KR20070011483A - Substantially pure 2-{[2-(2-methylamino-pyrimidin-4-yl)-1h-indole-5-carbonyl]-amino}-3-(phenylpyridin-2-yl-amino)-propionic acid as an ikb kinase inhibitor - Google Patents

Abstract

The present invention is directed to the substantially pure compound of formula (A), or pharmaceutically acceptable salt, or solvate of said compound; to a pharmaceutical composition comprising a pharmaceutically effective amount of the compound of formula (A), and a pharmaceutically acceptable carrier; and the use of a compound of formula (A) having activity as an inhibitor, preferably a selective inhibitor, of IkB (IKK), particularly IKK-2, and methods related thereto. ® KIPO & WIPO 2007

Description

Substantially pure 2-VII [2- (2-methylamino-pyrimidin-4-yl) -1H-indol-5-carbonyl] -amino｝ -3- (phenylpyridin-2-yl- as an Ib kinase inhibitor Amino) -propionic acid (SUBSTANTIALLY PURE 2-｛[2- (2-METHYLAMINO-PYRIMIDIN-4-YL) -1H-INDOLE-5-CARBONYL] -AMINO｝ -3- (PHENYLPYRIDIN-2-YL-AMINO) -PROPIONIC ACID AS AN IKB KINASE INHIBITOR}

The present invention relates to indole derivatives, their preparation, pharmaceutical compositions comprising them, their use and intermediate products thereof.

NF-κB is a heterodimeric transcription factor that regulates the expression of multiple inflammatory genes. The expression of more than 70 known proteins is regulated by transcription by binding NF-κB to specific sequence elements in the promoter portion of these genes. (Baeuerle and Baichwal, Advances in Immunology 65: 111-137, 1997). NF-κB is known for angiogenesis (Koch et al., Nature 376: 517-519, 1995); Atherosclerosis (Brand et al., J Clin Inv. 97: 1715-1722, 1996), endotoxin shock and sepsis (Bohrer et al., J. Clin. Inv. 100: 972-985, 1997), inflammatory Intestinal disease (Panes et al., Am J Physiol. 269: H1955-H1964, 1995), ischemia / reflux injury (Zwacka et al., Nature Medicine 4: 698-704, 1998), and allergic lung inflammation (Gosset et al. , Int Arch Allergy Immunol. 106: 69-77, 1995). Thus, targeting NF-κB by targeting regulatory proteins during NF-κB activation may be a beneficial strategy for inducing anti-inflammatory treatment due to the central role of NF-κB in inflammatory conditions.

IκB kinases (IKKs) are major regulatory signal producing molecules that regulate NF-κB activity. Two IKKs, namely IKK-1 (IKK-α) and IKK-2 (IKK-β), are N-terminal with a double serine activation loop, leucine zipper domain, C-terminal helix-loop-helix domain, and serine cluster It is a structurally unique kinase that contains a kinase domain. IKK enzymes have relatively low sequence homology with other kinases and have not identified compounds with surprising efficacy in the initial profile of known kinase inhibitors. In kinetics analysis, IKK-2 binds to and phosphorylates IκBα and IκB with high affinity and relatively equivalent affinity (Heilker et al. 1999). Recombinant IKK-2 phosphorylates IκBα peptide 26-42, which has an affinity almost equivalent to full length IκBα, but the unique IKK enzyme complex is 25,000 times more effective in phosphorylating full length IκBα, which is the C-terminal region of IκBα. This suggests that there are additional regulatory proteins in the IKK enzyme complex that are important regulatory sequences or that accelerate the rate of catalytic reactions (Burke et al., Journal of Biological Chemistry 274: 36146-36152, 1999). Phosphorylation of IκBα occurs through random sequential mechanics, meaning that ATP or IκBα first binds to IKK-2, both of which must be bound before the phosphorylation of IκBα occurs (Peet And Li, Journal of Biological Chemistry 274: 32655-32661, 1999). Compared to other serine-threonine kinases such as p38 and JNK, IKK-2 binds to ATP with a uniquely high affinity (Ki = 130 nM), which, when tested against IKK-2, is a broad region with many unique ATP binding pockets. It means that it has a relatively weak activity against the specific kinase inhibitor of. To date, there are no reports on the crystal structure of IKK-2. However, homology modeling has shown that serine-rich tails are likely to mediate the formation of N-terminal kinase domains with activation loops, homo / dimers of IKK-1 and IKK-2, and serine-rich tails. Three structural domains were identified, including the C-terminal helix-loop-helix having. Activation of IKK-2 depends on activation or phosphorylation of serine 177 and 181 in the T-loop. Alanine mutations eliminate activity, while glutamate mutations structurally produce active enzymes. (Mercurio et al. Science 278: 860-866, 1997; Delhase et al., Science 284: 30 313, 1999).

IKK-1 and IKK-2 are both produced as heterodimers, and the IKK-2 homodimer is associated with a 700-900 kDa cytoplasmic enzyme complex, termed "IKK Signalsome" (Mercurio et al., Science 278: 860-866, 1997). Another component, IKKAP-1 or NEMO / IKKγ, has no surface catalytic function, but can associate directly with IKK-2, which is essential for the full activation of NF-κB (Mercurio et al., Mol Cell Biol 19: 1526 -1538, 1999). Many immune and inflammatory mediators, including TNFα, lipopolysaccharide (LPS), IL-1β, CD3 / CD28 (antigen provided), CD40L, FasL, viral infection, oxidative stress, have been shown to induce NF-κB activation lost. Although receptor complexes that introduce these various stimuli are very different in these protein components, it is understood that each of these stimuli induces activation of IKKs and NF-κB.

The IKK complex appears to be the central integrator of various inflammatory signals that induce IκB phosphorylation. IKKs are double serine by upstream kinases, including NF-κB induced kinases, NIK (Malinin et al., Nature 385: 540-544, 1997) and MEKK-1 (Yujiri et al., Science 282: 1911-1914, 1998). Activated at the residue moiety. Initial data show that these kinases preferentially activate IKK-1 and IKK-2, respectively, but the differential activation of NIK and MEKK-1 is not yet clear. Activated IKK phosphorylates the cytoplasmic inhibitor protein IκB (which binds to NF-κB), blocking the nuclear localization signal present in the Rel protein (Cramer et al., Structure 7: R1-R6, 1999). IKK phosphorylation of IκB on serine 32 and 36 forms a structural motif recognized by E3 ligand, βTRcP (Yaron et al., Nature 396: 590-594, 1998). Docking of βTRcP results in the formation of a ligase complex that polyubiquitinates IκB, which is targeted for degradation by 26S proteosomes. Free NF-κB can then be identified by nuclear transport proteins, translocated into the nucleus and associated with sequence specific regulatory elements on the gene promoter at this site.

