CN1903835A - Compound, composition and method for preventing neurodegeneration in acute and chronic injury of central nervous system - Google Patents

Abstract

The present invention provides a compound capable of preventing and curing nervous system diseases due to neuronal death and its method. Said invention includes synthesis of 5-benzyl aminosalicylic acid (BAS) and its derivative. The BAS and its derivative can protect cortical neuron from toxic injury of N-methyl-D-aspartate, zinc ion and active oxygen cluster, so that said invention provides compound and method for curing the diseases of apoplexy, cerebral trauma, epilepsy, spinal cord injury and neurodegenerative disease, etc.

Description

Neurodegenerative compound, composition and method in the prevention central nervous system acute and chronic injury

Technical field

The present invention relates to new salicylic acid compound, composition, and method, described compound, composition and method are in order to prevent and prevent to be accompanied by the nervosa systemic disease of neuronal death.

Background technology

Exitotoxicity and disease of brain

The ionization glutamate receptor that overactivity is responsive to N-methyl-D-aspartate salt (nmda receptor) can cause neuronal death, and known its mediates multiple nervous system disorders [Choi, Neuron 1:623-634 (1988)].Glutaminate is an excitatory neurotransmitter, it accumulates in the brain in a large number, be subject to the hypoxemia ischemic injuries, hypoxemia ischemic injuries activating ion glutamate receptor, make it permeable to calcium ion and sodium ion, cause neuronal death [Choi and Rothman, Annu Rev Neurosci:13:171-182 (1990)] subsequently.The nmda receptor antagonist has significantly reduced the hypoglycemia that brain injury causes, hypoxemia, hypoxemia ischemic [Simon, Swan, Griffiths and Meldrum.Science 226:850-852 (1984); Park, Nehls, Graham, Teasdale, and McCulloch, Ann Neurol 24:543-551 (1988).; Wieloch, Science230:681-683 (1985); Kass, Chambers, and Cottrell, Exp.Neurol.103:116-122 (1989); Weiss, Goldberg, and Choi, Brain Res.380:186-190 (1986)].Therefore, the nmda receptor antagonist has the brain of making and avoids hypoglycemia, hypoxemia, the treatment potential of hypoxemia ischemic injuries.

As if exitotoxicity have effect to the neuronal degeneration behind the traumatic brain injury (TBI).Quinolinic acid is a kind of endogenous agonist of nmda receptor, its level raise in TBI patient's body 5 times of-50 times of [E.H.Sinz, P.M.Kochanek, M.P.Heyes, S.R.Wisniewski, M.J.Bell, R.S.Clark, S.T.DeKosky, A.R.Blight, and D.W.Marion.].Quinolinic acid raises in cerebrospinal fluid, and relevant with mortality ratio behind the people TBI.[.J.Cereb.Blood Flow Metab.18：610-615，(1998)]。In the cerebral trauma animal model, the level of glutaminate and aspartate significantly raise [Faden, Demediuk, Panter, and Vink, Science 244:798-800 (1989)].After the collision wound, in the spinal cord of rat, also observe glutaminate and discharge [Demediuk, Daly and Faden.J Neurochem J.Neurochem.52:1529-1536 (1989)].The nmda receptor antagonist reduces neuronal death [Faden, Lemke, the Simon, and Noble.J.Neurotrauma.5:33-45 (1988) after cerebral trauma and the Spinal injury; Okiyama, Smith, White, Richter and McIntosh.J Neurotrauma.14:211-222 (1997)].

Glutaminate epileptic seizures induce and propagate in play an important role [Dingledine, McBain, and McNamara, Trends.Pharmacol.Sci.11:334-338 (1990); Holmes.Cleve.Clin.J.Med.62:240-247 (1995)].The nmda receptor antagonist is used as spasmolytic medicine and antiepileptic drug [Anderson, Swartzwelder, and Wilson, JNeurophysiol:57:1-21 (1987) in various epilepsy models; Wong, Coulter, Choi, and Prince.Neurosci.Lett.85:261-266 (1988); McNamara, Russell, Rigsbee, and Bonhaus, Neuropharmacology 27:563-568 (1988)].

Amyotrophic lateral sclerosis (ALS) is accompanied by upper motor neuron and all sex change of lower motor neuron, it is characterized by neuropathic muscular atrophy, weak and spontaneous contraction.Although the pathogeny of ALS is not also resolved, exitotoxicity is considered to participate in the ALS process.Especially, ALS patient shows extracellular glutaminate level increase and glutaminate transportation defective.Pathological change [Rothstein.Clin.Neurosci.3:348-359 (1995) by excitotoxin administration simulation ALS patient spinal cord; Ikononiidou, Qin, Labruyere and Olney J.Neuropathol.Exp.Neurol.55:211-224 (1996)].

