks; or low dose of compound 1 (n = 16) for 4 weeks. Data shown are mean ± sem, where **P <0.05, ***P <0.01.
2B depicts the effect of compound 1 on the total number of visits in young and old C57B1/6 mice in new (N) versus familiar (F) cancers during the Y-maze trial. Two-month-old mice were treated with IgG for two weeks (n = 18). 18 month old mice were treated with IgG for 2 weeks (n = 18); high dose of compound 1 (n = 15) for 2 weeks; low dose of compound 1 (n = 31) for 2 weeks; or low dose of compound 1 (n = 16) for 4 weeks. Data shown are mean ± sem, *P <0.05, **P <0.01, ***P <0.001
3 depicts the effect of compound 1 on entry ("number of visits") of young and old C57B1/6 mice in new (N) versus familiar (F) cancers in the Y-maze test. The number of visits for each cancer was plotted for each treatment group and a paired T-test was performed. 3-month-old mice (n = 15) were subcutaneously injected with vehicle (veh) by Aljet mini-pump (0.5 μL/hr) for 4 weeks with one replacement. 18.5 month old mice were subcutaneously injected with vehicle (veh, n = 16) or 50 mg/mL Compound 1 (n = 16) by Aljet mini pump (0.5 μL/hr) for 4 weeks with one replacement. . Data shown are mean ± sem.; *P <0.05.
Figure 4A depicts the effect of compound 1 on the mean time taken in C57B1/6 mice to find a target hole in the Barnes maze test. The mean time for each treatment group was plotted. Three-month-old mice (n = 18) were administered 4 subcutaneous injections of vehicle (veh) for 4 weeks by Aljet mini pump (0.5 μL/hr) with one replacement. 18.5 month old mice were injected subcutaneously with vehicle (veh, n = 15) or 50 mg/mL Compound 1 (n = 15) by Aljet mini pump (0.5 μL/hr) for 4 weeks with one replacement. . Data shown are mean ± sem
Figure 4B depicts the effect of compound 1 on the difference in latency for finding target holes between the last test and the first test on the last day of the Barnes maze test in C57B1/6 mice. Differences were plotted for each treatment group and the data were subjected to an unpaired T-test. Three-month-old mice (n = 18) were administered 4 subcutaneous injections of vehicle (veh) for 4 weeks by Aljet mini pump (0.5 μL/hr) with one replacement. 18.5 month old mice were injected subcutaneously with vehicle (veh, n = 15) or 50 mg/mL Compound 1 (n = 15) by Aljet mini pump (0.5 μL/hr) with one replacement. Data shown are mean ± sem
Figure 5 depicts the effect of compound 1 on neurogenesis in C57B1/6 mice by detecting BrdU positive cells from all sections of the dental gyrus. The number of BrdU positive cells from the dental gyrus was plotted for each treatment group and the data were subjected to an unpaired T-test between the 16.5 month old groups. Three-month-old mice (n = 17) were subcutaneously injected with vehicle (veh) by Aljet mini pump (0.5 μL/hr) for 4 weeks with one replacement. 18.5 month old mice were injected subcutaneously with vehicle (veh, n = 17) or 50 mg/mL Compound 1 (n = 17) by Aljet mini pump (0.5 μL/hr) for 4 weeks with one replacement. Data shown are mean ± sem.; *P <0.05.
Figure 6 depicts the levels of Compound 1 detected in the CSF of C57BL/6 mice in both the young (n = 2) and old (n = 3) groups (the levels were all below 10 nM). Mass spectrometry was used to detect the level of compound 1.
7 depicts the distribution of Compound 1 (measured as area under the curve (AUC)) in C57BL/6JOlaHsd mouse tissues over time. Compounds were tagged with a carbon-14 label and measurements were performed at 1, 24 and 168 hours.
8 depicts the pharmacokinetic profile of Compound 1 after oral (PO) dosing. Plasma of male 2-month-old C57Bl/6 mice dosed at 30 mg/kg or 150 mg/kg by oral gavage was measured for Compound 1 at 20 minutes, 2 hours, 8 hours and 12 hours.
9 depicts the effect of compound 1 on search preference during open field testing in young C57B1/6 mice. Search preferences were expressed as the ratio of time spent in the periphery to the center, and the data were applied to one-way ANOVA. 2-month-old mice were treated with vehicle pobid (n = 11); compound 1 pobid 30 mg/kg (n = 12); vehicle pobid + recombinant mouse CCL11 (eotaxin-1 or "rmE") ip (n = 14); or Compound 1 pobid 30 mg/kg+recombinant mice CCL11 (n=14). Data shown are mean ± sem.; **P < 0.01.
10A depicts the effect of compound 1 on time spent in new (N) versus familiar (F) cancers in the Y-maze test in young C57B1/6 mice. Time spent on new versus familiar cancer was plotted for each treatment group and data were subjected to a paired t-test. 2-month-old mice were treated with vehicle pobid (n = 13); compound 1 pobid 30 mg/kg (n = 14); vehicle pobid and recombinant mouse CCL11 (eotaxin-1) ip (n = 15); or compound 1 pobid 30 mg/kg+recombinant mice CCL11 (n=15). Data shown are mean ± sem.; **P < 0.01.
10B depicts the effect of compound 1 on the ratio of time spent on novel versus familiar arms of the Y-maze test in young C57B1/6 mice. Proportions for each treatment group were plotted and data were subjected to post-analysis ANOVA Kruskal-Wallis tests. Two-month-old mice were treated with vehicle pobid (n = 12); compound 1 pobid 30 mg/kg (n = 14); vehicle pobid and recombinant mouse CCL11 (eotaxin-1) ip (n = 15); or Compound 1 pobid 30 mg/kg+recombinant mouse CCL11 (n=15). Data shown are mean ± sem.; *P <0.05.
10C depicts the effect of compound 1 on the ratio of the number of entries introduced into novel versus familiar cancers in the Y-maze test in young C57B1/6 mice. Proportions for each treatment group were plotted and data were t-tested between vehicle and recombinant mouse CCL11 treatment. 2-month-old mice were treated with vehicle pobid (n = 13); compound 1 pobid 30 mg/kg (n = 14); vehicle pobid and recombinant mouse CCL11 (eotaxin-1) ip (n = 15); or compound 1 pobid 30 mg/kg+recombinant mice CCL11 (n=15). Data shown are mean ± sem.; *P <0.05.
11A depicts the effect of compound 1 on mean delay time to find a target hole during Barnes' maze test in young C57B1/6 mice. The average delay time as a unit of time for each treatment group in each trial was plotted. 2-month-old mice were treated with vehicle pobid (n = 13); compound 1 pobid 30 mg/kg (n = 14); vehicle pobid and recombinant mouse CCL11 (eotaxin-1) ip (n = 15); or compound 1 pobid 30 mg/kg+recombinant mice CCL11 (n=15). Data shown are mean ± sem.
11B depicts the effect of compound 1 on the delay time to find a target hole during Barnes' maze test in young C57B1/6 mice. The latencies over the last 3 trials for each treatment group were averaged and the data were subjected to non-matched t-tests. 2-month-old mice were treated with vehicle pobid (n = 13); compound 1 pobid 30 mg/kg (n = 14); vehicle pobid and recombinant mouse CCL11 (eotaxin-1) ip (n = 15); or compound 1 pobid 30 mg/kg+recombinant mice CCL11 (n=15). Data shown are mean ± sem.; *P <0.05.
12A depicts the effect of compound 1 on memory on new cancers in the Y maze, by the number of entries made of new arms for total entries. 24 month old mice were treated with Compound 1 pobid30 mg/kg or vehicle. Data shown are mean ± sem.; **P < 0.01.
12B depicts the effect of compound 1 on total distance traveled in the Y maze as a measure of locomotor activity. 24 month old mice were treated with Compound 1 bid30 mg/kg or vehicle. Data shown are mean ± sem.; *P <0.05.
13A depicts the effect of compound 1 on entry and exit of new and familiar cancers (“number of visits”) by C57B1/6 mice in the Y-maze test in 24-month-old mice. The number of visits for each arm was plotted for each treatment group and subjected to a corresponding t-test. 23 month old mice were administered BID (twice daily) subcutaneously with vehicle control or Compound 1 for 21 days. After 3 weeks, mice were subjected to a Y-maze test. All mice received BrdU injections at 50 mg/kg IP for 5 consecutive days immediately before the start of treatment. Data shown are mean ± sem in independent sample t-test; *P<0.05, **P<0.01, ***P<0.001 new versus familiar cancer.
13B depicts the effect of compound 1 on the ratio of the number of entries introduced into novel versus familiar cancers in the Y-maze test in 24-month-old mice. Ratios for each treatment group were plotted and data were t-tested between vehicle and Compound 1 treatments. 23 month old mice were administered BID (twice daily) subcutaneously with vehicle control or Compound 1 for 21 days. After 3 weeks, mice were subjected to a Y-maze test. All mice received BrdU injections at 50 mg/kg IP for 5 consecutive days immediately before the start of treatment. Data shown are mean ± sem compared to control by independent sample t-test; *P<0.05, **P<0.01, ***P<0.001.
13C depicts the effect of compound 1 on time spent on novel versus familiar cancers in the Y-maze test in 24-month-old C57B1/6 mice. Time spent on new versus familiar cancers was plotted for each treatment group and data were subjected to a corresponding t-test. 23 month old mice were administered BID (twice daily) subcutaneously with vehicle control or Compound 1 for 21 days. After 3 weeks, mice were subjected to a Y-maze test. All mice received BrdU injections at 50 mg/kg IP for 5 consecutive days immediately before the start of treatment. Data shown are mean ± sem. by paired independent t-test; *P<0.05, **P<0.01, ***P<0.001 new versus familiar cancer.
13D depicts the effect of Compound 1 on the ratio of time spent on novel to familiar cancer (“duration”) of the Y-maze test in 24-month-old C57B1/6 mice. Proportions for each treatment group were plotted and data were subjected to a t-test. 23 month old mice were administered BID (twice daily) subcutaneously with vehicle control or Compound 1 for 21 days. After 3 weeks, mice were subjected to a Y-maze test. All mice received BrdU injections at 50 mg/kg IP for 5 consecutive days immediately before the start of treatment. Data shown are mean ± sem compared to control by independent sample t-test; *P<0.05, **P<0.01, ***P<0.001.
13E depicts the effect of compound 1 on mean velocity during the Y-maze test in 24-month-old C57B1/6 mice. Mean rates for each treatment group were plotted and data were t-tested. 23 month old mice were administered BID (twice daily) subcutaneously with vehicle control or Compound 1 for 21 days. After 3 weeks, mice were subjected to a Y-maze test. All mice received BrdU injections at 50 mg/kg IP for 5 consecutive days immediately before the start of treatment. Data shown are mean ± sem compared to control by independent sample t-test; *P<0.05, **P<0.01, ***P<0.001.
14A depicts the effect of Compound 1 on the mean time taken by 24-month-old C57B1/6 mice to find target holes in the Barnes' maze test. The mean time for each treatment group was plotted. 23 month old mice were administered BID (twice daily) subcutaneously with vehicle control or Compound 1 for 21 days. After 3 weeks, mice were subjected to the Barnes maze test. Data shown are mean ± sem compared to control by independent sample t-test; *P <0.05.
14B depicts the effect of Compound 1 on rate in 24-month-old C57B1/6 mice in the Barnes Maze test, averaged across all trials for each treatment group. 23 month old mice were administered BID (twice daily) subcutaneously with vehicle control or Compound 1 for 21 days. After 3 weeks, mice were subjected to the Barnes maze test. Data shown are mean ± sem compared to control by independent sample t-test; *P <0.05.
15A depicts the effect of compound 1 on distance traveled in an open field test in 24-month-old C57B1/6 mice. The average distance traveled was plotted for each treatment group. 23 month old mice were administered BID (twice daily) subcutaneously with vehicle control or Compound 1 for 21 days. After 3 weeks, mice were subjected to open field testing. Data shown are mean ± sem.
15B depicts the effect of compound 1 on rate in an open field test in 24-month-old C57B1/6 mice. The average speed of mice was plotted for each treatment group. 23 month old mice were administered BID (twice daily) subcutaneously with vehicle control or Compound 1 for 21 days. After 3 weeks, mice were subjected to open field testing. Data shown are mean ± sem.
16A depicts the effect of Compound 1 on TNF alpha cytokine levels in plasma of 24-month-old C57B1/6 mice. 23 month old mice were administered BID (twice daily) subcutaneously with vehicle control or Compound 1 for 21 days. The level of inflammatory cytokine propensity is lower in mice treated with Compound 1 than in vehicle treated mice.
16B depicts the effect of Compound 1 on IL6 cytokine levels in plasma of 24-month-old C57B1/6 mice. 23 month old mice were administered BID (twice daily) subcutaneously with vehicle control or Compound 1 for 21 days. The level of inflammatory cytokine propensity is lower in mice treated with Compound 1 than in vehicle treated mice.
16C depicts the effect of Compound 1 on IL1 beta cytokine levels in plasma of 24-month-old C57B1/6 mice. 23 month old mice were administered BID (twice daily) subcutaneously with vehicle control or Compound 1 for 21 days. The level of inflammatory cytokine propensity is lower in mice treated with Compound 1 than in vehicle treated mice.
16D depicts the effect of Compound 1 on IL5 cytokine levels in plasma of 24-month-old C57B1/6 mice. 23 month old mice were administered BID (twice daily) subcutaneously with vehicle control or Compound 1 for 21 days. The level of inflammatory cytokine propensity is lower in mice treated with Compound 1 than in vehicle treated mice.
16E depicts the effect of Compound 1 on IL17 cytokine levels in plasma of 24-month-old C57B1/6 mice. 23 month old mice were administered BID (twice daily) subcutaneously with vehicle control or Compound 1 for 21 days. The level of inflammatory cytokine propensity is lower in mice treated with Compound 1 than in vehicle treated mice.
Figure 17 depicts the effect of compound 1 on activated microglia in 24-month-old C57B1/6 mice. 23 month old mice were administered BID (twice daily) subcutaneously with vehicle control or Compound 1 for 21 days. There was a lower level of CD68+ activated microglia propensity in mice treated with compound 1 than in vehicle treated mice.
Figure 18 depicts the effect of Compound 1 on the percentage of eosinophils in complete white blood cell (WBC) counts in 24 month old C57BI/6 mice treated with BID, SQ for 3 weeks with either control saline or 30 mg/kg of Compound 1. Compound 1 reduced endogenous age-related blood eosinophilia.
19 depicts the effect of Compound 1 on the percentage of eosinophils in complete white blood cell (WBC) counts in 3 month old hairless mice treated with 2 weeks BID, PO with control saline or 30 mg/kg of Compound 1. Compound 1 reduced the oxazolone-induced increase in blood eosinophils.
20 shows the results of compound 1 in the rotarod test for motor coordination. 24 month old C57 mice were treated with continuous infusion of Compound 1 or vehicle by osmotic pump for 4 weeks. Treated mice were more successful than vehicle treated mice in the binomial test, *P <0.05.
21 shows the results of compound 1 in the T maze test for memory. 24 month old C57 mice were treated with continuous infusion of Compound 1 or vehicle by osmotic pump for 4 weeks. Treated mice were more successful than vehicle treated mice in the binomial test, *P <0.05.
22A shows the results of compound 1 on fecal excretion overnight. 24 month old C57 mice were treated with continuous infusion of Compound 1 or vehicle by osmotic pump for 4 weeks. Fecal pellets were weighed overnight. Mice treated with compound 1 had significantly lower stool excretion compared to vehicle-treated mice by an independent t-test, *P <0.05.
22B shows the results of compound 1 drinking water overnight. 24 month old C57 mice were treated with continuous infusion of Compound 1 or vehicle by osmotic pump for 4 weeks. Total water intake was measured overnight. Mice treated with compound 1 drank significantly more water by independent t-test compared to vehicle treated mice (*Po0.05).
22C shows the results of Compound 1 on overnight food intake. 24 month old C57 mice were treated with continuous infusion of Compound 1 or vehicle by osmotic pump for 4 weeks. Total food consumed was measured overnight. There was no difference in total food intake overnight between the two groups.
23 depicts the effect of compound 1 (Cmpd 1) on the number of CD68-positive (CD68 +) activated microglia in the brain of 3 month old mice treated with LPS and treated with compound 1 for 18 days. Compound 1-treated mice showed reduced CD68 + immunoreactivity and thus reduced gliosis.
24A and 24B depict the effect of Compound 1 on anxiety in an open field trial in 3-month-old mice treated with LPS IP for 4 weeks and compound 1 orally with BID (twice daily) for 1 week. LPS treatment increased preference around open fields, indicating increased anxiety. Compound 1 treatment reduced LPS-induced anxiety in the open field. Data shown are mean ± sem by independent sample t-test; *P <0.05.
25A and 25B depict the effect of compound 1 on the number of new and familiar cancer entry in the Y-maze test by 3-month-old C57B1/6 mice treated with LPS. The number of visits for each arm was plotted for each treatment group and subjected to a corresponding t-test. 3-month-old mice were administered vehicle or LPS IP for 6 weeks and BID (twice daily) orally administered vehicle control or Compound 1 for 3 weeks. Vehicle-treated mice did not, whereas Compound 1-treated mice showed a significant preference for novel cancer. Data shown are mean ± sem. of new versus familiar cancers by paired independent sample t-test; **P<0.01, *P<0.05.
Figures 26A and 26B depict the effect of Compound 1 on IL1 beta mRNA in the brain of 3-month-old C57B1/6 mice treated with LPS and/or Compound 1. Mice received vehicle control or LPS IP for 7 weeks. Vehicle or Compound 1 was administered orally BID (twice daily) for 4 weeks. Tissues were harvested and RNA was prepared from cortical brain tissue. Levels of IL1 beta mRNA were measured by qPCR and data are presented relative to vehicle control. The level of IL1 beta mRNA increased with LPS treatment and there was a significant decrease trend with compound 1 treatment. Data shown are mean ± sem; *P <0.05 by independent sample t-test.
Figures 27A, 27B and 27C depict the effect of Compound 1 on microglia activation in the hippocampus from 3-month-old C57B1/6 mice treated with LPS and/or Compound 1. Mice were dosed with vehicle control or LPS IP for 7 weeks, and vehicle or compound 1 orally with BID (twice daily) for 4 weeks. Tissues were harvested and brain sections were subjected to immunohistochemistry for CD68, a marker of activated microglia. There was a trend for increased CD68 levels with LPS treatment and a trend to decrease with compound 1 treatment. Data shown are mean ± sem.
Figures 28A, 28B and 28C depict the effect of Compound 1 on total microglia in the hippocampus from 3-month-old C57B1/6 mice treated with LPS and/or Compound 1. Mice were dosed with vehicle control or LPS IP for 7 weeks, and vehicle or compound 1 orally with BID (twice daily) for 4 weeks. Tissues were harvested and brain sections were subjected to immunohistochemistry for Iba1, a marker of microgliomas. There was a significant increase in Iba1 by LPS treatment and a tendency to decrease by compound 1 treatment. Data shown are mean ± sem by independent sample t-test; ***P<0.001, *P<0.05.
29A and 29B depict the effect of compound 1 on hippocampal total astrocytes from 3-month-old C57B1/6 mice treated with LPS and/or compound 1. Mice were dosed with vehicle control or LPS IP for 7 weeks and BID (twice daily) orally with vehicle or compound 1 for 4 weeks. Tissues were harvested and brain sections were subjected to immunohistochemistry for GFAP, a marker of astrocytes. There was a tendency to decrease with compound 1 treatment. Data shown are mean ± sem.
30 depicts dosing regimens for three groups of C57BL/6 mice treated as follows: (1) control for both LPS and compound 1; (2) vehicle control for LPS+compound 1; or (3) LPS + Compound 1. All three groups were treated simultaneously with control, LPS or Compound 1 for 3 consecutive days. The graph shows the results of histological analysis of the mouse brain in an acute model of LPS-induced inflammation. Microgliosis was measured by determining the percentage of Iba-1 positive areas in the hippocampus. Data shown are mean±sem; ^* P<0.05, ^*** P<0.001; A post-hoc Dunnett's multiple comparison test between treatment groups, testing for statistical significance using the usual one-way ANOVA.
31 shows vehicle control; vehicle treated LPS; or the dosing regimen of three groups of C57Bl/6J mice treated with Compound 1. The graph shows the results of histological analysis of the mouse brain in an acute model of LPS-induced inflammation. Microgliosis was measured by determining the percentage of Iba-1 positive areas in the hippocampus. Data shown are mean±sem. ^* P<0.05, ^**** P<0.0001; General one-way ANOVA was used to test for statistical significance, and Dunnett's multiple comparisons were post-hoc tests between treatment groups.
32 depicts a treatment study timeline for a mouse MPTP model of Parkinson's disease. There is a study of two arms, the first arm tested gait and fine motor function 10 days after treatment and the second arm was tested for immune cell infiltration 3 days after treatment
Figure 33A displays a correlation heat map illustrating the degree of correlation for 97 gait parameters in gait and microkinetic kinematic tests of C57Bl/6J mice on study day 11 after 10 days of compound 1 treatment. Ten major components are presented that indicate how the original parameters are correlated in the data set. The higher the intensity there is for each parameter, the stronger the parameter is related to its principal component. Red is positive correlation with respect to the 10 individual principal components (x-axis) and blue is negative correlation. The left y-axis shows the entire group for the individual variables on the right y-axis. For example, "ILC" means intra-limb coordination and "Tail B" means tail base.
33B shows the fine motor skills and gait characteristics of C57Bl/6J mice on study day 11 after 10 days of compound 1 treatment. Described by the overall gait analysis score of MPTP-treated C57Bl/6J mice. Differences between treatment groups for each of the 10 main components are summed up as a composite score to show differences from vehicle-treated controls. There was a significant difference in overall gait with MPTP treatment ( ^* p <0.05), and there was no further significant difference with compound 1 treatment.
FIG. 34 depicts forelimb toe spacing (one of the gait characteristics evaluated in principal component analysis) of C57Bl/6J mice on study day 11 after 10 days of Compound 1 treatment. Data are presented as mean±SEM (Group 1: Vehicle+Vehicle, n=15; Group 2: MPTP+Vehicle, n=14; Group 3: MPTP+Compound 1 (30 mg/kg), n=13). Statistical significance: ^* p<0.05, group 2: MPTP+vehicle vs. group 1: vehicle+vehicle (unmatched t-test).
35 depicts forelimb swing speed (one of the gait characteristics evaluated in principal component analysis) of C57Bl/6J mice on study day 11 after 10 days of compound 1 treatment. Data are presented as mean±SEM (Group 1: Vehicle+Vehicle, n=15; Group 2: MPTP+Vehicle, n=14; Group 3: MPTP+Compound 1 (30 mg/kg), n=13). Statistical significance: ^* p <0.05, group 2: MPTP+vehicle vs. group 1: vehicle+vehicle (unmatched t-test).
36 depicts ankle range of motion (one of the gait characteristics evaluated in principal component analysis) of C57Bl/6J mice at study day 11 after 10 days of compound 1 treatment. Data are presented as mean±SEM (Group 1: Vehicle+Vehicle, n=15; Group 2: MPTP+Vehicle, n=14; Group 3: MPTP+Compound 1 (30 mg/kg), n=13). Statistical significance: ^* p<0.05, group 2: MPTP+vehicle vs. group 1: vehicle+vehicle (unmatched t-test).
37 shows the acute effect of MPTP and compound 1 on T-cell transport to the brain 3 days after compound 1 treatment. The total number of CD3-positive T cells counted in the substantia nigra in three sections of 30 μm for each mouse is shown. Data shown are mean±sem. ^*** P <0.001; One-way ANOVA, Sidak's post hoc multiple comparison test.
38A and 38B show the acute effect of MPTP and compound 1 on microglia after 3 days of compound 1 treatment. 38A shows the extent of CD68-positive regions measured in the striatum (n=7 for saline+vehicle; n=8 for MPTP+vehicle; n=7 for MPTP+compound 1). 38B shows the extent of CD68-positive regions measured in the substantia nigra compaction (n=7 for saline+vehicle; n=8 for MPTP+vehicle; n=8 for MPTP+compound 1). Data shown are mean±sem. ^** P<0.01, ^*** P<0.001, ^**** P<0.0001; One-way ANOVA, Sidak's post hoc multiple comparison test.
Figure 39 depicts plasma eotaxin-1 levels in 6-month-old line 61 synuclein-overexpressing transgenic mice. Eotaxin-1 levels (CCL11) in non-transgenic (NTg) versus transgenic (Tg) line 61 synuclein mice were plotted as pg/mL concentrations.
40 shows the wire suspension test results for Line 61 synuclein mice (transgenic (Tg) and non-transgenic (nTg) age-matched littermates). Mean wire suspension times per group show significantly higher wire suspension times for animals in group C (non-transgenic, vehicle treated). Animals in group A (transgenic, treated with compound-1) showed significantly higher wire suspension times compared to group B (transgenic, treated with vehicle). Data are presented as mean±SEM of all animals per group. ^*** P <0.001;Dunn's post hoc test; ^* P<0.05 Mann Whitney test for groups A versus B).
41 shows the grip strength test results for Line 61 synuclein mice (transgenic (Tg) and non-transgenic (nTg) age-matched litters). The average maximum grip force [g] per group is indicated. Animals in groups A and C (transgenic, compound-1 treated and non-transgenic, vehicle treated, respectively) exhibited significantly higher grip force compared to group B (transgenic, vehicle treated). Data are presented as mean±SEM of all animals per group. Groups were compared to vehicle treated transgenic animals (Group B). One-way ANOVA followed by Bonferroni's post hoc test
Figure 42 shows the number of mice in each treatment group that were able to successfully traverse the beam in the beam walk test. N = 15 mice in group A, 14 mice in group B and 15 mice in group C. Mice (groups described in Figure 40) All mice were able to traverse the easiest beam in Experiment 1, whereas mice in treatment group B were unable to traverse the most difficult beam in Experiment 5 (Experiment 1 = 13 mm rectangular beam) , Trial 2 = 10 mm rectangular beam, Trial 3 = 28 mm cylindrical beam, Trial 4 = 16 mm cylindrical beam, Trial 5 = 11 mm cylindrical beam).
Figure 43: Five trials of beam walk slips for three groups of mice (Group A, transgenic compound 1 treated; Group B, transgenic vehicle treated; Group C, non-transgenic vehicle treated). show the results. Only mice that completely crossed the beam were included in the analysis. The graph represents the average number of slips per group [n] and each graph represents one trial (1-5). Data are mean±SEM of all animals per group. Groups were compared to group B. One-way ANOVA followed by Bonferroni's post hoc test. Statistics for experiment 5 could not be performed because the mice in group B could not cross the beam.
44A and 44B show eosinophil counts from peripheral blood. Figure 44A shows the percentage of eosinophils in peripheral blood (out of all leukocytes) of three groups of mice (Tg Cmpd 1 = transgenic compound 1 treated, Tg Veh = transgenic vehicle treated, nTG Veh = non-transgenic vehicle treated). Data were compared by t-test. Figure 44B shows the absolute eosinophil counts in peripheral blood from three groups of mice (Tg Cmpd 1 = transgenic compound 1 treated; Tg Veh = transgenic vehicle treated; nTG Veh = non-transgenic vehicle treated). Data were compared by t-test.
45A-45G show the effect of compound 1 on neuroinflammation. 45A shows quantified CD68 positive regions in the hippocampus: non-transgenic, vehicle treated mice; transgenic, vehicle treated mice; and transgenic, compound 1 treated mice (n = 14, 12 and 15, respectively). 45B shows quantified CD68 positive regions in the striatum of non-transgenic, vehicle treated mice; transgenic, vehicle treated mice; and transgenic, compound 1 treated mice (n = 15, 11 and 16, respectively). 45C shows quantified Iba-1 positive regions in the hippocampus: non-transgenic, vehicle treated mice; transgenic, vehicle treated mice; and transgenic, Compound 1 treated mice (n = 14, 13 and 16, respectively). 45D shows quantified Iba1-positive regions in the following striatum: non-transgenic, vehicle treated mice; transgenic, vehicle treated mice; and transgenic, compound 1 treated mice (n = 15, 11 and 16, respectively). Data are mean +/- sem. ^* P<0.05. 45E shows quantified GFAP positive regions in the hippocampus: non-transgenic, vehicle treated mice; transgenic, vehicle treated mice; and transgenic, Compound 1 treated mice (n = 14, 13 and 15, respectively). 45F shows quantified GFAP positive regions in the striatum of non-transgenic, vehicle treated mice. transgenic, vehicle treated mice; and transgenic, Compound 1 treated mice (n = 15, 13 and 15, respectively). 45G shows quantified Iba-1 positive regions in pars compacts of the substantia nigra of non-transgenic, vehicle-treated mice; transgenic, vehicle treated mice; and transgenic, compound 1-treated mice. Data are mean +/- sem. ^* P<0.05.
46A and 46B show the effect of Compound 1 on circulating levels of IL-4 and IL-6 cytokines in non-transgenic, vehicle-treated mice; transgenic, vehicle treated mice; and transgenic, Compound 1 treated mice (n = 14, 15 and 17, respectively). Figure 46A shows the levels of IL-4 measured in terminal cardiac plasma of all three groups. Figure 46B shows the levels of IL-6 measured in terminal cardiac plasma of all three groups. Data shown are mean +/- sem. ^* P<0.05, ^** P<0.01; One-way ANOVA, Dunnett's post hoc multiple comparison test.
47A-47D show the effect of compound 1 on EAE-inducing markers in the cerebellum of C57BL/6 mice. EAE resulted in an increase in CD3 and CD8 positive infiltrating T cells in the cerebellum. Figure 47A shows an increase in CD3 positive infiltrating T-cells in the cerebellum, which is significantly reduced after treatment with Compound 1 for 9 days. Figure 47B shows a significantly reduced increase in CD8 positive infiltrating cytotoxic T-cells in the cerebellum after treatment with Compound 1 for 9 days. Figure 47C shows a significant increase in Iba-1 positive areas in the cerebellum after EAE, which was significantly reduced in the cerebellum after treatment days. Figure 47D shows a significant increase in CD68 positive area in the cerebellum after EAE, which was significantly reduced in the cerebellum after 9 days of treatment. Data shown are mean +/- sem. ^* P<0.05, ^** P<0.01; One-way ANOVA, Dunnett's post hoc multiple comparison test.
Figure 48 depicts the concentration of human eotaxin-1 in a protein screen. Relative concentrations of human eotaxin-1 were determined by a commercially available affinity based assay (SomaLogic). Plasma samples from each of the 18, 30, 45, 55 and 66 year old donors were plotted.
49A depicts the effect of compound 1 on inhibition of eosinophil morphological changes. Whole blood from humans treated with compound 1 was incubated with recombinant eotaxin to induce eosinophil morphological changes. The inhibition of shape change was plotted against plasma Compound 1 concentration.
Figure 49B depicts the effect of compound 1 on CCR3 internalization. Whole blood from humans treated with compound 1 was incubated with recombinant eotaxin to induce CCR3 internalization and labeled with an anti-CCR3 antibody. Inhibition of CCR3 internalization by Compound 1 was plotted against plasma Compound 1 concentration.

Aspects of the present invention include methods of treating age-related disorders/neurodegenerative diseases. Age-related disorders can be in a number of different ways, e.g., age-associated cognitive and/or physiological disorders, e.g., forms of damage to central or peripheral organs of the body: cell damage, tissue damage, organ dysfunction, age-related shortening lifespan and carcinogenesis; Specific organs and tissues of interest herein include skin, neurons, muscle, pancreas, brain, kidney, lung, stomach, intestine, spleen, heart, adipose tissue, testis, ovary, uterus, liver and bone; reduced neurogenesis, and the like, but are not limited thereto.

In some embodiments, the aging-related disorder is an aging-related disorder of an individual's cognitive abilities, ie, an aging-related cognitive impairment. Cognitive ability, or “cognition,” refers to the mind including attention and concentration, learning complex tasks and concepts, memory (collecting, maintaining, and retrieving new information in the short and/or long term), and information processing (manipulating information collected by the five senses). means process. ), spatial perception skills (visual perception, depth perception, mental image use, copying pictures, constructing objects or shapes), language generation and understanding, language fluency (finding words), problem solving, decision-making and planning functions (planning and prioritizing) designation) is included. "Cognitive decline" means a gradual decline in one or more of these abilities, including a decrease in memory, language, thinking, and judgment. "Impairment of cognitive ability" and "cognitive impairment" refer to healthy individuals, eg, age-appropriate healthy individuals, or for eg 2 weeks, 1 month, 2 months, 3 months, 6 months, 1 year, 2 years, It refers to a decrease in cognitive ability relative to an individual's ability at a time earlier than 5 or 10 years. Age-related cognitive impairments include, for example, cognitive impairments associated with the natural aging process, eg, impairments in cognitive ability associated with aging, such as mild cognitive impairment (M.C.I.); and cognitive impairments associated with age-related disorders, i.e., disorders that appear to increase in frequency with increasing age, e.g., Alzheimer's disease, Parkinson's disease, Lewy body dementia, frontotemporal dementia, Huntington's disease, amyotrophic lateral sclerosis, multiple sclerosis, glaucoma, muscular dystrophy, vascular dementia, progressive supranuclear palsy, ataxia, weakness, and the like.

"Treatment" means that at least amelioration of one or more symptoms associated with a senescence-associated disorder afflicting a subject is achieved, wherein amelioration is achieved in a broad sense that amelioration of at least a parameter, eg, symptoms associated with the disorder being treated, is achieved it means. Thus, treatment may also include complete suppression, e.g., preventing the development or cessation of, e.g., cessation of, the pathological condition, or at least the symptoms associated therewith, such that the adult mammal suffers from symptoms indicative of the disorder or at least the disorder. It means not getting In some cases, "treatment", "treatment", and the like refer to obtaining a desired pharmacological and/or physiological effect. The effect may be prophylactic in terms of completely or partially preventing a disease or a symptom thereof and/or a partial or complete therapeutic for the disease and/or side effects caused by the disease. "Treatment" can be any treatment for a disease in a subject, (a) preventing the disease from developing in a subject susceptible to but not yet diagnosed with; (b) inhibiting the disease, i.e., slowing its progression. to hinder; or (c) alleviating the disease, ie causing regression of the disease. Treatment can result in a variety of physical symptoms, including modulation of gene expression, increased neurogenesis, and rejuvenation of tissues or organs. Treatment of a progressive disease in which treatment stabilizes or reduces undesirable clinical symptoms in a patient occurs in some embodiments. Such treatment may be performed before complete loss of function in the affected tissue. The subject therapy may be administered during and in some cases after the symptomatic stage of the disease.

In some instances where the age-related disorder is age-related cognitive decline, treatment by the methods of the present invention slows or reduces the progression of age-related cognitive decline. In other words, an individual's cognitive ability declines no more slowly after treatment with the disclosed method than before or without treatment with the disclosed method. In some instances, treatment by the methods of the present disclosure stabilizes the individual's cognitive abilities. For example, in an individual suffering from age-related cognitive decline, the progression of cognitive decline is stopped after treatment by the disclosed methods. As another example, cognitive decline in an individual expected to suffer age-related cognitive decline, eg, an individual 40 years of age or older, is prevented following treatment by the disclosed methods. In other words, cognitive impairment is no longer observed. In some instances, treatment by a method of the present disclosure reduces or reverses observed cognitive impairment, for example, by improving cognitive ability in an individual suffering from age-related cognitive decline. In other words, the cognitive ability of an individual suffering from age-related cognitive decline after treatment by the disclosed methods is better than before treatment by the disclosed methods. That is, it is improved upon treatment. In some examples, treatment by a method of the present disclosure eliminates cognitive impairment. In other words, the cognitive ability of an individual suffering from age-related cognitive decline, eg, as evidenced by improved cognition, after treatment by the disclosed methods, eg, when the individual is about 40 years of age or younger. restored to the level of

In some instances where the age-related disorder is an age-related movement disorder or reduction, treatment by the methods of the present invention slows or reduces the progression of age-related damage or reduction. In other words, an individual's motor capacity decreases more slowly after treatment with the disclosed methods compared to before or without treatment with the disclosed methods. In some examples, treatment by the methods of the present disclosure stabilizes the individual's motor ability. For example, the progression of motor loss in an individual suffering from age-related motor loss is halted after treatment by the disclosed methods. As another example, loss of movement in an individual expected to suffer age-related reduced movement, eg, an individual over the age of 40, is prevented following treatment by the disclosed methods. In other words, no (no longer) motor impairments are observed. In some instances, treatment by the methods of the present disclosure reduces or reverses motor impairment as observed, for example, by improving motor coordination or ability in an individual suffering from age-related motor decline. In other words, the motor performance of an individual suffering from age-related reduced movement after treatment by the disclosed method is better than before treatment by the disclosed method. That is, it is improved upon treatment. In some examples, treatment by the methods of the present disclosure eliminates movement disorders. In other words, the motor coordination or ability of an individual suffering from age-related motor decline can be reduced, for example, by treatment by the disclosed method to a level that, for example, the individual would be about 40 years of age or younger. is recovered

In some cases, treatment of an adult mammal according to a method results in a change in a central organ, eg, a central nervous system organ such as the brain, spinal cord, etc., where the change may be multiplied. For example, in the form of enhanced neurogenesis, including, but not limited to, molecular, structural and/or functional, as described in detail below.

A method of treating a dysfunction caused by a neurodegenerative disease is provided, the method comprising administering a compound of the formula discussed below. One embodiment of the present invention includes a method of improving cognitive or motor activity in a subject having a brain related, cognitive related or motor disorder, the method comprising administering a therapeutically effective amount of a compound of the formulas discussed below. A further embodiment of the present invention comprises a method of increasing neurogenesis in a subject suffering from a brain or cognitive related disease, said method comprising administering a therapeutically effective amount of a compound of the formula discussed below. A further embodiment of the present invention comprises a method of alleviating or treating a symptom of a brain or cognitive related disease, said method comprising administering a therapeutically effective amount of a compound of the formula A further embodiment of the present invention includes a method for alleviating the symptoms of a central nervous system related disease comprising administering a therapeutically effective amount of a primary peripheral agent A still further embodiment of the present invention comprises a method for improving motor activity in a subject having age-related motor dysfunction, said method comprising administering a therapeutically effective amount of a compound of the formula discussed below. The methods of the present invention may also include monitoring for improvement of an age-related disease, including, for example, improvement in cognition, motor activity, neurogenesis, etc., in a subject diagnosed with one or more such diseases or dysfunctions.

A further embodiment of the invention comprises administering a therapeutically effective amount of a compound, wherein the compound is in the form of a co-crystal or salt of the formula A further embodiment of the present invention comprises the step of administering a therapeutically effective amount of a compound, wherein the compound is in the form of an individual optical isomer, a mixture of individual enantiomers, a racemic or enantiomerically pure compound. A further embodiment of the invention also comprises administering a therapeutically effective amount of a compound, wherein the compound is in the form of pharmaceutical compositions and formulations discussed further below.

A further embodiment of the present invention for treating age-related movement disorders or decline comprises a modifier that inhibits the eotaxin/CCR3 pathway. Such modifiers include compounds of the formula: as well as other eotaxins and CCR3 inhibitors. Modifiers contemplated include, but are not limited to, compounds of the formula: and other CCR3 small molecule inhibitors (e.g., the bipiperdine derivatives described in U.S. Pat. No. 7,705,153, e.g., the cyclicamine derivatives described in U.S. Pat. No. 7,576,117 and Pease JE) and Horuk R, Expert Opin Drug Discov (2014) 9(5): 467-83, all of which are incorporated herein by reference in their entirety; anti-iotaxin antibody; anti-CCR3 antibody; eotaxin or aptamers that inhibit CCR3 expression or function (methods for making such aptamers include US Pat. Nos. 5,270,163, 5,840,867, 6,180,348); antisense oligonucleotides or siRNAs that inhibit the expression or function of eotaxin or CCR3 (eg, described in US Pat. No. 6,822,087); soluble CCR3 receptor protein (eg, decoy); etc.

Methods of using a diagnostic test or companion diagnostic test in connection with the described age-related disorder are also provided. One embodiment of such a diagnostic or companion diagnostic test is an in vitro diagnostic test. One embodiment of an in vitro diagnostic test is a companion device for use with a specific therapeutic agent. Embodiments of a particular therapeutic agent include, for example, a CCR3 small molecule inhibitor, an anti-CCR3 antibody, a small molecule inhibitor to the CCR3 ligand eotaxin-1, an anti-Eotaxin-1 antibody, and an antisense RNA to either eotaxin-1 or CCR3. However, it is not limited thereto.

By way of example and not limitation, such diagnosis or companion diagnosis includes determining or detecting the presence of a subset of leukocytes in a subject. A diagnosis or companion diagnosis may also be a determination of the presence, relative or absolute concentration, relative or absolute number of eosinophils from a subject's blood, tissue, or other such sample. Blood may be drawn, for example, by venipuncture or other similar methods. Other samples include, but are not limited to, sputum, cerebrospinal fluid, or tissue biopsies.