These kinases can phosphorylate IκB in vitro, but in earlier studies using genetic mutations, IKK-2, but not IKK-1, activates NF-κB by pro-inflammatory stimuli such as IL-1β and TNFα. Appeared to be essential. In addition, only catalytically inactive mutants of IKK-2 block the expression of NF-κB regulated genes such as monocyte chemical attracting proteins (MCP-1) and intracellular adsorption molecules (ICAM-1) (Mercurio et al., Science 278: 860-866, 1997). Initial findings have been demonstrated in studies of knockout animals on IKK-1 and IKK-2 (Hu et al., Science 284: 316-320, 1999; Li et al., Genes & Development 13: 1322-1328, 1999; Li et al. , Science 284: 321-324, 1999; Takeda et al., Science 84: 313-316, 1999; Tanaka et al., Immunity 10: 421-429, 1999). IKK-1 ^-/-The animal was alive at birth but died within hours. Puppies showed skin abnormalities due to deficient proliferation and differentiation, but no severe deficiency was observed in cytokine-induced activation of NF-κB. In contrast, IKK-2 ^{− / −} embryos died on days 14-16 of pregnancy due to surprisingly similar liver degeneration and apoptosis as observed in Rel A knockout animals (Beg et al., Nature 376: 167-170, 1995). . In addition, IKK-2 ^-/- embryonic fibroblasts of animals significantly reduced NF-κB activation after cytokine stimulation, but not IKK-1 ^-/- .

Thus, in cell and animal experiments, IKK-2 is a central regulator of the pro-inflammatory role of NF-κB, where IKK-2 is activated in response to immune and inflammatory stimuli and signal production pathways. Many of these immune and inflammatory mediators, including IL-1β, LPS, TNFα, CD3 / CD28 (antigen provided), CD40L, FasL, viral infections and oxidative stress, play an important role in respiratory diseases. In addition, ubiquitous expression of NF-κB in response to multiple stimuli means that almost all cell types present in the lung are potential targets of anti-NF-κB / IKK-2 therapy. This includes alveolar epithelium, mast cells, fibroblasts, vascular endothelial cells, invasive leukocytes (including neutrophils, macrophages, lymphocytes, eosinophilic leukocytes, basophilic leukocytes). Cyclooxygenase-2 and 12-lipooxygenase (inflammatory mediator synthesis), TAP-1 peptide carrier (antigen processing), MHC type I H-2K and type II constant chain (antigen provided), E-selectin And vascular cell adsorption molecules (leukocyte supplementation), interleukin-1, 2, 6, 8 (cytokines), RANTES, eotaxin, GM-CSF (chemokine), superoxide dismutase, NADPH quinone oxidoreductase (reactive) By inhibiting the expression of genes such as oxygen species), it is believed that inhibitors of IKK-2 exert a wide range of anti-inflammatory activities.

Patent application WO 94/12478 (incorporated by reference) describes indole derivatives that inhibit blood platelet aggregation. Patent applications WO 01/00610 and WO 01/30774 (incorporated by reference) describe indole derivatives and benzimidazole derivatives capable of modulating NF-κB, respectively. As described above, NF-κB is a heterodimer transcription factor capable of activating a number of genes encoding pro-inflammatory cytokines such as IL-1, IL-2, TNFα or IL-6. NF-κB is present in the cytosol of cells, where it complexes with the naturally occurring inhibitor IκB. Cellular stimulation by cytokines leads to, for example, IκB phosphorylation and consequently proteolysis. This proteolytic destruction leads to NF-κ B activation, which migrates to the nucleus of the cell, where it activates many pro-inflammatory genes.

In diseases such as rheumatoid arthritis, osteoarthritis, asthma, chronic obstructive pulmonary disease (COPD), rhinitis, multiple sclerosis, myocardial infarction, Alzheimer's disease, type II diabetes, inflammatory bowel disease or atherosclerosis, NF-κB is above normal Is activated. Inhibition of NF-κB may be used by itself or as a useful cancer therapy in addition to cytostatic therapy. Inhibiting NF-κB-activated signal chains at various sites or directly interfering with gene transcription by glucocorticoids, salicylates or gold salts has been shown to be useful in treating rheumatoids.

The first step in the above mentioned signal cascade is the collapse of IκB. Such phosphorylation is regulated by specific IκB kinases. Thus, inhibitors of IκB kinases are often known to have the disadvantage of being non-specific substances for inhibiting only one type of kinase. For example, most inhibitors of IκB kinases simultaneously inhibit several different kinases because the structures of the catalytic domains of these kinases are very similar. As a result, inhibitors work in an undesirable way for many enzymes, including those that possess important functions.

Chronic obstructive pulmonary disease (COPD) is a debilitating inflammatory disease of the lungs characterized by the gradual development of air flow restrictions that are not completely reversible (Pauwels et al., 2001). Airflow restriction is implicated in abnormal inflammatory responses of the lungs to non-toxic particles or gases (mostly due to tobacco smoke). While COPD affects the lungs, it can also produce major systemic consequences. The term COPD also includes chronic obstructive bronchitis such as small airway obstruction, emphysema, enlargement of the air space and destruction of lung parenchyma, loss of lung elasticity, obstruction of the small airways. In contrast, chronic bronchitis is characterized by a cough (due to excessive secretion of mucus) that causes phlegm for more than three months for two consecutive years. There is some epidemiologic evidence that excessive secretion of mucus involves blockage of air flow as a result of obstruction of the peripheral airways. Most patients with COPD have all three pathological conditions (chronic obstructive bronchitis, emphysema, mucus filling), but the relative degrees of emphysema and obstructive bronchitis in individual patients vary (Vestbo et al., 1996; Barnes, 2004a, Barnes, 2004b; Hogg, 2004).

In developed countries, smoking causes COPD, but in developing countries, other environmental pollutants (especially sulfur dioxide and particles) and certain occupational chemicals (such as cadmium) are important sources. Secondhand smoke is also a risk factor.

COPD patients tend to be aggravated, which rapidly exacerbates their respiratory symptoms. COPD exacerbation is an event of the natural course of the disease characterized by changes in the patient's baseline dyspnea, coughing and sputum beyond daily changes sufficient to warrant a change in management (Rodriguez-Roisin, 2000; Burge and Wedzicha, 2003).

Tracheal and bronchial infections are a major cause of COPD exacerbations, but the specific nature of infectious agents and their exact role is still controversial (Wedzicha, 2002; White et al., 2003). In addition, COPD exacerbation is clearly associated with levels of respirable particles and environmental air pollutants, which are also associated with hospital admission rates (Rennard and Farmer, 2004).