As if the nmda receptor antagonist be applicable to treatment Parkinson's disease (PD).Several NMD acceptor antagonist protection dopaminergic neurons are avoided neurotoxin MPTP (1-methyl 4-propyl group-1,2,3,6-tetrahydropyrimidine) effect [Lange, Loschmann, Sofic, Burg, Horowski, Kalveram, Wachtel and Riederer.Naunyn Schmiedebergs Arch.Pharmacol.348:586-592 (1993); Brouillet and Beal.Neuroreport.4:387-390 (1993)].The nmda receptor antagonist also improves the dyskinesia of levodopa inductive, therefore can improve therapeutic action [Papa and Chase, the Ann.Neurol.39:574-578 (1996) of levodopa; Marin, Papa, Engber, Bonastre, Tolosa, and Chase, Brain Res.736:202-205 (1996)].Two kinds of nmda receptor antagonists, memantine and Dextromethorphane Hbr are proved useful [Verhagen, Del Dotto, Natte, van den Munckhof, and Chase, Neurology51:203-206 (1998) in the treatment to PD patient; Merello, Nouzeilles, Cammarota, and Leiguarda.Clin.Neuropharmacol.22:273-276 (1999)].

Huntington (HD) is a kind of progressive neurodegenerative disease, mainly attacks small-sized and the neutral relay cell, but does not injure the NADPH-Lipoyl dehydrogenase neurone that contains tethelin inhibitor and neuropeptide in striatum.Striatum injection or the striatum cultivated are with after NMDA contacts in quinolinic acid, these pathological characters of HD in the striatum tissue, have been observed, possibility [the Koh that this neurotoxicity that has improved the nmda receptor mediation works to neurone selectivity death among the HD, Peters, and Choi, Science 234:73-76 (1986); .Beal, Kowall, Ellison, Mazurek, Swartz, and Martin, Nature 321:168-171 (1986); Beal, Ferrante, Swartz, and KowallhJ.Neurosci.11:1649-1659 (1991)].

Free radical and disease of brain

Hypoxemia ischemic or cerebral trauma and Spinal injury cause that free radical produces [Hall and Braughler, Free Radic.Biol.Med.6:303-313 (1989) in sex change brain zone; Anderson and Hall, Ann.Emerg.Med.22:987-992 (1993); Siefjo and Siesjo, Eur.J.Anaesthesiol.13:247-268 (1996); Love, Brain Pathol 9:119-131 (1999)].Antioxidant or the operation of removing free radical alleviate brain injury [Faden, the Pharmacol.Toxicol.78:12-17 (1996) that hypoxemia ischemic and wound cause; Zeidman, Ling, Ducker, and Ellenbogen, J.Spinal.Disord.9:367-380 (1996); Chan, Stroke 27:1124-1129 (1996); Hall, Neurosurg.Clin.N.Am.8:195-206 (1997)].Evidence support widely: be subjected to the brain zone of sex change in neurodegenerative disease, can produce free radical, this may be because the copper in ALS/zinc superoxide dismutase origination point sudden change, gsh reduces and the iron increase in PD, and iron accumulates and mitochondria dysfunction [Rosen in HD in AD, Siddique, Patterson, Figlewicz, Sapp, Hentati, Donaldson, Goto, O ' Regan and Deng.Nature 362:59-62 (1993); Jenner and Olanow, Neurology 47:S161-S170 (1996); Smith, Harris, Sayre, and Perry, Proc.Natl.Acad.Sci.U.S.A.94:9866-9868 (1997); Browne, Ferrante, and Beal, Brain Pathol.9:147-163 (1999)].Therefore, antioxidant is the neuroprotective of these neurodegenerative diseases.[Jenner, Pathol-Biol. (Paris.) 44:57-64 (1996); Beal, Ann.Neurol.38:357-366 (1995); Prasad, Cole, and Kumar.JAm.Coll.Nutr.18:413-423 (1999); Eisen and Weber, Drugs Aging 14:173-196 (1999); Grundman, Am.J.Clin.Nutr.71:630S.-636S (2000)].

Zinc and disease of brain

Zine ion is mediating observed neurodegenerative process in epileptic seizures, ischemic, wound and alzheimer's disease (AD).Kainate is a kind of excitotoxin of inducing epileptic seizures, the central administration of kainate, zine ion is displaced in the post-synapse sex change neurone in several forebrains zone [Frederickson, Hennandez and McGinty.Brain Res.480:317-321 (1989)].With calcium-EDTA blocking-up zine ion displacement, can alleviate neuron loss [Koh, Suh, Gwag, He, Hsu and Choi, Science 272:1013-1016 (1996) that preceding cerebral ischemia of transience or traumatic brain injury cause; Suh, Chen, Motamedi, Bell, Listiak, Pons, Danscher, and Frederickson, Brain Res.852:268-273 (2000)].In the extracellular of AD patch and sex change neurone, observed zine ion, its may work to the neuronal degeneration among the AD [Bush, Pettingell, Multhaup, Paradis, Vonsattel, Gusella, Beyreuther, Masters, and Tanzi, Science 265:1464-1467 (1994); Suh, Jensen, Jensen, Silva, Kesslak, Danscher and Frederickson.Brain Res.852:274-278852 (2000)].