Methods for detecting or determining the presence, absolute or relative concentration, or relative or absolute number of leukocytes such as eosinophils, neutrophils, lymphocytes, basophils and monocytes are known to those skilled in the art. Methods for determining the presence, absolute or relative concentration, or relative or absolute number of eosinophils are also known to those skilled in the art. See Walsh GM, Eosinophils-Methods and Protocols ISBN: 9781493910151. These methods include, for example, but are not limited to: (1) Counting the total blood count with a 5-part difference using a hematology analyzer (e.g., O'Neil et al., Laboratory Hematology 7: 116-124). (2001) and US Centers for Disease Control and Prevention Laboratory Methods (available at https://wwwn.cdc.gov/nchs/data/nhanes/2013-2014/labmethods/CBC_H_MET_COMPLETE_BLOOD_COUNT.pdf); (2) Hematoxylin and Eosin Quantification of eosinophil counts in sputum using smears stained with (H and E) (eg Bandyopadhyay A, et al. Lung India. 2013 Apr-Jun; 30(2): 117-123); (3) CD45, CD125; , flow cytometric quantification of eosinophils using antibodies specific for various immune cells, such as CD193, F4/80 and Siglec-8 (Abcam, San Francisco, CA) (e.g., Yu YRA et al., Am J Respir Cell Mol Biol). 2016 Jan; 54(1): 13–24 and Cossarizza A, et al., Eur. J. Immunol. 2017 47: 1584-1797);(4) quantified eosinophil autofluorescence cytometry using flow cytometry ( See e.g. Guenther G et al., J Immunol May 1, 2015, 194(1 Supplement) 206.12 and Thurau AM et al., Cytometry 23:159-58(1996)),(5) Dextran 70 in whole blood; Eosinophil Isolation Using Ficoll-Paque and Antibody-Based Magnetic Colloidal Cell Separation (e.g., Munoz NM, et al., Nature Protocols, 2613-20 (2006); Akuthota P, et al., Curr Protoc Immunol 98(1): pp. 7.31.1-7.31.8, 2012; and Akut hota P, et al., Methods Mol Biol 1178: 13-20 (2014)); (6) histological staining by normal staining (eosin or similar dye) or eosinophil-specific protein (major basic protein (MBP), eosinophil cation Antibody-based staining for sexual proteins (ECP), eosinophil peroxidase (EPO), eosinophil-derived neurotoxin (EDN), SiglecF (or Siglec8). (e.g., Koller DY et al., Allergy 54(10): 1094- 9 (1999) and Akuthota P., Capron K., Weller PF (2014) Eosinophil Purification from Peripheral Blood. In: Walsh G. (eds) Eosinophils. Methods in Molecular Biology (Methods and Protocols), vol 1178. See Humana Press, New York, NY); (7) degranulation, eosinophil extracellular trap (EET) and/or specific proteins of these (eg, galectin 10); Histological staining of processes spatially related to the same eosinophils.

A diagnostic or companion diagnostic device may incorporate such methods to determine the presence, absolute or relative concentration or relative or absolute number of eosinophils. By determining the presence, absolute or relative concentration, or relative or absolute number of eosinophils, the device can be used in conjunction with other methods of the present invention. By way of example and not limitation, such other methods may include methods of treating the age-related disorders/neurodegenerative diseases mentioned herein. In some embodiments, the aging-related disorder is an aging-related disorder of an individual's cognitive abilities, ie, an aging-related disorder. Such age-related disorders may include, but are not limited to, neurodegenerative conditions such as, for example, Alzheimer's disease, Parkinson's disease, Lewy body dementia, frontotemporal dementia, Huntington's disease, amyotrophic lateral sclerosis, multiple sclerosis, glaucoma, muscular dystrophy, vascular dementia does not Such age-related disorders may include cognitive impairment and/or motor impairment or decline.

One embodiment of the invention comprises treating a subject diagnosed with a senescence-associated disorder with a therapeutically effective amount of at least one of the compounds of the invention disclosed herein in conjunction with a diagnostic or companion diagnostic device. Another embodiment of the invention comprises treating said subject with a therapeutically effective amount of Compound 1 disclosed herein in combination with a diagnostic device. Embodiments of the device may be used, for example, to determine or detect the presence, absolute or relative concentration, or absolute or relative number of eosinophils in a blood or tissue sample of a subject. Another embodiment performs the step of detecting or determining the presence, absolute or relative concentration, or absolute or relative number of eosinophils in a blood or tissue sample of a subject prior to, during, or after treatment.

The experimental figures herein report that a mammalian model of Parkinson's disease of synuclein overexpression results in eosinophilia, which is restored to the level of non-transgenic mice through compound 1 treatment, suggesting beneficial immune modulation in this model of Parkinson's disease. This also suggests that determining the level of eosinophils in Parkinson's disease patients treated with Compound 1 can be a biomarker for disease, including determining the level of progression, stasis or regression, and therapeutic efficacy (e.g. : see FIGS. 44A and 44B).

Another embodiment performs the step of detecting or determining the presence, absolute or relative concentration, or absolute or relative number of eosinophils in a blood or tissue sample of a subject diagnosed with Parkinson's disease prior to, during, or after treatment. A further embodiment comprises determining the presence, absolute or relative concentration, or absolute or relative number in a blood or tissue sample of a subject diagnosed with Parkinson's disease prior to treatment with a compound of the invention, such as compound 1, to obtain a baseline. . A further embodiment subsequently performs the step of detecting or determining the level of eosinophils after treatment to compare this level to a baseline concentration or number of eosinophils to monitor the efficacy of the treatment. Comparison of these levels may show an increase or decrease in the number or concentration of eosinophils compared to baseline. Another embodiment of the invention comprises monitoring the progression of a disease or disorder by comparing the number or concentration of eosinophils, wherein an increase in the number after treatment improves the progression of the disease. In other embodiments, the improvement comprises a 1-5% increase, 5-10% increase over baseline in the number or concentration of eosinophils in the blood or tissue of the subject; 11-15% increase; 16-20% increase; 21-25% increase; 26-30% increase; 31-35% increase; 36-40% increase; 41-45% increase; 46-50% increase; 51-55% increase; 56-60% increase; 61-65% 66-70% increase; 71-75% increase; 76-80% increase; 81-85% increase; 86-90% increase; 91-95% increase; 96-100% increase; and 1-1.5 ×, 1.5-2 ×, 2-2.5 ×, 2.5-3 ×, 3-3.5 ×, 3.5-4 ×, 4-4.5 ×, 4.5-5 ×, 5-5.5 ×, 5.5-6 × , 6-6.5 ×, 6.5-7 ×, 7-7.5 ×, 7.5-8 ×, 8-8.5 ×, 8.5-9 ×, 9-9.5 ×, 9.5-10 ×, and 10 × or greater.

Another embodiment of the present invention is for diagnosing Parkinson's disease by determining a baseline of the number or concentration of eosinophils in the blood or tissue of a subject and comparing it with a standard number or concentration of eosinophils in a non-Parkinson's disease population with normal eosinophil count include that It is known in the art that eosinophil count is the number of eosinophils in the body. The normal number of eosinophils in adults is 30 to 350, but ^{can rise up to 500 cubic millimeters (mm 3} ) in the blood (see: Medical News Today, December 2018): https://www.medicalnewstoday.com/articles /323868.php, the entire contents of which are incorporated herein by reference). A blood level of 500 mm ³ or higher is considered eosinophilia. In some diseases, such as alcoholism and cortisol overprotection, the number of eosinophils is lower than normal. (Id.) One aspect of the invention detects or determines the number or concentration of eosinophils in a subject suspected of having Parkinson's disease. If the number or concentration of eosinophils in a subject is lower than normal (eg, 30 mm ^{3 in the} blood), the caregiver can use the results to determine a diagnosis of the disease.

a. compound

The methods of the present invention further comprise administering to the subject a compound In a group, radical or moiety defined in this "Compounds" section, the number of carbon atoms is often specified prior to that group, for example C 1-6 alkyl means an alkyl group or radical having 1 to 6 carbon atoms. do. In general, for groups comprising two or more subgroups disclosed in this "Compounds" section, the last group is the point of radical attachment, e.g. "thioalkyl" means a monovalent radical of the formula HS-Alk- . Unless otherwise specified below, conventional definitions of the valences of conventional stable atoms are assumed in all formulas and groups.

One embodiment of the present invention further comprises administration of a compound of formula 1 to a subject,

1

One

in the above formula

A is CH ₂ , O or NC _1-6 -alkyl;

R ¹ is

· NHR ^1.1 , NMeR ^1.1 ;

· NHR ^1.2 , NMeR ^1.2 ;

· NHCH ₂ -R ^1.3 ;

NH-C _3-6 -cycloalkyl, wherein optionally one carbon atom is replaced by a nitrogen atom and the ring is C _1-6 -alkyl, OC _1-6 -alkyl, NHSO ₂ -phenyl, NHCONH-phenyl, halogen, optionally substituted with 1 or 2 moieties selected from the group consisting of CN, SO ₂ -C _1-6 -alkyl, COO-C _{1-6 -alkyl;}

C _{9 or 10} -bicyclic-ring, wherein 1 or 2 carbon atoms are replaced by nitrogen atoms, the ring system is bonded to the basic structure of formula 1 via a nitrogen atom, the ring system is C _1-6 -alkyl, COO-C _1-6 -alkyl, C _1-6 -haloalkyl, OC _1-6 -alkyl, NO ₂ , halogen, CN, NHSO ₂ -C _1-6 -alkyl, m-methoxy-phenyl optionally substituted with 1 or 2 residues selected from;

a group selected from NHCH(pyridinyl)CH ₂ COO-C _1-6 -alkyl, NHCH(CH ₂ OC _1-6 -alkyl)-benzoimidazolyl, optionally substituted with halogen or CN;

· or 1-amino cyclopentyl, optionally substituted with methyl-oxadiazole;

R ^1.1 is C _1-6 -alkyl, C _2-6 -alkenyl, C _2-6 -alkynyl, C _1-6 -haloalkyl, C _1-6 -alkylene-OH, C _2-6 -alke nylene-OH, C _2-6 -alkynylene-OH, CH ₂ CON(C _1-6 -alkyl) ₂ , CH ₂ NHCONH-C _3-6 -cycloalkyl, CN, CO-pyridinyl, CONR ^1.1.1 R ^1.1.2 , COO-C _1-6 -alkyl, N(SO ₂ -C _1-6 -alkyl)(CH ₂ CON(C _1-4 -alkyl) ₂ ) OC _1-6 -alkyl, O-pyri dinyl, SO ₂ -C _1-6 -alkyl, SO ₂ -C _1-6 -alkylene-OH, SO ₂ -C _3-6 -cycloalkyl, SO ₂ -piperidinyl, SO ₂ NH-C _{1- 6} -alkyl, SO ₂ N(C _1-6 -alkyl) ₂ , halogen, CN, CO-morpholinyl, CH ₂ -pyridinyl, or C _1-6 -alkyl, NHC _1-6 -alkyl and =O phenyl, optionally substituted with 1 or 2 moieties selected from the group consisting of heterocyclic rings, optionally substituted with 1 or 2 moieties selected from the group consisting of;

^1.1.1 R is H, C _1-6 - optionally substituted with alkyl, C _1-6 - alkyl, C _3-6 - cycloalkyl, C _1-6 - haloalkyl, CH ₂ CON (C _1-6 - alkyl,) ₂ , CH ₂ CO-azetidinyl, C _1-6 -alkylene-C _3-6 -cycloalkyl, CH ₂ -pyranyl, CH ₂ -tetrahydro-furanyl, CH ₂ -furanyl, C _1-6 -alkylene-OH or thiadiazolyl;

R ^1.1.2 is H, C _1-6 -alkyl, SO ₂ C _1-6 -alkyl; or

R ^1.1.1 and R ^1.1.2 optionally contain one N or O, replacing a carbon atom of the ring, C _1-6 -alkyl, C _1-4 -alkylene-OH, OH, =O together form a 4-, 5- or 6-membered carbon ring optionally substituted with one or two moieties selected from the group consisting of;

or

R ^1.1 is phenyl, wherein two adjacent moieties together form a 5 or 6 membered carbocyclic aromatic or non-aromatic ring, optionally independently of one another, one or two N, S or SO replacing a carbon atom of the ring ₂ , wherein the ring is _{optionally substituted with C 1-4} -alkyl or =O;

R ^1.2 is

· C _1-6 -alkyl, C _2-6 -alkenyl, C _2-6 -alkynyl, C _3-6 -cycloalkyl, CH ₂ COO-C _1-6 -alkyl, CONR ^1.2.1 R ^{1.2. 2} , COR ^1.2.3 , COO-C _1-6 -alkyl, CONH ₂ , OC _1-6 -alkyl, halogen, CN, SO ₂ N(C _1-6 -alkyl) ₂ , or 1 or 2 C ₁ heteroaryl optionally substituted with one or two moieties selected from the group consisting of heteroaryl optionally substituted with _{-6 -alkyl moieties;}

Heteroaryl, optionally substituted with a 5- or 6-membered carbocyclic non-aromatic ring containing, independently of each other, two N, O, S or SO _{2 replacing carbon atoms;}

Aromatic or non-aromatic C _{9 or 10} -bicyclic-ring, wherein one or two carbon atoms are from the group consisting of _{N(C 1-6} -alkyl) ₂ , CONH-C _{1-6 -alkyl, =O} replaced by N, O or S, each optionally substituted with one or two residues selected from;

- a heterocyclic non-aromatic ring, optionally substituted with pyridinyl;

4,5-dihydronaphtho[2,1-d]thiazole, optionally substituted with NHCO-C _{1-6 -alkyl;}

R ^1.2.1 is H, C _1-6 -alkyl, C _1-6 -alkylene-C _3-6 -cycloalkyl, C _1-4 -alkylene-phenyl, C _1-4 -alkylene-furanyl , C _3-6 -cycloalkyl, C _1-4 -alkylene-OC _1-4 -alkyl, C _1-6 -haloalkyl or 5- or 6-membered carbocyclic non-aromatic ring, optionally independently of one another, contains 1 or 2 N, O, S or SO ₂ , replacing a carbon atom of the ring, optionally substituted with 4-cyclopropylmethyl-piperazinyl;

R ^1.2.2 is H, C _1-6 -alkyl;

R ^1.2.3 are optionally independently of one another a 5- or 6-membered carbocyclic non-aromatic ring containing 1 or 2 N, O, S or SO _{2 , replacing a carbon atom of the ring;}

R ^1.3 is selected from phenyl, heteroaryl or indolyl, each as C _1-6 -alkyl, C _3-6 -cycloalkyl, OC _1-6 -alkyl, OC _1-6 -haloalkyl, phenyl, heteroaryl optionally substituted with 1 or 2 residues selected from the group consisting of;

R ² is selected from C _1-6 -alkylene-phenyl, C _1-6 -alkylene-naphthyl, and C _1-6 -alkylene-heteroaryl; C _1-6 - alkyl, C _1-6 - haloalkyl, OC _1-6 - alkyl, OC _1-6 - a haloalkyl, one, two or three moieties selected from the group consisting of halogen and optionally substituted, respectively;

R ³ is H, C _1-6 -alkyl;

R ⁴ is H, C _1-6 -alkyl;

or R ³ and R ⁴ together form a CH ₂ —CH ₂ group.

Another embodiment of the present invention further comprises administration of a compound of formula 1 to a subject,

From here,

A is CH ₂ , O or NC _1-4 -alkyl;

R ¹ is

· NHR ^1.1 , NMeR ^1.1 ;

· NHR ^1.2 , NMeR ^1.2 ;

· NHCH ₂ -R ^1.3 ; and

R ^1.1 is C _1-6 -alkyl, C _2-6 -alkenyl, C _2-6 -alkynyl, C _1-6 -haloalkyl, C _1-6 -alkylene-OH, C _2-6 -alke nylene-OH, C _2-6 -alkynylene-OH, CH ₂ CON(C _1-6 -alkyl) ₂ , CH ₂ NHCONH-C _3-6 -cycloalkyl, CN, CO-pyridinyl, CONR ^1.1.1 R ^1.1.2 , COO-C _1-6 -alkyl, N(SO ₂ -C _1-6 -alkyl)(CH ₂ CON(C _1-4 -alkyl) ₂ ) OC _1-6 -alkyl, O-pyri dinyl, SO ₂ -C _1-6 -alkyl, SO ₂ -C _1-6 -alkylene-OH, SO ₂ -C _3-6 -cycloalkyl, SO ₂ -piperidinyl, SO ₂ NH-C _{1- 6} -alkyl, SO ₂ N(C _1-6 -alkyl) ₂ , halogen, CN, CO-morpholinyl, CH ₂ -pyridinyl, or C _1-6 -alkyl, NHC _1-6 -alkyl and =O phenyl, optionally substituted with 1 or 2 moieties selected from the group consisting of heterocyclic rings, optionally substituted with 1 or 2 moieties selected from the group consisting of;

^1.1.1 R is H, C _1-6 - optionally substituted with alkyl, C _1-6 - alkyl, C _3-6 - cycloalkyl, C _1-6 - haloalkyl, CH ₂ CON (C _1-6 - alkyl,) ₂ , CH ₂ CO-azetidinyl, C _1-6 -alkylene-C _3-6 -cycloalkyl, CH ₂ -pyranyl, CH ₂ -tetrahydro-furanyl, CH ₂ -furanyl, C _1-6 -alkylene-OH or thiadiazolyl;

R ^1.1.2 is H, C _1-6 -alkyl, SO ₂ C _1-6 -alkyl; or

R ^1.1.1 and R ^1.1.2 optionally contain one N or O, replacing a carbon atom of the ring, as C _1-6 -alkyl, C _1-4 -alkylene-OH, OH, =O together form a 4-, 5- or 6-membered carbocyclic ring optionally substituted with one or two moieties selected from the group consisting of;

^1.1.1 R is H, C _1-6 - optionally substituted with alkyl, C _1-6 - alkyl, C _3-6 - cycloalkyl, C _1-6 - haloalkyl, CH ₂ CON (C _1-6 - alkyl,) ₂ , CH ₂ CO-azetidinyl, C _1-6 -alkylene-C _3-6 -cycloalkyl, CH ₂ -pyranyl, CH ₂ -tetrahydro-furanyl, CH ₂ -furanyl, C _1-6 -alkylene-OH or thiadiazolyl;

R ^1.1.2 is H, C _1-6 -alkyl, SO ₂ C _1-6 -alkyl; or

R ^1.1.1 and R ^1.1.2 optionally contain one N or O, replacing a carbon atom of the ring, as C _1-6 -alkyl, C _1-4 -alkylene-OH, OH, =O together form a 4-, 5- or 6-membered carbocyclic ring optionally substituted with one or two moieties selected from the group consisting of;

R ^1.1 is phenyl, wherein two adjacent moieties together form a 5 or 6 membered carbocyclic aromatic or non-aromatic ring, optionally independently of one another, one or two N, S or SO replacing a carbon atom of the ring ₂ , wherein the ring is _{optionally substituted with C 1-4} -alkyl or =O;

R ^1.2 is

· C _1-6 -alkyl, C _2-6 -alkenyl, C _2-6 -alkynyl, C _3-6 -cycloalkyl, CH ₂ COO-C _1-6 -alkyl, CONR ^1.2.1 R ^{1.2. 2} , COR ^1.2.3 , COO-C _1-6 -alkyl, CONH ₂ , OC _1-6 -alkyl, halogen, CN, SO ₂ N(C _1-4 -alkyl) ₂ , or 1 or 2 C ₁ heteroaryl optionally substituted with one or two moieties selected from the group consisting of heteroaryl optionally substituted with _{-6 -alkyl moieties;}

heteroaryl, optionally substituted with a 5- or 6-membered carbocyclic non-aromatic ring containing, independently of each other, two N, O, S or SO _{2 replacing a carbon atom;}

R ^1.2.1 is H, C _1-6 -alkyl, C _1-6 -alkylene-C _3-6 -cycloalkyl, C _1-4 -alkylene-phenyl, C _1-4 -alkylene-furanyl , C _3-6 -cycloalkyl, C _1-4 -alkylene-OC _1-4 -alkyl, C _1-6 -haloalkyl or 5- or 6-membered carbocyclic non-aromatic ring, optionally independently of one another, contains 1 or 2 N, O, S or SO ₂ , replacing a carbon atom of the ring, optionally substituted with 4-cyclopropylmethyl-piperazinyl;

R ^1.2.2 is H, C _1-6 -alkyl;

R ^1.2.3 are optionally independently of one another a 5- or 6-membered carbocyclic non-aromatic ring containing 1 or 2 N, O, S or SO _{2 , replacing a carbon atom of the ring;}

R ^1.3 is selected from phenyl, heteroaryl or indolyl, each as C _1-6 -alkyl, C _3-6 -cycloalkyl, OC _1-6 -alkyl, OC _1-6 -haloalkyl, phenyl, heteroaryl optionally substituted with 1 or 2 residues selected from the group consisting of; In some cases R ^1.3 is selected from phenyl, pyrazolyl, isoxazolyl, pyrimidinyl, indolyl or oxadiazolyl, each C _1-6 -alkyl, C _3-6 -cycloalkyl, OC _1-6 - optionally substituted with 1 or 2 moieties selected from the group consisting of alkyl, OC _{1-6 -haloalkyl;}

R ² is selected from C _1-6 -alkenyl-phenyl or C _1-6 -alkenyl-naphthyl and C _1-6 -alkenyl-thiophenyl; C _1-6 - alkyl, C _1-6 - haloalkyl, OC _1-6 - alkyl, OC _1-6 - a haloalkyl, one or two moieties selected from the group consisting of halogen and optionally substituted, respectively;

R ³ is H, C _1-4 -alkyl;

R ⁴ is H, C _1-4 -alkyl;

or R ³ and R ⁴ together form a _{CH 2} -CH _{2 group.}

Another embodiment of the present invention further comprises administration of a compound of formula 1 to a subject,

From here,

A is CH ₂ , O or NC _1-4 -alkyl;

R ¹ is

· NHR ^1.1 , NMeR ^1.1 ;

R ^1.1 is C _1-6 -alkyl, C _2-6 -alkenyl, C _2-6 -alkynyl, C _1-6 -haloalkyl, C _1-6 -alkylene-OH, C _2-6 -alke nylene-OH, C _2-6 -alkynylene-OH, CH ₂ CON(C _1-6 -alkyl) ₂ , CH ₂ NHCONH-C _3-6 -cycloalkyl, CN, CO-pyridinyl, CONR ^1.1.1 R ^1.1.2 , COO-C _1-6 -alkyl, N(SO ₂ -C _1-6 -alkyl)(CH ₂ CON(C _1-4 -alkyl) ₂ ) OC _1-6 -alkyl, O-pyri dinyl, SO ₂ -C _1-6 -alkyl, SO ₂ -C _1-6 -alkylene-OH, SO ₂ -C _3-6 -cycloalkyl, SO ₂ -piperidinyl, SO ₂ NH-C _{1- 6} -alkyl, SO ₂ N(C _1-6 -alkyl) ₂ , halogen, CN, CO-morpholinyl, CH ₂ -pyridinyl, or C _1-6 -alkyl, NHC _1-6 -alkyl and =O phenyl, optionally substituted with 1 or 2 moieties selected from the group consisting of heterocyclic rings, optionally substituted with 1 or 2 moieties selected from the group consisting of;

^1.1.1 R is H, C _1-6 - optionally substituted with alkyl, C _1-6 - alkyl, C _3-6 - cycloalkyl, C _1-6 - haloalkyl, CH ₂ CON (C _1-6 - alkyl,) ₂ , CH ₂ CO-azetidinyl, C _1-6 -alkylene-C _3-6 -cycloalkyl, CH ₂ -pyranyl, CH ₂ -tetrahydro-furanyl, CH ₂ -furanyl, C _1-6 -alkylene-OH or thiadiazolyl;

R ^1.1.2 is H, C _1-6 -alkyl, SO ₂ C _1-6 -alkyl; or

R ^1.1.1 and R ^1.1.2 optionally contain one N or O, replacing a carbon atom of the ring, as C _1-6 -alkyl, C _1-4 -alkylene-OH, OH, =O together form a 4-, 5- or 6-membered carbocyclic ring optionally substituted with one or two moieties selected from the group consisting of;

or

R ^1.1 is phenyl, wherein two adjacent moieties together form a 5 or 6 membered carbocyclic aromatic or non-aromatic ring, optionally independently of one another, one or two N, S or SO replacing a carbon atom of the ring ₂ , wherein the ring is _{optionally substituted with C 1-4} -alkyl or =O;

R ² is selected from C _1-6 -alkylene-phenyl, C _1-6 -alkylene-naphthyl, and C _1-6 -alkylene-heteroaryl; C _1-6 - alkyl, C _1-6 - haloalkyl, OC _1-6 - alkyl, OC _1-6 - a haloalkyl, one, two or three moieties selected from the group consisting of halogen and optionally substituted, respectively;

R ³ is H, C _1-4 -alkyl;

R ⁴ is H, C _1-4 -alkyl;

or R ³ and R ⁴ together form a CH ₂ —CH ₂ group.

Another embodiment of the present invention further comprises administration of a compound of formula 1 to a subject,

From here,

A is CH ₂ , O or NC _1-4 -alkyl;

R ¹ is

· NHR ^1.2 , NMeR ^1.2 ;

R ^1.2 is

· C _1-6 -alkyl, C _2-6 -alkenyl, C _2-6 -alkynyl, C _3-6 -cycloalkyl, CH ₂ COO-C _1-6 -alkyl, CONR ^1.2.1 R ^{1.2. 2} , COR ^1.2.3 , COO-C _1-6 -alkyl, CONH ₂ , OC _1-6 -alkyl, halogen, CN, SO ₂ N(C _1-4 -alkyl) ₂ , or 1 or 2 C ₁ heteroaryl optionally substituted with one or two moieties selected from the group consisting of heteroaryl optionally substituted with _{-6 -alkyl moieties;}

Heteroaryl, optionally substituted with a 5- or 6-membered carbocyclic non-aromatic ring containing, independently of each other, two N, O, S or SO _{2 replacing carbon atoms;}

benzothiazolyl, indazolyl, dihydro-indole, each optionally substituted with one or two moieties selected from the group consisting of N(C _1-6 -alkyl) ₂ , CONH-C _{1-6 -alkyl, =O} lyl, indanyl, tetrahydro-quinolinyl;

• piperidinyl, optionally substituted with pyridinyl;

4,5-dihydronaphtho[2,1-d]thiazole, optionally substituted with NHCO-C _{1-6 -alkyl;}

R ^1.2.1 is H, C _1-6 -alkyl, C _1-6 -alkylene-C _3-6 -cycloalkyl, C _1-4 -alkylene-phenyl, C _1-4 -alkylene-furanyl , C _3-6 -cycloalkyl, C _1-4 -alkylene-OC _1-4 -alkyl, C _1-6 -haloalkyl or 5- or 6-membered carbocyclic non-aromatic ring, optionally independently of one another, contains 1 or 2 N, O, S or SO ₂ , replacing a carbon atom of the ring, optionally substituted with 4-cyclopropylmethyl-piperazinyl;

R ^1.2.2 is H, C _1-6 -alkyl;

R ^1.2.3 are optionally independently of one another a 5- or 6-membered carbocyclic non-aromatic ring containing 1 or 2 N, O, S or SO _{2 , replacing a carbon atom of the ring;}

R ² is selected from C _1-6 -alkylene-phenyl or C _1-6 -alkylene-naphthyl, and C _1-6 -alkylenethiophenyl; C _1-6 - alkyl, C _1-6 - haloalkyl, OC _1-6 - alkyl, OC _1-6 - a haloalkyl, one, two or three moieties selected from the group consisting of halogen and optionally substituted, respectively;

R ³ is H, C _1-4 -alkyl;

R ⁴ is H, C _1-4 -alkyl;

or R ³ and R ⁴ together form a _{CH 2} -CH _{2 group.}

Another embodiment of the present invention further comprises administration of a compound of formula 1 to a subject,

From here,

A is CH ₂ , O or NC _1-4 -alkyl;

R ¹ is

· NHR ^1.2 , NMeR ^1.2 ;

R ^1.2 is

· C _1-6 -alkyl, C _2-6 -alkenyl, C _2-6 -alkynyl, C _3-6 -cycloalkyl, CH ₂ COO-C _1-6 -alkyl, CONR ^1.2.1 R ^{1.2. 2} , COR ^1.2.3 , COO-C _1-6 -alkyl, CONH ₂ , OC _1-6 -alkyl, halogen, CN, SO ₂ N(C _1-4 -alkyl) ₂ , or 1 or 2 C ₁ heteroaryl optionally substituted with one or two moieties selected from the group consisting of heteroaryl optionally substituted with _{-6 -alkyl moieties;}

heteroaryl, optionally substituted with a 5- or 6-membered carbocyclic non-aromatic ring containing, independently of each other, two N, O, S or SO _{2 replacing a carbon atom;}

R ^1.2.1 is H, C _1-6 -alkyl, C _1-6 -alkylene-C _3-6 -cycloalkyl, C _1-4 -alkylene-phenyl, C _1-4 -alkylene-furanyl , C _3-6 -cycloalkyl, C _1-4 -alkylene-OC _1-4 -alkyl, C _1-6 -haloalkyl or 5- or 6-membered carbocyclic non-aromatic ring, optionally independently of one another, contains 1 or 2 N, O, S or SO ₂ , replacing a carbon atom of the ring, optionally substituted with 4-cyclopropylmethyl-piperazinyl;

R ^1.2.2 is H, C _1-6 -alkyl;

R ^1.2.3 are optionally independently of one another a 5- or 6-membered carbocyclic non-aromatic ring containing 1 or 2 N, O, S or SO _{2 , replacing a carbon atom of the ring;}

R ² is selected from C _1-6 -alkylene-phenyl or C _1-6 -alkylene-naphthyl, and C _1-6 -alkylenethiophenyl; C _1-6 - alkyl, C _1-6 - haloalkyl, OC _1-6 - alkyl, OC _1-6 - a haloalkyl, one, two or three moieties selected from the group consisting of halogen and optionally substituted, respectively;

R ³ is H, C _1-4 -alkyl;

R ⁴ is H, C _1-4 -alkyl;

or R ³ and R ⁴ together form a _{CH 2} -CH _{2 group.}

Another embodiment of the present invention further comprises administration of a compound of formula 1 to a subject,

From here,

A is CH ₂ , O or NC _1-4 -alkyl;

R ¹ is

· NHCH ₂ -R ^1.3 ; selected from,

R ^1.3 is selected from phenyl, pyrazolyl, isoxazolyl, pyrimidinyl, indolyl or oxadiazolyl, each C _1-6 -alkyl, C _3-6 -cycloalkyl, OC _1-6 -alkyl, OC optionally substituted with 1 or 2 moieties selected from the group consisting of _{1-6 -haloalkyl;}

R ² is selected from C _1-6 -alkylene-phenyl, C _1-6 -alkylene-naphthyl, and C _1-6 -alkylene-thiothiophenyl; C _1-6 - alkyl, C _1-6 - haloalkyl, OC _1-6 - alkyl, OC _1-6 - a haloalkyl, one, two or three moieties selected from the group consisting of halogen and optionally substituted, respectively;

R ³ is H, C _1-4 -alkyl;

R ⁴ is H, C _1-4 -alkyl;

or R ³ and R ⁴ together form a CH ₂ —CH ₂ group.

Another embodiment of the present invention further comprises administration of a compound of formula 1 to a subject,

From here,

A is CH ₂ , O or NC _1-4 -alkyl;

R ¹ is

· NHR ^1.1 , NMeR ^1.1 ;

· NHR ^1.2 , NMeR ^1.2 ;

· NHCH ₂ -R ^1.3 ;

NH-C _3-6 -cycloalkyl, wherein optionally one carbon atom is replaced by a nitrogen atom and the ring is C _1-6 -alkyl, OC _1-6 -alkyl, NHSO ₂ -phenyl, NHCONH-phenyl, halogen, optionally substituted with 1 or 2 moieties selected from the group consisting of CN, SO ₂ -C _1-6 -alkyl, COO-C _{1-6 -alkyl;}

C _{9 or 10} -bicyclic-ring, wherein 1 or 2 carbon atoms are replaced by nitrogen atoms, the ring system is bonded to the basic structure of formula 1 via a nitrogen atom, the ring system is C _1-6 -alkyl, COO-C _1-6 -alkyl, C _1-6 -haloalkyl, OC _1-6 -alkyl, NO ₂ , halogen, CN, NHSO ₂ -C _1-6 -alkyl, m-methoxy-phenyl optionally substituted with 1 or 2 residues selected from;

• a group selected from NHCH(pyridinyl)CH ₂ COO-C _1-6 -alkyl, NHCH(CH ₂ OC _1-6 -alkyl)-benzoimidazolyl, optionally substituted with Cl;

· or 1-amino cyclopentyl, optionally substituted with methyl-oxadiazole;

R ^1.1 is C _1-6 -alkyl, C _1-6 -haloalkyl, CH ₂ CON(C _1-6 -alkyl) ₂ , CH ₂ NHCONH-C _3-6 -cycloalkyl, CN, CONR ^1.1.1 R ^1.1.2 , COO-C _1-6 -alkyl, OC _1-6 -alkyl, SO ₂ -C _1-6 -alkyl, SO ₂ -C _1-6 -alkylene-OH, SO ₂ -C _3-6 - cycloalkyl, SO _{2-piperidinyl,} SO ₂ NH-C _1-6 - alkyl, SO ₂ N (C _1-6 - alkyl) _2, halogen, CN, CO- morpholino carbonyl, CH _2-pyridinyl , or _{phenyl optionally substituted with 1 or 2 moieties selected from the group consisting of C 1-6} -alkyl, NHC _1-6 -alkyl and a heterocyclic ring optionally substituted with 1 or 2 moieties selected from the group consisting of =O ;

^1.1.1 R is H, C _1-6 - optionally substituted with alkyl, C _1-6 - alkyl, C _3-6 - cycloalkyl, C _1-6 - haloalkyl, CH ₂ CON (C _1-6 - alkyl,) ₂ , CH ₂ CO-azetidinyl, C _1-6 -alkylene-C _3-6 -cycloalkyl, CH ₂ -pyranyl, CH ₂ -tetrahydro-furanyl, CH ₂ -furanyl, C _1-6 -alkylene-OH or thiadiazolyl;

R ^1.1.2 is H, C _1-6 -alkyl, SO ₂ C _1-6 -alkyl; or

R ^1.1.1 and R ^1.1.2 are 4, 5 or 6 membered, optionally containing one O replacing a carbon atom of the ring and optionally substituted with one or two moieties selected from _{CH 2 OH} together form a carbon ring;

R ^1.2 is

C _1-6 -alkyl, C _3-6 -cycloalkyl, CH ₂ COO-C _1-6 -alkyl, CONR ^1.2.1 R ^1.2.2 , COO-C _1-6 -alkyl, CONH ₂ , OC _{1 -6} -alkyl, halogen, CN, SO ₂ N(C _1-6 -alkyl) ₂ , CO-pyrrolidinyl, CO-morpholinyl or hetero optionally substituted with _{1 or 2 C 1-6 -alkyl moieties} heteroaryl, optionally substituted with one or two moieties selected from the group consisting of aryl;

benzothiazolyl, indazolyl, dihydro-indole, each optionally substituted with one or two moieties selected from the group consisting of N(C _1-6 -alkyl) ₂ , CONH-C _{1-6 -alkyl, =O} lyl, indanyl, tetrahydro-quinolinyl;

• piperidinyl, optionally substituted with pyridinyl;

4,5-dihydronaphtho[2,1-d]thiazole, optionally substituted with NHCO-C _{1-6 -alkyl;}

R ^1.2.1 is H, C _1-6 -alkyl;

R ^1.2.2 is H, C _1-6 -alkyl;

R ^1.3 is selected from phenyl, pyrazolyl, isoxazolyl, pyrimidinyl, indolyl or oxadiazolyl, each C _1-6 -alkyl, C _3-6 -cycloalkyl, OC _1-6 -alkyl, OC optionally substituted with 1 or 2 moieties selected from the group consisting of _{1-6 -haloalkyl;}

R ² is selected from C _1-6 -alkylene-phenyl or C _1-6 -alkylene-naphthyl; C _1-6 - alkyl, C _1-6 - haloalkyl, OC _1-6 - alkyl, OC _1-6 - a haloalkyl, one or two moieties selected from the group consisting of halogen and optionally substituted, respectively; _{or CH 2} -thiophenyl, each optionally substituted with 1 or 2 moieties selected from halogen;

R ³ is H, C _1-4 -alkyl;

R ⁴ is H, C _1-4 -alkyl;

or R ³ and R ⁴ together form a CH ₂ —CH ₂ group.

Another embodiment of the present invention further comprises administration of a compound of formula 1 to a subject,

From here

A is CH ₂ , O or N-Me;

R ¹ is

· NHR ^1.1 , NMeR ^1.1 ;

· NHR ^1.2 , NMeR ^1.2 ;

· NHCH ₂ -R ^1.3 ;

NH-cyclohexyl, optionally substituted with 1 or 2 moieties selected from the group consisting of C _1-4 -alkyl, NHSO _{2 -phenyl, NHCONH-phenyl, halogen;}

NH-pyrrolidinyl, optionally substituted with 1 or 2 moieties selected from the group consisting of SO ₂ -C _1-4 -alkyl, COO-C _{1-4 -alkyl;}

piperidinyl, optionally substituted with 1 or 2 moieties selected from the group consisting of NHSO ₂ -C _{1-4 -alkyl, m-methoxyphenyl;}

· C _1-4 - alkyl, COO-C _1-4 - alkyl, C _1-4 - haloalkyl, OC _1-4 - alkyl, NO _2, 1 or 2 moieties selected from the group consisting of halogen, optionally substituted , dihydro-indolyl, dihydro-isoindolyl, tetrahydro-quinolinyl or tetrahydro-isoquinolinyl;

• a group selected from NHCH(pyridinyl)CH ₂ COO-C _1-6 -alkyl, NHCH(CH ₂ OC _1-6 -alkyl)-benzoimidazolyl, optionally substituted with Cl;

· or 1-amino cyclopentyl, optionally substituted with methyl-oxadiazole;

R ^1.1 is C _1-4 -alkyl, C _1-4 -haloalkyl, CH ₂ CON(C _1-4 -alkyl) ₂ , CH ₂ NHCONH-C _3-6 -cycloalkyl, CN, CONR ^1.1.1 R ^1.1.2 , COO-C _1-4 -alkyl, OC _1-4 -alkyl, SO ₂ -C _1-4 -alkyl, SO ₂ -C _1-4 -alkylene-OH, SO ₂ -C _3-6 - cycloalkyl, SO _{2-piperidinyl,} SO ₂ NH-C _1-4 - alkyl, SO ₂ N (C _1-4 - alkyl) _2, halogen, CN, CO- morpholino carbonyl, CH _2-pyridinyl , or imidazolidinyl, piperidinyl, oxazinanyl, pyrazolyl, triazolyl optionally substituted with 1 or 2 moieties selected from the group consisting of _{C 1-4} -alkyl, NHC _{1-4 -alkyl and =O} , phenyl, optionally substituted with 1 or 2 moieties selected from the group consisting of tetrazolyl, oxazolyl, oxadiazolyl, thiazolyl, pyridinyl, pyrimidinyl;

^1.1.1 R is H, C _1-4 - optionally substituted with alkyl, C _1-6 - alkyl, C _3-6 - cycloalkyl, C _1-4 - haloalkyl, CH ₂ CON (C _1-4 - alkyl,) ₂ , CH ₂ CO-azetidinyl, C _1-4 -alkylene-C _3-6 -cycloalkyl, CH ₂ -pyranyl, CH ₂ -tetrahydro-furanyl, CH ₂ -furanyl, C _1-4 -alkylene-OH or thiadiazolyl;

R ^1.1.2 is H, C _1-4 -alkyl, SO ₂ C _1-4 -alkyl; or

R ^1.1.1 and R ^1.1.2 are 4, 5 or 6 membered, optionally containing one O replacing a carbon atom of the ring and optionally substituted with one or two moieties selected from _{CH 2 OH} together form a carbon ring;

R ^1.2 is

C _1-4 -alkyl, C _3-6 -cycloalkyl, CH ₂ COO-C _1-4 -alkyl, CONR ^1.2.1 R ^1.2.2 , COO-C _1-4 -alkyl, CONH ₂ , OC _{1 -4} -alkyl, halogen, CO-pyrrolidinyl, CO-morpholinyl or pyrazolyl, triazolyl, tetrazolyl, oxazolyl, oxadiazolyl, optionally substituted with _{1 or 2 C 1-4 -alkyl moieties} pyridinyl, pyridazinyl, pyrrolyl, pyrazolyl, isoxazolyl, thiazolyl, thiadiazolyl, optionally substituted with one or two moieties selected from the group consisting of;

benzothiazolyl, indazolyl, dihydro-indole, each optionally substituted with one or two moieties selected from the group consisting of N(C _1-4 -alkyl) ₂ , CONH-C _{1-4 -alkyl, =O} lyl, indanyl, tetrahydro-quinolinyl;

• piperidinyl, optionally substituted with pyridinyl;

4,5-dihydronaphtho[2,1-d]thiazole, optionally substituted with NHCO-C _{1-4 -alkyl;}

R ^1.2.1 is H, C _1-4 -alkyl;

R ^1.2.2 is H, C _1-4 -alkyl;

R ^1.3 is selected from phenyl, pyrazolyl, isoxazolyl, pyrimidinyl, indolyl or oxadiazolyl, respectively C _1-4 -alkyl, C _3-6 -cycloalkyl, OC _1-4 -alkyl, OC optionally substituted with 1 or 2 moieties selected from the group consisting of _{1-4 -haloalkyl;}

R ² is selected from C _1-6 -alkylene-phenyl or C _1-6 -alkylene-naphthyl; C _1-4 - alkyl, C _1-4 - haloalkyl, OC _1-4 - alkyl, OC _1-4 - a haloalkyl, one or two moieties selected from the group consisting of halogen and optionally substituted, respectively; _{or CH 2} -thiophenyl, each optionally substituted with 1 or 2 moieties selected from halogen;

R ³ is H;

R ⁴ is H;

or R ³ and R ⁴ together form a CH ₂ —CH ₂ group.