Exacerbation frequency is associated with disease severity in COPD. Exacerbations can also adversely affect the natural history of these diseases by contributing to reduced lung function, systemic effects and increased rates of premature mortality. Unfortunately, to date there is no widely accepted definition of what exacerbates COPD (Rodriguez-Roisin, 2000). It is difficult to determine the intensity and duration of the augmented symptom required to be recognized as "bad". In addition, several definitions coexist and do not explain the definitions used in many clinical trials to use substantially different criteria or to diagnose exacerbations. The most widely used clinical criteria for characterizing acute exacerbation of COPD has been described by Anthonisen et al., (1987). Deterioration in the study is divided into three groups, type 1 deterioration is characterized by an increase in the frequency of breathing, an increase in the amount of sputum and new or increased sputum purulent; Type 2 includes two of these symptoms; Type 3 includes at least one type of laryngitis or nasal discharge within the last 5 days, including an increase in stiffness, an increase in cough or a 20% increase in respiratory or heart rate when compared to fever and basal levels. It consists of any one of the above symptoms with additional features. These criteria have been used as benchmarks to date and should correlate all proposed etiologies of exacerbations with their key features.

Inhibition of NF-κB can also be used by itself or in addition to cell suppression therapy to treat hypoproliferative diseases such as solid tumors and leukemia. Inhibition of NF-κB-activated signal chains at various time points has been shown to be useful in the treatment of rheumatism by directly interfering with transcription of genes by glucocorticoids, salicylates or gold salts.

Patent application WO 01/30774 describes indole derivatives, and US patent application 10 / 642,970 describes indole and benzimidazole derivatives capable of modulating NF-κB and exerting a potent inhibitory effect on IκB kinase. In particular, US application 10 / 642,970 discloses such compounds as the following indole and benzimidazole derivatives of formula (I) and their preparation, pharmaceutical compositions comprising these compounds, and disease prevention and treatment associated with increased activity of IκB kinases. It demonstrates how to administer.

In addition, US application 10 / 642,970 describes compounds of formulas B, C, and D:

However, in US application 10 / 642,970 M is N; It does not specifically describe compounds of formula (I) wherein R 1 is hydrogen, R 2 is carboxyl (-COOH), R 3 is methyl, R 4 is pyridin-2-yl, R 11 is hydrogen and X is CH.

In view of the foregoing, there is a need for inhibitors of IκB kinase that act through selective inhibition of IKK, in particular IKK-2 inhibitors. Inhibitors that exhibit local activity as opposed to systemic activity may be preferred. Such inhibitors should be available to treat patients with IKK-2 mediated pathological (disease) conditions such as asthma or chronic obstructive pulmonary disease (COPD) that may be ameliorated by targeted administration of the inhibitor.

Summary of the Invention

The present invention relates to compounds and compositions thereof and methods associated therewith which are active as inhibitors, preferably as selective inhibitors of IκB (IKK), in particular IKK-2.

In particular, the invention relates to substantially pure compounds of formula A or pharmaceutically acceptable salts or solvates thereof:

The invention also relates to a pharmaceutical composition comprising a pharmaceutically effective amount of a compound of formula A and a pharmaceutically acceptable carrier.

The invention also relates to the use of an inhibitor of IκB kinase of formula (A).

FIG. 1 shows a mouse imaging study plot in a NF-κB-luciferase reporter mouse model, wherein animals were treated with vehicle / PBS solution (negative control animals) and animals without any compound [vehicle / IL-1β] Or has IL-1β-induced NF-κB activity with increasing dose of the compound of Formula A or Compound B (0.3 mpk, 1 mpk, 3 mpk, 10 mpk).

FIG. 2 shows a mouse imaging study graph in the NF-κB-luciferase reporter mouse model showing bioluminescence levels in animals treated with vehicle / PBS solution (negative control animals), wherein the animal is free of any compound IL-1β-induced NF-κB activation is shown with increasing dose (0.3 mpk, 1 mpk, 3 mpk, 10 mpk) of [Vehicle / IL-1β] or Compound A or Compound B.

In FIG. 3 , the left graph shows the level of the compound of formula A after it is injected into the lung and plasma tissue in an amount of 0.3 mg / kg of the compound of formula A, and the right graph shows the amount of 0.3 mg / kg of the compound of formula B. As it is injected into the lung and plasma tissue it shows the level of the compound of formula B.

In FIG. 4 , the left graph shows lung exposure of the compound of formula A after administration with increasing dose of the compound of formula A (0.01 mpk, 0.03 mpk, 0.10 mpk, 0.30 mpk), and the graph on the right shows the compound of formula A Plasma exposure of the compound of formula A is shown after administration with increasing amount of (0.01 mpk, 0.03 mpk, 0.10 mpk, 0.30 mpk).

In FIG. 5 , the left graph shows lung exposure of the compound of Formula A and the compound of Formula B after administration with increasing dose of the compound of Formula B (0.01 mpk, 0.03 mpk, 0.10 mpk, 0.30 mpk) The graph shows the plasma exposure of the compound of Formula A and the compound of Formula B after administration with increasing dose of the compound of Formula B (0.01 mpk, 0.03 mpk, 0.10 mpk, 0.30 mpk).

List of abbreviations

As used above, and throughout the present invention, unless otherwise stated, the following abbreviations should be understood as:

Boc ₂ O Di-tert-butyl dicarbonate

DIEA N, N -diisopropylethylamine

DMAP 4-dimethylaminopyridine

DMF Dimethylformamide

DMSO Dimethylsulfoxide

ESI-MS electrospray ionization mass spectrometry

FAB-MS fast-atom bombardment mass spectrometry

HATU O- (7-azabenzotriazol-1-yl) -N , N , N ' , N' -tetramethyluroninum hexafluorophosphate

HPLC High Performance Liquid Chromatography

PBS Phosphate Buffered Saline

i.n. Intranasally

PO oral

i.p. Intraperitoneal

i.t. Intratracheally

Liver toxin produced by specific cyanobacteria of the genus Microcystin-LR Anabaena and Oscillatoria

mbr millibar

mpk mg / kg

HEPES 4- (2-hydroxyethyl) -1-piperazineethanesulfonic acid

DTT Dithiothreitol

ATP Adenosine Triphosphate

streptavidin-HRP Streptavidin-Horsead Peroxidase Conjugate

TMB tetramethylbenzidine

pfu flake-forming unit

MDI Instrumented Dose Inhaler

DPI dry powder inhaler

Justice

As used above, it should be understood that, through the description of the present invention, the following terms have the following meanings unless otherwise indicated.

“Compounds of the present invention” and equivalent expressions thereof refer to compounds of formula A described herein, which include pharmaceutically acceptable salts and solvates such as hydrates. Similarly, it is to be understood that even in the case of intermediates, salts and solvates are included if the content permits, whether or not they are claimed. For the sake of clarity, although some specific examples are mentioned in the text, they are intended to be purely illustrative and should not be understood as excluding other examples to the extent permitted by the text.