Summary of the invention

The invention provides with following chemical formula (I) 5-phenmethyl aminosallcylic acid (BAS) and derivative thereof expression, new:

Wherein,

X is CO, SO ₂Or (CH ₂) n (wherein n is the integer of 1-5, comprises 1 and 5);

R ₁Be hydrogen, alkyl or alkyloyl;

R ₂It is hydrogen or alkyl;

R ₃Be hydrogen or a kind of ethanoyl; With

R4 is a phenyl, and this phenyl is not substituted or is replaced by following one or more groups: nitro, halogen, alkylhalide group, the alkoxyl group of C1-C5; Or a kind of its pharmacy acceptable salt.

The present invention also provides a kind of method; this method makes neurone avoid acute and chronic injury to central nervous system (CNS); method comprises that patient or the Mammals to being subjected to this CNS injury gives neuroprotective compounds an amount of on the therapeutics, and this compound is represented with chemical formula (I).

The present invention also provides a kind of composition, and this composition makes axoneuron avoid acute and chronic injury to central nervous system, and composition comprises neuroprotective compounds an amount of on the therapeutics, and this compound is with chemical formula (I) expression.

The present invention also provides a kind of method to treat and prevent nervous system disorders, described nervous system disorders and NMDA neurotoxicity, zine ion neurotoxicity or oxidative stress are associated, this method comprises the compound that the patient who suffers from these diseases and Mammals is given effective dose on the therapeutics, and this compound is with chemical formula (I) expression.

The purposes of the compound that the present invention also provides chemical formula (I) in medicine is made, described medicine are used for protecting relay cell to avoid damage to the acute and chronic of central nervous system (CNS).

From describe in detail, those of ordinary skill in the art can understand above-mentioned and further feature of the present invention.

The accompanying drawing summary

Fig. 1. will be separately or the mouse cortex cell culture (DIV 12-14) that contains 10-300 micromole 5-aminosalicylic acid (AS) contact 10 minutes (1a) with 300 micromole NMDA, continue to contact (1b) with 50 micromole's ferrous ions, or contact 30 minutes (1c) with 300 micromole's zine ions.After 24 hours, be discharged into the LDH horizontal analysis neuronal death of substratum in bathing by measurement, mean value ± SEM (culture hole n=9-12 of every kind of situation (la), n=4-8 (lb) or n=4 (lc)), after vacation in 24 hours is washed (=0) and is continued to contact 500 micromole NMDA (=100), with average LDH outflow value calibration. ^*ANOVA and Student-Neuman-Keuls test are used in expression, and there is significant difference p＜0.05 o'clock with relevant contrast.

Fig. 2. will be separately or the mouse cortex cell culture (DIV 12-14) that contains the 5-phenmethyl aminosallcylic acid (BAS) of dose indicating contact 10 minutes (2a) with 300 micromole NMDA, continue to contact with the BSO (2c) of 50 micromole's ferrous ions (2b) or 10 mmoles, or contact 30 minutes (2d) with 300 micromole's zine ions.After 24 hours, be discharged into LDH horizontal analysis neuronal death in the substratum, mean value ± SEM (the culture hole n=7-8 of every kind of situation (2a), n=3-6 (2b), n=4 (2c) or n=4 (2d)) by measurement. ^*ANOVA and Student-Neuman-Keuls test are used in expression, and there is significant difference p＜0.05 o'clock with relevant contrast.

Fig. 3. will be separately or the mouse cortex cell culture (DIV 12-14) that contains 5-(the 4-oil of mirbane methyl) aminosallcylic acid (NBAS) of dose indicating contact 10 minutes (3a) with 300 micromole NMDA, continue to contact (3b) with 50 micromole's ferrous ions, or contact 30 minutes (3c) with 300 micromole's zine ions.After 24 hours, be discharged into LDH horizontal analysis neuronal death in the substratum, mean value ± SEM (the culture hole n=7-8 of every kind of situation (3a), n=3-6 (3b) or n=4 (3c)) by measurement. ^*ANOVA and Student-Neuman-Keuls test are used in expression, and there is significant difference p＜0.05 o'clock with relevant contrast.

Fig. 4. will be separately or the mouse cortex cell culture (DIV 12-14) that contains 5-(4-chlorophenylmethyl) aminosallcylic acid (CBAS) of dose indicating contact 10 minutes (4a) with 300 micromole NMDA, continue to contact (4b) with 50 micromole's ferrous ions, or contact 30 minutes (4c) with 300 micromole's zine ions.After 24 hours, be discharged into LDH horizontal analysis neuronal death in the substratum, mean value ± SEM (the culture hole n=3-4 of every kind of situation (4a), n=3-12 (4b) or n=4 (4c)) by measurement. ^*ANOVA and Student-Neuman-Keuls test are used in expression, and there is significant difference p＜0.05 o'clock with relevant contrast.

Fig. 5. will be separately or the mouse cortex cell culture (DIV 12-14) that contains 5-(the 4-phenylfluoroform methyl) aminosallcylic acid (TBAS) of dose indicating contact 10 minutes (5a) with 300 micromole NMDA, continue to contact (5b) with 50 micromole's ferrous ions, or contact 30 minutes (5c) with 300 micromole's zine ions.After 24 hours, be discharged into LDH horizontal analysis neuronal death in the substratum, mean value ± SEM (the culture hole n=3-4 of every kind of situation (5a), n=3-11 (5b) or n=4 (5c)) by measurement. ^*ANOVA and Student-Neuman-Keuls test are used in expression, and there is significant difference p＜0.05 o'clock with relevant contrast.