Another embodiment of the present invention further comprises administration of a compound of formula 1 to a subject,

From here

A is CH ₂ , O or N-Me;

R ¹ is

· NHR ^1.1 , NMeR ^1.1 ;

· NHR ^1.2 , NMeR ^1.2 ;

· NHCH ₂ -R ^1.3 ;

NH-piperidinyl, optionally substituted with pyridinyl;

NH-cyclohexyl, optionally substituted with 1 or 2 moieties selected from the group consisting of t-butyl, NHSO _{2 -phenyl, NHCONH-phenyl, F;}

• NH-pyrrolidinyl, optionally substituted with 1 or 2 moieties selected from the group consisting of _{SO 2 -Me, COO-t-butyl;}

piperidinyl, optionally substituted with 1 or 2 moieties selected from the group consisting of NHSO _{2 -n-butyl, m-methoxyphenyl;}

Dihydro-indolyl, dihydro-isoindolyl, tetrahydro-qui, optionally substituted with 1 or 2 moieties selected from the group consisting of Me, COO-Me, CF ₃ , OMe, NO _{2 , F, Br} nolinyl or tetrahydro-isoquinolinyl;

• a group selected from NHCH(pyridinyl)CH ₂ COO-C _1-6 -alkyl, NHCH(CH ₂ OC _1-6 -alkyl)-benzoimidazolyl, optionally substituted with Cl;

· or 1-amino cyclopentyl, optionally substituted with methyl-oxadiazole;

R ^1.1 is Me, ethyl, t-butyl, CF ₃ , CH ₂ CONMe ₂ , CH ₂ NHCONH-cyclohexyl, CN, CONR ^1.1.1 R ^1.1.2 , COO-Me, COO-ethyl, O-Me, SO ₂ -Me, SO ₂ -ethyl, SO ₂ -CH ₂ CH ₂ -OH, SO ₂ -cyclopropyl, SO ₂ -piperidinyl, SO ₂ NH-ethyl, SO ₂ NMeEt, F, Cl, CO-morphopoly nyl, CH ₂ -pyridinyl, or imidazolidinyl, piperidinyl, oxazinanyl, pyrazolyl, triazolyl, tetrazolyl optionally substituted with 1 or 2 moieties selected from the group consisting of Me, NHMe and =O , phenyl optionally substituted with 1 or 2 moieties selected from the group consisting of oxazolyl, oxadiazolyl, thiazolyl, pyridinyl, pyrimidinyl;

R ^1.1.1 is methyl, ethyl, t-butyl, isopropyl, cyclopropyl, CH ₂ -isopropyl, CH ₂ -t-butyl, CH(CH ₃ )CH ₂ CH ₃ , optionally substituted with H, Me _{CH 2 CHF 2, CH 2 CONMe} 2, CH 2 CO- azetidinyl, CH ₂ - cyclopropyl, CH _{2-cyclo-butyl,} CH _2-pyranyl, CH _{2-tetrahydro-furanyl,} CH _2-furanyl , CH ₂ CH ₂ OH or thiadiazolyl;

R ^1.1.2 is H, methyl, ethyl, SO ₂ Me, SO ₂ ethyl; or

R ^1.1.1 and R ^1.1.2 are 4, 5 or 6 membered, optionally containing one O replacing a carbon atom of the ring and optionally substituted with one or two moieties selected from _{CH 2 OH} together form a carbon ring;

R ^1.2 is

methyl, ethyl, propyl, butyl, cyclopropyl, CH ₂ COO-ethyl, CONR ^1.2.1 R ^1.2.2 , COO-methyl, COO-ethyl, CONH ₂ , OMe, Cl, Br, CO-pyrrolidinyl, Pyridinyl, pyri, optionally substituted with one or two moieties selected from the group consisting of CO-morpholinyl or pyrazolyl, triazolyl, tetrazolyl, oxazolyl, oxadiazolyl optionally substituted with one or two methyl moieties dazinyl, pyrrolyl, pyrazolyl, isoxazolyl, thiazolyl, thiadiazolyl;

benzothiazolyl, indazolyl, dihydro-indolyl, indanyl, tetrahydro-quinolinyl, each optionally substituted with one or two moieties selected from the group consisting of NMe _{2 , CONH-Me, =O;}

4,5-dihydronaphtho[2,1-d]thiazole, optionally substituted with NHCO-Me;

R ^1.2.1 is H, or methyl;

R ^1.2.2 is H, or methyl;

R ^1.3 is selected from phenyl, pyrazolyl, isoxazolyl, pyrimidinyl, indolyl or oxadiazolyl, each 1 selected from the group consisting of _{methyl, ethyl, propyl, cyclopentyl, O-Me, O-CHF 2} or optionally substituted with two residues;

R ² is CH ₂ - or phenyl-CH ₂ - is selected from naphthyl; each optionally substituted with 1 or 2 moieties selected from the group consisting of CH ₃ , CF ₃ , OCF _{3 , F, Cl, Br, ethyl; or CH 2} -thiophenyl, each optionally substituted with 1 or 2 moieties selected from the group consisting of Cl, Br;

R ³ is H;

R ⁴ is H;

or R ³ and R ⁴ together form a CH ₂ —CH ₂ group.

Another embodiment of the present invention further comprises administration of a compound of formula 1 to a subject,

From here

A is CH ₂ , O or N-Me;

R ¹ is

· NHR ^1.1 ,

· NHR ^1.2 ,

R ^1.1 is Me, ethyl, butyl, CF ₃ , CH ₂ CONMe ₂ , CH ₂ NHCONH-cyclohexyl, CN, CONR ^1.1.1 R ^1.1.2 , COO-Me, COO-ethyl, O-Me, SO ₂ — Me, SO ₂ -ethyl, SO ₂ -CH ₂ CH ₂ -OH, SO ₂ -cyclopropyl, SO ₂ -piperidinyl, SO ₂ NH-ethyl, SO ₂ NMeEt, F, Cl, CO-morpholinyl, CH ₂ -pyridinyl, or imidazolidinyl, piperidinyl, oxazinanyl, pyrazolyl, triazolyl, tetrazolyl, oxa optionally substituted with 1 or 2 moieties selected from the group consisting of Me, NHMe and =O phenyl optionally substituted with 1 or 2 moieties selected from the group consisting of zolyl, oxadiazolyl, thiazolyl, pyridinyl, pyrimidinyl;

R ^1.1.1 is methyl, ethyl, t-butyl, isopropyl, cyclopropyl, CH ₂ -isopropyl, CH ₂ -t-butyl, CH(CH ₃ )CH ₂ CH ₃ , optionally substituted with H, Me _{CH 2 CHF 2, CH 2 CONMe} 2, CH 2 CO- azetidinyl, CH ₂ - cyclopropyl, CH _{2-cyclo-butyl,} CH _2-pyranyl, CH _{2-tetrahydro-furanyl,} CH _2-furanyl , CH ₂ CH ₂ OH or thiadiazolyl;

R ^1.1.2 is H, methyl, ethyl, SO ₂ Me, SO ₂ ethyl; or

R ^1.1.1 and R ^1.1.2 are 4, 5 or 6 membered, optionally containing one O replacing a carbon atom of the ring and optionally substituted with one or two moieties selected from _{CH 2 OH} together form a carbon ring;

R ^1.2 is

methyl, ethyl, propyl, butyl, cyclopropyl, CH ₂ COO-ethyl, CONR ^1.2.1 R ^1.2.2 , COO-methyl, COO-ethyl, CONH ₂ , OMe, Cl, Br, CO-pyrrolidinyl, Pyridinyl, pyri, optionally substituted with one or two moieties selected from the group consisting of CO-morpholinyl or pyrazolyl, triazolyl, tetrazolyl, oxazolyl, oxadiazolyl optionally substituted with one or two methyl moieties dazinyl, pyrrolyl, pyrazolyl, isoxazolyl, thiazolyl, thiadiazolyl;

benzothiazolyl, indazolyl, dihydro-indolyl, indanyl, tetrahydro-quinolinyl, each optionally substituted with one or two moieties selected from the group consisting of NMe _{2 , CONH-Me, =O;}

4,5-dihydronaphtho[2,1-d]thiazole, optionally substituted with NHCO-Me;

R ^1.2.1 is H, or methyl;

R ^1.2.2 is H, or methyl;

R ² is CH ₂ - or phenyl-CH ₂ - is selected from naphthyl; each optionally substituted with 1 or 2 moieties selected from the group consisting of CH ₃ , CF ₃ , OCF _{3 , F, Cl, Br, ethyl;}

R ³ is H;

R ⁴ is H.

Another embodiment of the present invention further comprises administration of a compound of formula 1 to a subject,

From here

A is CH ₂ , O or N-Me;

R ¹ is

· NHR ^1.1 , NMeR ^1.1 ;

· NHR ^1.2 , NMeR ^1.2 ;

· NHCH ₂ -R ^1.3 ; and

R ^1.1 is Me, ethyl, butyl, CF ₃ , CH ₂ CONMe ₂ , CH ₂ NHCONH-cyclohexyl, CN, CONR ^1.1.1 R ^1.1.2 , COO-Me, COO-ethyl, O-Me, SO ₂ — Me, SO ₂ -ethyl, SO ₂ -CH ₂ CH ₂ -OH, SO ₂ -cyclopropyl, SO ₂ -piperidinyl, SO ₂ NH-ethyl, SO ₂ NMeEt, F, Cl, CO-morpholinyl, CH ₂ -pyridinyl, or imidazolidinyl, piperidinyl, oxazinanyl, pyrazolyl, triazolyl, tetrazolyl, oxa optionally substituted with 1 or 2 moieties selected from the group consisting of Me, NHMe and =O phenyl optionally substituted with 1 or 2 moieties selected from the group consisting of zolyl, oxadiazolyl, thiazolyl, pyridinyl, pyrimidinyl;

R ^1.1.1 is methyl, ethyl, t-butyl, isopropyl, cyclopropyl, CH ₂ -isopropyl, CH ₂ -t-butyl, CH(CH ₃ )CH ₂ CH ₃ , optionally substituted with H, Me _{CH 2 CHF 2, CH 2 CONMe} 2, CH 2 CO- azetidinyl, CH ₂ - cyclopropyl, CH _{2-cyclo-butyl,} CH _2-pyranyl, CH _{2-tetrahydro-furanyl,} CH _2-furanyl , CH ₂ CH ₂ OH or thiadiazolyl;

R ^1.1.2 is H, methyl, ethyl, SO ₂ Me, SO ₂ ethyl; or

R ^1.1.1 and R ^1.1.2 are 4, 5 or 6 membered, optionally containing one O replacing a carbon atom of the ring and optionally substituted with one or two moieties selected from _{CH 2 OH} together form a carbon ring;

R ^1.2 is

methyl, ethyl, propyl, butyl, cyclopropyl, CH ₂ COO-ethyl, CONR ^1.2.1 R ^1.2.2 , COO-methyl, COO-ethyl, CONH ₂ , OMe, Cl, Br, CO-pyrrolidinyl, Pyridinyl, pyri, optionally substituted with one or two moieties selected from the group consisting of CO-morpholinyl or pyrazolyl, triazolyl, tetrazolyl, oxazolyl, oxadiazolyl optionally substituted with one or two methyl moieties dazinyl, pyrrolyl, pyrazolyl, isoxazolyl, thiazolyl, thiadiazolyl;

benzothiazolyl, indazolyl, dihydro-indolyl, indanyl, tetrahydro-quinolinyl, each optionally substituted with one or two moieties selected from the group consisting of NMe _{2 , CONH-Me, =O;}

4,5-dihydronaphtho[2,1-d]thiazole, optionally substituted with NHCO-Me;

R ^1.2.1 is H, or methyl;

R ^1.2.2 is H, or methyl;

R ^1.3 is selected from phenyl, pyrazolyl, isoxazolyl, pyrimidinyl, indolyl or oxadiazolyl, each 1 selected from the group consisting of _{methyl, ethyl, propyl, cyclopentyl, O-Me, O-CHF 2} or optionally substituted with two residues;

R ² is CH ₂ - or phenyl-CH ₂ - is selected from naphthyl; each optionally substituted with 1 or 2 moieties selected from the group consisting of CH ₃ , CF ₃ , OCF _{3 , F, Cl, Br, ethyl; or CH 2} -thiophenyl, each optionally substituted with 1 or 2 moieties selected from the group consisting of Cl, Br;

R ³ is H;

R ⁴ is H;

or R ³ and R ⁴ together form a CH ₂ —CH ₂ group.

Another embodiment of the present invention further comprises administration of a compound of formula 1 to a subject,

From here

A is CH ₂ , O or N-Me;

R ¹ is

· NHR ^1.1 , NMeR ^1.1 ;

R ^1.1 is Me, ethyl, butyl, CF ₃ , CH ₂ CONMe ₂ , CH ₂ NHCONH-cyclohexyl, CN, CONR ^1.1.1 R ^1.1.2 , COO-Me, COO-ethyl, O-Me, SO ₂ — Me, SO ₂ -ethyl, SO ₂ -CH ₂ CH ₂ -OH, SO ₂ -cyclopropyl, SO ₂ -piperidinyl, SO ₂ NH-ethyl, SO ₂ NMeEt, F, Cl, CO-morpholinyl, CH ₂ -pyridinyl, or imidazolidinyl, piperidinyl, oxazinanyl, pyrazolyl, triazolyl, tetrazolyl, oxa optionally substituted with 1 or 2 moieties selected from the group consisting of Me, NHMe and =O phenyl optionally substituted with 1 or 2 moieties selected from the group consisting of zolyl, oxadiazolyl, thiazolyl, pyridinyl, pyrimidinyl;

R ^1.1.1 is methyl, ethyl, t-butyl, isopropyl, cyclopropyl, CH ₂ -isopropyl, CH ₂ -t-butyl, CH(CH ₃ )CH ₂ CH ₃ , optionally substituted with H, Me _{CH 2 CHF 2, CH 2 CONMe} 2, CH 2 CO- azetidinyl, CH ₂ - cyclopropyl, CH _{2-cyclo-butyl,} CH _2-pyranyl, CH _{2-tetrahydro-furanyl,} CH _2-furanyl , CH ₂ CH ₂ OH or thiadiazolyl;

R ^1.1.2 is H, methyl, ethyl, SO ₂ Me, SO ₂ ethyl; or

R ^1.1.1 and R ^1.1.2 are 4, 5 or 6 membered, optionally containing one O replacing a carbon atom of the ring and optionally substituted with one or two moieties selected from _{CH 2 OH} together form a carbon ring;

R ² is CH ₂ - or phenyl-CH ₂ - is selected from naphthyl; each optionally substituted with 1 or 2 moieties selected from the group consisting of CH ₃ , CF ₃ , OCF _{3 , F, Cl, Br, ethyl; or CH 2} -thiophenyl, each optionally substituted with 1 or 2 moieties selected from the group consisting of Cl, Br;

R ³ is H;

R ⁴ is H;

or R ³ and R ⁴ together form a CH ₂ —CH ₂ group.

Another embodiment of the present invention further comprises administration of a compound of formula 1 to a subject,

From here

A is CH ₂ , O or N-Me;

R ¹ is

· NHR ^1.1 , NMeR ^1.1 ;

R ^1.1 is Me, ethyl, butyl, CF ₃ , CH ₂ CONMe ₂ , CH ₂ NHCONH-cyclohexyl, CN, CONR ^1.1.1 R ^1.1.2 , COO-Me, COO-ethyl, O-Me, SO ₂ — Me, SO ₂ -ethyl, SO ₂ -CH ₂ CH ₂ -OH, SO ₂ -cyclopropyl, SO ₂ -piperidinyl, SO ₂ NH-ethyl, SO ₂ NMeEt, F, Cl, CO-morpholinyl, CH ₂ -pyridinyl, or imidazolidinyl, piperidinyl, oxazinanyl, pyrazolyl, triazolyl, tetrazolyl, oxa optionally substituted with 1 or 2 moieties selected from the group consisting of Me, NHMe and =O phenyl optionally substituted with 1 or 2 moieties selected from the group consisting of zolyl, oxadiazolyl, thiazolyl, pyridinyl, pyrimidinyl;

R ^1.1.1 is methyl, ethyl, t-butyl, isopropyl, cyclopropyl, CH ₂ -isopropyl, CH ₂ -t-butyl, CH(CH ₃ )CH ₂ CH ₃ , optionally substituted with H, Me _{CH 2 CHF 2, CH 2 CONMe} 2, CH 2 CO- azetidinyl, CH ₂ - cyclopropyl, CH _{2-cyclo-butyl,} CH _2-pyranyl, CH _{2-tetrahydro-furanyl,} CH _2-furanyl , CH ₂ CH ₂ OH or thiadiazolyl;

R ^1.1.2 is H, methyl, ethyl, SO ₂ Me, SO ₂ ethyl; or

R ^1.1.1 and R ^1.1.2 are 4, 5 or 6 membered, optionally containing one O replacing a carbon atom of the ring and optionally substituted with one or two moieties selected from _{CH 2 OH} together form a carbon ring;

R ² is as defined in Table 1 below;

R ³ is H;

R ⁴ is H;

or R ³ and R ⁴ together form a CH ₂ —CH ₂ group.

Another embodiment of the present invention further comprises administration of a compound of formula 1 to a subject,

From here

A is CH ₂ , O or N-Me;

R ¹ is

· NHR ^1.1 , NMeR ^1.1 ;

R ^1.1 is Me, ethyl, butyl, CF ₃ , CH ₂ CONMe ₂ , CH ₂ NHCONH-cyclohexyl, CN, CONR ^1.1.1 R ^1.1.2 , COO-Me, COO-ethyl, O-Me, SO ₂ — Me, SO ₂ -ethyl, SO ₂ -CH ₂ CH ₂ -OH, SO ₂ -cyclopropyl, SO ₂ -piperidinyl, SO ₂ NH-ethyl, SO ₂ NMeEt, F, Cl, CO-morpholinyl, CH ₂ -pyridinyl, or imidazolidinyl, piperidinyl, oxazinanyl, pyrazolyl, triazolyl, tetrazolyl, oxa optionally substituted with 1 or 2 moieties selected from the group consisting of Me, NHMe and =O phenyl optionally substituted with 1 or 2 moieties selected from the group consisting of zolyl, oxadiazolyl, thiazolyl, pyridinyl, and pyrimidinyl;

R ^1.1.1 and R ^1.1.2 are 4, 5 or 6 membered, optionally containing one O replacing a carbon atom of the ring and optionally substituted with one or two moieties selected from _{CH 2 OH} together form a carbon ring;

R ² is as defined in Table 1 below;

R ³ is H;

R ⁴ is H;

Another embodiment of the present invention further comprises administration of a compound of formula 1 to a subject,

From here

A is CH ₂ , O or N-Me;

R ¹ is

· NHR ^1.1 , NMeR ^1.1 ;

R ^1.1 is Me, ethyl, butyl, CF ₃ , CH ₂ CONMe ₂ , CH ₂ NHCONH-cyclohexyl, CN, CONR ^1.1.1 R ^1.1.2 , COO-Me, COO-ethyl, O-Me, F, Cl phenyl, optionally substituted with 1 or 2 residues selected from the group consisting of;

R ^1.1.1 is methyl, ethyl, t-butyl, isopropyl, cyclopropyl, CH ₂ -isopropyl, CH ₂ -t-butyl, CH(CH ₃ )CH ₂ CH ₃ , optionally substituted with H, Me _{CH 2 CHF 2, CH 2 CONMe} 2, CH 2 CO- azetidinyl, CH ₂ - cyclopropyl, CH _{2-cyclo-butyl,} CH _2-pyranyl, CH _{2-tetrahydro-furanyl,} CH _2-furanyl , CH ₂ CH ₂ OH or thiadiazolyl;

R ^1.1.2 is H, methyl, ethyl, SO ₂ Me, SO ₂ ethyl;

R ² is as defined in Table 1 below;

R ³ is H;

R ⁴ is H;

Another embodiment of the present invention further comprises administration of a compound of formula 1 to a subject,

From here

A is CH ₂ , O or N-Me;

R ¹ is

· NHR ^1.1 , NMeR ^1.1 ;

R ^1.1 is selected from the group consisting of SO ₂ -Me, SO ₂ -ethyl, SO ₂ -CH ₂ CH ₂ -OH, SO ₂ -cyclopropyl, SO ₂ -piperidinyl, SO ₂ NH-ethyl, SO _{2 NMeEt} phenyl, optionally substituted with 1 or 2 moieties;

R ^1.1.1 is methyl, ethyl, t-butyl, isopropyl, cyclopropyl, CH ₂ -isopropyl, CH ₂ -t-butyl, CH(CH ₃ )CH ₂ CH ₃ , optionally substituted with H, Me _{CH 2 CHF 2, CH 2 CONMe} 2, CH 2 CO- azetidinyl, CH ₂ - cyclopropyl, CH _{2-cyclo-butyl,} CH _2-pyranyl, CH _{2-tetrahydro-furanyl,} CH _2-furanyl , CH ₂ CH ₂ OH or thiadiazolyl;

R ^1.1.2 is H, methyl, ethyl, SO ₂ Me, SO ₂ ethyl;

R ² is as defined in Table 1 below;

R ³ is H;

R ⁴ is H;

Another embodiment of the present invention further comprises administration of a compound of formula 1 to a subject,

From here

A is CH ₂ , O or N-Me;

R ¹ is

· NHR ^1.1 , NMeR ^1.1 ;

R ^1.1 is Me, ethyl, butyl, CF ₃ , CH ₂ CONMe ₂ , CH ₂ NHCONH-cyclohexyl, CN, CONR ^1.1.1 R ^1.1.2 , COO-Me, COO-ethyl, O-Me, SO ₂ — Me, SO ₂ -ethyl, SO ₂ -CH ₂ CH ₂ -OH, SO ₂ -cyclopropyl, SO ₂ -piperidinyl, SO ₂ NH-ethyl, SO ₂ NMeEt, F, Cl, CO-morpholinyl, CH ₂ -pyridinyl, or imidazolidinyl, piperidinyl, oxazinanyl, pyrazolyl, triazolyl, tetrazolyl, oxa optionally substituted with 1 or 2 moieties selected from the group consisting of Me, NHMe and =O phenyl optionally substituted with 1 or 2 moieties selected from the group consisting of zolyl, oxadiazolyl, thiazolyl, pyridinyl, and pyrimidinyl;

R ^1.1.1 is methyl, ethyl, t-butyl, isopropyl, cyclopropyl, CH ₂ -isopropyl, CH ₂ -t-butyl, CH(CH ₃ )CH ₂ CH ₃ , optionally substituted with H, Me _{CH 2 CHF 2, CH 2 CONMe} 2, CH 2 CO- azetidinyl, CH ₂ - cyclopropyl, CH _{2-cyclo-butyl,} CH _2-pyranyl, CH _{2-tetrahydro-furanyl,} CH _2-furanyl , CH ₂ CH ₂ OH or thiadiazolyl;

R ^1.1.2 is H, methyl, ethyl, SO ₂ Me, SO ₂ ethyl;

R ² is as defined in Table 1 below;

R ³ is H;

R ⁴ is H;

Another embodiment of the present invention further comprises administration of a compound of formula 1 to a subject,

From here

A is CH ₂ , O or N-Me;

R ¹ is

· NHR ^1.2 , NMeR ^1.2 ;

R ^1.2 is

methyl, ethyl, propyl, butyl, cyclopropyl, CH ₂ COO-ethyl, CONR ^1.2.1 R ^1.2.2 , COO-methyl, COO-ethyl, CONH ₂ , OMe, Cl, Br, CO-pyrrolidinyl, Pyridinyl, pyridazinyl, pyrrolyl, optionally substituted with one or two moieties selected from the group consisting of CO-morpholinyl or methyl optionally substituted pyrazolyl, triazolyl, tetrazolyl, oxazolyl, oxadiazolyl , pyrazolyl, isoxazolyl, thiazolyl, thiadiazolyl;

benzothiazolyl, indazolyl, dihydro-indolyl, indanyl, tetrahydro-quinolinyl, each optionally substituted with one or two moieties selected from the group consisting of NMe _{2 , CONH-Me, =O;}

4,5-dihydronaphtho[2,1-d]thiazole, optionally substituted with NHCO-Me;

R ^1.2.1 is H, or methyl;

R ^1.2.2 is H, or methyl;

R ² is CH ₂ - or phenyl-CH ₂ - is selected from naphthyl; each optionally substituted with 1 or 2 moieties selected from the group consisting of CH ₃ , CF ₃ , OCF _{3 , F, Cl, Br, ethyl; or CH 2} -thiophenyl, each optionally substituted with 1 or 2 moieties selected from the group consisting of Cl, Br;

R ³ is H;

R ⁴ is H;

or R ³ and R ⁴ together form a CH ₂ —CH ₂ group.

Another embodiment of the present invention further comprises administration of a compound of formula 1 to a subject,

From here

A is CH ₂ , O or N-Me;

R ¹ is

· NHR ^1.2 , NMeR ^1.2 ;

R ^1.2 is

methyl, ethyl, propyl, butyl, cyclopropyl, CH ₂ COO-ethyl, CONR ^1.2.1 R ^1.2.2 , COO-methyl, COO-ethyl, CONH ₂ , OMe, Cl, Br, CO-pyrrolidinyl, Pyridinyl, pyridazinyl, pyrrolyl, optionally substituted with one or two moieties selected from the group consisting of CO-morpholinyl or methyl optionally substituted pyrazolyl, triazolyl, tetrazolyl, oxazolyl, oxadiazolyl , pyrazolyl, isoxazolyl, thiazolyl, thiadiazolyl;

R ^1.2.1 is H, or methyl;

R ^1.2.2 is H, or methyl;

R ² is CH ₂ - or phenyl-CH ₂ - is selected from naphthyl; each optionally substituted with 1 or 2 moieties selected from the group consisting of CH ₃ , CF ₃ , OCF _{3 , F, Cl, Br, ethyl; or CH 2} -thiophenyl, each optionally substituted with 1 or 2 moieties selected from the group consisting of Cl, Br;

R ³ is H;

R ⁴ is H;

or R ³ and R ⁴ together form a CH ₂ —CH ₂ group.

Another embodiment of the present invention further comprises administration of a compound of formula 1 to a subject,

From here

A is CH ₂ , O or N-Me;

R ¹ is

· NHCH ₂ -R ^1.3 ;

R ^1.3 is selected from phenyl, pyrazolyl, isoxazolyl, pyrimidinyl, indolyl or oxadiazolyl, each 1 selected from the group consisting of _{methyl, ethyl, propyl, cyclopentyl, O-Me, O-CHF 2} or optionally substituted with two residues;

R ² is CH ₂ - or phenyl-CH ₂ - is selected from naphthyl; each optionally substituted with 1 or 2 moieties selected from the group consisting of CH ₃ , CF ₃ , OCF _{3 , F, Cl, Br, ethyl; or CH 2} -thiophenyl, each optionally substituted with 1 or 2 moieties selected from the group consisting of Cl, Br;

R ³ is H;

R ⁴ is H;

or R ³ and R ⁴ together form a CH ₂ —CH ₂ group.

Another embodiment of the present invention further comprises administration of a compound of formula 1 to a subject,

From here

A is CH ₂ , O or N-Me;

R ¹ is

NH-piperidinyl, optionally substituted with pyridinyl;

NH-cyclohexyl, optionally substituted with 1 or 2 moieties selected from the group consisting of t-butyl, NHSO _{2 -phenyl, NHCONH-phenyl, F;}

• NH-pyrrolidinyl, optionally substituted with 1 or 2 moieties selected from the group consisting of _{SO 2 -Me, COO-t-butyl;}

piperidinyl, optionally substituted with 1 or 2 moieties selected from the group consisting of NHSO _{2 -n-butyl, m-methoxyphenyl;}

Dihydro-indolyl, dihydro-isoindolyl, tetrahydro-qui, optionally substituted with 1 or 2 moieties selected from the group consisting of Me, COO-Me, CF ₃ , OMe, NO _{2 , F, Br} nolinyl or tetrahydro-isoquinolinyl;

• a group selected from NHCH(pyridinyl)CH ₂ COO-C _1-6 -alkyl, NHCH(CH ₂ OC _1-6 -alkyl)-benzoimidazolyl, optionally substituted with Cl;

· or 1-amino cyclopentyl, optionally substituted with methyl-oxadiazole;

R ² is CH ₂ - or phenyl-CH ₂ - is selected from naphthyl; each optionally substituted with 1 or 2 moieties selected from the group consisting of CH ₃ , CF ₃ , OCF _{3 , F, Cl, Br, ethyl; or CH 2} -thiophenyl, each optionally substituted with 1 or 2 moieties selected from the group consisting of Cl, Br;

R ³ is H;

R ⁴ is H;

or R ³ and R ⁴ together form a CH ₂ —CH ₂ group.

Another embodiment of the invention further comprises administration of a compound of formula 1 to a subject, wherein A is CH ₂ , O or N-Me; R ¹ is NHR ^1.2 , NMeR ^1.2 ; R ² is as defined in Table 1 below; R ³ is H; R ⁴ is H; and R ^1.2 is

methyl, ethyl, propyl, t-butyl, cyclopropyl, CONR ^1.2.1 R ^1.2.2 , COO-methyl, COO-ethyl, CONH ₂ , OMe, Cl, Br, CO-pyrrolidinyl, CO-morphopoly pyridinyl, optionally substituted with one or two moieties selected from the group consisting of pyrazolyl, triazolyl, tetrazolyl, oxazolyl, oxadiazolyl, optionally substituted with nyl or methyl;

· pyrrolyl, optionally substituted with one or two moieties selected from the group consisting of methyl, ethyl, COO-methyl, COO-ethyl;

· pyrazolyl, optionally substituted with one or two moieties selected from the group consisting of methyl, ethyl, propyl, butyl, cyclopropyl, COO-ethyl, CO-pyrrolidinyl;

· isoxazolyl, optionally substituted with one or two moieties selected from the group consisting of t-butyl, COO-ethyl;

thiazolyl optionally substituted with one or two moieties selected from the group consisting of methyl, n-propyl, i-propyl, butyl, COO-methyl, COO-ethyl, CONR ^1.2.1 R ^1.2.2;

,·COO-thiadiazolyl optionally substituted with one or two residues selected from the group consisting of ethyl;

benzothiazolyl, indazolyl, dihydro-indolyl, indanyl, tetrahydro-quinolinyl, each optionally substituted with one or two moieties selected from the group consisting of NMe _{2 , CONH-Me, =O;}

4,5-dihydronaphtho[2,1-d]thiazole, optionally substituted with NHCO-Me;

R ^1.2.1 is H, or methyl;

R ^1.2.2 is H, methyl.

Another embodiment of the invention further comprises administration of a compound of formula 1 to a subject, wherein A is CH ₂ , O or N-Me; R ¹ is NHR ^1.2 , NMeR ^1.2 ; R ² is as defined in Table 1 below; R ³ is H; R ⁴ is H; and R ^1.2 is

optionally with one or two residues selected from the group consisting of methyl, ethyl, i-propyl, n-butyl, CONR ^1.2.1 R ^1.2.2 , COO-methyl, COO-ethyl, CONH _{2 , OMe, Cl, Br} substituted, pyridinyl;

· pyrrolyl, optionally substituted with one or two moieties selected from the group consisting of methyl, ethyl, COO-methyl, COO-ethyl;

· pyrazolyl optionally substituted with one or two moieties selected from the group consisting of methyl, ethyl, cyclopropyl, COO-ethyl, CO-pyrrolidinyl;

· isoxazolyl, optionally substituted with one or two moieties selected from the group consisting of t-butyl, COO-ethyl;

thiazolyl optionally substituted with one or two moieties selected from the group consisting of methyl, n-propyl, i-propyl, butyl, COO-methyl, COO-ethyl, CONR ^1.2.1 R ^1.2.2;

,·COO-thiadiazolyl optionally substituted with one or two residues selected from the group consisting of ethyl;

benzothiazolyl, indazolyl, dihydro-indolyl, indanyl, tetrahydro-quinolinyl, each optionally substituted with one or two moieties selected from the group consisting of NMe _{2 , CONH-Me, =O;}

4,5-dihydronaphtho[2,1-d]thiazole, optionally substituted with NHCO-Me;

R ^1.2.1 is H, or methyl;

R ^1.2.2 is H, methyl.

Another embodiment of the present invention further comprises administration of a compound of formula 1 to a subject,

· A is CH ₂ , O or N-Me; R ¹ is NHR ^1.2 , NMeR ^1.2 ; R ² is as defined in Table 1 below; R ³ is H; R ⁴ is H; R ^1.2 is one or two residues selected from the group consisting of methyl, ethyl, i-propyl, n-butyl, CONR ^1.2.1 R ^1.2.2 , COO-methyl, COO-ethyl, CONH _{2 , OMe, Cl, Br} pyridinyl, optionally substituted with ; R ^1.2.1 is H, or methyl and R ^1.2.2 is H, methyl.

· A is CH ₂ , O or N-Me; R ¹ is NHR ^1.2 , NMeR ^1.2 ; R ² is as defined in Table 1 below; R ³ is H; R ⁴ is H; R ^1.2 is pyrrolyl, optionally substituted with one or two moieties selected from the group consisting of methyl, ethyl, COO-methyl, COO-ethyl; R ^1.2.1 is H, or methyl and R ^1.2.2 is H, methyl.

· A is CH ₂ , O or N-Me; R ¹ is NHR ^1.2 , NMeR ^1.2 ; R ² is as defined in Table 1 below; R ³ is H; R ⁴ is H; R ^1.2 is pyrazolyl, optionally substituted with one or two moieties selected from the group consisting of methyl, ethyl, cyclopropyl, COO-ethyl, CO-pyrrolidinyl; R ^1.2.1 is H, or methyl and R ^1.2.2 is H, methyl.

· A is CH ₂ , O or N-Me; R ¹ is NHR ^1.2 , NMeR ^1.2 ; R ² is as defined in Table 1 below; R ³ is H; R ⁴ is H; R ^1.2 is isoxazolyl, optionally substituted with one or two moieties selected from the group consisting of t-butyl, COO-ethyl; R ^1.2.1 is H, or methyl and R ^1.2.2 is H, methyl.

· A is CH ₂ , O or N-Me; R ¹ is NHR ^1.2 , NMeR ^1.2 ; R ² is as defined in Table 1 below; R ³ is H; R ⁴ is H; R ^1.2 is thiazolyl optionally substituted with one or two moieties selected from the group consisting of methyl, n-propyl, i-propyl, butyl, COO-methyl, COO-ethyl, CONR ^1.2.1 R ^1.2.2; R ^1.2.1 is H, or methyl and R ^1.2.2 is H, methyl.

· A is CH ₂ , O or N-Me; R ¹ is NHR ^1.2 , NMeR ^1.2 ; R ² is as defined in Table 1 below; R ³ is H; R ⁴ is H; R ^1.2 is thiadiazolyl optionally substituted with one or two moieties selected from the group consisting of COO-ethyl; R ^1.2.1 is H, or methyl and R ^1.2.2 is H, methyl.

· A is CH ₂ , O or N-Me; R ¹ is NHR ^1.2 , NMeR ^1.2 ; R ² is as defined in Table 1 below; R ³ is H; R ⁴ is H; R ^1.2 is benzothiazolyl, indazolyl, dihydro-indolyl, indanyl, tetrahydro-quinolinyl, each optionally substituted with one or two moieties selected from the group consisting of NMe _{2 , CONH-Me, =O} ; R ^1.2.1 is H, or methyl and R ^1.2.2 is H, methyl.

Another embodiment of the present invention further comprises administration of a compound of formula 1 to a subject, wherein all groups are as described above, except that ^{R 1.3 is selected from:}

phenyl, optionally substituted with O-CHF _{2 ;}

pyrazolyl optionally substituted with methyl or ethyl;

isoxazolyl, optionally substituted with propyl;

pyrimidinyl, optionally substituted with O-Me;

· Indolyl;

• oxadiazolyl, optionally substituted with cyclopentyl.

Another embodiment of the present invention further comprises administration of the compound of formula 1 to a subject, wherein all groups are as described above except for A is CH _{2 .}

Another embodiment of the present invention further comprises administration of the compound of formula 1 to a subject, wherein all groups are as described above except for A is O.

Another embodiment of the present invention further comprises administration of the compound of formula 1 to a subject, wherein all groups are as described above except for A is NMe.

Another embodiment of the present invention is a compound of Formula 1,

A is CH ₂ , O or N-Me;

R ¹ is

is selected from;

R ² is

is selected from;

R ³ is H;

R ⁴ is H;

or R ³ and R ⁴ together form a CH ₂ —CH ₂ group.

Another embodiment of the present invention is a compound of Formula 1, wherein A is as described above; R ³ is H; R ⁴ is H; R ² is as defined in Table 1 below;

R ¹ is selected from.

Another embodiment of the present invention is a compound of Formula 1, wherein A is as described above; R ³ is H; R ⁴ is H; R ² is as defined in Table 1 below; R ¹ is

is selected from

Another embodiment of the present invention is a compound of Formula 1, wherein A is as described above; R ³ is H; R ⁴ is H; R ² is as defined in Table 1 below; R ¹ is

is selected from

Another embodiment of the present invention is a compound of Formula 1, wherein A is as described above; R ³ is H; R ⁴ is H; R ² is as defined in Table 1 below; R ¹ is

is selected from

Another embodiment of the present invention is a compound of Formula 1, wherein A is as described above; R ³ is H; R ⁴ is H; R ² is as defined in Table 1 below; R ¹ is,

is selected from

Another embodiment of the present invention is a compound of Formula 1, wherein A is as described above; R ³ is H; R ⁴ is H; R ² is as defined in Table 1 below; R ¹ is,

is selected from

Table 1: R ² is one of definitions 1 to 4 of Table 1 below.

Another embodiment of the invention further comprises administering to a subject a compound of Formula 1, wherein the compound of Formula 1 is in the form of individual optical isomers, individual enantiomers or mixtures of racemates, e.g., enantiomers It exists in pure compound form.

Another embodiment of the present invention further comprises administering to a subject a compound of Formula 1, wherein the compound of Formula 1 is in the form of an acid addition salt thereof with a pharmacologically acceptable acid and optionally a solvate and/or hydrate exist in the form

b. co-crystals and salts

A further embodiment of the present invention further comprises the administration of a co-crystal of a compound of formula (2) (hereinafter) to a subject. In general, for groups comprising two or more subgroups in this "Co-Crystals and Salts" section, the first named subgroup is the point of radical attachment, the substituent "C _1-3 -alkyl-aryl" is C _{1 - 3} -means an aryl group attached to an alkyl group, the latter of which is attached to the core or to the group to which the substituent is attached.

2

2

in the above formula

R ¹ is C _1-6 -alkyl, C _1-6 -haloalkyl, OC _1-6 -haloalkyl, halogen;

m is 1, 2 or 3;

R ^2a and R ^2b are each independently H, C _1-6 -alkyl, C _1-6 -alkenyl, C _1-6 -alkynyl, C _3-6 -cycloalkyl, COO-C _1-6 -alkyl, OC _1-6 -alkyl, CONR ^2b.1 R ^2b.2 , halogen;

R ^2b.1 is H, C _1-6 -alkyl, C _0-4 -alkyl-C _3-6 -cycloalkyl, C _1-6 -haloalkyl;

R ^2b.2 is H, C _1-6 -alkyl; this

R ^2b.1 and R ^2b.2 _{together are a C 3-6} -alkylene group, forming a hetero ring with a nitrogen atom, a carbon atom or ring may optionally be replaced by an oxygen atom,

R ³ is H, C _1-6 -alkyl;

X is chloride, bromide, iodide, sulfate, phosphate, methane sulfonate, nitrate, maleate, acetate, benzoate, citrate, salicylate, fumarate, tartrate, dibenzoyl tartrate, oxalate, an anion selected from the group consisting of succinate, benzoate and p-toluene sulfonate;

j is 0, 0.5, 1, 1.5 or 2;

Orotic acid, hippuric acid, L-pyroglutamic acid, D-pyroglutamic acid, nicotinic acid, L-(+)-ascorbic acid, saccharin, piperazine, 3-hydroxy-2-naphthoic acid, mucinous (galactaric) acid , palm (embonic) acid, stearic acid, cholic acid, deoxycholic acid, nicotinamide, isonicotinamide, succinamide, uracil, L-lysine, L-proline, D-valine, L-arginine, selected from the group consisting of glycine It has a co-crystal former.