"Treatment" or "treatment" means disease prevention, partial alleviation or treatment. The compounds and compositions of the present invention are useful for treating diseases characterized by enhanced levels of cytokines and mediators regulated by NF-κB activation and / or NF-κB including but not limited to TNFα and IL-1β. Do. Inhibition or inhibition of NF-κB and NF-κB-regulated genes, such as TNFα, may occur locally in specific tissues of a patient, or may be more strongly occurring throughout patients treated with such a disease. . Inhibition or inhibition of NF-κB and / or NF-κB-regulated genes such as TNFα can occur by inhibiting or inhibiting any stage of the pathway (s) such as one or more mechanisms such as inhibition of IKK. "NF-κB-associated state" refers to a disease characterized by the activation of NF-κB in the cytoplasm (eg, upon phosphorylation of IκB). This "TNFα-associated state" is a state characterized by enhanced levels of TNFα. As used herein, NF-κB-associated symptoms include, but are not limited to, TNFα-associated diseases, because NF-κB is involved in the activity and upregulation of other inflammatory precursor proteins and genes. Because. “Inflammatory or immune disease or disease” includes any of those associated with NF-κB-associated diseases and TNFα-associated diseases such as NF-κB release and / or enhanced levels of TNFα, including the diseases described herein. Disease.

"Patient" includes humans and other mammals.

A "pharmaceutically effective amount" means an amount of a compound, composition, drug or other active ingredient to achieve the desired therapeutic effect.

"Substantially pure" refers to a compound that is substantially free of biological or chemical components, for example separated from biological or chemical compositions where the biological or chemical components are separated together, wherein the analytical purity of the compound is preferably At least 70%. More preferably the analytical purity is at least 90%, more preferably the analytical purity is at least 95%; "Substantially pure" refers to a compound substantially free of prodrugs, such as the compound of Formula B.

The present invention also relates to a process for preparing the compound of formula A as shown in the following scheme.

Starting compounds of a chemical reaction are known or can be prepared directly by methods known in the literature. US application 10 / 642,970 describes the preparation of indole carboxylic acid intermediates (compound 8 ) used in the coupling reaction step (vi). Compounds (Formulas B, C, D) are prepared by the method described in US Patent Application 10 / 642,970, also incorporated by reference.

Peptide chemistry binding methods well known to those skilled in the art for condensing compounds (Houben-Weyl, Methoden der Organischen Chemie [Methods of Organic Chemistry], Volumes 15/1 and 15/2, Georg Thieme Verlag, Stuttgart, 1974). , Attached as a reference). Carbonyldiimidazole, carbodiimide, for example dicyclohexylcarbodiimide or diisopropylcarbodiimide (DIC), O-((cyano (ethoxycarbonyl) -methylene) -amino) -N Compounds such as, N, N ', N'-tetramethyluronium tetrafluoroborate (TOTU) or polyphosphoric acid (PPA) are suitable for use as condensing or coupling reagents.

Condensation may also be carried out under standard conditions. In condensation it is essential to protect non-reactive amino groups with reversible protecting groups. The same applies to carboxyl groups not involved in the reaction, which are preferably present as (C ₁ -C ₆ ) -alkyl esters, benzyl esters or tert-butyl esters during the condensation process. The amino group does not necessarily need to be protected when these amino groups are present in the form of precursors such as nitro groups or cyano groups, and are formed only by hydrogenation after the condensation reaction. After the condensation reaction, the protecting groups present are removed by appropriate means. For example, benzyl groups in NO ₂ groups (guanidino protection in amino acids), benzyloxycarbonyl groups and benzyl esters can be removed by hydrogenation. The protecting group of the tert-butyl type is removed in the acidic state and the 9-fluoromethyloxy-carbonyl radical is removed using a secondary amine.

The present invention relates to a pharmaceutical composition comprising a pharmaceutically effective amount of a compound of formula A and a pharmaceutically acceptable carrier.

Embodiment

Because of the pharmacological properties of the compounds according to the invention, this is a condition that can be alleviated by targeted administration of an inhibitor of IκB kinase to a site where treatment is better performed by localized activity against systemic activity, for example asthma Or all patients with chronic obstructive pulmonary disease (COPD).

Indeed, the compounds of the present invention are orally, inhaled, rectal, nasal, buccal, sublingual, vaginal, colonic, parenteral (including subcutaneous, intramuscular, intravenous, intradermal, intratracheal, epidural), water baths, in humans and other animals. Administration is in pharmaceutically acceptable dosages by local or systemic administration, including intraracisternal and intraperitoneal administration. It will be appreciated that the appropriate route may vary with the subject's condition.

Specific methods of administering the compounds of the present invention include aerosolization as well as intranasal, intratracheal or inhaled administration.

Combination therapy can improve efficacy and reduce the risk of side effects when compared to increasing single agent doses. IKK inhibitors include bronchodilators, including but not limited to short-acting β2-agonists, long-acting β2-agonists such as salmeterol and formoterol; Anticholinergic agents, for example, can be combined with ipratropium bromide and tiotropium bromide. IKK inhibitors can be combined with methylxanthine, such as theophylline.

Inhibitors of IKK2 can also be combined with several anti-inflammatory therapies, including but not limited to immunomodulators, which can act at various stages of the inflammatory cascade to mitigate the inflammatory process. Such therapies include, but are not limited to:

(A) inhibitors of cellular recruitment and toxic inflammatory mediators such as phosphodiesterase-4 inhibitors; p38 mitogen-activated protein kinase inhibitors; Herbal drugs such as anti-tumor necrosis factor-α, anti-interleukin-8, anti-monocyte chemoattractant protein-1; Inhibitors of adsorptive molecules and chemotick factors, molecules that interfere with cell survival and clearance / apoptosis;

(B) inhibitors of proteolytic enzymes such as neutrophil-induced serine proteases such as neutrophil elastase and matrix metalproteinases (MMPs) inhibitors such as MMP-2, MMP-9, MMP- Inhibitors of proteases, including but not limited to 12;

(C) antioxidants Antioxidants, including but not limited to, for example, N-acetylcysteine, inhibitors or scavengers of reactive oxygen species, toxic peptides such as defensins that are a direct cause of cell damage;

(D) mucus gene inhibitors; And mucus production inhibitors including but not limited to expectorants, mucolytic agents, mucodynamic agents, and the like; And

.(E) antibiotic therapies such as ketolides, for example Ketek

.

The drug complex of the present invention may be provided in separate pharmaceutically acceptable formulations which may be administered simultaneously or sequentially to cell (s) or human patients, the formulations comprising one or more therapeutic agents or a single substance and It may be provided by multiple substance formulation classification. However, these drug complexes constitute a pharmaceutically effective amount of components.

The therapeutic regimen / dose schedule may be reasonably modified during the course of treatment to administer each minimum amount of a pharmaceutically effective amount of the compounds used in combination with a satisfactory pharmacological effect. Continue only if necessary to successfully treat the patient.