Fig. 6. will be separately or the mouse cortex cell culture (DIV 12-14) that contains 5-(the 4-fluorobenzene methyl) aminosallcylic acid (FBAS) of dose indicating contact 10 minutes (6a) with 300 micromole NMDA, continue to contact (6b) with 50 micromole's ferrous ions.After 24 hours, be discharged into LDH horizontal analysis neuronal death in the substratum, mean value ± SEM (culture hole n=7-8 of every kind of situation (6a) or n=4-8 (6b)) by measurement. ^*ANOVA and Student-Neuman-Keuls test are used in expression, and there is significant difference p＜0.05 o'clock with relevant contrast.

Fig. 7. will be separately or the mouse cortex cell culture (DIV 12-14) that contains 5-(4-methoxybenzyl) aminosallcylic acid (MBAS) of dose indicating contact 10 minutes (7a) with 300 micromole NMDA, continue to contact (7b) with 50 micromole's ferrous ions.After 24 hours, be discharged into LDH horizontal analysis neuronal death in the substratum, mean value ± SEM (culture hole n=7-8 of every kind of situation (7a) or n=4-8 (7b)) by measurement. ^*ANOVA and Student-Neuman-Keuls test are used in expression, and there is significant difference p＜0.05 o'clock with relevant contrast.

Fig. 8. will be separately or the mouse cortex cell culture (DIV 12-14) that contains 5-(pentafluoro-benzyl) aminosallcylic acid (PBAS) of dose indicating contact 10 minutes (8a) with 300 micromole NMDA, continue to contact with the BSO (8c) of 50 micromole's ferrous ions (8b) or 10 mmoles, or contact 30 minutes (8d) with 300 micromole's zine ions.After 24 hours, be discharged into LDH horizontal analysis neuronal death in the substratum, mean value ± SEM (the culture hole n=11-16 of every kind of situation (8a), n=3-6 (8b), n=4-11 (8c) or n=12 (8d)) by measurement. ^*ANOVA and Student-Neuman-Keuls test are used in expression, and there is significant difference p＜0.05 o'clock with relevant contrast.

Fig. 9. will be separately or the mouse cortex cell culture (DIV 12-14) that contains ethyl-5-(4-oil of mirbane methyl) amino-2-hydroxybenzoic acid ethyl ester (NAHE) of dose indicating contact 10 minutes (9a) with 300 micromole NMDA, continue to contact (9b) with 50 micromole's ferrous ions.After 24 hours, be discharged into LDH horizontal analysis neuronal death in the substratum, mean value ± SEM (culture hole n=10-12 of every kind of situation (9a) or n=3-4 (9b)) by measurement. ^*ANOVA and Student-Neuman-Keuls test are used in expression, and there is significant difference p＜0.05 o'clock with relevant contrast.

Figure 10. will be separately or the mouse cortex cell culture (DIV 12-14) that contains 5-(4-oil of mirbane the methyl)-N-acetamido-2 hydroxybenzoic acid ethyl ester (NNAHE) of dose indicating contact 10 minutes (10a) with 300 micromole NMDA, continue to contact (10b) with 50 micromole's ferrous ions.After 24 hours, be discharged into LDH horizontal analysis neuronal death in the substratum, mean value ± SEM (culture hole n=3-4 of every kind of situation (10a) or n=3-4 (10b)) by measurement. ^*ANOVA and Student-Neuman-Keuls test are used in expression, and there is significant difference p＜0.05 o'clock with relevant contrast.

Figure 11. will be separately or the mouse cortex cell culture (DIV 12-14) that contains 5-(4-oil of mirbane the methyl)-N-acetamido-2-acetoxy-benzoic acid ethyl ester (NNAAE) of dose indicating contact 10 minutes (11a) with 300 micromole NMDA, continue to contact (11b) with 50 micromole's ferrous ions.After 24 hours, be discharged into LDH horizontal analysis neuronal death in the substratum, mean value ± SEM (the culture hole n=10-12 of every kind of situation (11a), n=7-8 (11b) or n=4) by measurement. ^*ANOVA and Student-Neuman-Keuls test are used in expression, and there is significant difference p＜0.05 o'clock with relevant contrast.

Figure 12. will be separately or the mouse cortex cell culture (DIV 12-14) that contains 5-(4-nitro benzoyl) aminosallcylic acid (NBAA) of dose indicating contact 10 minutes (12a) with 300 micromole NMDA, continue to contact (12b) with 50 micromole's ferrous ions.After 24 hours, be discharged into LDH horizontal analysis neuronal death in the substratum, mean value ± SEM (culture hole n=7-8 of every kind of situation (12a) or n=3-4 (12b)) by measurement. ^*ANOVA and Student-Neuman-Keuls test are used in expression, and there is significant difference p＜0.05 o'clock with relevant contrast.