Another embodiment of the present invention further comprises the administration of a compound of formula 2 co-crystal to a subject,

From here

R ^2a is H, C _1-6 -alkyl, C _1-6 -alkenyl, C _1-6 -alkynyl, C _3-6 -cycloalkyl, OC _1-6 -alkyl, CONR ^2a.1 R ^{2a. 2} ;

R ^2a.1 is H, C _1-6 -alkyl, C _1-6 -haloalkyl;

R ^2a.2 is H, C _1-6 -alkyl;

R ^2b is H, C _1-6 -alkyl, C _1-6 -alkenyl, C _1-6 -alkynyl, C _3-6 -cycloalkyl, COO-C _1-6 -alkyl, OC _1-6 - alkyl, CONR ^2b.1 R ^2b.2 , halogen;

R ^2b.1 is H, C _1-6 -alkyl, C _0-4 -alkyl-C _3-6 -cycloalkyl, C _1-6 -haloalkyl;

R ^2b.2 is H, C _1-6 -alkyl; this

R ^2b.1 and R ^2b.2 _{together are a C 3-6} -alkylene group, forming a hetero ring with a nitrogen atom, a carbon atom or ring may optionally be replaced by an oxygen atom, the remaining residues being as defined above like a bar

Another embodiment of the present invention further comprises the administration of a compound of formula 2 co-crystal to a subject,

From here

R ^2a is H, C _1-4 -alkyl, C _1-4 ^-alkynyl , C _3-6 -cycloalkyl, OC _1-4 -alkyl, CONR ^2a.1 R 2a.2 ;

R ^2a.1 is H, C _1-4 -alkyl;

R ^2a.2 is H;

R ^2b is H, C _1-4 -alkyl, OC _1-4 -alkyl, CONR ^2b.1 R ^2b.2 ;

R ^2b.1 is H, C _1-4 -alkyl, C _0-4 -alkyl-C _3-6 -cycloalkyl, C _1-4 -haloalkyl;

R ^2b.2 is H, C _1-4 -alkyl; this

R ^2b.1 and R ^2b.2 _{together are a C 3-6} -alkylene group, forming a hetero ring with a nitrogen atom, a carbon atom or ring may optionally be replaced by an oxygen atom, the remaining residues being as defined above like a bar

Another embodiment of the present invention further comprises the administration of a compound of formula 2 co-crystal to a subject,

From here

R ^2a is H, C _1-4 -alkyl;

R ^2b is H, CONR ^2b.1 R ^2b.2 ;

R ^2b.1 is H, C _1-4 -alkyl, C _0-4 -alkyl-C _3-6 -cycloalkyl, C _1-4 -haloalkyl;

R ^2b.2 is H, C _1-4 -alkyl; this

R ^2b.1 and R ^2b.2 _{together are a C 3-6} -alkylene group, forming a hetero ring with a nitrogen atom, a carbon atom or ring may optionally be replaced by an oxygen atom, the remaining residues being as defined above like a bar

Another embodiment of the present invention further comprises the administration of a compound of formula 2 co-crystal to a subject,

From here

R ¹ is C _1-6 -alkyl, C _1-6 -haloalkyl, OC _1-6 -haloalkyl, halogen;

m is 1, or 2;

R ^2a is H, C _1-4 -alkyl;

R ^2b is H, CONR ^2b.1 R ^2b.2 ;

R ^2b.1 is H, C _1-4 -alkyl, C _0-4 -alkyl-C _3-6 -cycloalkyl, C _1-4 -haloalkyl;

R ^2b.2 is H, C _1-4 -alkyl; this

R ^2b.1 and R ^2b.2 _{together are a C 3-6} -alkylene group, forming a hetero ring with a nitrogen atom, a carbon atom or ring may optionally be replaced by an oxygen atom;

R ³ is H, C _1-6 -alkyl;

X is an anion selected from the group consisting of chloride, dibenzoyl tartrate;

j is 1 or 2;

Another embodiment of the present invention further comprises the administration of a compound of formula 2 co-crystal to a subject,

From here

R ^2a is H, C _1-4 -alkyl; For example methyl, ethyl, propyl,

R ^2b is H, CONR ^2b.1 R ^2b.2 ;

R ^2b.1 is H, C _1-4 -alkyl; For example methyl, ethyl, propyl,

R ^2b.2 is H, C _1-4 -alkyl; For example, methyl, ethyl, propyl, and the remaining residues are as defined above.

Another embodiment of the present invention further comprises the administration of a compound of formula 2 co-crystal to a subject,

From here

R ^2a is H, C _1-4 -alkyl; For example methyl, ethyl, propyl,

R ^2b is H, CONR ^2b.1 R ^2b.2 ;

R ^2b.1 is C _0-4 -alkyl-C _3-6 -cycloalkyl;

R ^2b.2 is H, C _1-4 -alkyl; For example, methyl, ethyl, propyl, and the remaining residues are as defined above.

Another embodiment of the present invention further comprises the administration of a compound of formula 2 co-crystal to a subject,

From here

R ^2a is H, C _1-4 -alkyl; For example methyl, ethyl, propyl,

R ^2b is H, CONR ^2b.1 R ^2b.2 ;

R ^2b.1 is C _1-4 -haloalkyl;

R ^2b.2 is H, C _1-4 -alkyl; For example, methyl, ethyl, propyl, and the remaining residues are as defined above.

Another aspect of the invention further comprises the administration to a subject of a co-crystal of a compound of formula 2, wherein

R ^2b.1 and R ^2b.2 _{together are a C 3-6} -alkylene group, forming a hetero ring with a nitrogen atom, a carbon atom or ring may optionally be replaced by an oxygen atom; The remaining residues are defined as above.

Another aspect of the invention further comprises the administration of a co-crystal of a compound of formula 2 to a subject, wherein R ¹ , m, R ^2a , R ^2b , R ³ , X and j are defined as above and co-crystal. The crystal forming agent is selected from the group consisting of ascorbic acid, mucoic acid, pamoic acid, succinamide, nicotinic acid, nicotinamide, isonicotinamide, 1-lysine, 1-proline or a hydrate or hydrochloride salt thereof.

Another aspect of the invention further comprises the administration of a co-crystal of a compound of formula 2a to a subject, wherein R ^2a , R ^2b , R ³ , X and j are as defined above:

Another aspect of the invention further comprises the administration of a co-crystal of a compound of formula 2a to a subject, wherein

R ^2a is H, C _1-4 -alkyl; for example methyl, ethyl, propyl;

R ^2b is H, CONR ^2b.1 R ^2b.2 ;

R ^2b.1 is C _1-4 -alkyl; for example methyl, ethyl, propyl;

R ^2b.2 is C _1-4 -alkyl; For example, methyl, ethyl, propyl, and the remaining residues are as defined above.

Another aspect of the invention further comprises the administration of a co-crystal of a compound of formula 2a to a subject, wherein

R ^2a is H, C _1-4 -alkyl; for example methyl, ethyl, propyl;

R ^2b is H, CONR ^2b.1 R ^2b.2 ;

R ^2b.1 is C _0-4 -alkyl-C _3-6 -cycloalkyl;

R ^2b.2 is H, C _1-4 -alkyl; For example, H, methyl, ethyl, propyl, and the remaining residues are as defined above.

Another aspect of the invention further comprises the administration of a co-crystal of a compound of formula 2a to a subject, wherein

R ^2a is H, C _1-4 -alkyl; for example methyl, ethyl, propyl;

R ^2b is H, CONR ^2b.1 R ^2b.2 ;

R ^2b.1 is C _1-4 -haloalkyl;

R ^2b.2 is H, C _1-4 -alkyl; For example, H, methyl, ethyl, propyl, and the remaining residues are as defined above.

Another aspect of the invention further comprises the administration of a co-crystal of a compound of formula 2a to a subject, wherein

R ^2b.1 and R ^2b.2 _{together are a C 3-6} -alkylene group, forming a hetero ring with a nitrogen atom, a carbon atom or ring may optionally be replaced by an oxygen atom; The remaining residues are as defined above.

The free base of the compound of formula 2 (j = 0) is often amorphous and used in processes for preparing co-crystals, although salts of compounds of formula 2 are nevertheless used in some cases in processes for preparing co-crystals. Accordingly, another aspect of the present invention is a salt of a compound of formula 2, wherein R ¹ , m, R ^2a , R ^2b , R ³ are as defined above and

X is chloride, bromide, iodide, sulfate, phosphate, methane sulfonate, nitrate, maleate, acetate, benzoate, citrate, salicylate, fumarate, tartrate, dibenzoyl tartrate, oxalate, an anion selected from the group consisting of succinate, benzoate and p-toluene sulfonate; For example chloride, or dibenzoyl tartrate

j is 0, 0.5, 1, 1.5 or 2; For example 1 or 2.

Another aspect of the invention further comprises the administration of a co-crystal of a compound of formula 2 to a subject, wherein R ¹ , m, R ^2a , R ^2b , R ³ are as defined above and X is chloride; or dibenzoyl tartrate and j is 1 or 2.

Another aspect of the invention further comprises the administration of a co-crystal of a compound of formula 2 to a subject, wherein R ¹ , m, R ^2a , R ^2b , R ³ are as defined above and X is chloride; j is 2.

Another aspect of the invention further comprises the administration of a co-crystal of a compound of formula 2 to a subject, wherein R ¹ , m, R ^2a , R ^2b , R ³ are as defined above and X is dibenzoyl tartrate and j is 1.

Another aspect of the invention further comprises administration to a subject of a salt of a compound of formula 2a, wherein R ^2a , R ^2b , R ³ , X and j are as defined above:

Another aspect of the invention further comprises administering to a subject a salt of a compound of formula 2a, wherein

R ^2a is H, C _1-4 -alkyl; for example methyl, ethyl, propyl;

R ^2b is H, CONR ^2b.1 R ^2b.2 ;

R ^2b.1 is C _1-4 -alkyl; for example methyl, ethyl, propyl;

R ^2b.2 is C _1-4 -alkyl; For example, methyl, ethyl, propyl, and the remaining residues are as defined above.

Another aspect of the invention further comprises administering to a subject a salt of a compound of formula 2a, wherein

R ^2a is H, C _1-4 -alkyl; for example methyl, ethyl, propyl;

R ^2b is H, CONR ^2b.1 R ^2b.2 ;

R ^2b.1 is C _0-4 -alkyl-C _3-6 -cycloalkyl;

R ^2b.2 is H, C _1-4 -alkyl; For example, H, methyl, ethyl, propyl, and the remaining residues are as defined above.

Another aspect of the invention further comprises administering to a subject a salt of a compound of formula 2a, wherein

R ^2a is H, C _1-4 -alkyl; for example methyl, ethyl, propyl;

R ^2b is H, CONR ^2b.1 R ^2b.2 ;

R ^2b.1 is C _1-4 -haloalkyl;

R ^2b.2 is H, C _1-4 -alkyl; For example, H, methyl, ethyl, propyl, and the remaining residues are as defined above.

Another aspect of the invention further comprises administering to a subject a salt of a compound of formula 2a, wherein

R ^2b.1 and R ^2b.2 _{together are a C 3-6} -alkylene group, forming a hetero ring with a nitrogen atom, a carbon atom or ring may optionally be replaced by an oxygen atom; The remaining residues are as defined above.

Another aspect of the invention further comprises administration to a subject of a salt of a compound of formula 2a, wherein R ¹ , m, R ^2a , R ^2b , R ³ are as defined above for salt, X is chloride and j is 2

Another aspect of the invention further comprises the administration of a co-crystal of a compound of formula 2 to a subject, wherein R ¹ , m, R ^2a , R ^2b , R ³ are as defined above for salts, and X is dibenzoyl tartrate and j is 1. Another aspect of the invention further comprises administration to a subject of a salt of a compound of formula 2a, wherein R ¹ , m, R ^2a , R ^2b , R ³ are as defined above for salt, and X is (S) -(S)-(+)-dibenzoyl tartrate and j is 1.

c. formulation

Another embodiment of the present invention further comprises administering to a subject a pharmaceutical composition comprising a compound of formula (3):

From here

R ¹ is H, C _1-6 -alkyl, C _0-4 -alkyl-C _3-6 -cycloalkyl, C _1-6 -haloalkyl;

R ² is H, C _1-6 -alkyl;

X is an anion selected from the group consisting of chloride or 1/2 dibenzoyl tartrate; j is 1 or 2;

Another embodiment of the present invention further comprises administering to a subject a pharmaceutical composition comprising a compound of formula (3), wherein

R ¹ is H, C _1-6 -alkyl;

R ² is H, C _1-6 -alkyl;

X is an anion selected from the group consisting of chloride or 1/2 dibenzoyl tartrate; j is 1 or 2;

Another embodiment of the present invention further comprises administering to a subject a pharmaceutical composition comprising a compound of formula (3), wherein

R ¹ is H, methyl, ethyl, propyl, butyl;

R ² is H, methyl, ethyl, propyl, butyl;

X is chloride or an anion selected from the group consisting of 1/2 dibenzoyl tartrate, for example chloride; j is 1, or 2, for example 2.

Another embodiment of the present invention further comprises administering to a subject a pharmaceutical composition comprising a compound of formula (3), wherein

R ¹ is H, methyl, ethyl, propyl, butyl;

R ² is H, methyl;

X is chloride or an anion selected from the group consisting of 1/2 dibenzoyl tartrate, for example chloride; j is 1, or 2, for example 2.

Embodiments of the present invention further comprise administering to the subject a pharmaceutical composition containing a compound described in the following table as a hydrochloride salt. A further embodiment of the present invention further comprises administering to the subject a pharmaceutical composition containing the compound described in Table 2 below as 2-hydrochloride salt.

table 2

Another object of the present invention is to administer to a subject a pharmaceutical formulation of a compound as described above, wherein the formulation is an orally deliverable formulation.

Another object of the present invention is to administer to a subject a pharmaceutical dosage form of a compound as described above, in the form of a tablet, capsule, pellet, powder or granule.

Another object of the present invention is to administer to a subject a pharmaceutical formulation as described above for use as a medicament.

Another object of the present invention is the manufacture of a medicament for the treatment of a neurodegenerative disease or condition selected from Alzheimer's disease, Parkinson's disease, frontotemporal dementia, Huntington's disease, amyotrophic lateral sclerosis, multiple sclerosis, glaucoma, muscular dystrophy, vascular dementia and progressive supranuclear palsy use of the pharmaceutical formulation for

Another object of the present invention is Alzheimer's disease, Parkinson's disease, frontotemporal dementia, Huntington's disease, characterized in that an effective amount of the above-defined pharmaceutical formulation is orally administered to a subject or patient once, twice, three times or several times daily. , amyotrophic lateral sclerosis, multiple sclerosis, glaucoma, muscular dystrophy, vascular dementia and progressive supranuclear palsy.

d. Dosage form/ingredient

Solid pharmaceutical compositions prepared from compounds of formula 3 and ready for use/ingestion include powders, granules, pellets, tablets, capsules, chewable tablets, dispersible tablets, troches and lozenges. Specifically:

Capsule formulations according to the invention are conventional wet, dry or hot-melt granulation or hot-powder intermediates of the compound of formula 3, suitable intermediate blends, filled in customary capsules, for example hard gelatin or HPMC capsules intermediate blends comprising powder intermediates, pellets or granules obtained by melt extrusion or spray-drying.

The capsule formulation may also contain a powder intermediate of the compound of formula 3 in compressed form.

The capsule formulation according to the present invention comprises a compound of formula 3 suspended or diluted in a liquid or liquid mixture.

Tablet formulations according to the invention may be prepared by direct compression of a suitable final blend, or by conventional wet, dry or hot-melt granulation or hot-melt extrusion or spray drying of a suitable intermediate blend for tableting of pellets or granules. tablets obtained by

Another object of the present invention is a formulation in which a pH adjuster or buffer is added to improve the stability of the active ingredient. The pH adjusting/buffering agent may be a basic amino acid, which has, for example, an amino group and alkaline properties (isoelectric point, pI: 7.59-10.76), for example L-arginine, L-lysine or L-histidine. The buffer within the meaning of the present invention is L-arginine. L-Arginine exhibits a particularly suitable stabilizing effect for the composition of the present invention, for example, by inhibiting chemical degradation of the compound of formula (3).

Accordingly, in one embodiment, the present invention provides a compound of formula 3 and in particular L-arginine for stabilizing the composition against chemical degradation; and pharmaceutical compositions (eg oral solid dosage forms, particularly tablets) comprising one or more pharmaceutical excipients; is about

Suitably the pharmaceutical excipients used in the present invention are cellulose and its derivatives, D-mannitol, corn starch, pregelatinized starch as filler, copovidone as binder, crospovidone as disintegrant, magnesium stearate as lubricant, glidant Common materials such as colloidal anhydrous silica as a film coating agent, hypromellose as a film coating agent, polyethylene glycol as a plasticizer, titanium dioxide as a pigment, red/yellow iron oxide and talc, etc.

Specifically, the pharmaceutical excipient may be first and second diluents, binders, disintegrants and lubricants; Additional disintegrants and additional glidants are additional options.

Suitable diluents for the pharmaceutical composition according to the present invention include cellulose powder, microcrystalline cellulose, lactose of various crystalline modifications, dibasic calcium phosphate anhydride, dibasic calcium phosphate dihydrate, erythritol, low-substituted hydroxypropyl cellulose, mannitol, starch or modified (eg, pregelatinized or partially hydrolyzed) starch or xylitol. Among these diluents, mannitol and microcrystalline cellulose are used in some cases.

The diluents used as the second diluent are the above-mentioned diluents mannitol and microcrystalline cellulose.

Suitable lubricants for the pharmaceutical composition according to the invention are talc, polyethylene glycol, calcium behenate, calcium stearate, sodium stearyl fumarate, hydrogenated castor oil or magnesium stearate. In some cases the lubricant is magnesium stearate.

Suitable binders for the pharmaceutical composition according to the present invention include copovidone (a copolymer of vinyl pyrrolidone and other vinyl derivatives), hydroxy propyl methylcellulose (HPMC), hydroxy propyl cellulose (HPC), polyvinyl pyrrolidone (povidone), pregelatinized starch, stearic-palmitic acid, low substituted hydroxypropyl (L-HPC), copovidone and pregelatinized starch used in some formulations. The aforementioned binders pregelatinized starch and L-HPC exhibit additional diluent and disintegrant properties and can also be used as secondary diluents or disintegrants.

Suitable disintegrants for the pharmaceutical composition according to the invention are corn starch, crospovidone, polacrylin potassium, croscarmellose sodium, low substituted hydroxy propyl cellulose (L-HPC) or pregelatinized starch; For example, croscarmellose sodium can be used.

· Any glidant colloidal silicon dioxide can be used.

An exemplary composition according to the present invention comprises a diluent mannitol, microcrystalline cellulose as a diluent with additional disintegrating properties, copovidone as a binder, croscarmellose sodium as a disintegrant and magnesium stearate as a lubricant.

A typical pharmaceutical composition comprises (% by weight):

10-50% active ingredient

20-88% diluent 1,

5-50% diluent 2,

1-5% binder,

1-15% disintegrant and

0.1-5% lubricant.

A pharmaceutical composition according to some embodiments comprises (% by weight):

10-50% active ingredient

20-75% diluent 1,

5-30% diluent 2,

2-30% binder,

1-12% disintegrant and

0.1-3% lubricant

A pharmaceutical composition according to some embodiments comprises (% by weight):

10-90% active ingredient

5-70% diluent 1,

5-30% diluent 2,

0-30% binder,

1-12% disintegrant and

0.1-3% lubricant

A pharmaceutical composition according to some embodiments comprises (% by weight):

10-50% active ingredient

20-75% diluent 1,

5-30% diluent 2,

2-30% binder,

0.5-20% buffer,

1-12% disintegrant and

0.1-3% lubricant

A pharmaceutical composition according to some embodiments comprises (% by weight):

30-70% active ingredient

20-75% diluent 1,

5-30% diluent 2,

2-30% binder,

0.5-20% buffer,

1-12% disintegrant and

0.1-3% lubricant

In some cases pharmaceutical compositions containing 10-90% active ingredient are used, such as 30-70% active ingredient (% by weight).

The tablet formulations according to the invention may be coated, for example, uncoated or coated, for example, with a film-coating with a suitable coating which is known not to adversely affect the dissolution properties of the final coating. For example, tablets include dissolving a high molecular weight polymer such as polyvinyl pyrrolidone or hydroxypropyl-methyl cellulose together with a plasticizer, lubricant and optionally pigment and surfactant in water or an organic solvent such as acetone and mixing the mixture A seal coat can be provided for protection of the patient environment and clinical staff and for moisture protection purposes by spraying onto the tablet core inside a coating equipment such as a pan coater or a fluid bed coater with a wash insert.

In addition, agents such as film forming polymers such as beeswax, shellac, cellulose acetate phthalate, polyvinyl acetate phthalate, zein, hydroxy propyl cellulose, ethyl cellulose and polymeric methacrylate can be used to disintegrate/dissolve the formulation and form the coated dosage form. If stability is not affected, it can be applied to tablets.

After the formulation is film-coated, the sugar coating can be applied to the sealed pharmaceutical formulation. The sugar coating may include sucrose, dextrose, sorbitol, and the like, or mixtures thereof. If desired, colorants or opacifying agents may be added to the sugar solution.

The solid formulation of the present invention tends to be hygroscopic. These include moisture-proof packaging such as PVC blisters, PVDC-blisters or aluminum foil blister packs, alu/alu blisters, clear or opaque polymer blisters with pouches, polypropylene tubes, glass bottles and optionally HDPE bottles with child protection. can be packaged using The main packaging material may contain a desiccant such as molecular sieves or silica gel to improve the chemical stability of the API. Opaque packaging materials such as colored blister materials, tubes, and brown glass bottles can be used to extend the shelf life of APIs by reducing photodegradation.

e. Dosage

The dosage range of the compound of formula 3 is generally from 100 to 1000 mg, in particular from 200 to 900 mg, from 300 to 900 mg or from 350 and 850 mg or from 390 to 810 mg. It is possible to provide one or two tablets, in some cases two tablets of 100, 200, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900 mg per day; 350, 400, 450, 750, 800, 850 are used in some cases.

Dosage ranges may be achieved by one tablet or two tablets; In some cases, two tablets are administered, each containing half the dose.

Application of the active ingredient may be up to three times a day, such as once or twice a day. Particular dosage strengths are 400 mg or 800 mg.

f. Terms and definitions used

Terms not specifically defined herein should be given the meanings that will be assigned to those skilled in the art in light of the present disclosure and context. However, as used herein, unless otherwise specified, the following terms have the meanings indicated below.

The term “about” means at least 5% or less than the stated value. Thus, about 100 minutes can be read as 95 to 105 minutes.

In the event of any discrepancy when a compound of the present invention is shown in the form of a chemical name and expressed as a formula, the formula shall prevail. An asterisk may be used in a sub-formula to indicate a bond linked to a core molecule as defined.

Unless specifically stated otherwise, throughout the specification and appended claims, given formulas or names refer to tautomers and all stereoscopic, optical and geometric isomers (e.g., enantiomers, diastereomers, E/Z isomers, etc.) and salts comprising their racemates and their separated enantiomers, mixtures of diastereomers in different proportions, or mixtures of any of the above forms in which such isomers and enantiomers exist, and pharmaceutically acceptable salts thereof. and solvates thereof, including hydrates, including solvates of free compounds, or solvates of salts of compounds.

As used herein, the term “substituted” means that any one or more hydrogens on the designated atom are replaced with a selected from the group indicated if the substitution does not exceed the normal valence of the designated atom and the substitution results in a stable compound.

The term "optionally substituted", within the scope of the present invention, refers to the aforementioned groups, which are optionally substituted by small molecular groups. An example of a small molecular group that is considered to be chemically significant is a group consisting of 1-200 atoms. Groups that do not adversely affect the pharmacological efficacy of the compound are important. For example, a group may include:

A straight or branched carbon chain, optionally interrupted by a heteroatom, optionally substituted with a ring, heteroatom or other common functional group.

· Aromatic or non-aromatic ring system consisting of carbon atoms and optional heteroatoms, which may be substituted with functional groups.

· Multiple aromatic or non-aromatic ring systems consisting of any heteroatom and carbon atoms, which may be joined by one or more carbon chains, optionally interrupted by a heteroatom, and optionally substituted with a heteroatom, or other common functional group.

The compounds disclosed herein may exist as therapeutically acceptable salts. The present invention includes the compounds enumerated above in salt form, including acid addition salts. Suitable salts include those formed with both organic and inorganic acids. Such acid addition salts will generally be pharmaceutically acceptable. However, salts of salts that are not pharmaceutically acceptable may be useful in the preparation and purification of the compound. Basic addition salts may also be formed and are pharmaceutically acceptable. For details on the preparation and selection of salts, see Pharmaceutical Salts: Properties, Selection, and Use (Stahl, P. Heinrich. Wiley-VCHA, Zurich, Switzerland, 2002).

As used herein, the term “therapeutically acceptable salt” refers to a salt or zwitterionic form of a compound disclosed herein that is water-soluble or dispersible and therapeutically acceptable as defined herein. Salts can be prepared separately during the final isolation and purification of the compound or by reacting the appropriate compound in its free base form with a suitable acid. Representative acid addition salts are acetate, adipate, alginate, L-ascorbate, aspartate, benzoate, benzenesulfonate (besylate), bisulfate, butyrate, camphorate, camphorsulfonate, citrate, di Gluconate, formate, fumarate, gentisate, glutarate, glycerophosphate, glycolate, hemisulphate, heptanoate, hexanoate, hyporate, hydrochloride, bromate, iodate, 2-hydroxyethanesulfo Nate (isotheonate), lactate, maleate, malonate, DL-mandelate, mesitylenesulfonate, methanesulfonate, naphthylenesulfonate, nicotinate, 2-naphthalenesulfonate, oxalate, 3- Phenylpropionate, phosphonate, picrate, pivalate, propionate, pyroglutamate, succinate, sulfonate, tartrate, L-tartrate, trichloroacetate, trifluoroacetate, phosphate, glutamate, bicarbonate , para toluene sulfonate (p-tosylate) and undecanoate. In addition, basic groups in the compounds disclosed herein include methyl, ethyl, propyl and butyl chlorides, bromides and iodides; dimethyl, diethyl, dibutyl and diamyl sulfates; decyl, lauryl, myristyl and steryl chlorides, bromides and iodides; and benzyl and phenethyl bromide. Examples of acids that can be used to form therapeutically acceptable addition salts include inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid and phosphoric acid, and organic acids such as oxalic acid, maleic acid, succinic acid and citric acid. Salts may also be formed by coordination of compounds with alkali or alkaline earth metals. Accordingly, the present invention contemplates the sodium, potassium, magnesium and calcium salts of the compounds disclosed herein, and the like.

Base addition salts can be prepared during the final isolation and purification of compounds by reaction with a suitable base such as a hydroxide, carbonate or bicarbonate of a carboxyl group and a metal cation, or with ammonia or an organic primary, secondary or tertiary amine. The cations of the therapeutically acceptable salts include lithium, sodium, potassium, calcium, magnesium and aluminum as well as ammonium, tetramethyl ammonium, tetraethyl ammonium, methylamine, dimethylamine, trimethylamine, triethylamine, diethylamine, ethylamine, Tributylamine, pyridine, N,N-dimethyl aniline, N-methyl piperidine, N-methylmorpholine, dicyclohexylamine, procaine, dibenzylamine, N,N-dibenzylphenethylamine, 1- non-toxic quaternary amine cations such as epenamine and N,P-dibenzylethylenediamine. Other representative organic amines useful for the formation of base addition salts include ethylenediamine, ethanolamine, diethanolamine, piperidine and piperazine.

Although it may be possible to administer the compounds of the present invention as the raw chemical, they may also be presented as pharmaceutical formulations. Accordingly, pharmaceutical formulations comprising one or more specific compounds disclosed herein or one or more pharmaceutically acceptable salts, esters, prodrugs, amides or solvates, optionally one or more pharmaceutically acceptable carriers and other therapeutic ingredients are provided herein. is provided on The carrier(s) must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not deleterious to the recipient. Proper formulation depends on the route of administration chosen. Any of the carriers and excipients of the known art, for example from Remington's Pharmaceutical Sciences, may be used as are suitable and understood in the art. The pharmaceutical compositions disclosed herein may be prepared in any manner known in the art, for example, by conventional mixing, dissolving, granulating, dragee preparation, leaching, emulsifying, encapsulating, encapsulating or compression processes.

A “hetero ring” (“het”) is a 5-, 6- or 7-membered, saturated or unsaturated heterocyclic ring or 5- to 10-membered, bicyclic ring which may contain 1, 2 or 3 heteroatoms selected from oxygen, sulfur. click heterocycles; The ring may be linked to the molecule by a carbon atom or, if present, a nitrogen atom. The following are examples of 5-, 6- or 7-membered, saturated or unsaturated heterocycles:

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Examples of 5-10 membered bicyclic heterocyclic rings include pyrrolizine, indole, indolizine, isoindole, indazole, purine, quinoline, isoquinoline, benzimidazole, benzofuran, benzopyran, benzothiazole, benzoisothia sol, pyridopyrimidine, pteridine, pyrimidopyrimidine,

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While the term heterocycle includes heteroaromatic ring groups, the term heterocyclic aromatic group ("hetaryl") refers to one, two or three heterocyclic groups selected from oxygen, sulfur and nitrogen containing sufficient conjugated double bonds to form an aromatic system. 5 or 6 membered heteroaromatic ring group which may contain atoms or a 5 to 10 membered bicyclic heteroaryl ring. The ring may be connected to the molecule through a carbon atom or through a nitrogen atom, if present. The following are examples of 5- or 6-membered heteroaromatic ring groups:

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Examples of 5-10 membered bicyclic heteroaryl rings include pyrrolizine, indole, indolizine, isoindole, indazole, purine, quinoline, isoquinoline, benzimidazole, benzofuran, benzopyran, benzothiazole, benzoiso Thiazole, pyridopyrimidine, pteridine, pyrimidopyrimidine.

As used herein, the term “halogen” refers to a halogen substituent selected from fluoro, chloro, bromo or iodo.

The term “C _1-6 -alkyl” (including those that are part of another group) refers to branched and unbranched alkyl groups having 1 to 6 carbon atoms, and the _{term “C 1-4} -alkyl” refers to 1 to 4 branched and unbranched alkyl groups having two carbon atoms. Alkyl groups having from 1 to 4 carbon atoms are present in some cases. Examples of these include methyl, ethyl, n-propyl, isopropyl, n-butyl, iso-butyl, sec-butyl, tert-butyl, n-pentyl, iso-pentyl, neo-pentyl or hexyl. The abbreviations Me, Et, n-Pr, i-Pr, n-Bu, i-Bu, t-Bu, etc. may also be used in the aforementioned groups. Unless otherwise stated, the definitions propyl, butyl, pentyl and hexyl include all possible isomeric forms of the group. Thus, for example, propyl includes n-propyl and isopropyl, butyl includes isobutyl, sec-butyl and tert-butyl, and the like.

The term “C _1-6 -alkylene” (including those that are part of another group) refers to branched and unbranched alkylene groups having from 1 to 6 carbon atoms, the _{term “C 1-4} -alkylene” means branched and unbranched alkylene groups of 1 to 4 carbon atoms. Alkyl groups having from 1 to 4 carbon atoms are present in some cases. Examples include methylene, ethylene, propylene, 1-methyl ethylene, butylene, 1-methyl propylene, 1,1-dimethyl ethylene, 1,2-dimethyl ethylene, pentylene, 1,1-dimethyl propylene, 2,2-dimethyl propylene, 1,2-dimethyl propylene 1,3-dimethyl propylene or hexylene. Unless otherwise stated, the definitions propylene, butylene, pentylene and hexylene also include all possible isomeric forms of the related groups having the same number of carbons. Thus, for example, propyl also includes 1-methyl ethylene and butylene includes 1-methyl propylene, 1,1-dimethyl ethylene, 1,2-dimethyl ethylene.

The term "C _2-6 -alkenyl" (including those that are part of another group) refers to branched and unbranched alkenyl groups having 2 to 6 carbon atoms, and the _{term "C 2-4} -alkenyl" refers to 2 It refers to branched and unbranched alkenyl groups having to 4 carbon atoms, provided that there is at least one double bond, in some cases alkenyl groups having 2 to 4 carbon atoms are used. Examples include ethenyl or vinyl, propenyl, butenyl, pentenyl or hexenyl. Unless otherwise stated, the definitions propenyl, butenyl, pentenyl and hexenyl include all possible isomeric forms of the group. Thus, for example, propenyl includes 1-propenyl and 2-propenyl, butenyl includes 1-, 2- and 3-butenyl, 1-methyl-1-propenyl, 1-methyl-2 -Including propenyl and the like.

The term “C _2-6 -alkenylene” (including those that are part of another group) refers to branched and unbranched alkenylene groups having 2 to 6 carbon atoms, and the term “C _2-4 -alkenylene” refers to 2 branched and unbranched alkylene groups of from to 4 carbon atoms. Alkenylene groups having 2 to 4 carbon atoms are present in some cases. Examples include: ethenylene, propenylene, 1-methyl ethylene, butenylene, 1-methyl propylene, 1,1-dimethyl ethylene, 1,2-dimethyl ethylene, pentenylene, 1,1-dimethyl propylene , 2,2-dimethyl propylene, 1,2-dimethyl propylene, 1,3-dimethyl propenylene or hexenylene. Unless otherwise stated, the definitions propenylene, butenylene, pentenylene and hexenylene include all possible isomeric forms of the respective groups having the same number of carbons. Thus, for example, propenyl also includes 1-methylethenylene and butenylene includes 1-methyl propenylene, 1,1-dimethylethenylene, 1,2-dimethylethenylene.

The term “C _2-6 -alkynyl” (including those that are part of another group) refers to branched and unbranched alkynyl groups having 2 to 6 carbon atoms, and the term “C _2-4 -alkynyl” refers to 4 Branched and unbranched alkynyl groups of at least one carbon atom and having at least one triple bond. Alkynyl groups having 2 to 4 carbon atoms are present in some cases. Examples include ethynyl, propynyl, butynyl, pentynyl or hexynyl. Unless otherwise stated, the definitions propynyl, butynyl, pentynyl and hexynyl include all possible isomeric forms of the respective groups. Thus, for example, propynyl includes 1-propynyl and 2-propynyl, butynyl includes 1-, 2- and 3-butynyl, 1-methyl-1-propynyl, 1-methyl-2 -Including propynyl and the like.

The term "C _2-6 -alkynylene" (including those that are part of another group) refers to branched and unbranched alkynylene groups having 2 to 6 carbon atoms, and the _{term "C 2-4} -alkynylene" means branched and unbranched alkylene groups of 24 carbon atoms. Alkynylene groups having 2 to 4 carbon atoms are present in some cases. Examples include ethynylene, propynylene, 1-methylethynylene, butynylene, 1-methyl propynylene, 1,1-dimethylethynylene, 1,2-dimethylethynylene, pentynylene, 1,1-dimethyl propynylene, 2,2-dimethylpropynylene, 1,2-dimethylpropynylene, 1,3-dimethylpropynylene or hexynylene. Unless otherwise stated, the definitions propylene, butynylene, pentynylene and hexylene include all possible isomeric forms of each group having the same number of carbons. Thus, for example, propynyl includes 1-methylethynylene and butynylene includes 1-methylpropynylene, 1,1-dimethylethynylene, 1,2-dimethylethynylene.

As used herein, the term “C _3-6 -cycloalkyl” (including those that are part of another group) refers to a cycloalkyl group having 3 to 8 carbon atoms, and in some cases such a group having 5 to 6 carbon atoms. having a cycloalkyl group. Examples include cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl.

The term "C _1-6 -haloalkyl" (including those that are part of other groups) refers to branched and unbranched alkyl groups having from 1 to 6 carbon atoms, wherein at least one hydrogen atom is fluorine, chlorine or bromine; For example fluorine and chlorine, for example substituted with fluorine. The term "C _1-4 -haloalkyl" refers to the corresponding branched and unbranched alkyl groups having from 1 to 4 carbon atoms, wherein one or more hydrogen atoms are replaced analogously to those mentioned above. In some cases C _1-4 -halo alkyl is present. Examples include: CH ₂ F, CHF ₂ , CF ₃ .

The term "C _{1 -n} -alkyl", wherein n alone or in combination with other radicals is an integer from 2 to n, denotes an acyclic, saturated, branched or linear hydrocarbon radical having 1 to n carbon atoms. Examples of C _1-5 -alkyl include: H ₃ C-, H ₃ C-CH ₂ -, H ₃ C-CH ₂ -CH ₂ -, H ₃ C-CH(CH ₃ )-, H ₃ C-CH ₂ -CH ₂ -CH ₂ -, H ₃ C-CH ₂ -CH(CH ₃ )-, H ₃ C-CH(CH ₃ )-CH ₂ -, H ₃ CC(CH ₃ ) ₂ - , H ₃ C-CH ₂ -CH ₂ -CH ₂ -CH ₂ -, H ₃ C-CH ₂ -CH ₂ -CH(CH ₃ )-, H ₃ C-CH ₂ -CH(CH ₃ )-CH ₂ -, H ₃ C-CH(CH ₃ )-CH ₂ -CH ₂ -, H ₃ C-CH ₂ -C(CH ₃ ) ₂ -, H ₃ CC(CH ₃ ) ₂ -CH ₂ -, H ₃ C -CH(CH ₃ )-CH(CH ₃ )- and H ₃ C-CH ₂ -CH(CH ₂ CH ₃ )-.

The term "C _{1 -n} -haloalkyl", wherein n is an integer from 2 to n, alone or in combination with other radicals, means an acyclic, saturated, branched or linear, wherein at least one hydrogen atom has 1 to n C atoms. represents a hydrocarbon radical. It may be replaced by a halogen atom selected from fluorine, chlorine or bromine, such as fluorine and chlorine, for example fluorine. Examples include: CH ₂ F, CHF ₂ , CF ₃ .

The term "C _{1 -n} -alkylene", wherein n is an integer from 2 to n, alone or in combination with other radicals, refers to an acyclic, straight or branched chain divalent alkyl radical containing from 1 to n carbon atoms. indicates Examples of C _{1 -n} -alkylene include: -CH ₂ -, -CH ₂ -CH ₂ -, -CH(CH ₃ )-, -CH ₂ -CH ₂ -CH ₂ -, -C( CH ₃ ) ₂ -, -CH(CH ₂ CH ₃ )-, -CH(CH ₃ )-CH ₂ -, -CH ₂ -CH(CH ₃ )-, -CH ₂ -CH ₂ -CH ₂ -CH ₂ -, -CH ₂ -CH ₂ -CH(CH ₃ )-, -CH(CH ₃ )-CH ₂ -CH ₂ -, -CH ₂ -CH(CH ₃ )-CH ₂ -, -CH ₂ -C( CH ₃ ) ₂ -, -C(CH ₃ ) ₂ -CH ₂ -, -CH(CH ₃ )-CH(CH ₃ )-, -CH ₂ -CH(CH ₂ CH ₃ )-, -CH(CH _{2 )} CH ₃ )-CH ₂ -, -CH(CH ₂ CH ₂ CH ₃ )- , -CH(CH(CH ₃ )) ₂ - and -C(CH ₃ )(CH ₂ CH ₃ )-.

The term "C _{2 -n -alkenyl" is used in a group as defined in the definition of "C 1 -n} -alkyl" having two or more carbon atoms, wherein two or more of the carbon atoms of the group are bonded to each other. form a double bond.

The term "C _{2 -n -alkynyl" is used in groups as defined in the definition of "C 1 -n} -alkyl" having at least two carbon atoms, wherein at least two of the carbon atoms of the group are bonded to each other. to form a triple bond.

The term “C _{3 -n} -cycloalkyl”, wherein n is an integer from 4 to n, alone or in combination with other radicals, denotes a cyclic saturated unbranched hydrocarbon radical having 3 to n carbon atoms. Examples of C _3-n -cycloalkyl include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and cycloheptyl.

It should be noted that, as used herein and in the appended claims, the singular forms "a", "an" and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to "a cell" includes a plurality of such cells and reference to "a peptide" includes one or more peptides and equivalents thereof, such as polypeptides known to those skilled in the art, and the like.