The pharmaceutical composition according to the invention is preferably administered in unit dosage form, each unit containing the active ingredient of the compound, a particular dosage. Pharmaceutically acceptable salts of compounds of Formula A are within the scope of this invention. "Salt" refers to an acid or base salt formed with an acid and a base. "Salts" also include zwitterions salts (internal salts), for example, compounds of formula A include basic residues such as amines or pyridine or imidazole rings and acidic residues such as carboxylic acids do. Pharmaceutically acceptable (eg non-toxic, physiologically acceptable) salts are preferred, for example, acceptable metal and amine salts in which the cation does not contribute significantly to the toxicity or biological activity of the salt. However, other salts may be useful, for example in separation and purification steps that may be used during manufacture, which is also within the scope of the present invention. Salts of compounds of formula A can be made by reacting an equivalent amount of an acid or base with a compound of formula A in a medium in which salt precipitation occurs or in an aqueous medium and lyophilized.

Acid addition salts may be formed with compounds of the present invention having basic residue (s), such as imino nitrogen, with amino or modo or substituted groups present. Certain acid addition salts are pharmaceutically acceptable acid addition salts, for example, their anions are nontoxic to patients with pharmacological dosages of the salts, and the inherent beneficial effects of the free form of the compound are struck by side effects caused by anions. Do not receive. The salts selected are comparable to conventional pharmacological vehicles and are chosen to be optimally suitable for the mode of administration employed. Acid addition salts of the compounds of the present invention can be prepared by reacting an acid suitable for freeform of a molecule having a basic moiety according to a known method. For example, acid addition salts of the compounds of the present invention dissolve a free-form molecule having a basic moiety in water or a water soluble alcohol solution or other suitable solvent containing an appropriate acid, and vaporize the solution to separate the salt or have a basic moiety. The free form molecule and the acid can be reacted in an organic solvent, where the salt can be separated directly or the solution can be concentrated to obtain a salt. Some suitable acids that may be used to prepare such salts include hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, various organic carboxylic acids and sulfonic acids, such as acetic acid, citric acid, propionic acid, succinic acid, benzoic acid, tartaric acid, fumaric acid, bay Delic acid, ascorbic acid, malic acid, methanesulfonic acid, toluenesulfonic acid, mandelic acid, ascorbic acid, malic acid, fatty acids, adipate alginate, ascorbate, aspartate, benzenesulfonate, benzoate, cyclopentanepro Cypionate, digluconate, dodecyl sulfate, bisulfate, butyrate, lactate, laurate, lauryl sulfate, maleate, hydroiodide, 2-hydroxy-ethanesulfonate, glycerophosphate, Picrate, pivalate, palmoate, pectinate, persulfate, 3-phenylpropionate, thiocyanate, 2-naph Butylene and the like sulfonate, undecanoate, nicotinate, hemisulfate, heptanoate, hexanoate, camphor rate, camphor sulfonate and the like.

Acid addition salts of the compounds of the present invention can be used to regenerate the parent compounds of the present invention from salts by applying or employing known methods. For example, the parent compounds of the present invention can be regenerated from their acid addition salts by treatment with an alkali, for example, a water soluble sodium bicarbonate salt or a water soluble ammonia solution.

Base addition salts are made of compounds of the present invention having a carboxy moiety. Certain base addition salts are pharmaceutically acceptable base addition salts, for example, the cation of the salt is non-toxic to the patient at the pharmacological dose of the salt, and the inherent beneficial effects of the free form of the compound are eliminated by side effects due to anions Do not. The salt selected is comparable to conventional pharmacological vehicles and is chosen to be suitable for the mode of administration employed. The base addition salt of the present invention can be prepared by reacting a base suitable for freeform of a molecule having an acid residue by applying or employing a known method. For example, base addition salts of the compounds of the present invention dissolve free form molecules having acid residues in water or a water soluble alcohol solution or other suitable solvent containing an appropriate base, and evaporate the solution to separate salts or remove acid residues. The eggplant can be reacted with a free form molecule and a base in an organic solvent, where the salt can be separated directly or the solution can be concentrated to obtain a salt. Suitable bases that can be used to prepare such salts can be derived from alkali and alkaline earth metal salts or amines, for example sodium hydride, sodium hydroxide, potassium hydroxide, calcium hydroxide, aluminum hydroxide, lithium hydroxide, magnesium hydroxide, Zinc hydroxide, ammonia, ethylenediamine, N-methyl-glucamine, lysine, arginine, ornithine, choline, N, N'-dibenzylethylenediamine, chloroprocaine, diethanolamine, procaine, N-benzylphen Ethylamine, diethylamine, piperazine, tris (hydroxymethyl) -aminomethane, tetramethylammonium hydroxide and the like.

Base addition salts of the compounds of the invention are used to regenerate the parent compound of the invention from salts by applying or employing known methods. For example, the parent compounds of the present invention can be regenerated from their base addition salts by treatment with an acid such as hydrochloric acid.

Indeed, the compounds of the present invention are administered to patients in the form of suitable preparations, thereby particularly localizing their activity. It will be appreciated that the route of administration may vary depending upon the site of symptoms administered.

Pharmaceutically acceptable dosage forms refer to dosage forms of the compounds of the invention, including, for example, powders, suspensions, sprays, inhalants, tablets, emulsions, and solutions, especially those suitable for inhalation. Techniques and formulations are generally referred to Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, PA, latest publication.

If desired and for more effective distribution, the compounds may be released in a slow release or targeted delivery system such as a biocompatible, biodegradable polymer matrix (e.g., poly (d, l-lactide co-glycolide) ), Liposomes, and microspheres can be microencapsulated or attached and injected into the skin subcutaneously or intramuscularly using a technique called subcutaneous or intramuscular depot to release the compound slowly over a period of 2 weeks or longer.

The compound may be sterilized, for example, by combining the bactericide in the form of a sterile solid composition that can be filtered through a bacterial maintenance filter or dissolved in sterile water, or some other sterile medium can be used just before use.

Nasal or tracheal formulations refer to formulations in a form suitable for administration to a patient nasal or via inhalation. Formulations may include powdered carriers having a particle size in the range of 1 to 500 μm (particle sizes increased by 5 μm within the range of 20 to 500 μm, eg, 30 μm, 35 μm, etc.). Suitable formulations comprising liquid carriers for administration by nasal spray or nasal drop include aqueous or oily solutions of the active ingredient. Formulations suitable for aerosol administration may be prepared according to conventional methods and may be delivered with other therapeutic agents. MDI and DPI are suitable means effective for inhalation therapy by administering a dosage form of a compound of the invention.

The actual dosage level of the active ingredient in the compositions of the present invention can be varied to achieve the amount of active ingredient (s) to be effective in obtaining the desired therapeutic response to the particular composition and method of administration for the patient. Thus, the dosage levels selected for any particular patient may include the desired therapeutic effect, route of administration, duration of treatment, cause and severity of disease, patient's condition, weight, sex, diet, age, type and intensity of each active ingredient. , Rate of absorption, metabolism and / or excretion of other factors.