Figure 13. will be separately or the mouse cortex cell culture (DIV 12-14) that contains 5-(the 4-oil of mirbane alkylsulfonyl) aminosallcylic acid (NBSAA) of dose indicating contact 10 minutes (13a) with 300 micromole NMDA, continue to contact (13b) with 50 micromole's ferrous ions.After 24 hours, be discharged into LDH horizontal analysis neuronal death in the substratum, mean value ± SEM (culture hole n=3-4 of every kind of situation (13a) or n=2-8 (13b)) by measurement. ^*ANOVA and Student-Neuman-Keuls test are used in expression, and there is significant difference p＜0.05 o'clock with relevant contrast.

Figure 14. will be separately or contain 5-[2-(4-the nitrophenyl)-ethyl of dose indicating] the mouse cortex cell culture (DIV 12-14) of aminosallcylic acid (NPAA) contacts 10 minutes (14a) with 300 micromole NMDA, continue to contact (14b) with 50 micromole's ferrous ions.After 24 hours, be discharged into LDH horizontal analysis neuronal death in the substratum, mean value ± SEM (culture hole n=4 of every kind of situation (14a) or n=4-8 (14b)) by measurement. ^*ANOVA and Student-Neuman-Keuls test are used in expression, and there is significant difference p＜0.05 o'clock with relevant contrast.

Figure 15. will be separately or contain 5-[3-(4-nitrophenyl) n-propyl of dose indicating] the mouse cortex cell culture (DIV 12-14) of aminosallcylic acid (NPPAA) contacts 10 minutes (15a) with 300 micromole NMDA, continue to contact (15b) with 50 micromole's ferrous ions.After 24 hours, analyze neuronal death: be discharged into the LDH level in the substratum, mean value ± SEM (culture hole n=4 of every kind of situation (15a) or n=3-8 (15b)) by measuring following value. ^*ANOVA and Student-Neuman-Keuls test are used in expression, and there is significant difference p＜0.05 o'clock with relevant contrast.

Detailed Description Of The Invention

We have synthesized BAS (BAS) and derivative thereof, and prove that these synthetic compounds have multiple neuroprotection. At first, BAS and derivative thereof are at 100-1, and 000 micromolar dosage alleviates the NMDA neurotoxicity. Secondly, BAS and derivative thereof are antioxidants, and in the micromolar dosage blocking-up of 1-300 free radical neurotoxicity. At last, BAS and derivative thereof alleviate the zinc ion neurotoxicity. These of BSA and derivative thereof new with compound neuroprotection, be of value to the treatment apoplexy, cerebral trauma and spinal cord injury, epilepsy, and follow exitotoxicity, the neurodegenerative disease of zinc ion neurotoxicity and free radical toxicity.

BAS and derivative thereof can be by synthesizing 5-aminosalicylic acid and the reaction of a kind of suitable compound. Following reaction schematic diagram illustrates the synthetic of BAS and derivative thereof.

Reaction schematic diagram 1

Reagent (a): benzyl bromine, 4-nitrobenzene methyl bromine, 4-chlorophenylmethyl chloride, 4-(trifluoromethyl) benzyl chloride, 4-fluorobenzene MB, 4-mehtoxybenzyl chloride, N, N-dimethyl formamide (DMF), triethylamine, room temperature, 4 hours.

The preferred classes of compound comprises those compounds in the chemical formula (I), and the X in them is CO, SO₂Or (CH₂) n (wherein n is the integer between the 1-5, comprises 1 and 5); R₁Hydrogen, C₁-C ₅Alkyl or C₂-C ₅Alkanoyl; R₂Hydrogen or C₁-C ₅Alkyl; R₃Hydrogen or acetyl group; And R₄Be phenyl, this phenyl is not substituted or is replaced by following one or more groups: nitro, halogen, alkylhalide group, C₁-C ₅Alkoxyl; Or a kind of its pharmaceutically acceptable salt.

Reaction schematic diagram 2.

Reagent (a): DMF (DMF), triethylamine, room temperature, 4 hours.

Reaction schematic diagram 3.

Reagent (a) EtOH, H₂SO ₄, minute gold-plating, 6 hours.

(b) anhydrous acetic acid, MeOH, 0 ℃, 30 minutes.

(c) anhydrous acetic acid, H ₂SO ₄, 0 ℃, 30 minutes.

The more preferably classification of compound comprises those compounds in the chemical formula (I), and the X in them is CO, SO ₂Or (CH ₂) n (wherein n=1,2,3); R ₁Be hydrogen, C ₁-C ₃Alkyl or C ₂-C ₃Alkyloyl; R ₂Be hydrogen or C ₁-C ₃Alkyl; R ₃Be hydrogen or a kind of ethanoyl; And R ₄Be phenyl, this phenyl is not substituted or is replaced by following one or more groups: nitro, halogen, halogen (C ₁-C ₃) alkyl, C ₁-C ₃Alkoxyl group; Or a kind of its pharmacy acceptable salt.

Embodiment 5.

Reagent (a) N, N-methylformamide (DMF), triethylamine, room temperature, 4 hours.