An “individual suffering from or at risk of suffering from an age-related cognitive impairment” is defined as greater than about 50%, such as greater than 60%, such as greater than 70%, such as 75%, 80%, 85% over life expectancy. , 90%, 95% or 99% or more individuals. The age of an individual depends on the species. Therefore, this ratio is based on the expected lifespan of the species. For example, in humans, such an individual is 50 years of age or older, such as 60 years old or older, 70 years old or older, 80 years old or older, 90 years old or older, about 50 to 100 years old, eg 90 years old, and generally Not over 100 years old, ie 50 for example. . 55. . . 60. . . 65. . . 70. . . 75. . . 80. . . 85. . . 90. . . 95. . . individuals 100 years of age or older, or between 50 and 1000 years of age, suffering from aging-related symptoms, such as cognitive impairment associated with the natural aging process, as further described below; About 50 years old or older, such as 60 years old or older, 70 years old or older, 80 years old or older, 90 years old or older, generally not 100 years old or older, ie about 50 to 100 years old, eg 50 years old. . . 55. . . 60. . . 65. . . 70. . . 75. . . 80. . . 85. . . 90. . . 95. . . Individuals at the age of 100 who have not yet exhibited symptoms of age-related symptoms (eg cognitive impairment); Individuals of any age suffering from cognitive impairment due to age-related diseases, as further described below, and any age diagnosed with age-related diseases that have not yet begun to exhibit symptoms of cognitive impairment but are generally associated with cognitive impairment. is the individual of The age for non-human subjects is known and intended to apply.

As summarized elsewhere, in some cases the subject is a mammal. Mammalian species that can be treated with the present method include dogs and cats; Words; bull; cow; Primates, including humans. The methods, compositions and reagents of the present invention may also be applied to animal models including, for example, small mammals, such as mice, rabbits, and the like, in experimental studies.

As used herein and described above, “treatment” refers to any of (i) preventing a disease or disorder or (ii) reducing or eliminating symptoms of a disease or disorder. Treatment can be carried out prophylactically (before the onset of the disease) or therapeutically (after the onset of the disease). The effect may be prophylactic in terms of completely or partially preventing a disease or symptom thereof and/or may be therapeutic in terms of partial or complete treatment of a side effect to and/or caused by a disease. Accordingly, the term “treatment,” as used herein, includes any treatment of a senescence-associated disease or disorder in a mammal, and includes (a) preventing the disease from developing in a subject susceptible to but not yet afflicted with the disease; (b) inhibiting the disease, ie, arresting its progression; or (c) alleviating the disease, ie causing regression of the disease. Treatment can result in a variety of different physical symptoms, such as modulation of gene expression, rejuvenation of tissues or organs, and the like. The therapeutic agent may be administered before, during, or after the onset of the disease. Of particular importance is the treatment of progressive disease, in which the treatment stabilizes or reduces the patient's undesirable clinical symptoms. Such treatment may be performed before complete loss of function in the affected tissue. The subject therapy may be administered during and in some cases after the symptomatic stage of the disease.

In some embodiments, the condition being treated is an age-related disorder of the individual's cognitive abilities. Cognitive ability, or “cognition,” refers to the mind including attention and concentration, learning complex tasks and concepts, memory (collecting, maintaining, and retrieving new information in the short and/or long term), and information processing (manipulating information collected by the five senses). means process. ), spatial perception skills (visual perception, depth perception, mental image use, copying pictures, constructing objects or shapes), language generation and understanding, language fluency (finding words), problem solving, decision-making and planning functions (planning and prioritizing) designation) is included. "Cognitive decline" means a gradual decline in one or more of these abilities, such as a decrease in memory, language, thinking, or judgment. "Impairment of cognitive ability" and "cognitive impairment" refer to a healthy individual, eg, an age-appropriate healthy individual, or eg, 2 weeks, 1 month, 2 months, 3 months, 6 months, 1 year, 2 years, It refers to a decrease in cognitive ability relative to an individual's ability at a time earlier than 5 or 10 years. Age-related cognitive impairments include, for example, cognitive impairments associated with the natural aging process, eg, impairments in cognitive ability associated with aging, such as mild cognitive impairment (M.C.I.); and cognitive impairments associated with age-related disorders, i.e., disorders that appear to increase in frequency with increasing age, e.g., Alzheimer's disease, Parkinson's disease, Lewy body dementia, frontotemporal dementia, Huntington's disease, amyotrophic lateral sclerosis, multiple sclerosis, glaucoma, muscular dystrophy, vascular dementia, progressive supranuclear palsy, ataxia, weakness, and the like.

g. Combination

The compounds of formula (1) can be used alone or in combination with other active substances of formula (1) according to the invention. The compound of formula 1 may optionally be combined with other pharmacologically active substances. These include ß2-adrenergic receptor agonists (short- and long-acting), anticholinergics (short and long-acting), anti-inflammatory steroids (oral and topical corticosteroids), croglycates, methylxanthine, dissociated glucocorticoid mimetics, PDE3 inhibitors, PDE4-inhibitors, PDE7-inhibitors, LTD4 antagonists, EGFR-inhibitors, dopamine agonists, PAF antagonists, lipoxin A4 derivatives, FPRL1 modulators, LTB4-receptor (BLT1, BLT2) antagonists, histamine H1 receptor antagonists, histamine H4 receptor antagonists , dual histamine H1/H3-receptor antagonists, PI3-kinase inhibitors such as inhibitors of non-receptor tyrosine kinases such as LYN, LCK, SYK, ZAP-70, FYN, BTK or ITK, such as p38, ERK1, ERK2 , inhibitors of MAP kinases such as JNK1, JNK2, JNK3 or SAP, such as NF-κB signaling pathway inhibitors such as IKK2 kinase inhibitors, iNOS inhibitors, MRP4 inhibitors such as 5-lipoxygenase (5-LO) Inhibitors, cPLA2 inhibitors, leukotriene A4 hydrolase inhibitors or leukotriene biosynthesis inhibitors such as FLAP inhibitors, nonsteroidal anti-inflammatory drugs (NSAIDs), CRTH2 antagonists, DP1-receptor modulators, thromboxane receptor antagonists, additional CCR3 antagonists, CCR4 antagonists, CCR1 antagonists , CCR5 antagonist, CCR6 antagonist, CCR6 antagonist CCR7 antagonist CCR7 antagonist, CCR9 antagonist, CCR30 antagonist, CXCR3 antagonist, CXCR4 antagonist, CXCR2 antagonist, CXCR1 antagonist, CXCR5 antagonist, NK1, CXCR3 antagonist, CXCR3 antagonist Antagonists, sphingosine-1-phosphate receptor modulators, sphingosine monophosphatase inhibitors, such as adenosine receptor modulators such as A2a-agonists, such as modulators of purinergic receptors such as P2X7 inhibitors, histone deacetylase (HDAC) ) activator, bradykinin (BK1, BK2) antagonists, TACE inhibitors, PPAR gamma modulators, Rho-kinase inhibitors, interleukin 1-beta converting enzyme (ICE) inhibitors, toll-like receptor (TLR) modulators, HMG-CoA reductase inhibitors, VLA-4 antagonists, ICAM- 1 inhibitor, SHIP agonist, GABAa receptor antagonist, ENaC-inhibitor, melanocortin receptor (MC1R, MC2R, MC3R, MC4R, MC5R) modulator, CGRP antagonist, endothelin antagonist, TNFα antagonist, anti-TNF antibody, anti-GMF antibody , anti-CD46 antibody, anti-IL-1 antibody, anti-IL-2 antibody, anti-IL-4 antibody, anti-IL-5 antibody, anti-IL-13 antibody, anti-IL-4/IL-13 antibody, anti-TSLP antibody, anti-OX40 antibody, mucosal coagulant, immunotherapeutic agent, compound against airway dilatation, compound against cough, VEGF inhibitor, or a combination of two or three active substances.

In some embodiments, the other active agent is a beta agonist, anticholinergic agent, corticosteroid, PDE4-inhibitor, LTD4-antagonist, EGFR-inhibitor, CRTH2 inhibitor, 5-LO-inhibitor, histamine receptor antagonist and SYK-inhibitor, or both or It is a combination of three active substances, namely:

Beta agonists and corticosteroids, PDE4 inhibitors, CRTH2 inhibitors or LTD4 antagonists;

Anticholinergics and beta agonists, corticosteroids, PDE4-inhibitors, CRTH2-inhibitors or LTD4-antagonists;

Corticosteroids and PDE4-inhibitors, CRTH2-inhibitors or LTD4-antagonists;

PDE4 inhibitors and CRTH2 inhibitors or LTD4 antagonists;

· CRTH2 inhibitors and LTD4 antagonists.

In these embodiments, the compounds that make up the combination are co-administered to the subject. The terms "co-administration" and "combination with" include the administration of two or more therapeutic agents simultaneously, at the same time, or sequentially, without specific time limit. In one embodiment, the agent is present in a cell or subject simultaneously or exerts a biological or therapeutic effect simultaneously. In one embodiment, the therapeutic agents are of the same composition or unit dosage form. In other embodiments, the therapeutic agent is a separate composition or unit dosage form. In certain embodiments, the first agent is administered prior to administration of the second therapeutic agent (e.g., 5 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, 24 hours, 48 hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 8 weeks, or 12 weeks before), at the same time or after (e.g., 5 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, 24 hours, 48 hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks , after 8 or 12 weeks). "Concomitant administration" of a known therapeutic drug with a pharmaceutical composition of the present disclosure means administration of a compound and a second agent such that both the known drug and the composition of the present invention have a therapeutic effect. Such concomitant administration may include simultaneous (ie, simultaneous), prior or subsequent administration of the drugs with respect to administration of the subject compound. The routes of administration of the two agents may vary, and representative routes of administration are described in more detail below. One of ordinary skill in the art will not have difficulty determining the appropriate timing, sequence, and dosage for administration for particular drugs and compounds of the present disclosure. In some embodiments, compounds (e.g., a compound of the invention and one or more additional compounds) are administered within 24 hours of each other, such as within 12 hours of each other, within 6 hours of each other, within 3 hours of each other, or within 1 hour of each other. In an embodiment, the compounds are administered within 1 hour of each other. In certain embodiments, the compounds are administered substantially simultaneously. By substantially simultaneous administration it is meant that the compounds are administered to the subject within about 10 minutes or less of each other, such as 5 minutes or less, or 1 minute or less of each other.

"Companion diagnostics" or "companion diagnostics device" means an in vitro diagnostic device or imaging tool that provides information essential for the safe and effective use of a given therapeutic product. The use of an in vitro diagnostic companion device with a specific therapeutic agent is specified in the guidance for use in the labeling of the device and that therapeutic agent, as well as generic equivalents and biosimilar equivalents of the therapeutic agent.

Companion diagnostic testing may take many forms, including, but not limited to, for example: tests that screen for genetic patterns in the family and test for difficult-to-diagnose symptoms; prognostic tests to predict the future course of the disease; therapeutic tests that indicate the patient's response to the prescribed treatment; monitoring tests to evaluate the effectiveness and appropriate dosage of prescribed therapy; and relapse testing, which analyzes the patient's risk for recurrence of the disease. See: Agarwal A, et al., Pharmgenomics Pers Med. 8:99-110 (2015), which is incorporated herein by reference in its entirety.

h. formulation

Formulations suitable for administering the compound of Formula 1 and the co-crystal or salt form of Formulas 2 and 2a include, for example, tablets, capsules, suppositories, solutions, powders and the like. The content of the pharmaceutically active compound(s) ranges from 0.05 to 90% by weight, such as 0.1 to 50% by weight of the total composition. Suitable tablets include, for example, the active substance(s) with known excipients, for example inert diluents such as calcium carbonate, calcium phosphate or lactose, disintegrants such as corn starch or alginic acid, binders such as starch or gelatin, magnesium stearate or by mixing with a lubricant such as talc and/or a release retardant such as carboxymethyl cellulose, cellulose acetate phthalate or polyvinyl acetate. The tablet may also contain several layers.

Coated tablets may be prepared similarly to tablets by coating the prepared core with substances commonly used to coat tablets, for example, kollidone or shellac, gum arabic, talc, titanium dioxide or sugar. The core may also consist of multiple layers to achieve delayed release or to prevent incompatibility. Similarly, tablet coatings may be composed of multiple or layers to achieve delayed release, possibly using the excipients mentioned above for tablets.

Syrups or elixirs containing the active substances or combinations thereof according to the invention may additionally contain sweeteners such as saccharin, cyclamate, glycerol or sugar and flavor enhancers such as vanillin or orange extract. They may also contain suspension aids or thickening agents, for example sodium carboxymethyl cellulose, wetting agents, for example condensation products of fatty alcohols with ethylene oxide or preservatives such as p-hydroxy benzoate.

Solutions are prepared in the usual way, using, for example, isotonic agents, preservatives such as p-hydroxy benzoate or alkali metal salts of ethylenediaminetetraacetic acid, optionally emulsifying and/or dispersing agents, but water is the diluent For example, an organic solvent may optionally be used as a solubilizing agent or solubilizing aid, and the solution may be transferred into an injection vial or ampoule or infusion bottle.

Capsules containing one or more active substances or combinations of active substances can be prepared, for example, by mixing the active substances with an inert carrier such as lactose or sorbitol and packaging them in gelatin capsules.

Suitable suppositories can be prepared by mixing with carriers provided for this purpose, such as, for example, neutral fats or polyethylene glycol or derivatives thereof.

Excipients that may be used include, for example, water, pharmaceutically acceptable organic solvents such as paraffin (eg petroleum fraction), vegetable oils (eg peanut or sesame oil), monofunctional or polyfunctional alcohols (eg ethanol or glycerol), carriers such as natural mineral powders (such as kaolin, clay, talc, chalk), synthetic mineral powders (such as highly disperse silicic acids and silicates), sugars (such as cane sugar, lactose and glucose), emulsifiers (eg lignin, waste sulfite liquor, methyl cellulose, starch and polyvinyl pyrrolidone) and lubricants (eg magnesium stearate, talc, stearic acid and sodium lauryl sulfate).

For oral use, tablets may contain, in addition to the carriers specified, additives such as sodium citrate, calcium carbonate and dicalcium phosphate, together with various additional substances such as starches, for example potato starch, gelatin and the like. Lubricating agents such as magnesium stearate, sodium lauryl sulfate and talc may also be used to make the tablets. In the case of aqueous suspensions, the active substance may be combined with various flavor enhancers or colorants in addition to the above excipients.

In order to administer the compound of Formula 1 or the co-crystal or salt form of Formulas 2 and 2a, an agent or pharmaceutical formulation suitable for inhalation may be used. Inhalable formulations include inhalable powders, propellants containing metered doses of aerosols, or inhalable solutions without propellants. Within the scope of the present invention, the term propellant-free inhalation solution also includes ready-to-use concentrates or sterile inhalation solutions. Formulations that may be used within the scope of the present invention are described in more detail in the following sections of this specification.

The inhalable powders that can be used according to the invention may contain the compound of formula (1) or the co-crystal or salt form of formulas (2) and (2a), alone or in combination with suitable physiologically acceptable excipients.

When the active substance of the compound of formula 1 or the co-crystal or salt form of formulas 2 and 2a is present in admixture with physiologically acceptable excipients, the following physiologically acceptable excipients may form these inhalable powders according to the present invention. Can be used to prepare: monosaccharides (e.g. glucose or arabinose), disaccharides (e.g. lactose, saccharose, maltose), oligosaccharides and polysaccharides (e.g. dextran), polyalcohols (e.g. sorbitol, mannitol, xylitol), salts (eg sodium chloride, calcium carbonate) or mixtures thereof. In some cases, monosaccharides or disaccharides such as lactose or glucose are used, for example in the form of a hydrate, for example lactose, for example, in the form of lactose monohydrate.

Excipients within the scope of the inhalable powder according to the present invention include a maximum average particle size of up to 250 μm, such as 10 to 150 μm, and 15 to 80 μm. Sometimes it may be appropriate to add a finer fraction of excipients having an average particle size of 1 to 9 μm to the above-mentioned excipients. These finer excipients are also selected from the group of possible excipients listed above. Finally, in order to prepare the inhalable powder according to the present invention, a micronized active substance in the form of a compound of formula 1 or co-crystals or salts of formulas 2 and 2a, for example, comprising an average particle size of 1 to 5 μm Active substances of 0.5 to 10 μm are added to the excipient mixture. Methods for preparing inhalable powders according to the invention by mixing the components together by grinding and micronizing are known from the prior art.

The inhalable powder according to the present invention can be administered using an inhaler known from the prior art.

Inhalation aerosols containing a propellant gas according to the present invention may contain the compounds of formula (1) or formulas (2) and (2a) in the form of co-crystals or salts dissolved in the propellant gas or in dispersed form. The compound of formula 1 or the co-crystal or salt form of formulas 2 and 2a may be contained in separate formulations or in a common formulation, both of which are dissolved, dispersed, or in each case only one component is dissolved and the other is It is dispersed. Propellant gases that can be used to prepare inhalation aerosols are known from the prior art. Suitable propellant gases are selected from hydrocarbons such as n-propane, n-butane or isobutane and halo hydrocarbons such as fluorinated derivatives of methane, ethane, propane, butane, cyclopropane or cyclobutane. The propellant gases may be used alone or in combination. In some cases, the propellant gas is a halogenated alkane derivative selected from TG134a and TG227 and mixtures thereof.

A propellant-driven inhalation aerosol may also contain other ingredients such as co-solvents, stabilizers, surfactants, antioxidants, lubricants and pH adjusting agents. All of these ingredients are known in the art.

The aforementioned propellant-driven inhalation aerosol according to the present invention can be administered using an inhaler known in the art (MDI = metered dose inhaler).

In addition, the active substance in the form of the compound of formula (1) or co-crystals or salts of formulas (2) and (2a) according to the present invention can be administered in the form of inhalable solutions and suspensions without propellants. The solvent used may be aqueous or alcoholic such as an ethanol solution. The solvent may be water itself or a mixture of water and ethanol. The relative proportions of ethanol to water are not limited, but in some cases the maximum volume includes up to 70% by volume, for example up to 60% by volume, up to 30% by volume. The remainder of the volume consists of water. A solution or suspension containing the compound of formula 1 or the co-crystal or salt form of formulas 2 and 2a is adjusted to a pH of 2 to 7, such as 2 to 5, using a suitable acid. The pH may be adjusted using an acid selected from inorganic or organic acids. Examples of particularly suitable inorganic acids include hydrochloric acid, hydrobromic acid, nitric acid, sulfuric acid and/or phosphoric acid. Examples of particularly suitable organic acids include ascorbic acid, citric acid, malic acid, tartaric acid, maleic acid, succinic acid, fumaric acid, acetic acid, formic acid and/or propionic acid and the like. In some cases, the inorganic acids are hydrochloric acid and sulfuric acid. It is also possible to use acids which have already formed acid addition salts with one of the active substances. Among the organic acids, ascorbic acid, fumaric acid and citric acid are used in some cases. If desired, mixtures of these acids can be used, especially in the case of acids having properties other than acidifying properties, for example as flavoring agents, antioxidants or complexing agents, for example as citric acid or ascorbic acid. According to the present invention, in some cases hydrochloric acid is used to adjust the pH.

If desired, the addition of edetic acid (EDTA) or sodium edetate, one of the known salts thereof, as a stabilizing or complexing agent may be omitted in these formulations. Other embodiments may contain this compound or these compounds. In one embodiment, sodium edetate based content includes less than 100 mg/100 ml, such as less than 50 mg/100 ml, and less than 20 mg/100 ml. Inhalable solutions with a content of sodium edetate from 0 to 10 mg/100 ml are used in some cases. Cosolvents and/or other excipients may be added to solutions for inhalation without propellants, such as those containing hydroxyl groups or other polar groups. Examples are alcohols - in particular isopropyl alcohol, glycol - in particular propylene glycol, polyethylene glycol, poly propylene glycol, glycol ethers, glycerol, polyoxyethylene alcohol and polyoxyethylene fatty acid esters. The terms excipients and additives in this context mean pharmacologically acceptable substances that are not active substances, but which can be formulated with the active substances or substances in physiologically suitable solvents to improve the qualitative properties of the active substance formulation. In some embodiments, these substances have no pharmacological effect or, with respect to the desired therapy, no significant or at least no undesirable pharmacological effect. Excipients and additives are, for example, surfactants such as soybean lecithin, oleic acid, sorbitan esters such as polysorbate, polyvinylpyrrolidone, other stabilizers, complexing agents, prolonging the shelf life of the final pharmaceutical formulation or antioxidants and/or preservatives, flavorings, vitamins and/or other additives known in the art. Additives also include pharmacologically acceptable salts such as sodium chloride as isotonic agents.

In some embodiments, excipients include, for example, antioxidants such as ascorbic acid, vitamins A, vitamin E, tocopherols, and similar vitamins and provitamins occurring in the human body that have not already been used to adjust pH.

Preservatives can be used to protect the formulation from contamination by pathogens. Suitable preservatives are those known in the art, in particular acetyl pyridinium chloride, benz alkonium chloride or benzoic acid or sodium benzoate, for example sodium benzoate in concentrations known from the prior art. The aforementioned preservatives may be present in concentrations of up to 50 mg/100 ml, such as 5 to 20 mg/100 ml.

In some embodiments, the formulation contains only benzalkonium chloride and sodium edetate in addition to the solvent water and the compound of Formula 1 or the co-crystal or salt form of Formulas 2 and 2a. In one embodiment, no sodium edetate is present.

The dosage of the compounds according to the invention is naturally highly dependent on the method of administration and the condition being treated. When administered by inhalation, the compound of Formula 1 or the co-crystal or salt form of Formulas 2 and 2a is characterized by high potency even at doses in the μg range. Compounds of formula (1) or co-crystals or salts of formulas (2) and (2a) can also be used effectively in the μg range or more. Dosages may range, for example, in grams.

In another aspect, the present invention relates to the aforementioned pharmaceutical formulation, characterized in that it contains a compound of formula 1 or co-crystal or salt form of formulas 2 and 2a, in particular the aforementioned pharmaceutical formulation. It can be administered by inhalation.

The following examples of formulations illustrate the invention without limiting the scope of the invention:

i. Examples of pharmaceutical formulations

A) per tablet tablet

100 mg of active substance 1, 2 or 2a

lactose 140 mg

240 mg corn starch

15 mg polyvinyl pyrrolidone

5 mg magnesium stearate

500 mg

Finely ground active substance, lactose and some corn starch are mixed together. After screening the mixture, it is wetted with a solution of polyvinyl pyrrolidone in water, kneaded, wet granulated and dried. The granules, the remaining corn starch and magnesium stearate were screened and mixed together. The mixture is compressed into tablets of suitable shape and size.

B) per tablet tablet

80 mg of active substance 1, 2 or 2a

Lactose 55 mg

190 mg corn starch

35 mg of microcrystalline cellulose

15 mg polyvinyl pyrrolidone

23 mg sodium carboxymethyl starch

2 mg magnesium stearate

400 mg

Finely ground active substance, some corn starch, lactose, microcrystalline cellulose and polyvinyl pyrrolidone are mixed together, the mixture is screened and processed with the remaining corn starch and water to form granules which are dried and screened. Add sodium carboxymethyl starch and magnesium stearate, mix and compress the mixture to form tablets of suitable size.

C) ampoule solution

50 mg of active substance 1, 2 or 2a

50 mg sodium chloride

5 ml of infusion water

The active substance is dissolved in water at its own pH or optionally pH 5.5 to 6.5 and the solution is made isotonic by addition of sodium chloride. The resulting solution is filtered to remove the pyrogen, and the filtrate is transferred to an ampoule under aseptic conditions, sterilized and heat sealed. Ampoules contain 5 mg, 25 mg and 50 mg of active substance.

D) metered aerosol

Active substance 1, 2 or 2a 0.005

Sorbitan Trioleate 0.1

monofluorotrichloromethane and

TG134a : TG227 2 : 1 ad 100

The suspension is transferred to an existing aerosol container with a metering valve. Preferably 50 μl suspension is discharged with each actuation. The active substance may also be released in higher doses if desired (eg 0.02% by weight).

E) solution (mg/100ml)

333.3 mg of active substance 1, 2 or 2a

10.0 mg benzalkonium chloride

EDTA 50.0 mg

HCl (1N) and pH 2.4

This solution can be prepared in the usual way.

F) Inhalable powder

12 μg of active substance 1, 2 or 2a

Lactose monohydrate ad 25 mg

Inhalable powders are prepared in the usual manner by mixing the individual ingredients.

j. indication

Methods are provided for improving cognition or other symptoms of a cognitive disorder by treating a subject/patient diagnosed with the cognitive disorder. Aspects of the method use a CCR3 modulator in a manner sufficient to modulate CCR3, eg, treat a patient for a cognitive-related disease. The method comprises treating a cognitively related disorder with an orally administered and bioavailable composition comprising a composition of a compound of Formula 1, a co-crystal or salt of Formula 2 or 2a, or a formulation of Formula 3 as described above. As summarized above and described in more detail below, a variety of age-related disorders, eg, cognitive-related diseases, can be treated by embodiments of the present invention. In some cases, the target condition is a cognitive-related disease state associated with neurodegeneration, e.g., evidenced by one or more decreased neurogenesis, e.g., a reduced number of BrdU or EdU positivity as compared to unaffected tissue. There is nerve damage exhibited by cells, Ki67 positive cells and Dcx positive cells. Compositions that modulate CCR3 may be used in cognitive-related diseases such as, but not limited to: mild cognitive impairment (MCI); Alzheimer's disease; Parkinson's disease; frontotemporal dementia (FTD); Huntington's disease; amyotrophic lateral sclerosis (ALS); multiple sclerosis (MS); glaucoma; muscular dystrophy; idiot; progressive supranuclear palsy (PSP); ataxia; bundle-system atrophy; weakness; Others are administered to a patient/subject diagnosed with a cognitive-related disorder as further described below. The methods of the present invention may further comprise monitoring improvement in progression of a neurodegenerative disease by measuring cognitive or physical improvement.

Methods are provided for improving motor coordination, function, or other symptoms of a movement disorder by treating a subject/patient diagnosed with the movement disorder. Aspects of the method include modulating CCR3 in a manner sufficient to treat a patient for movement disorders, eg, with a CCR3 modulator. The method comprises treating a movement disorder with an orally administered and bioavailable composition comprising a composition of a compound of Formula 1, a co-crystal or salt of Formula 2 or 2a, or a formulation of Formula 3, as described above. As summarized above and described in more detail below, various age-related disorders, such as movement disorders, can be treated by embodiments of the present invention. In some cases, the target condition is a motor disorder associated with neurodegeneration, e.g., evidenced by reduced neurogenesis in one or more, e.g., a reduced number of BrdU or EdU positive cells when compared to unaffected tissue; There is nerve damage exhibited by Ki67-positive cells and Dcx-positive cells. Compositions that modulate CCR3 can be used for movement disorders such as Parkinson's disease; Parkinsonism; Lewy body dementia; ataxia; dystonia; cervical dystonia; chorea; Huntington's disease, multiple-system atrophy; convulsion; progressive supranuclear palsy; perceptual dyskinesia; Tourette's syndrome; and tremors; Others may be administered to a patient/subject diagnosed with a movement disorder as further described below. The methods of the present invention may further comprise monitoring improvement in progression of a neurodegenerative disease by measuring cognitive or physical improvement.

Mild cognitive impairment (MCI) is a moderate impairment of cognition that presents with problems with memory or other mental functions, such as making decisions, planning, directing, or making decisions that deteriorate over time while overall mental function and daily activities remain intact. . Thus, although significant neuronal death does not typically occur, neurons in the aging brain are susceptible to lethal age-related alterations in structure at synapses, synaptic integrity, and molecular processing at synapses, all of which impair cognitive function. For example, individuals suffering from or at risk of developing an age-related cognitive impairment who would benefit from treatment with a compound of the invention by the methods disclosed herein also include individuals of any age suffering from a cognitive impairment due to do. age-related disorders; and individuals of any age diagnosed with an age-related disorder that generally accompanies cognitive impairment, individuals who have not yet begun to show symptoms of cognitive impairment. Examples of such age-related disorders include, without limitation, those listed below.

Alzheimer's disease. Alzheimer's disease is characterized by a gradual and indivisible loss of cognitive function associated with senile plaques in excess numbers of the brain cortex and cortical gray matter, in addition to excessive β-amyloid and nerve fiber entanglement composed of tau protein. The common form affects people over the age of 60, and the incidence increases with age. It accounts for more than 65% of dementia in the elderly.

The cause of Alzheimer's disease is unknown. The disease occurs in about 15-20% of families. In the rest, so-called sporadic cases, there are some genetic links. The disease has an autosomal dominant genetic pattern in most early onset and some onset cases, but with variable lifetime permeability. Environmental factors are the focus of important investigations.

In the course of the disease, synapses and ultimately neurons are lost in the cerebral cortex, hippocampus and subcortical structures (including selective cell loss at the base of the Meinert nucleus), locus, and dorsal sac nucleus. Decreased brain glucose use and perfusion in some areas of the brain (parietal and temporal cortex in early disease, prefrontal cortex in late disease). Neurogenic or senile plaques (composed of neurites, astrocytes and glial cells around the amyloid core) and nerve fiber entanglement (composed of paired helical filaments) play a role in the pathogenesis of Alzheimer's disease. Senile plaque and nerve fiber tangles occur with normal aging, but are much more common in people with Alzheimer's disease.

Parkinson's disease . Parkinson's disease (PD) is an idiopathic, slowly progressing, degenerative CNS disorder characterized by slow and diminished movement (kinesis), muscle stiffness, resting tremor (dystonia), muscle freezing, and postural instability. Originally considered primarily a movement disorder, PD is now recognized to cause depression and emotional changes. PD can also affect cognitive, behavioral, sleep, autonomic and sensory functions. The most common cognitive impairments include attention and concentration disorders, working memory, management functions, speech production, and spatial perception. A characteristic of PD is that symptoms associated with impaired motor function take precedence over symptoms associated with cognitive impairment, which is helpful in diagnosing the disease.

In primary Parkinson's disease, pigment neurons in the substantia nigra, locus, and other groups of brain stem dopaminergic cells degenerate. The cause is unknown. Loss of substantia nigra neurons projecting to the caudate nucleus and conch nucleus depletes the neurotransmitter dopamine in these areas. The onset is usually after the age of 40, and the incidence increases in the elderly.

Parkinson's disease is newly diagnosed in about 60,000 Americans each year and currently affects about 1 million Americans. Although PD is not fatal in itself, complications are the 14th leading cause of death in the United States. Currently, PD cannot be cured, treatment is usually prescribed to control symptoms, and later surgery is prescribed in severe cases.

Treatment options for PD include administering medications to help manage exercise deficits. These options either increase or replace dopamine, a neurotransmitter that has low concentrations in the brains of people with PD. These drugs include carbidopa/levodopa (which makes more dopamine in the brain); apomorphine, pramipexolo, ropinirole and rotingotine (dopamine agonists); selegiline and rasagiline (MAO-B inhibitors that prevent the breakdown of dopamine); entacapone and tolcapone (catechol-O-methyl transferase [COMT] inhibitors that give more levodopa in the brain); benztropine and trihexyphenidyl (anticholinergics); and amantadine (modulates tremor and spasticity). Exercise/physical therapy is usually prescribed to maintain physical and mental function.

However, current treatment options treat the symptoms of PD but are not curative and do not prevent disease progression. In addition, current drugs tend to lose efficacy in late PD. Levodopa, the most commonly prescribed drug, usually causes side effects within 5-10 years of taking the drug. These side effects can be severe and can result in motor fluctuations and unpredictable tremors in motor control between doses, as well as sudden movements/tremors (dyskinesias) that are difficult to manage and even as disturbing as the symptoms of PD itself. Therefore, there remains a need for new therapies with novel mechanisms of action that can be administered with or in combination with current PD drugs.

Parkinsonism . Secondary Parkinsonism (also called atypical Parkinson's disease or Parkinsonism) results from loss or interference with the action of dopamine in the basal ganglia due to other idiopathic degenerative diseases, drugs, or exogenous toxins. The most common cause of secondary parkinsonism is the ingestion of antipsychotics or reserpine, which blocks dopamine receptors, causing parkinsonism. Less common causes include carbon monoxide or manganese poisoning, hydrocephalus, structural lesions (tumors, infarcts affecting the midbrain or basal ganglia), subdural hematomas and degenerative disorders, including nigrostriatal degeneration. Certain disorders, such as progressive supranuclear palsy (PSP), multiple-system atrophy (MSA), cortical basal ganglia (CBD), and Lewy body dementia (DLB), may show symptoms of Parkinsonism before the main symptoms required for a specific diagnosis appear. It is marked as "Parkin's Syndrome."

Assessment of progression in PD

Several grading scales were used to assess the progression of PD. The most widely used scales are the UPDRS (Unified Parkinson's Disease Rating Scale) introduced in 1987 (J. Rehabil Res. Dev., 2012 49(8): 1269-76) and the Horn and Night Scale ((Neruology) , 1967 17(5): 427-42) Additional scales include the Movement Disorder Society's (MDS) updated UPDRS scale (MDS-UPDRS) and the Schwab and England's Activities of Daily Living (ADL) scale.

The UPDRS scale evaluates 31 items contributing to three subscales: (1) psychological state, behavior, and mood; (2) activities of daily living; and (3) movement tests. The Horn and Night scale classifies PD into 5 stages with distinct substages: 0 - no signs of disease; 1 - symptoms only on one side; 1.5 - symptoms on one side, but also includes the neck and spine; 2-symptoms on both sides without balance disturbance; 2.5 - mild symptoms on both sides that recover when undergoing a pull test; 3-balance disorders of mild to moderate disease; 4 - severe disability, but the ability to stand and walk without assistance; and 5- Needs a wheelchair or bed without assistance. The Schwab and England scale classifies PD into several percentages (100%-completely independent to 10%-completely dependent).

frontotemporal dementia . Frontotemporal dementia (FTD) is a symptom of progressive deterioration of the frontal lobe of the brain. Over time, degeneration may progress to the temporal lobe. FTD accounts for 20% of senile dementia, second only to common Alzheimer's disease (AD). Symptoms are divided into three groups according to the function of the frontal and temporal lobes affected.

behavioral variant FTD (bvFTD), on the one hand lethargy and apathy, on the other hand, disinhibition; In progressive nonfluent aphasia, difficulty in pronunciation, disruption of verbal fluency due to phonological and/or syntax errors is observed but word comprehension is maintained, and semantic dementia (SD), where the patient is fluent in normal phonology and syntax, but I'm having trouble understanding names and words. Other cognitive symptoms common to all FTD patients include impaired management function and ability to concentrate. Other cognitive abilities, including perception, spatial skills, memory, and practice, are generally intact. FTD can be diagnosed by observing frontal and/or frontotemporal atrophy on a structural MRI scan.

There are many forms of FTD, any of which can be treated or prevented using the subject methods and compositions. For example, one form of frontotemporal dementia is SD (Semantic Dementia). SD is characterized by loss of semantic memory in both the verbal and non-verbal domains. SD patients often complain of difficulty finding words. Clinical signs include fluency aphasia, name aphasia, impaired comprehension of words, and associative visual aphasia (not matching semantically related pictures or objects). As the disease progresses, behavioral and personality changes often appear similar to those seen in frontotemporal dementia, although cases have been described for 'pure' semantic dementia, often with few symptoms of behavioral delay. Structural MRI images show a characteristic pattern of atrophy in the temporal lobe (mainly on the left), with a lower than superior and a greater than posterior temporal atrophy.

As another example, another form of frontotemporal dementia is Pick's disease (PiD, PcD). A hallmark of the disease is the accumulation of the tau protein in neurons, which accumulates in silver-colored spherical aggregates known as "pic bodies." Symptoms include loss of speech (aphasia) and dementia. Patients with frontal dysfunction become respectful and socially maladjusted.

They may have or steal obsessive or repetitive stereotypes. Patients with prefrontal dysfunction may show a lack of worry, apathy, or spontaneous decline. Patients may lose self-monitoring, abnormal self-awareness, and ability to understand meaning.

Patients with gray matter loss in the bilateral posterior orbital prefrontal cortex and right anterior chamber may show pathological changes in eating habits, such as a preference for sweets. Patients with more focal gray matter loss in the anterior orbital prefrontal cortex may develop bulimia. Some of the symptoms may be relieved at first, but the disease progresses and patients often die within 2-10 years.

Huntington's disease . Huntington's disease (HD) is characterized by emotional, behavioral and mental disorders; loss of intellectual or cognitive function; and a hereditary progressive neurodegenerative disorder characterized by the development of motor abnormalities (motor disorders). Classic signs of HD include involuntary, rapid, erratic, oscillating movements that may affect the face, arms, legs, or torso, and progressive cognitive decline, including progressive loss of thought processing and intellectual abilities. impairment of memory, abstract thinking and judgment; Inappropriate perception of time, place, or identity (disorientation); increased confusion; There may be personality changes (personality breakdown). Symptoms usually become evident during the first 40 or 50 years of life, but the age of onset is variable and varies from infancy to late adulthood (eg, 70s or 80s).

HD is an autosomal dominant trait that is transmitted within families. This disorder occurs as a result of an abnormally long sequence or "repeat" of coded instructions within the gene on chromosome 4 (4p16.3). The progressive loss of nervous system function associated with HD results from loss of neurons in certain areas of the brain, including the basal ganglia and cerebral cortex.

Amyotrophic Lateral Sclerosis . Amyotrophic lateral sclerosis (ALS) is a rapidly progressing, always fatal, neurological disease that attacks motor neurons. Signs of muscle weakness and atrophy and anterior horn cell dysfunction are initially most common in the hands and less frequently in the feet. The site of onset is random and the progression is asymmetric. Convulsions are common, followed by weakness. Rarely, patients survive for 30 years. 50% die within 3 years of onset, 20% survive 5 years, and 10% survive 10 years.

Onset during middle or late adult life and includes progressive, general motor involvement without paresthesia. Nerve conduction velocity is normal until late in the disease. Recent studies have suggested cognitive impairments, particularly reductions in immediate verbal memory, visual memory, and language and management functions.

Decrease in cell volume, synapse number, and total synapse length have also been reported in normal-looking neurons from ALS patients. It has been suggested that when the plasticity of the active region reaches its limit, synapses continue to be lost and dysfunction may occur. Preventing the formation or promotion of new synapses or loss of synapses in these patients can preserve neuronal function.

Multiple Sclerosis . Multiple sclerosis (MS) is characterized by a variety of symptoms and signs of CNS dysfunction with remission and recurrent exacerbations. The most common symptoms are numbness in one or more extremities, torso, or one side of the face, weakness or blunting in the legs or hands; or visual impairment, such as partial blindness and pain in one eye (posterior optic neuritis), darkening of vision or scotomas. Common cognitive impairments include memory impairment (gathering, retaining, and retrieving new information), attention and concentration (particularly distributed attention), information processing, management functions, spatial perception functions, and language fluency. Common early symptoms are double vision (double vision) caused by ocular palsy, temporary weakness in one or more extremities, slight stiffness or abnormal obesity in the extremities, mild gait disturbance, difficulty in bladder control, dizziness, and mild emotional disturbances; All show CNS involvement and often occur months or years before disease is recognized. Excessive heat can accentuate symptoms and signs.

This process is highly variable and unpredictable and varies more and less in most patients. At first, months or years of remission can separate episodes. This is especially true when the disease begins with retroocular optic neuritis. However, some patients have frequent seizures and are rapidly disabling. Some processes can be fast.

glaucoma . Glaucoma is a common neurodegenerative disease that affects retinal ganglion cells (RGCs). This is indicated by the presence of compartmentalized degeneration programs at synapses and dendrites, including RGCs. Recent evidence also indicates a correlation between cognitive impairment in the elderly and glaucoma (Yochim BP, et al. Prevalence of cognitive impairment, depression, and anxiety symptoms among older adults with glaucoma. J Glaucoma. 2012;21(4): 250-254).

muscular dystrophy . Muscular dystrophy (DM) is an autosomal dominant multiple system disorder characterized by dystrophic muscle weakness and myositis. The molecular defect is an extended trinucleotide (CTG) repeat in the 3' untranslated region of the myotonin protein kinase gene on chromosome 19q. Symptoms can occur at any age and have a wide range of clinical severity. Myotonia is prominent in the hand muscles, and ptosis is common even in mild cases. In severe cases, peripheral muscle weakness appears markedly, and it may be accompanied by cataracts, premature baldness, slanted face, cardiac arrhythmias, testicular atrophy, and endocrine abnormalities (eg diabetes). Mental retardation is common in severe congenital forms, and aging-related declines in frontal and temporal cognitive functions, particularly language and management functions, are observed in moderate adult forms of disability. Severely affected people die in their early 50s.

dementia . Dementia is a type of disorder with symptoms that severely affect thinking and social skills to the extent that it interferes with daily functioning. In addition to the dementia observed in the later stages of the age-related disorders discussed above, other cases of dementia include vascular dementia, and dementia with Lewy bodies as described below.

In vascular dementia, or "multi-infarct dementia," cognitive impairment is caused by problems with blood supply to the brain, usually a series of minor strokes, or sometimes one large stroke followed by another small stroke. Vascular lesions may be the result of diffuse cerebrovascular disease, such as small blood vessel disease, focal lesions, or both. Patients with vascular dementia have cognitive impairment either acutely or subacutely after an acute cerebrovascular accident, after which progressive cognitive decline is observed. Cognitive impairments are similar to those observed in Alzheimer's disease, including impairments in speech, memory, complex visual processing or management functions, and brain-related changes are due to chronic reduced blood flow to the brain and not due to AD pathology, but dementia develops. Single photon emission computed tomography (SPECT) and positron emission tomography (PET) neuroimaging can be used to confirm the diagnosis of multi-infarct dementia with assessments related to mental state testing.