The total daily dose of a compound of the invention administered in a single or divided dose to a patient is about 1000 mg, more particularly about 50 mg to 300 mg, and more particularly about 10 mg to 100 mg. However, higher or lower dosages may also be appropriate. Daily dosages may be administered in a single unit form only once, in several small unit dosage forms, or by means of multiple doses at divided intervals. The proportion of active ingredient in the composition may vary, but should be comprised of such proportions that an appropriate dosage will be obtained. Obviously, several unit dosage forms can be administered at about the same time. Dosage forms may be administered as often as necessary to obtain the desired therapeutic effect. Some patients may respond to higher or lower doses and a much weaker maintenance dose may be appropriate. In some other patients it may be necessary to maintain long-term treatment at a rate of 1 to 4 doses daily, depending on the physiological requirements of each particular patient. For other patients, one or two daily doses may be prescribed.

The formulations may be in unit dosage form using any method known in the pharmacopoeia. Such methods include the step of bringing into association the active ingredient with the carrier consisting of one or more minor ingredients. In general, formulations are made homogeneously and intimately associated with the active ingredient and the liquid carrier or with the finely divided solid carrier or both, and by molding the product, if necessary.

Experiment

Mass-spectroscopic methods (FABMS, ESIMS) are used for analysis. Temperature is expressed in degrees Celsius, and RT means room temperature (22 ° C. to 26 ° C.). Abbreviations used correspond to those described or common to those skilled in the art.

The invention is illustrated by the following examples.

Manufacture 1

Synthesis of methyl (3- ( N -phenyl- N -2-pyridyl) amino) -2- (di-tert-butoxycarbonylamino) propionate (Scheme 1, compound 3 )

Methyl 2- (di-tert -butoxycarbonylamino ) acrylate (Scheme 1, compound 2 )

50 g (0.228 mol) of (tert-butoxycarbonyl) serine ( 1 ) were dissolved in 300 mL of acetonitrile. 107 g (0.493 mol) of di-tert-butyl bicarbonate and 2.64 g (22 mmol) of 4- (dimethylamino) pyridine (DMAP) were added. The mixture was stirred overnight at room temperature, then the solvent was removed under reduced pressure and the residue was taken up in 500 mL of ethyl acetate. The organic phase was washed with 500 mL of 1 N HCl, dried over magnesium sulfate and the organic solvent was removed under reduced pressure. 23 g of acrylate 2 was obtained by crystallizing the residue from 200 mL of heptane at -30 ° C and suction filtration. The mother liquor was concentrated and the residue was dissolved in 140 mL of acetonitrile. 31 g (0.142 mol) of di-tert-butylbicarbonate and 1.26 g (10 mmol) of DMAP were added. After heating the mixture at 50 ° C. for 8 hours, the solvent was removed in vacuo and the residue was taken up in 500 mL of ethyl acetate. The organic phase was washed with 400 mL of 1 N HCl and dried over magnesium sulfate. The solvent was removed in vacuo and crystallized from heptane to afford an additional 31.5 g of acrylate 2 . Yield: 54.5 g (0.181 mol) 79%. Experimental Formula C ₁₄ H ₂₃ NO ₆ ; MW = 301.34; MS ((2M *) + Na < + >) 625.7. _{^{1 H NMR (DMSO- d 6)}} 1.40 (s, 18 H), 3.74 (s, 3 H), 5.85 (s, 1 H), 6.28 (s, 1 H).

Methyl (3- ( N - phenyl - N -2 -pyridyl ) amino) -2- (di-tert -butoxycarbonylamino ) -propionate (Scheme 1, compound 3 )

4.96 g (16.5 mmol) of acrylate 2 was mixed with 5.6 g (33 mmol) of 2-anilinopyridine and 32.16 g (98.7 mmol) of cesium carbonate salt. 50 mL of acetonitrile were added and the mixture was stirred at 45 ° C. for 2 days. The solid was suction filtered through diatomaceous earth and washed three times with 100 mL portions of acetonitrile. The combined organic phases are evaporated and the residue is chromatographed on silica gel using 1: 1 heptane / diethyl ether. 5.66 g (73%) of ester 3 are obtained. Experimental Formula C ₂₅ H ₃₃ N ₃ O ₆ ; MW = 471.56; MS (M + H) 472.2.

Enantiomer separation ( Scheme 1, Compound 3 (S) and Compound 3 (R) )

Racemic amino ester 3 was prepared from the corresponding acrylic acid ester 2 and enantiomer 3 (S) according to preparative HPLC using a chiral stationary phase such as Chiralpak AD (Daicel) 100 × 380, RT, flow rate 300 mL / min . And 3 (R) . The purity of the enantiomer was determined using analytical HPLC such as Chiralpak-ADH (Daicel) 4.6 × 250, 30 ° C., flow rate 1 ml / min, room temperature.

Manufacture 2

Synthesis of 2- (2-methylaminopyrimidin-4-yl) -1 H -indole-5-carboxylic acid ( 8 ) (Scheme 2, compound 8 )

1-dimethylamino-4,4 -dimethoxypent- 1-en-3-one (Scheme 2, compound 6 )

Stir 100 g (0.76 mol) of 3,3-dimethoxy-2-butanone ( 4 ) with 90.2 g (0.76 mol) of N , N -dimethylformamide dimethylacetal ( 5 ) at 120 ° C. for 48 hours. I was. Methanol formed in the reaction was continuously removed from the reaction solution through distillation. Crystals form spontaneously when cooling the solution and a small amount of heptane is added to complete the crystallization. This resulted in 128.24 g of crude 6 (yield 90%), which was reacted without further purification. Experimental Formula C ₉ H ₁₇ NO ₃ ; MW = 187.24; MS (M + H) 188.2. ¹ H NMR (DMSO d ₆ ) 1.22 (s, 3 H), 2.80 (s, 3 H), 3.10 (s, 9 H), 5.39 (d, J = 15 Hz, 1 H), 7.59 (d, J = 15 Hz, 1 H).

[4- (1,1 -dimethoxyethyl ) pyrimidin-2-yl] methylamine (Scheme 2, compound 7 )

1.22 g (53 mmol) of sodium were dissolved in 100 mL of absolute ethanol. While stirring, 5.8 g (53 mmol) of methylguanidine hydrochloride and 10 g (53 mmol) of 1-dimethylamino-4,4-dimethoxypent-1-en-3-one ( 6 ) were added to this solution. The whole was heated under reflux for 4 hours. To terminate the reaction, ethanol was evaporated. Product 7 was used in a continuous reaction without further purification. Yield: 11.5 g (58 mmol, quantitative); Experimental Formula C ₉ H ₁₅ N ₃ O ₂ ; MW = 197.24; MS (M + H) 198.2. ¹ H NMR (DMSO d ₆ ) 1.45 (s, 3 H), 2.78 (s, 3 H), 3.10 (s, 6 H), 6.75 (d, J = 3 Hz, 1 H), 7.0-7.1 (s (b), 1 H), 8.30 (d, J = 3 Hz, 1 H).