Interested particular compound is as follows in the chemical formula (I):

5-phenmethyl aminosallcylic acid

5-(4-oil of mirbane methyl) aminosallcylic acid (NBAS)

5-(4-chlorophenylmethyl) aminosallcylic acid (CBAS)

5-(4-phenylfluoroform methyl) aminosallcylic acid (TBAS)

5-(4-fluorobenzene methyl) aminosallcylic acid (FBAS)

5-(4-methoxybenzyl) aminosallcylic acid (MBAS)

5-(pentafluoro-benzyl) aminosallcylic acid (PBAS)

5-(4-oil of mirbane methyl) amino-2-hydroxybenzoic acid ethyl ester (NAHE)

5-(4-oil of mirbane methyl)-N-acetamido-2 hydroxybenzoic acid ethyl ester (NNAHE)

5-(4-oil of mirbane methyl)-N-acetamido-2-acetoxyl group-ethyl benzoate (NNAAE)

5-(4-nitro benzoyl) aminosallcylic acid (NBAA)

5-(4-oil of mirbane alkylsulfonyl) aminosallcylic acid (NBSAA)

5-[2-(4-nitrophenyl)-ethyl] aminosallcylic acid (NPAA)

5-[3-(4-nitrophenyl) n-propyl] aminosallcylic acid (NPPAA), or a kind of its pharmacy acceptable salt.

Term " pharmacy acceptable salt " comprises the additive salt that is commonly used to form alkali-metal salt and forms free acid or free alkali.So long as acceptable pharmaceutically, the character of salt is inessential, and for those those skilled in the art, and the bronsted lowry acids and bases bronsted lowry that is used for forming these salt is well-known.Can be used for forming the pharmaceutically acid of acceptable acid salt, the example comprises the mineral acid example hydrochloric acid, sulfuric acid and phosphoric acid; Organic acid such as toxilic acid, succsinic acid and citric acid.Other salt comprises the salt of alkali-metal salt or alkaline-earth metal, sodium for example, potassium, calcium and magnesium, the perhaps salt of organic bases, for example dicyclohexylamine.All these salt can be by ordinary method by the preparation of the respective compound in the chemical formula (I), and by reaction, for example, the compound in suitable acid or alkali and the chemical formula (I) prepares.

Synthetic embodiment has shown the representational method of preparation typical compound.

The preparation of synthetic embodiment 1:5-phenmethyl aminosallcylic acid (BAS)

Under room temperature and nitrogen protection atmosphere; the exsiccant DMF solution (25 milliliters) of 5-aminosalicylic acid (2.0 grams, 13 mmoles are bought from U.S. Aldrich Chemical Company) and triethylamine, add phenmethyl bromine (2.68 grams; 1.90 milliliter, 15.6 mmoles).Reaction mixture was at room temperature stirred 4 hours.Borneol is added reaction mixture, then get rid of solvent in a vacuum.The dilute with water reaction mixture is then used the ethyl acetate extracting.With organic layer water and normal saline washing, then use anhydrous magnesium sulfate drying.After solvent evaporated, use the column chromatography purification residuum, recrystallization is to obtain 3.6 gram (73% output) white solid-phenmethyl aminosallcylic acids in methanol/ethyl acetate/hexane (1: 3: 1).Mp:173.5-174.5 ℃ (decomposition).

To C ₁₄H ₁₃NO ₃Ultimate analysis.

	％C	％H	％N
				The notional result actual result	69.12 69.30	5.39 5.18	5.76 5.63

The preparation of synthetic embodiment 2:5-(4-oil of mirbane methyl) aminosallcylic acid (NBAS)

By imitateing the similar step among the synthetic embodiment 1, by using 5-aminosalicylic acid (2.00 grams, 13.0 mmole) and 4-oil of mirbane methyl bromine (3.38 gram, 15.6 mmoles), 2.90 gram (79% output) yellow solid 5-(4-oil of mirbane methyl) aminosallcylic acids have been obtained.Mp:211-212 ℃ (decomposition).

To C ₁₄H ₁₃N ₂O ₅Ultimate analysis.

	％C	％H	％N
				The notional result actual result	58.33 58.38	4.20 4.21	9.72 9.71

The preparation of synthetic embodiment 3:5-(4-chlorophenylmethyl) aminosallcylic acid (CBAS)

By imitateing the similar step among the synthetic embodiment 1, by using (500 milligrams of 5-aminosalicylic acids, 3.26 mmole) and 4-chlorophenylmethyl muriate (630 milligrams, 3.91 mmoles), obtained 480 milligrams of (53% output) white solid 5-(4-chlorophenylmethyl) aminosallcylic acid.mp：227-228℃。

To C ₁₄H ₁₂ClNO ₃Ultimate analysis.

	％C	％H	％N
				The notional result actual result	60.55 60.43	4.36 4.21	5.04 5.02

The preparation of synthetic embodiment 4:5-(4-phenylfluoroform methyl) aminosallcylic acid (TBAS)

By imitateing the similar step among the synthetic embodiment 1, by using (500 milligrams of 5-aminosalicylic acids, 3.26 mmole) and (760 milligrams of 4-trifluoromethylbenzene Methochlorides, 3.92 mmole), obtained 510 milligrams of (50% output) white solid 5-(4-phenylfluoroform methyl) aminosallcylic acid.Mp＞188 ℃ (decomposition).