Lewy body dementia . Lewy body dementia (DLB), known by a variety of other names including Lewy body dementia, diffuse Lewy body dementia, cortical Lewy body disease, and senile dementia of the Lewy body type, is a neurodegenerative disease of Lewy bodies (alpha- It is characterized by the presence of clumps of synuclein and ubiquitin proteins. The main characteristic is a decline in cognitive function, especially in management function. Agility and short-term memory will go up and down.

Persistent or recurrent visual hallucinations with vivid, detailed pictures are often an early diagnosis symptom. DLB is sometimes confused with Alzheimer's disease and/or vascular dementia in its early stages. While Alzheimer's disease usually begins very gradually, DLB often has a rapid or acute onset. DLB symptoms include motor symptoms similar to Parkinson's symptoms. DLB, with respect to Parkinson's disease, is distinguished from the dementia that occasionally occurs in Parkinson's disease, depending on how long dementia symptoms appear. Parkinson's disease due to dementia (PDD) will be a diagnosis when the onset of dementia is more than one year after the onset of Parkinson's disease. DLB is diagnosed when cognitive symptoms begin concurrently with Parkinson's symptoms or within 1 year.

DLB processing is a complex process and requires a multifaceted approach (Neurology, 2017 89:1-13). Typical Parkinsonism therapies, such as dopaminergic and anticholinergic drugs, can worsen cognitive and behavioral symptoms. Optimal treatment generally uses both pharmacological (exercise, cognitive training and caregiver-centered training) and non-pharmacological approaches. For cognitive symptoms, acetylcholinesterase inhibitors such as the NMDA receptor antagonist, memantine may be administered (eg rivastigmine, donepezil). For neuropsychiatric symptoms, acetylcholinesterase inhibitors can improve apathy and hallucinations. Antipsychotics unfortunately increase the risk of death in people with DLB. Motor symptoms are less responsive to dopamine therapy in people with DLB and may exacerbate the risk of psychosis. Levodopa can be used, but the threshold dose is low, so there is a need to treat DLB with new drugs.

Progressive supranuclear palsy . Progressive supranuclear palsy (PSP) is a brain disorder that causes severe and progressive problems with gait and balance control, along with complex eye movement and thinking problems. One of the classic signs of the disease is an inability to aim the eye correctly due to lesions in the brain area that coordinates eye movements. Some describe this effect as blurry. Affected people often show changes in mood and behavior, including depression, apathy, and progressive mild dementia. The long name for the disorder is that the disease starts slowly, continues to get worse (progressive), and causes weakness (paralysis) by damaging certain parts of the brain above the pea-sized structures called the nuclei that control eye movement (the brain nucleus). indicates. PSP was first described as a distinct disorder in 1964 when three scientists published a paper distinguishing symptoms from Parkinson's. It is sometimes referred to as Still-Richardson-Olzewski syndrome, reflecting the combined names of the scientists who defined the disorder. The PSP gets progressively worse, but the PSP itself doesn't kill anyone.

ataxia . People with ataxia have coordination problems because the part of the nervous system that controls movement and balance is affected. Ataxia can affect movements of the fingers, hands, arms, legs, body, speech, and eyes. Ataxia is often used to describe the conditioning symptoms that may be associated with infections, injuries, other diseases, or degenerative changes in the central nervous system. Ataxia is also used to denote a group of specific degenerative disorders of the nervous system called hereditary and sporadic ataxia, a major emphasis of the National Ataxia Foundation.

Multiple system atrophy . Multiple system atrophy (MSA) is a neurodegenerative disorder. MSA is associated with the degeneration of nerve cells in certain areas of the brain. This cellular degeneration causes problems with the body's movement, balance, and other autonomic functions, such as bladder control or blood pressure control.

The cause of MSA is unknown and no specific risk factors have been identified. In men, it occurs in about 55% of the typical age of late 50s to early 60s. MSA often presents the same symptoms as Parkinson's. However, MSA patients usually have minimal responses to dopaminergic drugs used for Parkinson's disease.

dystonia . Dystonia is a condition in which involuntary muscle contractions continue. Such contractions can lead to twisting and repetitive movements. The disorder can affect the whole body or specific parts of the body referred to as generalized dystonia or focal dystonia (respectively). Cervical dystonia may be prolonged or intermittent contractions in the neck muscles. There is no cure for dystonia. Current treatments include carbidopa-levodopa, trihexyphenidyl, benztropine, tetrabenazine, diazepam, clonazepam, baclofen, physiotherapy, speech therapy, stretching, massage, and invasive surgery.

weakening . Weakness, one of the geriatric syndromes, is a geriatric syndrome characterized by functional and physical decline, including reduced mobility, muscle weakness, physical slowing, poor endurance, low physical activity, malnutrition and involuntary weight loss. This decline is often accompanied by comorbid dysfunction and the consequences of diseases such as cancer. However, weakness can occur even in the absence of disease. Individuals with frailty are at increased risk of adverse prognosis from fractures, accidental falls, disability, comorbidities, and premature death (C. Buigues, et al. Effect of a Prebiotic Formulation on Frailty Syndrome: A Randomized, Double-Blind Clinical Trial, Int. J. Mol. Sci. 2016, 17, 932). In addition, individuals suffering from infirmity have an increased incidence of higher medical costs (Id.).

Common symptoms of weakness can be determined by certain types of tests. For example, unintentional weight loss involves losing 10 pounds or more. 5% or more of the previous year's body weight; Muscle weakness can be determined by a reduced grip strength (adjusted for gender and BMI) of at least 20% from baseline; Physical slowing can be based on the time required to walk a distance of 15 feet. Endurance weakness can be determined by an individual's report of exhaustion. Low physical activity can be measured using standardized questionnaires (Z. Palace et al., The Frailty Syndrome, Today's Geriatric Medicine 7(1), 18(2014)).

In some embodiments, the methods and compositions of the present invention are used to slow the progression of age-related cognitive, motor or other age-related disorders. In other words, the individual's cognitive, motor, or other ability will decrease more slowly after treatment with the disclosed method compared to before or without treatment with the disclosed method. In some such cases, the method of treatment includes measuring the progression of cognitive, motor, or other age-related decline after treatment, and determining that the progression of regression is reduced. In some such instances, the determination is made, for example, by measuring and comparing the individual's rate of regression, exercise, or other age-related ability prior to treatment at a point prior to administration of the subject blood product, eg, at two or more time points.

The methods and compositions of the present invention are also used to stabilize cognitive, motor or other abilities in an individual, eg, an individual suffering from age-related cognitive decline or an individual at risk of suffering from age-related cognitive decline. For example, an individual has some age-related cognitive impairment, and the progression of cognitive impairment observed prior to treatment with the disclosed methods will stop after treatment with the disclosed methods. As another example, an individual may be at risk of developing an age-related cognitive decline (eg, the individual may be over 50 years of age or have been diagnosed with an age-related disorder), and the individual's cognitive abilities do not change substantially, i.e., , no cognitive decline was detected after treatment with the disclosed method compared to before treatment with the disclosed method.

The methods and compositions of the present invention are also used to reduce cognitive, motor or other age-related disorders in individuals suffering from age-related disorders. In other words, the ability to be affected in an individual after treatment by the subject method is improved. For example, the cognitive ability of the individual is, for example, at least 2 fold, at least 5 fold, at least 10 fold, at least 15 fold, at least 20 fold, at least 30 fold, compared to the cognitive ability observed in the individual prior to treatment by the subject method; 50 times or more, 60 times or more, 70 times or more, 80 times or more, 90 times or more, or 100 times or more.

In some instances, treatment with the methods and compositions of the present invention restores the cognitive, motor or other ability of an individual suffering from age-related cognitive or motor decline, eg, to the level it would have been when the individual was about 40 years of age or younger. In other words, there is no cognitive or motor impairment.

k. Diagnosis and monitoring methods for improvement of neurodegenerative diseases

One of ordinary skill in the art will recognize that, among the various methods of diagnosing and monitoring disease progression and amelioration of neurodegenerative-related diseases, the following types of assessment may be used alone or in combination with a subject suffering from a neurodegenerative disease. Methods of the following types are given by way of example and are not limited to the recited methods. One of ordinary skill in the art will recognize that other methods of monitoring for disease will be useful in practicing the present invention. These methods are also contemplated by the methods of the present invention.

i. general perception

The methods of the present invention further comprise a method of monitoring the effect of a drug or treatment on a subject for treating cognitive impairment and/or age-related dementia, the method comprising comparing cognitive function before and after treatment. One of ordinary skill in the art recognizes that there are known methods for assessing cognitive function. For example, but not limited to, the method may include assessment of cognitive function based on medical history, family history, physical and neurological examination, laboratory testing, and neuropsychological assessment by a clinician specializing in dementia and cognitive function. have. Additional embodiments contemplated by the present invention include: assessment of consciousness, such as using the Glasgow Coma Scale (EMV); Mental status tests, including abbreviated mental test scores (AMTS) or mini mental status tests (MMSE) (Folstein et al., J. Psychiatr. Res 1975; 12: 1289-198); global assessment of higher functioning; For example, intracranial pressure estimation by fundus endoscopy.

In one embodiment, examination of the peripheral nervous system includes odor sensation, visual field and vision, eye movement and pupil (sympathetic and parasympathetic), sensory function of the face, strength of facial and shoulder girdle muscles, hearing, taste, pharyngeal movement and reflexes, tongue It includes exercise, etc., and can be individually tested. Example: Visual acuity can be tested with a Snellen acuity table, a reflex hammer can be used to test the masses, biceps, triceps, knee tendons, ankle sway and plantar reflexes (i.e. Babinski reflex; muscle strength is measured on the MRC scale 1 to 5; muscle tone and signs of stiffness may be tested.

ii. multiple sclerosis

In addition to monitoring for improvement in symptoms related to cognition, the progression or improvement of neurodegeneration associated with multiple sclerosis (MS) can be monitored using techniques well known to those skilled in the art. By way of example and not limitation, monitoring may include cerebrospinal fluid (CSF) monitoring; magnetic resonance imaging (MRI) to detect lesions and development of demyelinating plaques; evoked potential studies; There is gait monitoring.

CSF analysis can be performed, for example, via lumbar puncture to obtain pressure, appearance and CSF content. The range of normal values is generally: pressure (70-180 mm H ₂ 0); Appearance is clear and colorless; total protein (15-60 mg/100 ml); IgG is 3-12% of total protein; Glucose is 50-80mg/100ml; Cell count is 0-5 white blood cells and no red blood cells; Chloride (110-125 mEq/L). Abnormal results may indicate the presence or progression of MS.

MRI is another technique that can be performed to monitor disease progression and improvement. Typical criteria for monitoring MS by MRI include the appearance of uneven areas of abnormal white matter in the brain hemispheres and ventricles, and lesions present in the cervical or thoracic regions of the cerebellum and/or brainstem and spinal cord.

Evoked potentials can be used to monitor the progression and improvement of MS in a subject. Evoked potential measures the blunting of electrical impulses such as Visual Evoked Response (VER), Brain Stem Auditory Evoked Responses (BAER), and Somatosensory Evoked Responses (SSER). Abnormal responses indicate decreased conduction velocity in the central sensory pathway.

Gait monitoring can also be used to monitor disease progression and improvement in MS patients. MS is often accompanied by impaired mobility and abnormal gait due to fatigue. For example, monitoring may be performed using a mobile monitoring device worn by the subject. (Moon, Y., et al. , Monitoring gait in multiple sclerosis with novel wearable motion sensors , PLOS One, 12(2): e0171346 (2017)).

iii. huntington

In addition to monitoring for improvement in symptoms related to cognition, progression or amelioration of neurodegeneration associated with Huntington's disease (HD) can be monitored using techniques well known to those skilled in the art. By way of example and not limitation, monitoring may be performed through techniques such as: motor function; behavior; functional evaluation; and imaging.

Examples of motor function that can be monitored as signs of disease progression or improvement include chorea and dystonia, spasticity, twitching, ocular movement disorders, and gait/balance changes. Techniques for performing monitoring of these metrics are well known to those skilled in the art (see Tang C, et al. , Monitoring Huntington's disease progression through preclinical and early stages , Neurodegener Dis Manag 2(4):421-35 (2012)).

The psychiatric effects of HD provide an opportunity to monitor disease progression and improvement. For example, a psychiatric diagnosis may be performed to determine whether a subject suffers from psychosis of depression, irritability, confusion, anxiety, apathy, and paranoia. (Id.)

Functional assessments can also be used to monitor disease progression or improvement. A total function score technique ((Id.) has been reported, decreasing by 1 point per year in some HD groups.

MRI or PET may also be used to monitor disease progression or improvement. For example, in HD, striatal projection neurons are lost, and changes in the number of these neurons can be monitored in a subject. Techniques for determining neuronal changes in HD subjects include imaging of dopamine D2 receptor binding (Id.).

iv. ALS

In addition to monitoring for improvement in symptoms related to cognition, the progression or improvement of neurodegeneration associated with amyotrophic lateral sclerosis (ALS) can be monitored using techniques well known to those skilled in the art. By way of example and not limitation, monitoring may include functional evaluation; determination of strength; measurement of respiratory function; Measure low motor neuron (LMN) loss; and measurement of upper motor neuron (UMN) dysfunction.

Functional assessment may be performed using functional measures well known to those of skill in the art, such as the ALS Functional Rating Scale (ALSFRS-R), which evaluates symptoms related to medulla oblongata, extremities and respiratory function. Rate of change is useful for predicting survival as well as disease progression or improvement. Another measure includes the Combined Assessment of Function and Survival (CAFS), which combines changes in survival time and ALSFRS-R to rank patients' clinical outcomes (Simon NG, et al. , Quantifying Disease Progression in Amyotrophic). Lateral Sclerosis , Ann Neurol 76:643-57 (2014)).

Muscle strength can be tested and quantified using Complex Manual Muscle Test (MMT) scoring. This includes average measurements taken from different muscle groups using the Medical Research Council (MRC) strength rating scale (Id.). Manual dynamometry (HHD) may be used, among other techniques (Id.).

Respiratory function can be performed using a portable spirometry device and is used to obtain Forced Vital Capacity (FVC) at baseline to predict disease progression or improvement. In addition, peak inspiratory pressure, sniff nasal inspiratory pressure (SNIP) and surf FVC can be used to monitor disease progression/improvement. (Id.)

Low motor neuron loss is another indicator that can be used to monitor disease progression or improvement in ALS. Neurophysiological indices can be determined by measuring complex muscle action potentials (CMAPs) in motor nerve conduction studies, the parameters of which include CMAP amplitude and F-wave frequency ( Id. and de Carvalho M, et al. , Nerve conduction studies in amyotrophic lateral sclerosis . Muscle Nerve 23:344-352,(2000)). A lower motor neuron unit count (MUNE) can also be estimated. In MUNE, estimates of the contribution of individual motor units to the maximal CMAP response to estimate the number of residual motor axons supplying the muscle are used to determine disease progression or improvement (Simon NG, et al., supra ). . Additional techniques for determining loss of LMN include detecting changes in muscle thickness using nerve excitability, electrical impedance electromyography, and muscle ultrasound (Id.).

Upper motor neuron dysfunction is another measure that can be used to monitor disease progression or improvement of ALS. Techniques to determine dysfunction include MRI or PET scans of the brain and spinal cord, transcranial magnetic stimulation; Determination of levels of biomarkers in cerebrospinal fluid (CSF).

v. glaucoma

In addition to monitoring for improvement in symptoms related to cognition, the progression or amelioration of neurodegeneration associated with glaucoma can be monitored using techniques well known to those skilled in the art. By way of example and not limitation, monitoring may include: measurement of intraocular pressure; assessment of damage to the optic disc or optic nerve head; visual field testing for peripheral vision loss; and imaging of the optical disc and retina for topographic analysis.

vi. Progressive supranuclear palsy (PSP)

In addition to monitoring for improvement in symptoms related to cognition, the progression or improvement of neurodegeneration associated with progressive supranuclear palsy (PSP) can be monitored using techniques well known to those skilled in the art. By way of example and not limitation, monitoring may include functional assessment (daily living or ADL); exercise evaluation; determination of psychiatric symptoms; There are volumetric and functional magnetic resonance imaging (MRI).

A subject's level of functioning with respect to independence, partial dependence, or complete dependence on others may be useful in determining disease progression or amelioration (Duff, K, et al. , Functional impairment in progressive supranuclear palsy , Neurology 80:380). -84, (see 2013) The Progressive Supranuclear Palsy Rating Scale (PSPRS) is a rating scale consisting of 28 metrics in six categories: daily activity (by record); behavior; years; The result is a score ranging from 0 to 100. 6 items are rated 0-2 and 22 items are rated 0-4 for a total score of 100. The PSPRS score is a practical measure and patient survival It is also a sensitive predictor of disease progression and useful for monitoring disease progression or improvement ( Golbe LI, et al. , A clinical rating scale for progressive supranuclear palsy , Brain 130:1552-65, (2007). )).

The ADL section of the Unified Parkinson's Disease Rating Scale (UPDRS) can also be used to quantify functional activity in PSP subjects (Duff K, et al. , supra). Similarly, the Schwab & England Daily Living Activity Score (SE-ADL) can be used to assess independence. In addition, the motor function section of UPDRS is useful as a reliable measure for assessing disease progression in PSP patients. The exercise section may include, for example, 27 different measures to quantify motor function in PSP patients. Examples of these include tremors, stiffness, finger tapping, posture, and gait. Behavioral abnormalities (e.g., delusions, hallucinations, agitation, depression, anxiety, euphoria, apathy, disinhibition, irritability and The frequency and severity of abnormal motor behavior) can be measured (Id.).

Functional MRI (fMRI) can also be used to monitor disease progression and improvement. fMRI is a technique that uses MRI to measure changes in brain activity in a specific area of the brain, usually based on blood flow to that area. Blood flow is thought to be involved in brain region activation. Patients with neurodegenerative disorders such as PSP may have physical or mental testing before or during a scan with an MRI scanner. By way of example and not limitation, the test may be a well-established force control paradigm in a patient who is asked to generate a force with the hand most affected by the PSP, and maximal spontaneous contraction (MVC) is measured by fMRI immediately after the test is performed. (Burciu, RG, et al. , Distinct patterns of brain activity in progressive supranuclear palsy and Parkinson's disease , Mov. Disord. 30(9):1248-58(2015)).

Volumetric MRI is a technique in which MRI scanners measure volume differences in regional brain volumes. This can be done, for example, by contrasting different disorders, or by measuring the difference in volume of a patient's brain regions over time. Volumetric MRI can be used to determine disease progression or amelioration of neurodegenerative diseases such as PSP. This technique is well known to those skilled in the art. (Messina D, et al. , Patterns of brain atrophy in Parkinson's disease, progressive supranuclear palsy and multiple system atrophy , Parkinsonism and Related Disorders, 17(3):172-76(2011)) . Examples of brain regions that can be measured include, but include, intracranial volume, cerebral cortex, cerebellar cortex, thalamus, caudate, conch nucleus, pallor nucleus, hippocampus, amygdala, lateral ventricle, third ventricle, fourth ventricle and brainstem Not limited.

vii. neurogenesis

A non-invasive technique for assessing neurogenesis has been reported (Tamura Y. et al ., J. Neurosci. (2016) 36(31):8123-31). Positron emission tomography (PET) used in conjunction with the tracer, [ ¹⁸ F] FLT, in combination with the BBB transporter inhibitor probenecid, allows tracers to accumulate in neurogenic regions of the brain. Such imaging allows the assessment of neurogenesis in patients being treated for neurodegenerative diseases.

viii. Parkinson's disease and motor function

Several grading scales were used to assess the progression of PD. The most widely used scales are the Unified Parkinson's Disease Rating Scale (UPDRS), introduced in 1987 (J. Rehabil. Res. Dev., 2012 49(8): 1269-76) and the Horn and Night Scale (Neurology, 1967 17(5): 427-42). Additional scales include the Movement Disorder Society's (MDS) updated UPDRS scale (MDS-UPDRS) and the Schwab and England's Activities of Daily Living (ADL) scale.

The UPDRS scale evaluates 31 items contributing to three subscales: (1) mental, behavioral, and mood; (2) activities of daily living; and (3) movement tests. The Horn and Night scale classifies PD into five distinct sub-stages. 0 - no signs of disease; 1 - symptoms only on one side; 1.5 - symptoms on one side, but also includes the neck and spine; 2-symptoms on both sides without balance disturbance; 2.5 - mild symptoms on both sides that recover when undergoing a pull test; 3-balance disorders of mild to moderate disease; 4 - severe disability, but the ability to stand and walk without assistance; and 5- Needs a wheelchair or bed without assistance. The Schwab and England scale classifies PD into several percentages (100%-completely independent to 10%-completely dependent).

General motor function can be assessed using widely used scales, including the General Motor Function Scale (GMF). Three components of dependence, pain and anxiety are tested (Aberg A.C., et al. (2003) Disabil. Rehabil. 2003 May 6; 25(9):462-72.). Home monitoring or wearable sensors can also be used to assess motor function. For example, gait (speed of movement, variability, leg stiffness) can be detected by an accelerometer. Posture (trunk tilt) by gyroscope; Leg movements are driven by an accelerometer; Hand movements are by accelerometer and gyroscope; Tremors (amplitude, frequency, duration, asymmetry) are measured by an accelerometer; The fall is by an accelerometer; Gait freeze is by accelerometer; Dyskinesias are indicated by accelerometers, gyroscopes and inertial sensors; West movement (duration and frequency) is detected by an accelerometer and gyroscope, and aphasia (pitch) is detected using a microphone (Pastorino M, et al., Journal of Physics: Conference Series 450 (2013) 012055).

1. Reagents, Devices and Kits

Also provided are reagents, devices, and kits thereof for practicing one or more of the methods described above. The reagents, devices and kits of the present invention can be very diverse. Reagents of interest and devices of interest include those mentioned above with respect to methods of administering a compound of Formula 1 to a subject.

In addition to the above components, the subject kit will further comprise instructions for practicing the subject method. These instructions may exist in various forms in the subject kit, and one or more of them may be present in the kit. One form in which such instructions may exist is a suitable medium or substrate, for example, the paper or paper on which the information is printed, as information printed on the packaging of a kit, package insert, etc. Another means is a computer readable medium, such as a diskette on which information is recorded, a CD, a portable flash drive, and the like. Another means that may exist is a web site address that can be used to access information at a remote site via the Internet. The kit may have any convenient means.

VII. Example

The following examples are provided by way of illustration and not limitation.

a. pharmaceutical preparations

Pharmaceutical compositions administered to a subject suffering from a cognitive or neurodegenerative disease comprising the compounds, co-crystals and salts described above are disclosed in U.S. Patent Application Publication Nos. 2013/0266646, 2016/0081998, U.S. Patent Nos. 8,278,302, 8,653,075, RE 45323, 8,742,115, 9,233,950 and 8,680,280, which are incorporated herein by reference in their entirety. In addition, pharmaceutical compositions can be prepared as described in the Examples below:

1. Tablet Formulation - Wet Granulation

Copovidone is dissolved in ethanol at ambient temperature to produce a granulation liquid. A premix is prepared by blending a portion of the active CCR3 antagonist component, lactose and crospovidone in a suitable mixer. The premix is wetted with the granulation liquid and then granulated. The wet granules are optionally sieved through a sieve having a mesh size of 1.6-3.0 mm. The granules are dried at 45° C. in a suitable dryer to a residual moisture content corresponding to a loss of 1-3% on drying. The dried granules were sieved through a sieve with a mesh size of 1.0 mm. The granules are blended with crospovidone and a portion of microcrystalline cellulose in a suitable mixer. Magnesium stearate is added to this blend after passing through a 1.0 mm sieve to remove tangles. The final mixture is then produced by final mixing in a suitable mixer and compressed into tablets. The following tablet compositions can be obtained:

ingredient mg/
refine %/refine active ingredient 28.500 30.0 crospovidone 1.500 1.6 lactose 28.000 29.5 copovidone 3.000 3.2 Total (granules) 61.000 64.3 microcrystalline cellulose 31.000 32.6 crospovidone 2.500 2.6 magnesium stearate 0.500 0.5 Sum 95.000 100.000

2. Tablet Formulation - Melt Granulation

A pre-mix is prepared by blending the active CCR3 antagonist component, lactose, a portion of mcc, polyethylene glycol, lactose and a portion of crospovidone in a suitable mixer. The premix is heated in a high shear mixer and then granulated. The hot granules were cooled to room temperature and sieved through a sieve with a mesh size of 1.0 mm. The granules are blended with crospovidone and a portion of microcrystalline cellulose in a suitable mixer. Magnesium stearate is added to this blend after passing through a 1.0 mm sieve to detangle. The final mixture is then produced by final mixing in a suitable mixer and compressed into tablets. The following tablet compositions can be obtained:

ingredient mg/tablet %/refine active ingredient 28.500 30.0 crospovidone 1.500 1.6 lactose 11.000 11.6 polyethylene glycol 14.300 15.1 MCC 5.700 6.0 Total (granules) 61.000 64.3 microcrystalline cellulose 31.000 32.6 crospovidone 2.500 2.6 magnesium stearate 0.500 0.5 Sum 95.000 100.000

3. Tablet Formulation - Hot Melt Granulation

A premix is prepared by blending a portion of the active CCR3 antagonist component, mannitol, polyethylene glycol and crospovidone in a suitable mixer. The premix is heated in a high shear mixer and then granulated. The hot granules were cooled to room temperature and sieved through a sieve with a mesh size of 1.0 mm. The granules are blended with a portion of crospovidone and mannitol in a suitable mixer. Magnesium stearate is added to this blend after passing through a 1.0 mm sieve to detangle. The final mixture is then produced by final mixing in a suitable mixer and compressed into tablets. The following tablet compositions can be obtained:

ingredient mg/tablet %/refine active ingredient 28.500 30.0 crospovidone 1.500 1.6 mannitol 16.700 17.6 polyethylene glycol 14.300 15.1 Total (granules) 61.000 64.3 mannitol 31.000 32.6 crospovidone 2.500 2.6 magnesium stearate 0.500 0.5 Sum 95.000 100.000

4. Tablet Formulation - Hot Melt Extrusion

A premix is prepared by blending the active CCR3 antagonist component and stearic-palmitic acid in a suitable mixer. The premix is extruded in a twin screw extruder and then granulated. The granules are sieved through a sieve having a mesh size of 1.0 mm. The granules are blended with mannitol and crospovidone in a suitable mixer. Magnesium stearate is added to this blend after passing through a 1.0 mm sieve to detangle. The final mixture is then produced by final mixing in a suitable mixer and compressed into tablets. The following tablet compositions can be obtained:

ingredient mg/tablet %/refine active ingredient 28.500 30.0 Stear-Palmitic Acid 27.500 28.9 Total (granules) 56.000 58.9 mannitol 32.600 34.3 crospovidone 5.600 5.9 magnesium stearate 0.800 0.9 Sum 95.000 100.000

5. Tablet Formulation - Hot Melt Extrusion

A premix is prepared by blending the active CCR3 antagonist component and stearic-palmitic acid in a suitable mixer. The premix is extruded in a twin screw extruder and then granulated. The granules are sieved through a sieve having a mesh size of 1.0 mm. Granules are filled directly into hard capsules. The following capsule compositions can be obtained:

ingredient mg/tablet %/refine active ingredient 70.000 70.0 Stear-Palmitic Acid 30.000 30.0 Total (granules) 100.000 100.0 capsule 90.000 - Sum 190.000 100.000

6. Tablet Formulation - Roller Compression

A premix is prepared by blending mannitol and the active CCR3 antagonist component, which is part of crospovidone, and magnesium stearate in a suitable mixer. The premix is compressed with a roller compactor and then granulated. Optionally, the granules are sieved through a sieve having a mesh size of 0.8 mm. The granules are blended with a portion of mannitol and crospovidone in a suitable mixer. Magnesium stearate is added to this blend after passing through a 1.0 mm sieve to detangle. The final mixture is then produced by final mixing in a suitable mixer and compressed into tablets. The following tablet compositions can be obtained:

ingredient mg/tablet %/refine active ingredient 28.500 30.0 crospovidone 1.400 1.5 mannitol 34.600 36.4 magnesium stearate 0.500 0.5 Total (granules) 65.000 68.4 mannitol 27.000 28.4 copovidone 1.600 1.7 crospovidone 0.950 1.0 magnesium stearate 0.450 0.5 Sum 95.000 100.000

7. Tablet Formulation - Roller Compression

A premix is prepared by blending the active CCR3 antagonist component and magnesium stearate in a suitable mixer. The premix is compressed with a roller compactor and then granulated. Optionally, the granules are sieved through a sieve having a mesh size of 0.8 mm. The granules are blended with mannitol and croscarmellose sodium in a suitable mixer. Magnesium stearate is added to this blend after passing through a 1.0 mm sieve to detangle. The final mixture is then produced by final mixing in a suitable mixer and compressed into tablets. The following tablet compositions can be obtained:

ingredient mg/tablet %/refine active ingredient 114.200 66.0 magnesium stearate 1.800 1.0 Total (granules) 116.000 67.0 mannitol 51.000 29.5 Croscarmellose Sodium 3.500 2.0 magnesium stearate 2.500 1.5 Sum 173.000 100.000

8. Tablet Formulation - Roller Compression

A premix is prepared by blending the active CCR3 antagonist component and magnesium stearate in a suitable mixer. The premix is compressed with a roller compactor and then granulated. Optionally, the granules are sieved through a sieve having a mesh size of 0.8 mm. The granules are blended with microcrystalline cellulose and crospovidone in a suitable mixer. After passing through a 1.0 mm sieve for desulfurization, magnesium stearate is added to this state. The final mixture is then produced by final mixing in a suitable mixer and compressed into tablets. The following tablet compositions can be obtained:

ingredient mg/tablet %/refine active ingredient 114.200 66.0 magnesium stearate 1.800 1.0 Total (granules) 116.000 67.0 MCC 51.000 29.5 crospovidone 3.500 2.0 magnesium stearate 2.500 1.5 Sum 173.000 100.000

9. Coated Tablet Formulation

Film-coated tablets can be prepared using tablet cores according to the formulations mentioned above. The coating suspension is prepared by suspending hydroxypropyl methyl cellulose, polyethylene glycol, talc, titanium dioxide and iron oxide in purified water in a suitable mixer at ambient temperature. Film-coated tablets are prepared by coating tablet cores with a coating suspension at a weight gain of about 3%. The following film coating compositions can be obtained:

ingredient mg/tablet %/refine hydroxypropyl methyl cellulose 2.40 48.0 Polyethylene Glycol 6000 0.70 14.0 titanium dioxide 0.90 18.0 talc 0.90 18.0 red iron oxide 0.10 2.0 Purified water (volatile components) -- -- Sum 5.00 100.0

b. Drug formulation and administration

ratio. Drug formulation and administration

The research product of the present invention (compound 1) followed the following chemical structure:

compound 1

One of ordinary skill in the art will recognize that the compounds, co-crystals, salts and formulations described above in the above sections can also be used in these Examples.

Compound 1 was available as 100 mg, 200 mg and 400 mg film-coated tablets with double convex, round or oval and dull red color. Tablets were prepared by dry granulation process and contained microcrystalline cellulose, hydrogen phosphate, croscarmellose sodium, magnesium stearate, polyvinyl alcohol, titanium dioxide, polyethylene glycol, talc, red iron oxide and yellow iron oxide as inactive ingredients. . Placebo tablets consistent with the study product were manufactured by a direct compression process and contained the same inactive ingredients.

c. Preclinical example

1. Materials and Methods

(a) subcutaneous osmotic pump implantation

Aljet mini pumps were filled, prepared, and numbered by mouse ID the day before injection to allow priming and blinding of treatment at 37°C. The pump was implanted posteriorly, slightly posterior to the scapula and slightly lateral to the midline. Anesthesia was maintained with 1-3% isoflurane by anesthetizing the mice with 3-5% isoflurane using a vaporizer and controller in the induction chamber, then moving them to the procedure area and mounting a nose cone. Apply ophthalmic ointment to your eyes to prevent dryness. Mice were subcutaneously injected with meloxicam 5 mg/kg. Hair was removed from the incision site using small sharp scissors, and the site was cleaned with alternating applications of 70% isopropanol and betadine. A 0.5-1 cm incision was made and a hemostatic agent was inserted to spread the subcutaneous tissue and create a pump pocket. The pump was inserted into the pocket and the wound was closed with a wound clip. All surgical instruments were autoclaved prior to first use on the day of surgery. The instrument was then sterilized between animals with a glass bead sterilizer. Mice were fully recovered and placed in a clean recovery cage partially placed over a warming pad until taut. Mice were tested for induction of anesthesia by the toe pinch method and respiratory monitoring. Mice were monitored post-surgery every 15 minutes until recovery. The next day mice were administered a second dose of meloxicam. When signs of infection were observed, mice were administered 5 mg/kg Baytril subcutaneously daily until the infection disappeared.

(b) open field test

The Open Field is used to assess general motor activity and exploratory behavior in a new environment. Mice are brought to the laboratory for at least 30 min to acclimatize to laboratory conditions (dimmed lighting) prior to testing. The test field consists of a 50 cm x 50 cm square field. The mouse is placed in the center of the field and followed for 15 min. Time spent in the peripheral and central regions is analyzed along with parenting behavior. Clean all surfaces between tests with 70% ethanol.

(c) Y-maze

The large-scale Y-maze test assesses short-term memory of familiarity with a particular situation. Mice are brought to the laboratory to acclimatize for at least 30 min to laboratory conditions (dimmed light) prior to testing. For the initial training trial, mice are placed on one arm end of a large Y-maze designated as "start arm" (arm length: 15 inches). The third arm of the maze is blocked, allowing the mouse to freely explore two of the three arms (the “initiating arm” and the “familiar arm”) for 5 min. Each arm has a spatial cue. After 1 h, mice are placed back into the maze in the “start arm” and all three arms can be explored without blocking the third arm (“new arm”). Movement in and out of each arm is tracked using automatic tracking software (CleverSys). Perform the tests in dark lighting and clean the device with 70% ethanol between tests.

(d) Barnes' maze

The modified Barnes maze was used for spatial tasks/rational assessments such as learning and memory. The Barnes maze apparatus consists of a 122 cm diameter circular platform, each with 40 exits, each 5 cm in diameter, placed along three rings of varying distances from the center of the platform. An escape box is attached to one of the holes and all holes are unblocked. The maze is trained with bright lights and fans to provide an unfavorable stimulus to encourage escape. Visual cues are placed on all four sides of the maze. Mice are presented with a series of 4 or 5 trials, with an interval of approximately 10 minutes, with a maximum duration of 90 or 120 seconds for each trial. In each test, the mouse is placed in the center of the maze. After 10 seconds, the mouse can be explored, and the test ends if the mouse finds an escape box and escapes before the end of the test. Mice unable to find an escape box were allowed to enter the escape box and remain trapped in the house for 30 seconds. The training takes place over 4 days. Data recorded and analyzed includes speed, escape delay time and distance traveled.

Mice are divided into groups of 4-5 mice each in a balanced treatment group. For example, 4 trials are run on group 1 mice, 4 trials are run on group 2 mice, and so on until all groups have completed the tests. 70% ethanol is used to clean the field between tests.

(e) Rotarod

Mice were trained on a rotarod for 3 trials up to 100 seconds each, and the rotarod was a motor control test. Success or failure was recorded in the last trial and the delay time is defined as >90 s. A binomial test was performed to compare success rates between control and compound-treated mice.

(f) T maze

The water maze was filled with water at least 24 hours prior to reaching room temperature. The water was dyed with white latex paint, allowing the animals to be tracked and a hidden platform to be used. Two distinct visual cues were placed at either end of the T-arm of the T-maze insert. On Day 1, animals were given 4 trials, each with a visible platform and a trial test interval of 30 minutes. Animals were given 60 seconds to reach the platform. If they did not reach the platform at that time, they were guided to the platform and left there for 5 seconds before being removed from the tank. The target arm was changed every 3 mice, and both treatment groups had the same number of left and right rotation target arms. After each test, mice were placed in empty cages with blue pads, dried under a red cage, and then placed back in their home cages. Day 2 is the test day, animals are subjected to the same test between each of the 4 trials and the test interval of 30 min, but using a hidden platform. Animals were scored for right and wrong selection and delay time to reach the platform, and a binomial test was performed to compare success rates between control and compound-treated mice. All trials were recorded using TopScan.

(g) protein quantification

CCL11 protein levels were measured from mouse plasma by sandwich ELISA plasma and diluted 1:10 for analysis. (Mouse CCL11/Eotaxin Duo Set ELISA Kit, R&D Systems, Minneapolis, Minn.).

Human CCL11 from human plasma was determined by SOMAscan in a SomaLogic aptamer-based assay (SomaLogic, Inc., Boulder, CO). (Gold L, et al. (2010). Aptamer-Based Multiplexed Proteomic Technology for Biomarker Discovery. PLOS ONE 5(12): e15004.

A panel of circulating cytokines from mouse plasma was analyzed by Luminex. Luminex analysis services performed by Eve Technologies (Calgary, Alberta, Canada).

(h) mRNA quantification

Snap-frozen hemibrains were dissected into cortex, hippocampus, striatum, and thalamus. The cortex was divided into three homogenous sections and one section was used for RNA isolation. RNA was isolated using the Prolink RNA Mini Kit (Thermo Fisher Scientific # 12183025). cDNA was prepared using the Taqman RT kit (Thermo Fisher Sci # N8080234). qPCR was run in Quant Studio 6 using custom Taqman Multiplex Primers for IL-1beta and GAPDH. All samples were run together on a single plate and analyzed for ddCT first against endogenous controls and then against controls.

(i) HPLC/MS quantification of compound 1

Compound levels were determined by MS/MS by Quintara (Hayward, CA).

(j) flow analysis

number of eosinophils

200 uL of whole blood was collected during perfusion and shipped to Charles River Clinical Pathology Services (Shrewsbury, MA) for hematological analysis of eosinophil counts by FACS (fluorescence activated cell sorter).

Eosinophil morphology change (ESC)

ESCs determine morphological changes in human eosinophils activated by human eosinophils-1 (PreProTech, Rocky Hill, New Jersey) compared to human eosinophils. Changes are detected as changes in forward scatter as measured by FACS (Fluorescence Activated Cell Sorter). The mean forward scatter of the autofluorescence (eosinophil) population for each sample was determined with respect to the mean of each set of three samples. Methods for determining ESC have been previously described and known in the art.

CCR3 receptor internalization

The CCR3 receptor internalization assay is FACS-based, using anti-human CCR3 antibodies labeled with human Eotaxin-1 (PreProTech, Rocky Hill, New Jersey) and APC (R&D Systems, Minneapolis, Minnesota) or isotype control IgG2A antibody. CCR3 receptor internalization in human eosinophils induced by human eosinophil-1 compared to naive eosinophils is monitored. Median APC fluorescence intensity units for each sample were determined along with the average of triplicate sets of each sample. Methods for monitoring CCR3 receptor internalization have been described previously and are known in the art.

(k) histology

The mice were removed the next day after the end of the behavioral test. Anesthesia was induced by 2,2,2-tribromoethanol, and then the mice were extracardially perfused with 0.9% saline. The brain was dissected and sagittal cut in two halves. One half was flash frozen in dry ice for later use, and the other half was fixed in 4% paraformaldehyde in PBS for use in immunohistochemistry. After two days of fixation, the hemibrains were transferred to 30% sucrose in PBS solution and changed after two days. Hemibrains were cut to 30 μm in a microtome at -22°C. Brain sections were stored in cryoprotective medium at -20°C until needed for staining. Blocking was performed in free-floating sections in appropriate serum in 10% serum in 0.5% PBST. Primary antibodies were incubated overnight at 4°C. For light microscopy, antibodies of DCX, 1:200, Santa Cruz BioTech, CD68, 1:1000 were used at the given concentrations. AbD Serotec. Secondary biotinylated antibody was applied the next day at a concentration of 1:300. Staining visualization was achieved by reaction with ABC kit (Vector) and diaminobenzidine (Sigma). Dehydration of the mounted slides was achieved using ethanol and xylene dips. Images were acquired at 5x magnification on a Leica optical microscope.

For fluorescence microscopy, the following concentrations of antibody were used at the given concentrations: GFAP, 1:500, DAKO; Iba1, 1:1000, Wako; and BrdU, 1:500, AbCam. An antigen retrieval protocol was required for BrdU (2N HCl, 37C, 30 min) prior to blocking. An appropriate fluorescent secondary antibody was applied the next day at a concentration of 1:300 for 1 hour at room temperature. Prolong Gold encapsulant was used to cover slip the slides. Images were acquired with an optical microscope at 5 magnification.

2. Experimental group

1st experimental group (see FIGS. 1-2) : 2 or 18 month old C57BL/6 mice were subcutaneously administered with IgG antibody control by IP injection or Compound 1 by Aljet osmotic pump for 2 or 4 weeks. During the last week of treatment, mice underwent behavioral testing prior to perfusion on the last day of treatment. All mice received BrdU injections at 150 mg/kg IP for 5 consecutive days immediately before the start of treatment.

drug formulation. Control rat IgG2A clone 54447 (MAB006, R & D Systems) was administered at 50 ug/kg in sterile saline. Compound 1 was formulated with 40% HP-β-cyclodextrin and adjusted to pH 6.5 with NaOH (1M). A fresh solution was prepared weekly and stored at 4°C.