2- (2- Methylaminopyrimidin-4- yl) -1 H -indole-5 -carboxylic acid (Scheme 2, compound 8 )

150 mL of 50% sulfuric acid at room temperature with stirring 5 g (25 mmol) of [4- (1,1-dimethoxyethyl) pyrimidin-2-yl] methylamine ( 7 ) and 3.85 g of 4-hydrazinebenzoic acid Was added and the mixture was heated at 130 ° C. for 4 hours. Methanol formed in the reaction was continuously removed from the reaction solution using distillation. After cooling to 10 ° C., the reaction mixture was poured into 200 mL of ice and the pH was adjusted to about 5.5 with concentrated sodium hydroxide solution. The sodium sulfate precipitate and the formed product mixture were filtered off and the filtered residue was extracted several times with methanol. The combined methanol extracts were concentrated and the product 8 was purified using flash chromatography (CH ₂ Cl ₂ / methanol 9: 1). Yield: 0.76 g (11%); Experimental Formula C ₁₄ H ₁₂ N ₄ O ₂ ; MW = 268.28; MS (M + H) 269.1. ¹ H NMR (DMSO d ₆ ) 2.95 (s, 3 H), 6.90-7.10 (s (b), 1 H), 7.18 (d, J = 3 Hz, 1 H), 7.4 (s, 1 H), 7.58 (d, J = 4.5 Hz, 1 H), 7.80 (d, J = 4.5 Hz, 1 H), 8.30 (s, 1 H), 8.38 (d, J = 3 Hz, 1 H), 11.85 (s , 1 H), 12.40-12.60 (s (b), 1H).

Example  One

2-{[2- (2-Methylamino-pyrimidin-4-yl) -1 H -indole-5-carbonyl] -amino} -3- (phenyl-pyridin-2-yl-amino) -propionic acid ( Synthesis of A )

2-{[2- (2- Methylamino- pyrimidin-4-yl) -1 H -indole-5-carbonyl] -amino} -3- ( phenyl -pyridin-2-yl-amino) -propionic acid, Methyl ester (Scheme 3, compound 10 )

2.9 g of S 3 enantiomer ( 3 (S) ) of compound 3 was dissolved in 30 mL of dioxane and the solution was cooled to 0 ° C. 30 mL of 4N HCl dioxane was added and the mixture was allowed to come to room temperature and stirred for 12 hours. The solvent was removed in vacuo. The residue was taken up in 30 mL of DMF (solution A). 2.47 g (9.2 mmol) of acid 8 were dissolved in 50 mL of DMF and cooled to 0 ° C. 4.21 g HATU and 6.4 mL DIEA were added. The mixture was stirred at 0 ° C. for 45 min, then cooled to rt and solution A was added. The mixture was stirred at RT for 12 h. The solvent was removed in vacuo and the residue was partitioned between 300 mL of saturated NaHCO ₃ solution and 300 mL of ethyl acetate. The aqueous phase was washed three times with 100 mL portions of ethyl acetate and the combined organic phases were washed with 400 mL of saturated NaCl solution. The organic phase was dried over magnesium sulfate. The solvent was removed under reduced pressure and the residue was chromatographed on silica gel with 1: 3 heptane / ethyl acetate. 1.78 g (55%) of ester compound 10 were obtained. Experimental Formula C ₂₉ H ₂₇ N ₇ O ₃ ; MW = 521.58; MS (M + H) 522.2.

2-{[2- (2- Methylamino- pyrimidin-4-yl) -1 H -indole-5-carbonyl] -amino} -3- ( phenyl -pyridin-2-yl-amino) -propionic acid ( Scheme 3, compound A )

2.0 g (3.8 mmol) of methyl ester 10 were dissolved in 200 mL of methanol. To 1 mL of 2 N water soluble NaOH, the mixture was stirred for 12 hours at room temperature. The solvent was evaporated and the residue dissolved in water and the pH was NaH ₂ PO ₄ It was adjusted to ˜5 using a saturated solution. The resulting precipitate was filtered off and washed with water. After drying under reduced pressure of 1 mbar at 40 ° C., 1.95 g (quantitative yield) of acid compound A was separated. Experimental Formula C ₂₈ H ₂₅ N ₇ O ₃ ; MW = 507.56; MS (M + H) 508.3. _{^{1 H NMR (DMSO- d 6)}} 2.95 (s, 3 H), 4.22-4.50 (m, 2 H), 4.65-4.72 (m, 1 H), 6.29 6.36 (d, 1 H), 6.70 6.79 (m , 1 H), 6.90 7.10 (sb, 1 H), 7.13-7.19 (m, 1 H), 7.22-7.38 (m, 5 H), 7.40-7.48 (m, 3 H), 7.50-7.55 (m, 1 H), 7.57-7.60 (m, 1 H), 7.96 (bs, 1 H), 8.34 8.40 (m, 2 H), 8.80 8.90 (d, 1 H), 11.80 (s, 1 H).

In vitro ( IN VITRO A) test process

IKK -enzyme ELISA

The test buffer has the following composition (50 mM HEPES, 10 mM MgCl 2, 10 mM β-glycerophosphate, 2 M Microcystin-LR, 0,01% NP-40, 5 mM DTT).

The IKK enzyme preparation was diluted to 1:50 (hand made formulation) + test compound / DMSO (2% final concentration in wells).

The inspection process is as follows:

Incubate the enzyme and compound for 30 minutes;

Addition of 1 mM ATP or 50 μM ATP;

pSer36-IkB peptide (substrate): 40 μΜ;

Incubate for 45 minutes, add anti-pSer32-pSer36-IkB peptide-antibody;

Incubate for 45 minutes, transfer to protein-G-coated plates;

3x wash after incubation for 90 minutes;

Add streptavidin-HRP, 45 min incubation, 6 × wash;

TMB addition and incubation for 15 minutes; And

Reading with termination solution and photometer.

In vitro profile results are shown in Table 1 below.

At 1 mM ATP in the IKK-enzyme ELISA described above, the compound of Formula A has κB kinase IC ₅₀ activity 705 times, 4,725 times, 47.5 times greater than the compounds of Formulas B, C, and D, respectively. This data demonstrates that the activity of the compound of Formula A is better than expected for the compounds of Formulas B, C, and D.

Between the compound of formula A and the compound of formula B In vivo  Comparison test

NF-κB-induced gene expression contributes significantly to the development of inflammatory diseases such as asthma and arthritis. IκB Kinases (IKKs) are a converging point for the activation of NF-κB by a broad spectrum of inflammatory agonists.

IKK is a multi-subunit complex comprising two catalytic subunits IKK-1 (also known as IKK-α) and IKK-2 (also known as IKK-β) and a regulatory subunit IKK-γ. In gene knock out studies, IKK-2 or IKK-β subunits of the IKK complex are required to activate NF-κB by all known proinflammatory inflammation stimuli including lipopolysaccharide (LPS) and IL-1β. It is clearly explained. Thus, IKK-β-deficient cells are insufficient to activate IKK and NF-κB in response to tumor necrosis factor α (TNFα) or interleukin-1β (IL-1β).