To C ₁₄H ₁₂F ₃NO ₃Ultimate analysis.

	％C	％H	％N
				The notional result actual result	57.88 57.61	3.89 3.98	4.50 4.44

The preparation of synthetic embodiment 5:5-(4-fluorobenzene methyl) aminosallcylic acid (FBAS)

By imitateing the similar step among the synthetic embodiment 1, by using (500 milligrams of 5-aminosalicylic acids, 3.26 mmole) and 4-fluorobenzene MB (740 milligrams, 3.92 mmoles), obtained 480 milligrams of (44% output) white solid 5-(4-fluorobenzene methyl) aminosallcylic acid (FBAS).Mp＞210 ℃ (decomposition).

To C ₁₄H ₁₂FNO ₃Ultimate analysis.

	％C	％H	％N
				The notional result actual result	64.36 64.10	4.63 4.42	5.36 5.07

The preparation of synthetic embodiment 6:5-(4-methoxybenzyl) aminosallcylic acid (MBAS)

By imitateing the similar step among the synthetic embodiment 1, by using 5-aminosalicylic acid (1.00 grams, 6.53 mmole) and 4-mehtoxybenzyl muriate (1.23 gram, 7.84 mmoles), obtained 890 milligrams of (50% output) white solid 5-(4-methoxybenzyl) aminosallcylic acid.Mp=205-206 ℃ (decomposition).

To C ₁₅H ₁₅NO ₄Ultimate analysis.

	％C	％H	％N
				The notional result actual result	65.92 65.83	5.53 5.45	5.13 5.07

The preparation of synthetic embodiment 7:5-(pentafluoro-benzyl) aminosallcylic acid

By imitateing the similar step among the synthetic embodiment 1, by using (500 milligrams of 5-aminosalicylic acids, 3.26 mmole) and pentafluoro-benzyl bromide (1.02 gram, 3.92 mmoles), obtained 650 milligrams of (60% output) white solid 5-(pentafluoro-benzyl) aminosallcylic acid.Mp＞190 ℃ (decomposition).

To C ₁₄H ₈F ₅NO ₃Ultimate analysis.

	％C	％H	％N
				The notional result actual result	50.46 50.53	2.42 2.19	4.20 4.23

The preparation of synthetic embodiment 8:5-(4-oil of mirbane methyl) amino-2-hydroxybenzoic acid ethyl ester

In the ethanolic soln (35 milliliters) of 5-(4-oil of mirbane methyl) aminosallcylic acid (1.0 grams, 3.4 mmoles), add Conc.H carefully at 0 ℃ ₂SO ₄(3.5 milliliters).80 ℃ of stirring reaction mixtures 6 hours, cool to room temperature.After getting rid of solvent in a vacuum, with ethyl acetate extractive reaction mixture.With organic layer water, 10%NaHCO ₃Solution, 5%HCL solution, and normal saline washing.After with the anhydrous magnesium sulfate drying organic layer, it is concentrated in a vacuum.Use the column chromatography purification residuum, yellow solid-5-(4-oil of mirbane methyl) the amino-2-hydroxybenzoic acid ethyl ester of recrystallization in ethyl acetate/hexane (1: 2) to obtain 500 milligrams (46% output).Mp:106.5-107.5 ℃ (decomposition).

To C ₁₆H ₁₆N ₂O ₅Ultimate analysis.

	％C	％H	％N
				The notional result actual result	60.75 60.68	5.10 5.24	8.86 9.04

The preparation of synthetic embodiment 9:5-(4-oil of mirbane methyl)-N-acetamido-2 hydroxybenzoic acid ethyl ester

Protect under the atmosphere with nitrogen at 0 ℃, in the dry methanol solution (50 milliliters) of 5-(4-oil of mirbane methyl) amino-2-hydroxybenzoic acid ethyl ester (500 milligrams, 1.58 mmoles), add anhydrous acetic acid carefully.10 ℃ of stirring reaction mixtures 2 hours.After joining borneol in the reaction mixture lentamente, get rid of solvent in the vacuum.With ethyl acetate and water extractive reaction mixture, water, 10%NaHCO ₃(30 milliliters * 3) solution, 5%HCL (30 milliliters * 2) solution and normal saline washing organic layer.With anhydrous magnesium sulfate drying organic layer and dehydration.Use the column chromatography purification residuum, light yellow solid 5-(4-oil of mirbane the methyl)-N-acetamido-2 hydroxybenzoic acid ethyl ester of recrystallization in ethyl acetate/hexane (1: 3) to obtain 400 milligrams (70% output).mp：105.5-106.0℃。

To C ₁₈H ₁₈N ₂O ₆Ultimate analysis.