Treatment group:

Treatment group 1, young control group: young C57BL/6 mice (n = 18) aged 1-2 months old received 5 injections of control IgG by intraperitoneal (IP) injection, once every 3 days, for 14 days. received. .

Treatment group 2, old control: 18-month-old aged C57BL/6 mice (n = 18) received 5 injections of control IgG by IP injection for 14 days, once every 3 days.

Treatment group 3, compound 1 (dose 1): 18-month-old aged C57BL/6 mice (n = 16), model 2001 (1 uL/hr) injected with ~50 mg/ml compound 1 by Aljet mini-pump) , for 2 weeks with one replacement.

Treatment group 4, compound 1 (dose 2): 18-month-old aged C57BL/6 mice (n = 16) by model 2002 (Aljet mini-pump, model 2002 (0.5uL //) ~ 50mg/ml Received an infusion of compound 1, for 2 weeks.

Treatment group 5, compound 1 (dose 2): 18-month-old aged C57BL/6 mice (n = 16) by model 2002 (Aljet mini-pump, model 2002 (0.5uL //) ~ 50mg/ml Received an infusion of compound 1, for 4 weeks with one replacement.

Subcutaneous administration of Compound 1 for 4 weeks in 18-month-old mice increased the number of doublecortin-positive cells in the hippocampus, an indicator of neurogenesis (see FIG. 1A ). Higher administration of Compound 1 for 2 weeks resulted in a trend of increased BrdU-positive cells in the hippocampus, a marker of cell proliferation (see Figure 1B). Low or high dose infusion of Compound 1 for 2 or 4 weeks improved the signaling Y-maze, a memory test (Figure 2 reporting the percentage of time normalized to total interaction time and number of visits when assessing total time spent). Reference). Compared with the control group treated with the IgG antibody without the compound (treatment groups 1 and 2), administration and behavior were performed simultaneously.

Thus, administration of compound 1 increased the number of Dcx and BrdU positive cells, indicating that compound 1 increased neurogenesis and cell survival, respectively. Compound 1 was able to improve memory (cognition) as evidenced by its performance in the Y-Maze test.

Second experimental group (see FIGS. 3-6 ): 3 month old or 16.5 month old C57B1/6 mice were subcutaneously administered with vehicle control or Compound 1 by Aljett osmotic pump for 4 weeks. During the last week of treatment, mice underwent behavioral testing prior to perfusion on the last day of treatment. All mice received BrdU injections at 150 mg/kg IP for 5 consecutive days immediately before the start of treatment.

drug formulation. Compound 1 was formulated with 40% HP-β-cyclodextrin and adjusted to pH 6.5 with NaOH (1M). Vehicle solutions were formulated and pH adjusted similarly. A fresh solution was prepared weekly and stored at 4°C.

Treatment group:

Treatment group 1, young control group: 3-month-old young C57BL/6 mice (n = 19) received vehicle infusion with one replacement for 4 weeks by Aljet mini-pump, model 2002 (0.5 uL/hr).

Treatment group 2, old control group: 16-month-old aged C57BL/6 mice (n = 19) received vehicle infusion by Aljet mini-pump, model 2002 (0.5 uL/hr) for 4 weeks with one replacement.

Treatment group 3, prior compound 1 (dose 2): 16-month-old aged C57BL/6 mice (n = 19), ˜50 mg by model 2002 (Aljet mini-pump, model 2002 (0.5uL //)) /ml received an injection of compound 1, for 4 weeks with one replacement).

Subcutaneous administration of Compound 1 for 4 weeks in 16.5 month old mice improved cognitive performance in the signal Y-maze test for memory (see FIG. 3 ). 4-week dosing of Compound 1 tended to improve performance in Barnes' maze, a hippocampal-dependent spatial memory test (see FIG. 4 ). Trends in memory improvement were also observed by analyzing the difference between the average delay time of the Day 4 trial and the delay time of trials 13 and 16. In addition, 4-week administration of Compound 1 significantly increased the number of BrdU-positive cells in the hippocampus, hippocampal cells, and markers of neurogenesis (see FIG. 5 ). Thus, compound 1 was able to improve cell survival and improve memory (cognition) as evidenced by the results obtained using the Y-maze and Barnes' maze tests.

Complex cerebrospinal fluid levels in mice

Cerebrospinal fluid (CSF) from both the 2-month-old "young" and 15.6 month old "old" groups were collected and the level of Compound 1 was determined by mass spectrometry. Figure 6 depicts the levels of compounds of the invention detected in mouse CSF for both young and old groups (less than 10 nM). These CSF levels do not approach the Ki for compounds in mice (124 nM, determined by cell line receptor binding) and do not cross the blood-brain barrier (BBB) at significant concentrations. For comparison, at 50 μg/hr of a 50 mg/mL solution, the levels of the compounds of the present invention measured in the plasma of young (2-month-old) and old (18-month-old mice) perfused 2 and 4 weeks, respectively, were significantly higher. (352 ± 31 nM and 355 ± 43 nM, respectively; values are mean ± sem), further indicating that the compounds of the present invention do not cross the BBB in significant amounts.

These data show that compound 1 acts peripherally because it does not act directly in the CNS. Also, there is not enough concentration effective to cross the BBB. In addition, there was no difference in BBB penetration between young and old mice, indicating that the effect of compound 1 on cognitive and neurodegeneration was not due to differences in BBB between the two groups.

Compound tissue distribution level

The distribution of Compound 1 in mouse tissues was determined after oral administration of [ 14 C]-radiolabeled compound (“labeled compound”) to male stained C57BL/6JOlaHsd mice (Harlan Labs, BV). The labeled compound was administered at 10 mg/kg body weight, corresponding to 17 μmol/kg. One animal was sacrificed 1, 24 and 168 hours after dosing. Blood, plasma and ocular radioactivity concentrations were determined using liquid scintillation counting (LSC). Tissue and organ concentrations were measured by whole body automated radiography technique (QWBA). Preparation of whole-body animal sections was performed using a cristat microtome Reichert-Jung CRYO MACROCUT or CRYO MACROCUT LEICA CM 36003 according to known techniques (S. Ullberg, et al., Autoradiography in Pharmacology in: Int. Encyclopedia of Pharmacology). , J. Cohen (Ed.), 1 (78): 221) -39 (1971)).

To enable quantitative assessment of radioactivity, visceral animals and body parts were selected at 5-7 levels, and the following sections were taken at different levels: adrenal glands; blood; marrow; brain; eye (lens); epididymis; fat (white and brown); Harder Sun; Heart; kidney; liver; lungs; muscle; pituitary; kidneys; prostate, spinal cord; spleen; salivary glands; skin; testicle; thyroid; thymus; vineyard. Two sections of each selected level were taken per animal and these sections were lyophilized in a microtome at -20 to -25°C for a minimum of 48 hours.

7 depicts a chart of the area under the curve (AUC) values for all three time points, quantifying the exposure of each tissue to a labeled POI after administration of the compound. 7 shows that compound 1 does not cross the blood-brain barrier (BBB) at a significant level.

Again, these results show that compound 1 acts in a peripheral manner because it cannot cross the BBB to a significant level. Thus, the effect of compound 1 does not act directly on the central nervous system and overcomes the difficulties that have caused the failure of many CNS disease-targeting drug candidates.

P.O. Pharmacokinetic Profile of Administration

Plasma from male 2-month-old C57B1/6 mice was measured for compound 1 concentrations at 20 min, 2 h, 8 h and 12 h time points after oral gavage at two doses: 30 mg/kg and 150 mg/kg. A dose of 30 mg/kg was found to be sufficient to achieve a compound 1 greater than 100 nM for 8 hours ( FIG. 8 ).

Third experimental group (see FIGS. 9-11 ) : 2-month-old C57B1/6 mice were administered either vehicle control or Compound 1 by oral gavage twice daily for 18 days. During the last week of treatment, mice underwent behavioral testing prior to perfusion on the last day of treatment. All mice received BrdU injections at 150 mg/kg IP for 5 consecutive days immediately before the start of treatment.

Drug formulation: Compound 1 was formulated with 40% HP-β-cyclodextrin and adjusted to pH 6.5 with NaOH (1M). Vehicle solutions were formulated and pH adjusted similarly. A fresh solution was prepared weekly and stored at 4°C.

Treatment group:

Treatment group 1a, vehicle treatment: 7-week-old young C57BL/6 mice (n = 15) received twice daily (BID) oral (PO) treatment of vehicle solution for 18 days with one injection on the last day, total 35 injections.

Treatment group 1b, compound 1 treatment: 7-week-old young C57BL/6 mice (n = 15) were treated orally (PO) with 30 mg/kg compound 1 twice daily (BID) for 18 days as a single injection on the last day. , and a total of 35 injections.

Treatment group 2a, vehicle treatment with rmCCL11: 7-week-old young C57BL/6 mice (n = 15) received twice daily (BID) oral (PO) treatment of vehicle solution for 18 days with the last single injection; A total of 35 injections. Mice simultaneously received peripheral injections (IP) of rmCCL11 every 3 days on day 5 of treatment for a total of 5 injections.

Treatment group Treatment group 2b, Compound 1 treatment with rmCCL11: 7-week-old young C57BL/6 mice (n = 15) received oral (PO) treatment of Compound 1, 30 mg/kg twice daily (BID) for 18 days. received for 18 days. On the last day, only one injection was administered, for a total of 35 injections. Mice simultaneously received peripheral injections (IP) of rmCCL11 every 3 days on day 5 of treatment for a total of 5 injections.

Recombinant mouse CCL11 (“rmE”) treatment significantly exacerbated anxiety in the open field, whereas compound 1 treatment for 2 weeks showed improved anxiety with twice daily oral administration (see FIG. 9 ). rmCCL11 corrupted memory in the Y-maze; However, mice treated with compound 1 were no longer significantly different from control mice (see FIG. 10 ). rmCCL11 impaired the memory of Barnes' maze, and treatment with compound 1 significantly improved the memory (see FIG. 11 ). Thus, rmE exacerbated anxiety through both the open field test and memory, as evidenced by the performance measured in the Y-maze and Barnes' maze tests. However, compound 1 treatment could attenuate this effect.

Fourth experimental group (see FIG. 12 ) : 23 month old C57B1/6 mice were administered vehicle control or Compound 1 by oral gavage twice daily for 19 days. After 11 days of treatment, mice were subjected to a Y maze test.

Drug formulation: Compound 1 was formulated with 40% HP-β-cyclodextrin and adjusted to pH 6.5 with NaOH (1M). Vehicle solutions were formulated and pH adjusted similarly. Kolliphor was added to each solution at a rate of 10% weekly and stored at 4°C.

Treatment group:

Treatment group 1, vehicle treatment: 23-month-old aged C57B1/6 mice (n = 8) received oral gavage (PO) treatment with vehicle solution twice daily (BID) for 19 days.

Treatment group 2, compound 1 treatment: 23-month-old aged C57B1/6 mice (n = 11) received oral gavage (PO) treatment with compound 1, 30 mg/kg twice a day (BID) for 19 days.

Compound 1 treatment significantly improved the memory of the Y-maze. Mice treated with Compound 1 showed intact memory for the new cancer at the number of visits ( FIG. 12A ). Treatment with compound 1 also significantly increased the distance traveled by mice during the test ( FIG. 12B ).

Experimental group 5 (see FIGS. 13-17 ): 23-month-old C57B1/6 mice were subcutaneously administered with vehicle control or compound 1 twice daily for 21 days. Three weeks later, mice were subjected to a behavioral test and sacrificed the day after the last behavioral test.

Drug formulation: Compound 1 was formulated with 40% HP-β-cyclodextrin and adjusted to pH 6.5 with NaOH (1M). Vehicle solutions were formulated and pH adjusted similarly. Kolliphor was added to each solution at a rate of 10% weekly and stored at 4°C.

Treatment group:

Treatment group 1b, vehicle treatment: 23-month-old aged C57B1/6 mice (n = 9) were treated with vehicle solution twice a day (BID) subcutaneously (SQ) for 21 days.

Treatment group 4, compound 1 treatment: 23-month-old aged C57B1/6 mice (n = 17) were treated with compound 1, 30 mg/kg twice daily (BID) subcutaneously (SQ) for 21 days.

Compound 1 treatment significantly improved memory in the Y-maze and Barnes' maze. Mice treated with Compound 1 exhibited intact memory for new cancers in both the number of visits ( FIGS. 13A-B ) and time spent in new cancers ( FIGS. 13C-D ). Treatment with compound 1 also significantly increased the speed of mice during the test ( FIG. 13E ). Mice treated with Compound 1 performed significantly better in the Barnes Maze for spatial memory ( FIG. 14A ) and exhibited increased rates during the test ( FIG. 14B ). Motor activity was also improved in the open field, and both movement distance and speed showed a strong tendency to improve ( FIGS. 15A and 15B , respectively).

Levels of inflammatory cytokines were determined from mouse plasma by Luminex assay ( FIG. 16 ). There was a strong tendency to decrease several inflammatory markers including TNFa, IL6, IL1beta, IL5 and IL17. Activated microglia were quantified by IHC staining in the hippocampus of mice ( FIG. 17 ). Compound 1 treatment resulted in a strong trend towards reduction of microglia.

6th experimental group (see Fig. 18) : 23-month-old C57B1/6 mice were subcutaneously administered with vehicle control or Compound 1 twice daily for 30 days, and then sacrificed on the day of the last injection.

Drug formulation: Compound 1 was formulated with 40% HP-β-cyclodextrin and adjusted to pH 6.5 with NaOH (1M). Vehicle solutions were formulated and pH adjusted similarly. Kolliphor was added to each solution at a rate of 10% weekly and stored at 4°C.

Treatment group:

Treatment group 1a, vehicle treatment: 23 month old aged C57B1/6 mice (n = 9) received subcutaneous (SQ) treatment of vehicle solution twice a day (BID) for 30 days.

Treatment group 3, compound 1 treatment: 23 month old aged C57B1/6 mice (n = 18) received subcutaneous (SQ) treatment of 30 mg/kg compound 1 twice daily (BID) for 30 days.

Compound 1 treatment showed a strong trend towards reduction of blood eosinophils in blood by FACS analysis in a subset of animals (n = 2, 5) ( FIG. 18 ).

7th experimental group (see FIG. 19) : 3 month old hairless mice were treated with oxazolone (Ox) every other day, and treated with saline or 30 mg/kg Compound 1 for BID and PO for 2 weeks.

Drug formulation: Compound 1 was formulated with 40% HP-β-cyclodextrin and adjusted to pH 6.5 with NaOH (1M). Vehicle solutions were formulated and pH adjusted similarly. Kolliphor was added to each solution at a rate of 10% weekly and stored at 4°C.

Treatment group:

Treatment group 1, vehicle treatment: 3 month old hairless mice (n = 3) received oral gavage (PO) orally twice daily (BID) of vehicle solution for 2 weeks.

Treatment group 2, vehicle treatment and oxazolone treatment: 3 month old hairless mice (n = 6) received twice daily (BID) oral gavage (PO) treatment of vehicle solution for 2 weeks.

Treatment group 3, compound 1 treatment and oxazolone treatment: 3 month old hairless mice (n = 6) received oral gavage (PO) treatment of compound 1, 30 mg/kg twice daily (BID) for 2 weeks.

Compound 1 reduced oxazolone-induced eosinophilia by complete blood count (CBC) analysis. N = 3, 6, 8. *p <0.05, **p <0.01. Compound 1 dramatically reduced blood eosinophil counts in an eosinophilia model of eosinophilia ( FIG. 19 ). This demonstrates that inhibition of CCR3 may be sufficient to normalize eosinophil levels and function particularly in diseased states, as a second mechanism by which Compound 1 may be effective in treating neuronal loss and associated motor and cognitive deficits (brain inflammation). in addition to the reduction).

Eighth experimental group (see FIGS. 20 to 22) : 24 month old C57 mice were treated for 4 weeks by continuous infusion of Compound 1 or vehicle by Aljett osmotic pump, and transplanted to deliver two treatments in succession.

Drug formulation: Compound 1 was formulated with 40% HP-β-cyclodextrin and adjusted to pH 6.5 with NaOH (1M). Vehicle solutions were formulated and pH adjusted similarly. Kolliphor was added to each solution at a rate of 10% weekly and stored at 4°C.

Treatment group:

Treatment group 1, old control group: 23-month-old aged C57BL/6 mice (n = 15) received vehicle infusion with one replacement for 4 weeks by Aljet mini-pump, model 2002 (0.5uL/hr).

Treatment group 2, compound 1 (dose 2): 23-month-old aged C57BL/6 mice (n = 15) by model 2002 (Aljet mini-pump, model 2002 (0.5uL //) ~ 50mg/ml Received an infusion of compound 1, for 4 weeks with one replacement.

Mice were tested on a rotarod for motor coordination. Success or failure was recorded in the last trial and the delay time is defined as >90 s. Compound 1 treated mice were more successful than vehicle treated mice by the binomial test. N = 15, respectively, *p <0.05. A greater proportion of mice treated with Compound 1, 47% were able to stay on the bar for more than 90 seconds, a threshold at which only 20% of control-treated mice could be maintained after three consecutive trials ( FIG. 20 ). These results suggest that there is a consistent effect of compound 1 on motor function in the eotaxin elevation model.

Mice were tested in the T maze for cognitive function ( FIG. 21 ). The number of successes or failures in reversing the correct arm was recorded. Compound 1 treated mice were more successful than vehicle treated mice by the binomial test. *p <0.05.

The fecal output was measured by weighing the dry weight of the fecal pellets overnight (FIG. 22). Gastrointestinal function is a well-known symptom of Parkinson's disease. Water and food intakes were measured over the same period. Mice treated with Compound 1 had significantly lower stool output compared to control mice. They also had significantly higher overnight water intake compared to control mice. *p <0.05. These results indicate that the deficit in gastric function can be altered by compound 1 treatment.

Experimental group 9 (see FIG. 23 ) : 3-month-old C57 mice were given a single dose of 10 mg/kg IP of lipopolysaccharide (LPS) to induce inflammation and were treated with Compound 1 for 18 days.

Drug formulation: Compound 1 was formulated with 40% HP-β-cyclodextrin and adjusted to pH 6.5 with NaOH (1M). Vehicle solutions were formulated and pH adjusted similarly. Kolliphor was added to each solution at a rate of 10% weekly and stored at 4°C.

Treatment group:

Treatment group 1, vehicle treatment: 3-month-old young C57BL/6 mice (n = 15) received a single injection of LPS and were BID-treated with vehicle by oral gavage for 18 days.

· Treatment group 2, compound 1 treatment: 3-month-old young C57BL/6 mice (n = 15) received a single injection of LPS and were treated with compound 1, 30 mg/kg, BID by oral gavage for 18 days. .

Brain sections were immunostained for detection of CD68+ activated microglia. Compound 1-treated mice significantly reduced CD68+ immune reactivity (reduction of activated microglia) in contrast to vehicle (saline)-treated mice ( FIG. 23 ). N = 9, 10, 8 by one-way ANOVA. *p <0.05.

These data indicate the potent anti-neuroinflammatory effect of compound 1 and indicate therapeutic potential for reducing neuroinflammatory-induced toxicity to neurons in neurodegeneration or diseases exhibiting cognitive or motor decline.

Experimental group 10 (see FIGS. 24-29) : 3-month-old C57 mice were given a dose of 0.5 mg/kg IP of lipopolysaccharide (LPS) once daily to induce inflammation and continued with Compound 1 for up to 4 weeks. processed.

Drug formulation: Compound 1 was formulated with 40% HP-β-cyclodextrin and adjusted to pH 6.5 with NaOH (1M). Vehicle solutions were formulated and pH adjusted similarly. Kolliphor was added to each solution at a rate of 10% weekly and stored at 4°C.

Treatment group:

Treatment group 1, vehicle treatment: 3-month-old C57 mice (n = 10) received IP injections of LPS daily for 7 weeks, followed by twice-daily (BID) oral gavage (PO) treatment of vehicle solution for up to 4 weeks. received.

Treatment group 2, vehicle treatment and oxazolone treatment: 3-month-old C57 mice (n = 10) received IP injections of LPS daily for 7 weeks, followed by twice daily (BID) oral gavage of vehicle solution (BID) for up to 4 weeks. PO) was treated.

Treatment group 3, compound 1 treatment and oxazolone treatment: 3-month-old C57 mice (n = 9) received IP injections of LPS daily for 7 weeks, compound 1, 30 mg/kg twice daily (BID) for up to 4 weeks. ) received oral gavage (PO) treatment.

24A illustrates the dosing paradigm for the tenth experimental group. Sections of the paradigm highlighted with squares indicate the time points at which the analysis in Figure 24B was performed. Anxiety was tested in an open field test after 1 week of compound 1 treatment ( FIG. 24B ). LPS treatment significantly increased anxiety in the open field, and compound 1 significantly reduced increased anxiety, *p<0.05.

25A illustrates the dosing paradigm for the tenth experimental group. Sections of the paradigm highlighted by squares indicate the time points at which the analysis in Figure 25B was performed. Cognitive tests were performed in the Y maze after 3 weeks of compound 1 treatment ( FIG. 25B ). Mice treated with LPS showed no significant preference for novel cancers. However, mice treated with compound 1 showed a significant preference for novel cancers similar to vehicle treated mice, * p <0.05, ** p < 0.01.

26A illustrates the dosing paradigm for the tenth experimental group. Sections of the paradigm highlighted by squares indicate the time points at which the analysis in Figure 26b was performed. The mRNA level of the inflammatory cytokine IL-1 beta was measured by quantitative PCR after 4 weeks of compound 1 treatment ( FIG. 26B ). LPS-treated mice showed a trend for increased levels of IL-1 beta, and mice treated with Compound 1 showed a significant decrease in IL-1 beta expression (* p <0.05).

27A illustrates the dosing paradigm for the tenth experimental group. Sections of the paradigm highlighted by squares indicate the time points at which the analysis in Figure 27b was performed. Brain sections were immunostained for detection of CD68+ activated microglia. Compound 1-treated mice dramatically reduced CD68+ immune reactivity (reduction of activated microglia) in contrast to LPS-only-treated mice ( FIG. 27B ).

28A illustrates the dosing paradigm for the tenth experimental group. Sections of the paradigm highlighted with squares indicate the time points at which the analysis in Figure 28b was performed. Brain sections were immunostained for detection of Iba1-positive microglia. Compound 1-treated mice significantly reduced Iba1+ immune reactivity (reduction of total microglia) compared to LPS-only-treated mice ( FIG. 28B ).

29A illustrates the dosing paradigm for the tenth experimental group. Sections of the paradigm highlighted by squares indicate the time points at which the analysis in Figure 29B was performed. Brain sections were immunostained for detection of GFAP-positive astrocytes. Compound 1-treated mice significantly reduced GFAP+ immune reactivity (reduction of total astrocytes) in contrast to LPS-only-treated mice ( FIG. 29B ).

These data indicate the potent anti-neuroinflammatory effect of compound 1 and indicate therapeutic potential for reducing neuroinflammatory-induced toxicity to neurons in neurodegeneration or diseases exhibiting cognitive or motor decline.

Experimental group 11 (see Fig. 30) : 45 2-month-old C57BL/6 mice were assigned to three groups: vehicle-treated control, vehicle-treated LPS mice and compound 1 treated LPS mice. Vehicle or compound was administered orally twice daily (PO BID) for a total of 6 doses. Histological evaluation was performed in the hippocampus using the microglia marker Iba-1.

Drug formulation: Compound 1 was formulated with 40% HP-β-cyclodextrin and adjusted to pH 6.5 with NaOH (1M). Vehicle solutions were similarly formulated and adjusted for pH. Solutions were prepared fresh weekly and stored at 4°C.

LPS formulation: LPS was formulated at 0.55 mg/ml in saline and administered at 5 mg/kg.

Treatment group:

Treatment group 1, vehicle-treated control group: 2-month-old young C57BL/6 mice (n = 13) were injected with vehicle PO BID for 3 days after 1 IP injection of saline for a total of 6 injections.

Treatment group 2, vehicle-treated LPS: 2-month-old young C57BL/6 mice (n = 19) received a single IP injection of LPS (5 mg/kg) followed by a total of 6 injections of vehicle PO BID for 3 days.

Treatment group 3, compound 1 treatment LPS: 2 month old young C57BL/6 mice (n = 18) were given a total of 6 compound 1 (30 mg/kg) PO BID for 3 days after a single IP injection of LPS (5 mg/kg). injected twice.

Tissue processing and histology: Mouse hemibrains were sectioned into 30 µm in a microtome at -22 °C. Brain slices were sequentially collected into 12 tubes so that all 12 sections of the hippocampus were displayed in a given tube. Brain sections were stored in cryoprotective medium at -20°C until staining was required. Free-flowing sections were blocked in appropriate serum with 10% serum in 0.5% PBST. Primary antibody Iba1 (Wako, 1:1000) was incubated overnight at 4°C. The appropriate fluorescent secondary antibody was applied the next day at a concentration of 1:300 for 1 h at room temperature. Slides were coverslips using Prolong Gold Mounting Media.

Neuroinflammation Quantification: Iba-1-positive areas were quantified as a percentage of the total hippocampal periphery ROI using thresholding in Image Pro Premier v9.2 software. Iba-1 positive percentage areas were averaged in approximately 5 sections for each mouse. General one-way ANOVA was used to test for statistical significance, and Dunnett's multiple comparisons were post-hoc tests between treatment groups.

30 shows the results of histological analysis of the mouse brain in an acute model of LPS-induced inflammation. It shows a significant reduction in LPS-induced microgliosis in the hippocampus of mice treated with Compound 1 for 3 days when treatment with Compound 1 and LPS were both started on the same day. Microgliosis was measured by determining the percentage of Iba-1 positive areas in the hippocampus. General one-way ANOVA was used to test for statistical significance, and Dunnett's multiple comparisons were post-hoc tests between treatment groups ( ^* P <.05; ^*** P <.001).

Twelfth experimental group (see FIG. 31 ) : 15 2-month-old C57BL/6 mice were assigned to three groups: vehicle-treated control, vehicle-treated LPS mice and compound 1 treated LPS mice. Vehicle or compound was administered orally twice daily (PO BID) for a total of 6 doses. Histological evaluation was performed in the hippocampus using the microglia marker Iba-1.

Drug formulation: Compound 1 was formulated with 40% HP-β-cyclodextrin and adjusted to pH 6.5 with NaOH (1M). Vehicle solutions were similarly formulated and adjusted for pH. Solutions were prepared fresh weekly and stored at 4°C.

LPS formulation: LPS was formulated at 0.55 mg/ml in saline and administered at 5 mg/kg.

Treatment group:

Treatment group 1, vehicle-treated control group: 2-month-old young C57BL/6 mice (n = 13) were injected with saline IP after IP injection, followed by vehicle PO BID injections 72 hours later for a total of 6 injections for 3 days.

Treatment group 2, vehicle-treated LPS: 2-month-old young C57BL/6 mice (n = 19) received a single IP injection of LPS (5 mg/kg) followed by a total of 6 injections of vehicle PO BID over 3 days 72 hours late. did.

Treatment group 3, Compound 1 treated LPS: 2-month-old young C57BL/6 mice (n = 18) received a single IP injection of LPS (5 mg/kg) followed by Compound 1 (30 mg/kg) PO BID 72 hours late. A total of 6 injections were made over 3 days.

Tissue processing and histology: Mouse hemibrains were sectioned into 30 µm in a microtome at -22 °C. Brain slices were sequentially collected into 12 tubes so that all 12 sections of the hippocampus were displayed in a given tube. Brain sections were stored in cryoprotective medium at -20°C until staining was required. Free-flowing sections were blocked in appropriate serum with 10% serum in 0.5% PBST. Primary antibody Iba1 (Wako, 1:1000) was incubated overnight at 4°C. The appropriate fluorescent secondary antibody was applied the next day at a concentration of 1:300 for 1 h at room temperature. Slides were coverslips using Prolong Gold Mounting Media.

Neuroinflammation Quantification: Iba-1-positive areas were quantified as a percentage of the total hippocampal periphery ROI using thresholding in Image Pro Premier v9.2 software. Iba-1 positive percentage areas were averaged in approximately 5 sections for each mouse. General one-way ANOVA was used to test for statistical significance, and Dunnett's multiple comparisons were post-hoc tests between treatment groups.

31 shows the results of histological analysis of the mouse brain in an acute model of LPS-induced inflammation. When compound 1 administration was started after LPS was administered 3 days before, the hippocampus of mice treated with compound 1 for 3 days showed a significant reduction in LPS-induced microgliosis. Microgliosis was measured by determining the percentage of Iba-1 positive areas in the hippocampus. General one-way ANOVA was used to test for statistical significance, and Dunnett's multiple comparisons were post-hoc tests between treatment groups ( ^* P <.05; ^*** P <.001).

3. Mouse MPTP Model of Parkinson's Disease

A mouse model using MPTP (1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine) was used to determine the level of improvement in MPTP-induced Parkinson's syndrome. MPTP is a prodrug of MPP+ and can cause permanent Parkinson's symptoms. MPP+ works by killing dopaminergic neurons in the substantia nigra region of the brain.

A total of 74 8-week-old male C57Bl/6J mice were used in the experiment. Animals were divided into two separate study groups, the 12- and 4-day study groups, and treated similarly in both groups. Mice were divided into three groups within the study group, with one group receiving MPTP vehicle and combined vehicle serving as a control group, the other two receiving MPTP twice daily on days 1 and 2, plus an additional combination vehicle or Compound 1 Daily (30 mg/kg) on day 12, and only one dose on day 12 (FIG. 32). On study day 11 after 10 days of treatment, motor function of the 12-day study cancer mice was assessed by microkinetic kinematic analysis. Endpoint tissue treatments were performed on study 12 and 4 days. Samples were processed for hematology, immunohistochemistry and HPLC measurements. Striatal levels of dopamine (DA), 3,2-dihydroxyphenylacetic acid (DOPAC) and homovanic acid (HVA) were assessed by HPLC on day 12. Tyrosine hydroxylase (TH) immunoreactivity was assessed from brain sections collected through the substantia nigra.

Principal component analysis (PCA) was performed because there are many 97 individual parameters. PCA combines all parametric data to reveal correlations between different parameters, providing a holistic view of fine motor and gait characteristics. Figure 33A shows a visualization of the 10 main components or PCs, showing how the original parameters are correlated in the data set. The more intensity (blue or red) for each parameter, the more strongly a particular parameter is related to that PC. The heat map consists of a PCA of all available parametric data. PC scores are presented as overall gait analysis scores in FIG. 33B based on the differences between Group 2: MPTP+Vehicle and Group 1: Vehicle+Vehicle mice at all PC scores.

Figure 34 shows the gait characteristics of forefoot toe spacing on study day 11. The MPTP+vehicle treated group showed a statistically significant change in forelimb toe spacing compared to the vehicle+vehicle treated group. And the MPTP+Compound 1 treatment group showed a positive trend compared to the MPTP+vehicle treatment group. Data are presented as mean±SEM (Group 1: Vehicle+Vehicle, n=15; Group 2: MPTP+Vehicle, n=14; Group 3: MPTP+Compound 1, n=13). Statistical significance: *p <0.05, group 2: MPTP+vehicle vs. group 1: vehicle+vehicle (unmatched t-test)

35 shows the gait characteristics of forefoot swing speed on study day 11. The MPTP+vehicle treated group showed a statistically significant change in forefoot swing speed compared to the vehicle+vehicle treated group. And the MPTP+Compound 1 treatment group showed a positive trend compared to the MPTP+vehicle treatment group. Data are presented as mean±SEM (Group 1: Vehicle+Vehicle, n=15; Group 2: MPTP+Vehicle, n=14; Group 3: MPTP+Compound 1 (30 mg/kg), n=13). Statistical significance: *p <0.05, group 2: MPTP+vehicle vs. group 1: vehicle+vehicle (unmatched t-test)

Figure 36 shows the gait characteristics of the ankle range of motion on study day 11. The MPTP+vehicle treated group showed a statistically significant change in ankle range of motion compared to the vehicle+vehicle treated group. And the MPTP+Compound 1 treatment group showed a positive trend compared to the MPTP+vehicle treatment group. Data are presented as mean±SEM (Group 1: Vehicle+Vehicle, n=15; Group 2: MPTP+Vehicle, n=14; Group 3: MPTP+Compound 1 (30 mg/kg), n=13). Statistical significance: *p <0.05, group 2: MPTP+vehicle vs. group 1: vehicle+vehicle (unmatched t-test)

37 shows the short-term effects of MPTP and Compound 1 on T cell infiltration into the brain. The total number of CD3 positive T cells counted in the substantia nigra in 3 30 μm thick sections for each mouse on study day 4 is presented. Data shown are mean±sem. ^*** P <0.001; One-way ANOVA, Sidak's post hoc multiple comparison test. Mice treated with MPTP+Compound 1 tended to have fewer CD3-positive cells, indicating less T-cell infiltration into the brain.

38 shows the short-term effects of MPTP and Compound 1 on microblastosis at study day 4; 38A depicts the extent of CD68-positive areas measured in the striatum in three 30 μm thick sections for each mouse on study day 4; 38B depicts the extent of CD68 positive areas measured in the substantia nigra from three 30 μm thick sections for each mouse from day 4 of the study. These data show that there is pronounced microgliosis in the substantia nigra, the major site of neuronal cell loss observed in Parkinson's disease, and not the striatum, the main brain region with neurons in the substantia nigra project, suggesting that treatment affects a key site of disease pathology. , but not the quadratic projection area.

4. Synuclein Transgenic Mouse Model of Parkinson's Disease

A synuclein transgenic mouse model was used to measure the chronic CCR3 inhibitory effect of Compound 1 and its ability to slow, stop, or reverse the progression of Parkinson's disease symptoms. This model is an overexpressed human α-synuclein test to show whether interventional treatments can prevent α-synuclein-induced behavioral and pathological effects. To select the optimal model for testing, plasma eotaxin levels were measured as biomarkers in multiple synuclein mouse models of different ages. Models included A53T, DxJ9M and line 61. Plasma from 6 month old line 61 synuclein transgenic mice was measured by ELISA assay to detect plasma eotaxin levels as biomarkers. 6-month-old line 61 mice (QPS Neuro, Grambach, Austria) showed transgene-induced increases in eotaxin levels ( FIG. 39 ), and plasma eotaxin levels were an appropriate clinical trial for selecting treatment populations. It has been shown that it can act as a biomarker.

Thirty-three 4.5 month old male alpha-synuclein transgenic mice (Line 61) were assigned to two groups (group A, n = 17, compound 11 mg/mL; group B, n = 16, vehicle). And 15 non-transgenic littermates of the same age (group C, vehicle) were treated with drinking water for 6 weeks. Animals received either vehicle or compound 1. Animal behavior was assessed at the end of the treatment period. After that, tissue processing was performed.

40 shows the results of the wire suspension test. Average wire suspension times per group are shown, animals in group C (non-transgenic, vehicle treated) show much higher wire suspension times. Animals in group A (transgenic, treated with compound-1) showed significantly higher wire suspension times compared to group B (transgenic, treated with vehicle). Data are presented as mean±SEM of all animals per group. ^*** P <0.001;Dunn's post hoc test; *P <0.05 Mann Whitney test for group A versus group B). These data demonstrate that the dramatic loss of ability to hang on wires in synuclein mice is at least partially rescued by Compound 1 treatment.

41 shows the results of the grip strength test. The average maximum grip force [g] per group is indicated. Animals in group A and group C (transgenic, compound 1 treated and non-transgenic, vehicle treated, respectively) exhibited significantly higher grip force compared to group B (transgenic, vehicle treated). Data are presented as mean±SEM of all animals per group. Groups were compared to vehicle treated transgenic animals (Group B). One-way ANOVA followed by Bonferroni's post hoc test. These data demonstrate that a statistically significant deficit in forelimb strength in synuclein mice was completely rescued by Compound 1 treatment.

42 shows the results of the beam walk test. The number of animals per group for each trial that was able to fully traverse the beam is indicated. None of the animals in group B (transgenic, vehicle treated) crossed the fifth beam (the most difficult), indicating that group A transgenic mice treated with compound 1 performed better than group B transgenic mice treated with vehicle. performed, and was closer to the non-transgenic control group in the beam walk test.

Figure 43 shows three groups of mice (Group A, transgenic compound 1 treated; Group B, transgenic vehicle treated; Group C, non-transgenic vehicle treated). The graphs represent the average number of slips per group [n] and each graph represents one trial (1-5). Data are mean±SEM of all animals per group. group was compared to group B. One-way ANOVA followed by Bonferroni's post hoc test. These data clearly show that synuclein-overexpressing mice have significantly impaired ability to cross the beam, and treatment with compound 1 rescued this effect, at least in part, showing a dramatic improvement in motor function.

44 shows the number of eosinophils in peripheral blood. Figure 44A shows the percentage of eosinophils in peripheral blood from three groups of mice (Tg Cmpd 1 = transgenic compound 1 treated; Tg Veh = transgenic vehicle treated; nTG Veh = non-transgenic vehicle treated). Data were compared by t-test. Figure 44B shows the absolute eosinophil counts in peripheral blood from three groups of mice (Tg Cmpd 1 = transgenic compound 1 treated; Tg Veh = transgenic vehicle treated; nTG Veh = non-transgenic vehicle treated). Data were compared by t-test. These data show that a Parkinson's disease model of synuclein overexpression results in eosinophilia, which was restored to non-transgenic mouse levels through Compound 1 treatment, suggesting beneficial immune modulation in this Parkinson's disease model. This also suggests that determining the level of eosinophils in Parkinson's disease patients treated with Compound 1 could be a biomarker for the disease, including determining the level of progression, stasis or regression, and therapeutic efficacy.

45 shows the effect of compound 1 on neuroinflammation. 45A shows quantified CD68 positive regions in the hippocampus: non-transgenic, vehicle treated mice; transgenic, vehicle treated mice; and transgenic, compound 1 treated mice (n = 14, 12 and 15, respectively). 45B shows quantified CD68 positive regions in the striatum of the following; non-transgenic, vehicle treated mice; transgenic, vehicle treated mice; and transgenic, compound 1 treated mice (n = 15, 11 and 16, respectively). Figure 45C shows quantified Iba1 positive regions in the hippocampus: non-transgenic, vehicle treated mice; transgenic, vehicle treated mice; and transgenic, Compound 1 treated mice (n = 14, 13 and 16, respectively). 45D shows quantified Iba1-positive regions in the following striatum: non-transgenic, vehicle treated mice; transgenic, vehicle treated mice; and transgenic, compound 1 treated mice (n = 15, 11 and 16, respectively). Data are mean +/- sem. ^* P<0.05. 45E shows GFAP positive astrocytes quantified in the hippocampus of: non-transgenic, vehicle treated mice; transgenic, vehicle treated mice; and transgenic, Compound 1 treated mice (n = 14, 13 and 15, respectively). 45F shows quantified GFAP positive regions in the striatum: non-transgenic, vehicle treated mice; transgenic, vehicle treated mice; and transgenic, Compound 1 treated mice (n = 15, 13 and 15, respectively). Data are mean +/- sem. ^* P<0.05. Figure 45G shows quantified Iba-1 positive regions in the compaction of the substantia nigra; non-transgenic, vehicle treated mice; transgenic, vehicle treated mice; and transgenic, compound 1-treated mice.

These data show that synuclein overexpression does not lead to dramatic microgliosis as evidenced by CD68 and Iba1 or astrocytosis as evidenced by GFAP immune reactivity, however, treatment with Compound 1 is linked to microgliosis and GFAP by both markers. By reducing astrocytes caused by astrocytosis, we demonstrate that it potentially has a global anti-inflammatory effect, i.e., an effect that is not simply directed to one inflammatory cell type. The striatum is also more affected than the hippocampus, demonstrating that areas associated with Parkinson's disease are particularly reduced in microgliosis and astrocytosis markers. This is further evidenced by the effect of compound 1 on reversing microgliosis on the dense part of the substantia nigra.

Figure 46 reports the effect of compound 1 on circulating levels of IL-4 and IL-6 cytokines in; non-transgenic, vehicle treated mice; transgenic, vehicle treated mice; and transgenic, Compound 1 treated mice (n = 14, 15 and 17, respectively). Figure 46A shows the levels of IL-4 measured in terminal cardiac plasma of all three groups. Figure 46B shows the levels of IL-6 measured in terminal cardiac plasma of all three groups. Data shown are mean +/- sem. ^* P<0.05, ^** P<0.01; One-way ANOVA, Dunnett's post hoc multiple comparison test. These changes in key cytokines indicate the general effect of compound 1 treatment on proteins involved in immune function and inflammation.

5. T Cell Infiltration into the Brain

(a) Tissue processing and histology

On the 9th day after administration, mice were sacrificed 2 hours after the last PO administration. Anesthesia was induced by 2,2,2-tribromoethanol. Mice were then perfused carotidally with 1% EDTA in PBS followed by 4% PFA in PBS. Brains were dissected, sagittal cut in two halves and fixed in 4% PFA in PBS. After 2 days of fixation, the hemibrains were transferred to 30% sucrose in PBS solution and replaced 2 days later. Plasma was collected and stored on dry ice.