In house bio-imaging data confirmed that IL-1β-induced NF-κB activity in the lungs was inhibited by administration of the predominant negative form of IKKβ (Adv-IKK-2 DN). Thus, selective IKK-β inhibitors are of considerable interest as potential anti-inflammatory agents and can be a valuable means of understanding the mechanism by which these inflammatory agonists modulate NF-κB activation.

A. NF -κB- Luciferase  Reporter Mouse Model

Of formula A Compound and  Mice Using Compounds of Formula B Imaging  How to use research.

summary

Compounds were administered via the intranasal route using nanomilled suspension in 0.2% Tween-80 in PBS.

Intranasal Drugs and Inflammatory Stimulation Administration: Mice were anesthetized with oxygen containing 4% isoflurane gas. 25 μl of each was added to the nostrils of the mice, allowing the mice to inhale the suspension.

Balb / c female mice (6-8 weeks old) were used in the study, where AdV-NFκB-luciferase reporter was injected into the lungs. To image the mice, mice were anesthetized with oxygen containing 4% isoflurane. Luciferin was administered at a dose of 150 mg / kg ip. Ten minutes after luciferin injection, mice were photographed with the IVIS200 system (Xenogen), which was exposed to 1 minute bioluminescence. Alternatively 10-15 minutes after luciferin injection, mice are rapidly euthanized and internal tissues are excised and taken ex vivo .

B. Dose Response Measurement

1-2 × 10 ⁸ pfu adenovirus-NFκB-luciferase was administered intranasally after 3-5 days and then stimulated in with an inflammatory stimulant (IL-1β or LPS). Animals were stimulated with 0.5 μg LPS or 50 ng IL-1β after receiving 30 min-1 h of compound of 0.3-10 mg / kg in. Animals were taken once or several times for 1 to 24 hours after inflammatory irritant administration.

The results of the in vivo process are as follows.

The effects of compounds of Formula A and compounds of Formula B on IL-1β-induced NF-κB activation are shown in FIGS. 1 and 2. Both compounds inhibited NF-κB activity in a dose dependent manner, with the estimated ED ₅₀ of the compound of Formula A showing a better effect at about 1 mg / kg.

C. Pharmacokinetic Process

Male Hartley guinea pigs (450-550 g) were preferentially sensitized with ovalbumin and used to determine compound levels in lung and plasma. Nanomill suspensions of compounds of Formula A and compounds of Formula B were injected intratracheally in amounts of 0.01, 0.03, 0.1, 0.3 mg / kg. One hour after dosing, animals were euthanized (Euthasol), and a 1 mL blood sample was taken out of the heart and collected with a heparin-coated syringe. Plasma was separated from the cellular components of blood by centrifugation and stored at -80 ° C until used for testing. Lungs were incised, blotted dry, weighed and individually stored in 20-25 mL glass vials at −80 ° C. until compound levels were examined.

The main differences in the pharmacokinetic profiles of the compounds of Formula A and the compounds of Formula B are illustrated in FIG. 3 using those found from the maximum dose group (0.3 mg / kg).

3 shows i.t. Lung exposure to the compound of Formula A or the compound of Formula B after infusion was determined in i.t. It is relatively lower than the compound of formula A after administration, indicating that the compound of formula B is rapidly absorbed from the lungs.

3 to 5 show that the compound of formula B may be a candidate group vulnerable to inhalation, for 1) when administered intratracheally it is significantly distributed systemically upon exposure; 2) The compound of formula B is a prodrug of a compound of formula A having a different exposure profile; 3) The compound of formula B has a reduced exposure to the compound of formula A in the lung than is obtained by direct administration of the compound of formula A.

Compounds of formula (A) become stronger candidates for inhalation than compounds of formula (B), because: 1) Compounds of formula (A) are i.t. And systemic exposure was low after PO administration; 2) The compound of formula A must have a longer lung residence time.

It is also found that the compounds of formula B are very well used systemically upon oral administration.

For compounds of formula A, the ratio of lung to plasma is 143 to 284 (depending on dosage), and for compounds of formula B, the ratio of lung to plasma is 13 to 44 (depending on dosage) to be. These ratios were obtained by dividing the corresponding lung compound levels by plasma levels at the same dose level.

Claims (14)

Substantially pure compound of formula A or a pharmaceutically acceptable salt or solvate thereof. Formula A

A pharmaceutical composition comprising a pharmaceutically effective amount of a compound of formula A and a pharmaceutically acceptable carrier. A method of treating a patient in a morbid condition that can be alleviated by inhibiting IKK-2, comprising administering to the patient a pharmaceutically effective amount of the compound of claim 1. The method of claim 3, wherein administration is performed to localize activity. The method of claim 3, wherein the pathological condition is asthma, rhinitis, chronic obstructive pulmonary disease or exacerbation of chronic obstructive pulmonary disease. The method of claim 3, wherein the administration is intratracheal, intranasal, inhalation or aerosol administration. A method of treating an asthma patient comprising administering to the patient a pharmaceutically effective amount of the compound of claim 1. A method of treating a patient with rhinitis comprising administering to a patient a pharmaceutically effective amount of the compound of claim 1. A method of treating a patient with chronic obstructive pulmonary disease comprising administering to a patient a pharmaceutically effective amount of the compound of claim 1. A method of treating a patient with chronic obstructive pulmonary disease exacerbation, comprising administering to the patient a pharmaceutically effective amount of the compound of claim 1. The compound of claim 2 further comprising a pharmaceutically effective amount of a compound selected from the group consisting of bronchodilators, long-acting β-2 agonists, anticholinergics, methylxanthines and anti-inflammatory therapies, mixed with pharmaceutically acceptable carriers. Pharmaceutical compositions comprising. 12. The bronchodilator of claim 11, wherein the bronchodilator is a short acting β2-agonist; Long-acting β 2-agonists are selected from salmeterol and formoterol; Anticholinergic agents are selected from ipratropium bromide and tiotropium bromide; Methylxanthine is theophylline; The anti-inflammatory therapy is a pharmaceutical composition selected from cellular recruitment and toxic inflammatory mediator inhibitors, protease inhibitors, antioxidants, mucus production inhibitors and antibiotic therapy. The pharmaceutically effective amount of the compound according to claim 3, which is mixed with a pharmaceutically acceptable carrier, is selected from the group consisting of bronchodilators, long-acting β-2 agonists, anticholiners, methylxanthines and anti-inflammatory therapies. How to further include. The method of claim 13, wherein the bronchodilator is a short acting β2-agonist and the long-acting β2-agonist is selected from salmeterol and formoterol; Anticholinergic agents are selected from ipratropium bromide and tiotropium bromide; Methylxanthine is theophylline; Anti-inflammatory therapy is selected from cell recruitment and toxic inflammatory mediator inhibitors, protease inhibitors, antioxidants, mucus production inhibitors and antibiotic therapy.

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