Hemibrains were sectioned sagittal at 35 μm in a microtome at -22°C. Brain slices were sequentially collected into 12 tubes so that all 12 sections of the brain were displayed in a given tube. Brain sections were stored in cryoprotective medium at -20°C until staining was required. Free flow sections were blocked in appropriate serum in 10% serum in 0.5% PBST. Primary antibodies were incubated overnight at 4°C or room temperature as described below.

Rat anti-CD8a (63-0081-80, Thermo Fisher Scientific) was used at a concentration of 1:100 at room temperature, and murine anti-CD3 (555273, BD Biosciences) was used at a concentration of 1:100 at room temperature, and rabbit anti-CD3 (555273, BD Biosciences) was used at a concentration of 1:100 at room temperature. Iba-1 (016-26721, Wako) was used at a concentration of 1:1000 at 4°C, murine anti-CD68 (MCA1957, Bio-Rad) was used at 1:1000 at 4°C, Dylight 448/594 label Lycopersicon Esculentum (Tomato) Lectin (DL-1177, Fisher Scientific) was used at a concentration of 1:200 at room temperature. An appropriate fluorescent secondary antibody (Alexa-488/555/647, Invitrogen) was applied the next day at a concentration of 1:300 at room temperature for 1 hour. Slides were coverslips using Prolong Gold Mounting Media. Images were acquired at 20x on a Hamamatsu Nanozoomer 2.0HT slide scanner.

(b) T cell infiltration quantification

CD3 and CD8 positive cells were counted from images acquired with a Hamamatsu slide scanner from the cerebellum. The total number of CD3 and CD8 positive cells in the cerebellum was summed in approximately 5 sections of 35 m each. General one-way ANOVA was used to test for statistical significance, and Dunnett's multiple comparisons were post-hoc tests between treatment groups.

(c) quantification of neuroinflammation

Iba-1 and CD68 positive areas were quantified as percentages of ROIs around the whole cerebellum using thresholds in Image Pro Premier v9.2 software. Iba-1 and CD68 positive percent areas were averaged in approximately 5 sections for each mouse. General one-way ANOVA was used to test for statistical significance, and Dunnett's multiple comparisons were post-hoc tests between treatment groups.

processing group

Two month old C57Bl/6 mice were subjected to three treatments: vehicle control, vehicle treated EAE and compound 1 treated EAE (see Experimental Autoimmune Encephalomyelitis, Methods Mol Biol. 2012; 900: 381-401, incorporated herein by reference in its entirety). divided into groups. EAE was induced on day 0 by subcutaneous (SQ) injection of MOG + CFA emulsification and intravenous injection (IV) of pertussis toxin (PT). Additional injections of PT occurred on day 2. Vehicle or Compound 1 administration was via oral gavage (PO) on day 0 and continued twice daily (BID) until day 9. Mice were removed 2 hours after the last PO administration.

Drug formulation: Compound 1 was formulated with 40% HP-β-cyclodextrin and adjusted to pH 6.5 with NaOH (1M). Vehicle solutions were similarly formulated and adjusted for pH. Solutions were prepared fresh weekly and stored at 4°C.

EAE Emulsion: MOG 35-55 (Anaspec AS-60130-5) was reconstituted in PBS at 4 mg/ml. M. Tuberculosis H37 Ra (Fisher Scientific DF3114-33-8) was dissolved in Incomplete Freud's adjuvant at 4 mg/ml. These two solutions were emulsified using a glass syringe (Thermo Scientific Male Luer-LOK Priming Syringes 03-170-301) and a 3-way stop cock (Cadence 6001). The solution was prepared immediately prior to dosing.

Pertussis toxin: Pertussis toxin (Sigma P7208-50UG) was dissolved in saline at 0.002 mg/ml, and 100 μl IV was injected again on the day and 2 days after induction.

Treatment group:

Treatment group 1, vehicle treatment: 2.5-month-old young C57BL/6 mice (n = 5) received 100 μl of vehicle PO BID for 9 days starting 2 hours after SQ saline injection and only once on the first and last days. injected A total of 19 therapeutic injections.

Treatment group 2, vehicle treatment with EAE: young C57BL/6 mice (n = 10), 2.5 months old, received 100 μl of vehicle PO BID for 9 days starting 2 hours after EAE induction and only once at the beginning and at the end injected A total of 19 therapeutic injections.

Treatment group 3, compound 1 treatment with EAE: 2.5-month-old young C57BL/6 mice (n = 10) received 100 μl of compound 1 (30 mg/kg) PO BID for 9 days, starting 2 hours after EAE induction. was administered, and only one injection was administered at the beginning and at the end. A total of 19 therapeutic injections.

EAE induction resulted in an increase in CD3 and CD8 positive infiltrating T cells in the cerebellum. The number of these T-cells was significantly reduced when treated with compound 1. EAE induction also resulted in a significant increase in microglia in the cerebellum, which was also significantly rescued by compound 1 treatment after 9 days. Figure 47A shows an increase in CD3 positive infiltrating T cells in the cerebellum significantly reduced after treatment with Compound 1 for 9 days. Figure 47B shows an increase in CD8 positive infiltrating T-cells in the cerebellum, which is significantly reduced after treatment with Compound 1 for 9 days. Figure 47C shows a significant increase in Iba-1 positive areas in the cerebellum after EAE, which was significantly reduced in the cerebellum after 9 days of treatment. Figure 47D shows a significant increase in CD68 positive area in the cerebellum after EAE, which was significantly reduced in the cerebellum after 9 days of treatment.

d. human example

1. Eotaxin Levels and Aging

Levels of human eotaxin-1 were determined using a commercially available affinity based assay (SOMAscan, SomaLogic, Inc., Boulder, Colorado). Plasma samples were collected from 18, 30, 45, 55 and 66 year old donors and tested by SomaLogic using a SOMAscan aptamer-based affinity assay specifically tested for different levels of human eotaxin-1. Eotaxin-1 levels were determined and plotted by age group ( FIG. 45 ). Relative concentrations of eotaxin-1 increased with age, indicating that the eotaxin-1 pathway, including the primary receptor CCR3, is a target for treating age-related diseases such as neurodegenerative diseases and cognitive decline.

2. Human Biomarker Analysis

Whole blood from humans treated with Compound 1 was incubated with human recombinant eotaxin-1 to trigger eosinophil morphological changes (FIG. 49A) or CCR3 receptor internalization (FIG. 49B). Both assays showed strong concentration-dependent effects of compound 1 on their respective functional biomarker readouts.

Together, the results obtained from ESC and CCR3 receptor internalization assays confirm that Compound 1 acts as an inhibitor of the human eotaxin-1 pathway. In particular, compound 1 can act as a potent inhibitor of CCR3 by binding to its receptor.

In addition, the data obtained in FIG. 44 show that eosinophils were expressed at lower levels than non-transgenic animals in the mammalian transgenic Parkinson's disease model, which was reversed by compound 1 administration. Therefore, this data applied to patients with Parkinson's disease could help diagnose, monitor, and determine the prognosis of the disease.

3. Treatment of Parkinson's Disease Patients with Compound 1

Subjects diagnosed with Parkinson's disease receive 400 mg of compound or a placebo orally twice daily (BID). Subjects will receive treatment for 12 weeks and follow-up for a subsequent 2 weeks. Subjects are assessed for the effect of Compound 1 on motor function in a substantially defined off-drug state away from levodopa for at least 12 hours. Subjects are also assessed by flow cytometry, pharmacogenomics, and biomarker analysis performed on blood and plasma samples, including eosinophil levels. Gait analysis is assessed using the Zeno Walkway. Kinetics, tremors, general activity, and sleep are assessed using a wearable device.

Changes in baseline (day 1) motor function during a substantially defined no-dose state at Week 12 are determined using the Movement Disorders Association's Unified Parkinson's Disease Rating Scale (MDS-UPDRS), Part 3. Baseline (day 1) changes in clinical function, motor function, and activities of daily living at week 12 during drug administration are assessed by: MDS-UPDRS Part 1-4; Montreal Cognitive Assessment (MoCA); Schwab and UK Activities of Daily Living (SE-ADL) Scale; Clinical representation of severity index-PD (CISI-PD); PD Quality of Life Questionnaire-39 (PDQ-39); Sheehan-Suicide Rate Tracking Scale (S-STS); and walking 10 meters apart (also evaluated on dosing).

It is to be understood that the present invention is not limited to the specific embodiments that may be described, as modifications are possible. Furthermore, the terminology used herein is for the purpose of describing particular aspects and is not intended to be limiting, the scope of the invention being limited only by the appended claims.

Where a range of values is provided, unless the context dictates otherwise, the value intervening between the upper and lower limits of that range is, unless the pulse clearly dictates otherwise, each intervening value to the tenth of the unit of the lower limit. , or any of the above-mentioned intervening values are encompassed by the present invention. The upper and lower limits of these smaller ranges may independently be included in the smaller ranges and are also encompassed by the invention according to any specifically excluded limit in the stated range. Where the stated range includes one or both of the limits, ranges excluding either or both of those included limits are also included in the invention.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, representative exemplary methods and materials are now described.

All publications and patents cited herein are expressly and individually indicated to be incorporated by reference and each individual publication or patent is incorporated by reference to disclose and describe methods and/or materials related to the cited publication. As such, it is incorporated by reference. Citation of any publication is for publication prior to the filing date and is not to be construed as an admission that the present invention is not entitled to supersede such publication by virtue of prior invention. In addition, the disclosure dates provided may differ from the actual disclosure dates, which may need to be independently verified.

It should be noted that, as used herein and in the appended claims, the singular forms "a", "an" and "the" include plural referents unless the context clearly dictates otherwise. It is further noted that claims may be written to exclude any optional element. Accordingly, this statement is intended to serve as an antecedent basis for the use of exclusive terms such as "solely," "only," and the like in connection with the recitation of a claim or use of a "negative" limitation.

As will be apparent to one of ordinary skill in the art upon reading this disclosure, each individual aspect described and illustrated herein has distinct elements and features, which elements may be readily separated from or easily separated from the features of several other aspects without departing from the features. can be combined. All methods recited from the scope or spirit of the present invention may be performed in the recited order of events or any other order logically possible.

While the foregoing invention has been described in some detail by way of illustration and example for purposes of clarity of understanding, it will be apparent to those skilled in the art that certain changes and modifications may be made in light of the teachings of the invention. It is included herein without departing from the spirit or scope of the appended claims.

Accordingly, the foregoing merely illustrates the principles of the invention. It will be understood by those skilled in the art that, although not explicitly described or shown herein, they can devise various arrangements which embody the principles of the invention and are included within the spirit and scope of the invention. Further, all examples and conditional language recited herein are intended to aid the reader in understanding the principles of the invention and the concepts that the inventors have contributed to the advancement of the present technology, and are not to be construed as limited to the examples specifically recited. do. Moreover, the principles, aspects, and all examples of the specification and specific examples thereof recited herein are intended to encompass both structural and functional equivalents thereof.

Furthermore, it is intended that such equivalents include both currently known equivalents and equivalents developed in the future, ie, all elements that perform the same function, regardless of structure. Therefore, it is not intended that the scope of the invention be limited to the exemplary embodiments shown and described herein. Rather, the scope and spirit of the invention is embodied by the appended claims.

Claims (15)

A method of treating a neurodegenerative disease in a subject diagnosed with a neurodegenerative disease comprising administering a therapeutically effective amount of a compound of formula (1):

One
in the above formula
A is CH ₂ , O or NC _1-6 -alkyl;
R ¹ is
· NHR ^1.1 , NMeR ^1.1 ;
· NHR ^1.2 , NMeR ^1.2 ;
· NHCH ₂ -R ^1.3 ;
NH-C _3-6 -cycloalkyl, wherein optionally one carbon atom is replaced by a nitrogen atom and the ring is C _1-6 -alkyl, OC _1-6 -alkyl, NHSO ₂ -phenyl, NHCONH-phenyl, halogen, optionally substituted with 1 or 2 moieties selected from the group consisting of CN, SO ₂ -C _1-6 -alkyl, COO-C _{1-6 -alkyl;}
C _{9 or 10} -bicyclic-ring, wherein 1 or 2 carbon atoms are replaced by nitrogen atoms, the ring system is bonded to the basic structure of formula 1 via a nitrogen atom, the ring system is C _1-6 -alkyl, COO-C _1-6 -alkyl, C _1-6 -haloalkyl, OC _1-6 -alkyl, NO ₂ , halogen, CN, NHSO ₂ -C _1-6 -alkyl, m-methoxy-phenyl optionally substituted with 1 or 2 residues selected from;
a group selected from NHCH(pyridinyl)CH ₂ COO-C _1-6 -alkyl, NHCH(CH ₂ OC _1-6 -alkyl)-benzoimidazolyl, optionally substituted with halogen or CN;
or 1-amino cyclopentyl, optionally substituted with methyl-oxadiazole;
R ^1.1 is C _1-6 -alkyl, C _2-6 -alkenyl, C _2-6 -alkynyl, C _1-6 -haloalkyl, C _1-6 -alkylene-OH, C _2-6 -alke nylene-OH, C _2-6 -alkynylene-OH, CH ₂ CON(C _1-6 -alkyl) ₂ , CH ₂ NHCONH-C _3-6 -cycloalkyl, CN, CO-pyridinyl, CONR ^1.1.1 R ^1.1.2 , COO-C _1-6 -alkyl, N(SO ₂ -C _1-6 -alkyl)(CH ₂ CON(C _1-4 -alkyl) ₂ ) OC _1-6 -alkyl, O-pyri dinyl, SO ₂ -C _1-6 -alkyl, SO ₂ -C _1-6 -alkylene-OH, SO ₂ -C _3-6 -cycloalkyl, SO ₂ -piperidinyl, SO ₂ NH-C _{1- 6} -alkyl, SO ₂ N(C _1-6 -alkyl) ₂ , halogen, CN, CO-morpholinyl, CH ₂ -pyridinyl, or C _1-6 -alkyl, NHC _1-6 -alkyl and =O phenyl, optionally substituted with 1 or 2 moieties selected from the group consisting of heterocyclic rings, optionally substituted with 1 or 2 moieties selected from the group consisting of;
^1.1.1 R is H, C _1-6 - optionally substituted with alkyl, C _1-6 - alkyl, C _3-6 - cycloalkyl, C _1-6 - haloalkyl, CH ₂ CON (C _1-6 - alkyl,) ₂ , CH ₂ CO-azetidinyl, C _1-6 -alkylene-C _3-6 -cycloalkyl, CH ₂ -pyranyl, CH ₂ -tetrahydro-furanyl, CH ₂ -furanyl, C _1-6 -alkylene-OH or thiadiazolyl;
R ^1.1.2 is H, C _1-6 -alkyl, SO ₂ C _1-6 -alkyl; or
R ^1.1.1 and R ^1.1.2 optionally contain one N or O, replacing a carbon atom of the ring, as C _1-6 -alkyl, C _1-4 -alkylene-OH, OH, =O together form a 4-, 5- or 6-membered carbocyclic ring optionally substituted with one or two moieties selected from the group consisting of;
or
R ^1.1 is phenyl, wherein two adjacent moieties together form a 5 or 6 membered carbocyclic aromatic or non-aromatic ring, optionally independently of one another, one or two N, S or SO replacing a carbon atom of the ring ₂ , wherein the ring is _{optionally substituted with C 1-4} -alkyl or =O;
R ^1.2 is
· C _1-6 -alkyl, C _2-6 -alkenyl, C _2-6 -alkynyl, C _3-6 -cycloalkyl, CH ₂ COO-C _1-6 -alkyl, CONR ^1.2.1 R ^{1.2. 2} , COR ^1.2.3 , COO-C _1-6 -alkyl, CONH ₂ , OC _1-6 -alkyl, halogen, CN, SO ₂ N(C _1-6 -alkyl) ₂ , or 1 or 2 C ₁ heteroaryl optionally substituted with one or two moieties selected from the group consisting of heteroaryl optionally substituted with _{-6 -alkyl moieties;}
Heteroaryl, optionally substituted with a 5- or 6-membered carbocyclic non-aromatic ring containing, independently of each other, two N, O, S or SO _{2 replacing carbon atoms;}
Aromatic or non-aromatic C _{9 or 10} -bicyclic-ring, wherein one or two carbon atoms are from the group consisting of _{N(C 1-6} -alkyl) ₂ , CONH-C _{1-6 -alkyl, =O} replaced by N, O or S, each optionally substituted with one or two residues selected from;
a heterocyclic non-aromatic ring, optionally substituted with pyridinyl;
4,5-dihydronaphtho[2,1-d]thiazole, optionally substituted with NHCO-C _{1-6 -alkyl, and}
R ^1.2.1 is H, C _1-6 -alkyl, C _1-6 -alkylene-C _3-6 -cycloalkyl, C _1-4 -alkylene-phenyl, C _1-4 -alkylene-furanyl , C _3-6 -cycloalkyl, C _1-4 -alkylene-OC _1-4 -alkyl, C _1-6 -haloalkyl or 5- or 6-membered carbocyclic non-aromatic ring, optionally independently of one another, contains 1 or 2 N, O, S or SO ₂ , replacing a carbon atom of the ring, optionally substituted with 4-cyclopropylmethyl-piperazinyl;
R ^1.2.2 is H, C _1-6 -alkyl;
R ^1.2.3 are optionally independently of one another a 5- or 6-membered carbocyclic non-aromatic ring containing 1 or 2 N, O, S or SO _{2 , replacing a carbon atom of the ring;}
R ^1.3 is selected from phenyl, heteroaryl or indolyl, each as C _1-6 -alkyl, C _3-6 -cycloalkyl, OC _1-6 -alkyl, OC _1-6 -haloalkyl, phenyl, heteroaryl optionally substituted with 1 or 2 residues selected from the group consisting of;
R ² is selected from C _1-6 -alkylene-phenyl, C _1-6 -alkylene-naphthyl, and C _1-6 -alkylene-heteroaryl; C _1-6 - alkyl, C _1-6 - haloalkyl, OC _1-6 - alkyl, OC _1-6 - a haloalkyl, one, two or three moieties selected from the group consisting of halogen and optionally substituted, respectively;
R ³ is H, C _1-6 -alkyl;
R ⁴ is H, C _1-6 -alkyl;
or R ³ and R ⁴ together form a CH ₂ —CH ₂ group. According to claim 1,
A is CH ₂ , O or NC _1-4 -alkyl;
R ¹ is
· NHR ^1.1 , NMeR ^1.1 ;
· NHR ^1.2 , NMeR ^1.2 ;
· NHCH ₂ -R ^1.3 ;
NH-C _3-6 -cycloalkyl, wherein optionally one carbon atom is replaced by a nitrogen atom and the ring is C _1-6 -alkyl, OC _1-6 -alkyl, NHSO ₂ -phenyl, NHCONH-phenyl, halogen, optionally substituted with 1 or 2 moieties selected from the group consisting of CN, SO ₂ -C _1-6 -alkyl, COO-C _{1-6 -alkyl;}
C _{9 or 10} -bicyclic-ring, wherein 1 or 2 carbon atoms are replaced by nitrogen atoms, the ring system is bonded to the basic structure of formula 1 via a nitrogen atom, the ring system is C _1-6 -alkyl, COO-C _1-6 -alkyl, C _1-6 -haloalkyl, OC _1-6 -alkyl, NO ₂ , halogen, CN, NHSO ₂ -C _1-6 -alkyl, m-methoxy-phenyl optionally substituted with 1 or 2 residues selected from;
• a group selected from NHCH(pyridinyl)CH ₂ COO-C _1-6 -alkyl, NHCH(CH ₂ OC _1-6 -alkyl)-benzoimidazolyl, optionally substituted with Cl;
or 1-amino cyclopentyl, optionally substituted with methyl-oxadiazole;
R ^1.1 is C _1-6 -alkyl, C _1-6 -haloalkyl, CH ₂ CON(C _1-6 -alkyl) ₂ , CH ₂ NHCONH-C _3-6 -cycloalkyl, CN, CONR ^1.1.1 R ^1.1.2 , COO-C _1-6 -alkyl, OC _1-6 -alkyl, SO ₂ -C _1-6 -alkyl, SO ₂ -C _1-6 -alkylene-OH, SO ₂ -C _3-6 - cycloalkyl, SO _{2-piperidinyl,} SO ₂ NH-C _1-6 - alkyl, SO ₂ N (C _1-6 - alkyl) _2, halogen, CN, CO- morpholino carbonyl, CH _2-pyridinyl , or _{phenyl optionally substituted with 1 or 2 moieties selected from the group consisting of C 1-6} -alkyl, NHC _1-6 -alkyl and a heterocyclic ring optionally substituted with 1 or 2 moieties selected from the group consisting of =O ;
^1.1.1 R is H, C _1-6 - optionally substituted with alkyl, C _1-6 - alkyl, C _3-6 - cycloalkyl, C _1-6 - haloalkyl, CH ₂ CON (C _1-6 - alkyl,) ₂ , CH ₂ CO-azetidinyl, C _1-6 -alkylene-C _3-6 -cycloalkyl, CH ₂ -pyranyl, CH ₂ -tetrahydro-furanyl, CH ₂ -furanyl, C _1-6 -alkylene-OH or thiadiazolyl;
R ^1.1.2 is H, C _1-6 -alkyl, SO ₂ C _1-6 -alkyl; or
R ^1.1.1 and R ^1.1.2 are 4, 5 or 6 membered, optionally containing one O replacing a carbon atom of the ring and optionally substituted with one or two moieties selected from _{CH 2 OH} together form a carbon ring;
R ^1.2 is
C _1-6 -alkyl, C _3-6 -cycloalkyl, CH ₂ COO-C _1-6 -alkyl, CONR ^1.2.1 R ^1.2.2 , COO-C _1-6 -alkyl, CONH ₂ , OC _{1 -6} -alkyl, halogen, CN, SO ₂ N(C _1-6 -alkyl) ₂ , CO-pyrrolidinyl, CO-morpholinyl or hetero optionally substituted with _{1 or 2 C 1-6 -alkyl moieties} heteroaryl, optionally substituted with one or two moieties selected from the group consisting of aryl;
benzothiazolyl, indazolyl, dihydro-indole, each optionally substituted with one or two moieties selected from the group consisting of N(C _1-6 -alkyl) ₂ , CONH-C _{1-6 -alkyl, =O} lyl, indanyl, tetrahydro-quinolinyl;
• piperidinyl, optionally substituted with pyridinyl;
4,5-dihydronaphtho[2,1-d]thiazole, optionally substituted with NHCO-C _{1-6 -alkyl;}
R ^1.2.1 is H, C _1-6 -alkyl;
R ^1.2.2 is H, C _1-6 -alkyl;
R ^1.3 is selected from phenyl, pyrazolyl, isoxazolyl, pyrimidinyl, indolyl or oxadiazolyl, each C _1-6 -alkyl, C _3-6 -cycloalkyl, OC _1-6 -alkyl, OC optionally substituted with 1 or 2 moieties selected from the group consisting of _{1-6 -haloalkyl;}
R ² is CH ₂ - or phenyl-CH ₂ - is selected from naphthyl; C _1-6 - alkyl, C _1-6 - haloalkyl, OC _1-6 - alkyl, OC _1-6 - a haloalkyl, one or two moieties selected from the group consisting of halogen and optionally substituted, respectively; _{or CH 2} -thiophenyl, each optionally substituted with 1 or 2 moieties selected from halogen;
R ³ is H, C _1-4 -alkyl;
R ⁴ is H, C _1-4 -alkyl;
or R ³ and R ⁴ together form a _{CH 2} -CH _{2 group.} According to claim 1,
A is CH ₂ , O or N-Me;
R ¹ is
· NHR ^1.1 , NMeR ^1.1 ;
· NHR ^1.2 , NMeR ^1.2 ;
· NHCH ₂ -R ^1.3 ;
NH-cyclohexyl, optionally substituted with 1 or 2 moieties selected from the group consisting of C _1-4 -alkyl, NHSO _{2 -phenyl, NHCONH-phenyl, halogen;}
NH-pyrrolidinyl, optionally substituted with 1 or 2 moieties selected from the group consisting of SO ₂ -C _1-4 -alkyl, COO-C _{1-4 -alkyl;}
piperidinyl, optionally substituted with 1 or 2 moieties selected from the group consisting of NHSO ₂ -C _{1-4 -alkyl, m-methoxyphenyl;}
· C _1-4 - alkyl, COO-C _1-4 - alkyl, C _1-4 - haloalkyl, OC _1-4 - alkyl, NO _2, 1 or 2 moieties selected from the group consisting of halogen, optionally substituted , dihydro-indolyl, dihydro-isoindolyl, tetrahydro-quinolinyl or tetrahydro-isoquinolinyl;
• a group selected from NHCH(pyridinyl)CH ₂ COO-C _1-6 -alkyl, NHCH(CH ₂ OC _1-6 -alkyl)-benzoimidazolyl, optionally substituted with Cl;
or 1-amino cyclopentyl, optionally substituted with methyl-oxadiazole;
R ^1.1 is C _1-4 -alkyl, C _1-4 -haloalkyl, CH ₂ CON(C _1-4 -alkyl) ₂ , CH ₂ NHCONH-C _3-6 -cycloalkyl, CN, CONR ^1.1.1 R ^1.1.2 , COO-C _1-4 -alkyl, OC _1-4 -alkyl, SO ₂ -C _1-4 -alkyl, SO ₂ -C _1-4 -alkylene-OH, SO ₂ -C _3-6 - cycloalkyl, SO _{2-piperidinyl,} SO ₂ NH-C _1-4 - alkyl, SO ₂ N (C _1-4 - alkyl) _2, halogen, CN, CO- morpholino carbonyl, CH _2-pyridinyl , or imidazolidinyl, piperidinyl, oxazinanyl, pyrazolyl, triazolyl optionally substituted with 1 or 2 moieties selected from the group consisting of _{C 1-4} -alkyl, NHC _{1-4 -alkyl and =O} , phenyl, optionally substituted with 1 or 2 moieties selected from the group consisting of tetrazolyl, oxazolyl, oxadiazolyl, thiazolyl, pyridinyl, pyrimidinyl;
^1.1.1 R is H, C _1-4 - optionally substituted with alkyl, C _1-6 - alkyl, C _3-6 - cycloalkyl, C _1-4 - haloalkyl, CH ₂ CON (C _1-4 - alkyl,) ₂ , CH ₂ CO-azetidinyl, C _1-4 -alkylene-C _3-6 -cycloalkyl, CH ₂ -pyranyl, CH ₂ -tetrahydro-furanyl, CH ₂ -furanyl, C _1-4 -alkylene-OH or thiadiazolyl;
R ^1.1.2 is H, C _1-4 -alkyl, SO ₂ C _1-4 -alkyl; or
R ^1.1.1 and R ^1.1.2 are 4, 5 or 6 membered, optionally containing one O replacing a carbon atom of the ring and optionally substituted with one or two moieties selected from _{CH 2 OH} together form a carbon ring;
R ^1.2 is
C _1-4 -alkyl, C _3-6 -cycloalkyl, CH ₂ COO-C _1-4 -alkyl, CONR ^1.2.1 R ^1.2.2 , COO-C _1-4 -alkyl, CONH ₂ , OC _{1 -4} -alkyl, halogen, CO-pyrrolidinyl, CO-morpholinyl or pyrazolyl, triazolyl, tetrazolyl, oxazolyl, oxadiazolyl, optionally substituted with _{1 or 2 C 1-4 -alkyl moieties} pyridinyl, pyridazinyl, pyrrolyl, pyrazolyl, isoxazolyl, thiazolyl, thiadiazolyl, optionally substituted with one or two moieties selected from the group consisting of;
benzothiazolyl, indazolyl, dihydro-indole, each optionally substituted with one or two moieties selected from the group consisting of N(C _1-4 -alkyl) ₂ , CONH-C _{1-4 -alkyl, =O} lyl, indanyl, tetrahydro-quinolinyl;
• piperidinyl, optionally substituted with pyridinyl;
4,5-dihydronaphtho[2,1-d]thiazole, optionally substituted with NHCO-C _{1-4 -alkyl;}
R ^1.2.1 is H, C _1-4 -alkyl;
R ^1.2.2 is H, C _1-4 -alkyl;
R ^1.3 is selected from phenyl, pyrazolyl, isoxazolyl, pyrimidinyl, indolyl or oxadiazolyl, respectively C _1-4 -alkyl, C _3-6 -cycloalkyl, OC _1-4 -alkyl, OC optionally substituted with 1 or 2 moieties selected from the group consisting of _{1-4 -haloalkyl;}
R ² is CH ₂ - or phenyl-CH ₂ - is selected from naphthyl; C _1-4 - alkyl, C _1-4 - haloalkyl, OC _1-4 - alkyl, OC _1-4 - a haloalkyl, one or two moieties selected from the group consisting of halogen and optionally substituted, respectively; _{or CH 2} -thiophenyl, each optionally substituted with 1 or 2 moieties selected from halogen;
R ³ is H;
R ⁴ is H;
or R ³ and R ⁴ together form a _{CH 2} -CH _{2 group.} According to claim 1,
A is CH ₂ , O or N-Me;
R ¹ is
· NHR ^1.1 , NMeR ^1.1 ;
· NHR ^1.2 , NMeR ^1.2 ;
· NHCH ₂ -R ^1.3 ;
NH-piperidinyl, optionally substituted with pyridinyl;
NH-cyclohexyl, optionally substituted with 1 or 2 moieties selected from the group consisting of t-butyl, NHSO _{2 -phenyl, NHCONH-phenyl, F;}
• NH-pyrrolidinyl, optionally substituted with 1 or 2 moieties selected from the group consisting of _{SO 2 -Me, COO-t-butyl;}
piperidinyl, optionally substituted with 1 or 2 moieties selected from the group consisting of NHSO _{2 -n-butyl, m-methoxyphenyl;}
Dihydro-indolyl, dihydro-isoindolyl, tetrahydro-qui, optionally substituted with 1 or 2 moieties selected from the group consisting of Me, COO-Me, CF ₃ , OMe, NO _{2 , F, Br} nolinyl or tetrahydro-isoquinolinyl;
• a group selected from NHCH(pyridinyl)CH ₂ COO-C _1-6 -alkyl, NHCH(CH ₂ OC _1-6 -alkyl)-benzoimidazolyl, optionally substituted with Cl;
or 1-amino cyclopentyl, optionally substituted with methyl-oxadiazole;
R ^1.1 is Me, ethyl, t-butyl, CF ₃ , CH ₂ CONMe ₂ , CH ₂ NHCONH-cyclohexyl, CN, CONR ^1.1.1 R ^1.1.2 , COO-Me, COO-ethyl, O-Me, SO ₂ -Me, SO ₂ -ethyl, SO ₂ -CH ₂ CH ₂ -OH, SO ₂ -cyclopropyl, SO ₂ -piperidinyl, SO ₂ NH-ethyl, SO ₂ NMeEt, F, Cl, CO-morphopoly nyl, CH ₂ -pyridinyl, or imidazolidinyl, piperidinyl, oxazinanyl, pyrazolyl, triazolyl, tetrazolyl optionally substituted with 1 or 2 moieties selected from the group consisting of Me, NHMe and =O , phenyl optionally substituted with 1 or 2 moieties selected from the group consisting of oxazolyl, oxadiazolyl, thiazolyl, pyridinyl, pyrimidinyl;
R ^1.1.1 is methyl, ethyl, t-butyl, isopropyl, cyclopropyl, CH ₂ -isopropyl, CH ₂ -t-butyl, CH(CH ₃ )CH ₂ CH ₃ , optionally substituted with H, Me _{CH 2 CHF 2, CH 2 CONMe} 2, CH 2 CO- azetidinyl, CH ₂ - cyclopropyl, CH _{2-cyclo-butyl,} CH _2-pyranyl, CH _{2-tetrahydro-furanyl,} CH _2-furanyl , CH ₂ CH ₂ OH or thiadiazolyl;
R ^1.1.2 is H, methyl, ethyl, SO ₂ Me, SO ₂ ethyl; or
R ^1.1.1 and R ^1.1.2 are 4, 5 or 6 membered, optionally containing one O replacing a carbon atom of the ring and optionally substituted with one or two moieties selected from _{CH 2 OH} together form a carbon ring;
R ^1.2 is
methyl, ethyl, propyl, butyl, cyclopropyl, CH ₂ COO-ethyl, CONR ^1.2.1 R ^1.2.2 , COO-methyl, COO-ethyl, CONH ₂ , OMe, Cl, Br, CO-pyrrolidinyl, Pyridinyl, pyri, optionally substituted with one or two moieties selected from the group consisting of CO-morpholinyl or pyrazolyl, triazolyl, tetrazolyl, oxazolyl, oxadiazolyl optionally substituted with one or two methyl moieties dazinyl, pyrrolyl, pyrazolyl, isoxazolyl, thiazolyl, thiadiazolyl;
benzothiazolyl, indazolyl, dihydro-indolyl, indanyl, tetrahydro-quinolinyl, each optionally substituted with one or two moieties selected from the group consisting of NMe _{2 , CONH-Me, =O;}
4,5-dihydronaphtho[2,1-d]thiazole, optionally substituted with NHCO-Me;
R ^1.2.1 is H, or methyl;
R ^1.2.2 is H, or methyl;
R ^1.3 is selected from phenyl, pyrazolyl, isoxazolyl, pyrimidinyl, indolyl or oxadiazolyl, each 1 selected from the group consisting of _{methyl, ethyl, propyl, cyclopentyl, O-Me, O-CHF 2} or optionally substituted with two residues;
R ² is CH ₂ - or phenyl-CH ₂ - is selected from naphthyl; each optionally substituted with 1 or 2 moieties selected from the group consisting of CH ₃ , CF ₃ , OCF _{3 , F, Cl, Br, ethyl; or CH 2} -thiophenyl, each optionally substituted with 1 or 2 moieties selected from the group consisting of Cl, Br;
R ³ is H;
R ⁴ is H;
or R ³ and R ⁴ together form a _{CH 2} -CH _{2 group.} According to claim 1,
A is CH ₂ , O or N-Me;
R ¹ is
· NHR ^1.1 ,
· NHR ^1.2 ,
R ^1.1 is Me, ethyl, butyl, CF ₃ , CH ₂ CONMe ₂ , CH ₂ NHCONH-cyclohexyl, CN, CONR ^1.1.1 R ^1.1.2 , COO-Me, COO-ethyl, O-Me, SO ₂ — Me, SO ₂ -ethyl, SO ₂ -CH ₂ CH ₂ -OH, SO ₂ -cyclopropyl, SO ₂ -piperidinyl, SO ₂ NH-ethyl, SO ₂ NMeEt, F, Cl, CO-morpholinyl, CH ₂ -pyridinyl, or imidazolidinyl, piperidinyl, oxazinanyl, pyrazolyl, triazolyl, tetrazolyl, oxa optionally substituted with 1 or 2 moieties selected from the group consisting of Me, NHMe and =O phenyl optionally substituted with 1 or 2 moieties selected from the group consisting of zolyl, oxadiazolyl, thiazolyl, pyridinyl, pyrimidinyl;
R ^1.1.1 is methyl, ethyl, t-butyl, isopropyl, cyclopropyl, CH ₂ -isopropyl, CH ₂ -t-butyl, CH(CH ₃ )CH ₂ CH ₃ , optionally substituted with H, Me _{CH 2 CHF 2, CH 2 CONMe} 2, CH 2 CO- azetidinyl, CH ₂ - cyclopropyl, CH _{2-cyclo-butyl,} CH _2-pyranyl, CH _{2-tetrahydro-furanyl,} CH _2-furanyl , CH ₂ CH ₂ OH or thiadiazolyl;
R ^1.1.2 is H, methyl, ethyl, SO ₂ Me, SO ₂ ethyl; or
R ^1.1.1 and R ^1.1.2 are 4, 5 or 6 membered, optionally containing one O replacing a carbon atom of the ring and optionally substituted with one or two moieties selected from _{CH 2 OH} together form a carbon ring;
R ^1.2 is
methyl, ethyl, propyl, butyl, cyclopropyl, CH ₂ COO-ethyl, CONR ^1.2.1 R ^1.2.2 , COO-methyl, COO-ethyl, CONH ₂ , OMe, Cl, Br, CO-pyrrolidinyl, Pyridinyl, pyri, optionally substituted with one or two moieties selected from the group consisting of CO-morpholinyl or pyrazolyl, triazolyl, tetrazolyl, oxazolyl, oxadiazolyl optionally substituted with one or two methyl moieties dazinyl, pyrrolyl, pyrazolyl, isoxazolyl, thiazolyl, thiadiazolyl;
benzothiazolyl, indazolyl, dihydro-indolyl, indanyl, tetrahydro-quinolinyl, each optionally substituted with one or two moieties selected from the group consisting of NMe _{2 , CONH-Me, =O;}
4,5-dihydronaphtho[2,1-d]thiazole, optionally substituted with NHCO-Me;
R ^1.2.1 is H, or methyl;
R ^1.2.2 is H, or methyl;
R ² is CH ₂ - or phenyl-CH ₂ - is selected from naphthyl; each optionally substituted with 1 or 2 moieties selected from the group consisting of CH ₃ , CF ₃ , OCF _{3 , F, Cl, Br, ethyl;}
R ³ is H;
R ⁴ is H. According to claim 1,
A is CH ₂ , O or NMe;
R ¹ is

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is selected from ;

R ² is

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is selected from ;
R ³ is H;
R ⁴ is H;
or R ³ and R ⁴ together form a _{CH 2} —CH _{2 group.} The method of claim 1 , wherein the compound is a co-crystal of the formula:

in the above formula
R ¹ is C _1-6 -alkyl, C _1-6 -haloalkyl, OC _1-6 -haloalkyl, halogen;
m is 1, 2 or 3;
R ^2a and R ^2b are each independently H, C _1-6 -alkyl, C _1-6 -alkenyl, C _1-6 -alkynyl, C _3-6 -cycloalkyl, COO-C _1-6 -alkyl, OC _1-6 -alkyl, CONR ^2b.1 R ^2b.2 , halogen;
R ^2b.1 is H, C _1-6 -alkyl, C _0-4 -alkyl-C _3-6 -cycloalkyl, C _1-6 -haloalkyl;
R ^2b.2 is H, C _1-6 -alkyl; this
R ^2b.1 and R ^2b.2 _{together are a C 3-6} -alkylene group, forming a hetero ring with a nitrogen atom, a carbon atom or ring may optionally be replaced by an oxygen atom,
R ³ is H, C _1-6 -alkyl;
X is chloride, bromide, iodide, sulfate, phosphate, methane sulfonate, nitrate, maleate, acetate, benzoate, citrate, salicylate, fumarate, tartrate, dibenzoyl tartrate, oxalate, an anion selected from the group consisting of succinate, benzoate and p-toluene sulfonate;
j is 0, 0.5, 1, 1.5 or 2;
Orotic acid, hippuric acid, L-pyroglutamic acid, D-pyroglutamic acid, nicotinic acid, L-(+)-ascorbic acid, saccharin, piperazine, 3-hydroxy-2-naphthoic acid, mucus (galactaric) acid , palm (embonic) acid, stearic acid, cholic acid, deoxycholic acid, nicotinamide, isonicotinamide, succinamide, uracil, L-lysine, L-proline, D-valine, L-arginine, selected from the group consisting of glycine It has a co-crystal former. The method of claim 1 , wherein the compound is a crystalline salt of the formula:

8. A method according to claim 1, characterized in that the compound comprises at least one co-crystal of the compound of the formula according to claim 7 and a pharmaceutically acceptable carrier. Method according to claim 1, characterized in that the compound of formula (1) is administered in the form of the individual optical isomer, mixture of individual enantiomers, racemate or enantiomerically pure compound. Method according to claim 1, characterized in that the compound is a pharmaceutical composition comprising as active ingredients at least one compound of the formula: a first diluent, a second diluent, a binder, a disintegrant and a lubricant:

in the above formula
R ¹ is H, C _1-6 -alkyl, C _0-4 -alkyl-C _3-6 -cycloalkyl, C _1-6 -haloalkyl;
R ² is H, C _1-6 -alkyl;
X is an anion selected from the group consisting of chloride or ½ dibenzoyl tartrate;
j is 1 or 2. 12. The method of claim 11, wherein the pharmaceutical composition further comprises an additional disintegrant. 13. The method according to claim 11 or 12, wherein the pharmaceutical composition further comprises an additional lubricant. 14. The method according to claim 11, 12 or 13, wherein the diluent of the pharmaceutical composition is cellulose powder, dibasic calcium phosphate anhydride, dibasic calcium phosphate dehydrate, erythritol, low substituted hydroxypropyl cellulose, mannitol, pregelatinization Method, characterized in that it further comprises starch or xylitol. According to any one of the preceding claims, wherein the neurodegenerative disease is from the group consisting of Alzheimer's disease, Parkinson's disease, frontotemporal dementia, Huntington's disease, amyotrophic lateral sclerosis, multiple sclerosis, glaucoma, amyotrophic dystrophy, vascular dementia, progressive supranuclear palsy A method characterized in that selected.

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