	％C	％H	％N
				The notional result actual result	60.33 60.54	5.06 5.35	7.82 8.12

The preparation of synthetic embodiment 10:5-(4-oil of mirbane methyl)-N-acetamido-2-acetoxyl group-ethyl benzoate

Protect under the atmosphere with nitrogen at 0 ℃, in the anhydrous acetic acid solution (10 milliliters) of 5-(4-oil of mirbane methyl) amino-2-hydroxybenzoic acid ethyl ester (500 milligrams, 1.58 mmoles), add conc.H carefully ₂SO ₄(0.5 milliliter).10 ℃ of stirring reaction mixtures 30 minutes.After joining borneol in the reaction mixture lentamente, get rid of solvent in the vacuum.Dilute with water reaction mixture and with ethyl acetate extractive reaction mixture.Water, 10%NaHCO ₃(30 milliliters * 3) solution, 5%HCL (30 milliliters * 2) solution and normal saline washing organic layer are then used anhydrous magnesium sulfate drying.After the solvent evaporated, with light yellow oil 5-(4-oil of mirbane the methyl)-N-acetamido-2-acetoxyl group-ethyl benzoate of column chromatography purification residuum to obtain 450 milligrams (71% output).

To C ₂₀H ₂₀N ₂O ₇Ultimate analysis.

	％C	％H	％N
				The notional result actual result	60.00 59.96	5.03 4.84	7.00 7.15

The preparation of synthetic embodiment 11:5-(4-nitro benzoyl) aminosallcylic acid

By imitateing the similar step among the synthetic embodiment 1; by using (500 milligrams of 5-aminosalicylic acids; 3.26 mmole) and (700 milligrams of 4-nitro benzoyl muriates; 3.77 mmole), obtained 550 milligrams of (56% output) light yellow solid 5-(4-nitro benzoyl) aminosallcylic acid.：mp＝270-271℃。

To C ₁₄H ₁₀N ₂O ₆Ultimate analysis.

	％C	％H	％N
				The notional result actual result	55.63 55.82	3.33 3.43	9.27 9.08

The preparation of synthetic embodiment 12:5-(4-oil of mirbane alkylsulfonyl) aminosallcylic acid

By imitateing the similar step among the synthetic embodiment 1; by using (500 milligrams of 5-aminosalicylic acids; 3.26 mmole) and (720 milligrams of 4-oil of mirbane methyl alkylsulfonyl muriates; 3.26 mmole), obtained yellow solid 5-(the 4-oil of mirbane alkylsulfonyl) aminosallcylic acid of 390 milligrams (35% output).：mp＝239-240℃。

To C ₁₃H ₁₀N ₂O ₇The ultimate analysis of S.

	％C	％H	％N	S％
					The notional result actual result	46.15 46.27	2.98 2.92	8.28 8.34	9.48 9.51

Synthetic embodiment 13:5-[2-(4-nitrophenyl)-ethyl] preparation of aminosallcylic acid

By imitateing the similar step among the synthetic embodiment 1, by using (500 milligrams of 5-aminosalicylic acids, 3.26 mmole) and (900 milligrams of 4-p-ethyl bromide things, 3.92 mmole), obtained 890 milligrams of (50% output) light yellow solid 5-[2-(4-nitrophenyl)-ethyl] aminosallcylic acid.：mp＝234-236℃。

To C ₁₅H ₁₄N ₂O ₅Ultimate analysis.

	％C	％H	％N
				The notional result actual result	59.60 59.77	4.67 4.79	9.27 9.24

Synthetic embodiment 14:5-[3-(4-nitrophenyl) n-propyl] preparation of aminosallcylic acid

By imitateing the similar step among the synthetic embodiment 1, by using (500 milligrams of 5-aminosalicylic acids, 3.26 mmole) and (950 milligrams of 3-(4-nitrophenyl) propyl group bromides, 3.92 mmole), obtained 520 milligrams of (50% output) light yellow solid 5-[3-(4-nitrophenyl) n-propyl] aminosallcylic acid.：mp＝229-231℃。

To C ₁₆H ₁₆N ₂O ₅Ultimate analysis.

	％C	％H	％N
				The notional result actual result	60.75 60.77	5.10 5.07	8.86 8.89

EXPERIMENTAL EXAMPLE

Preparation is from the elementary tegumental cell culture of fetal mice, is used for the neuroprotective of detection compound.Mouse cortex cell cultures objects system can be widely used for studying the mechanism and the drug intervention of neuronal death in the nervous system disease.Briefly, be concerned about the draft that mechanism of the council is approved, from the brain of the tire mouse in 15 day age, take out pallium according to our animal.Neopallium gently ground and is layered on be coated with in advance 10 mcg/ml poly--24 orifice plates (5 hemisphere/plate) of D-Methionin and 4 mcg/ml laminines on.Plate culture medium is to have replenished 5% horse serum, 5% foetal calf serum, the minimum base substance substratum of the Eagles of 2 mmole glutamine and 21 mmole glucose (MEM, Earles salt, no glutamine).Culture is kept at 37 degrees centigrade, and in the moist atmosphere of 5% carbonic acid gas, vitro culture is (DIV 7) after 7 days, and culture is transferred on the growth medium, and this substratum is except lacking tire serum, and all the other compositions and plate culture medium are together.When dividing 7-9 time, the cytosine(Cyt) arbinofuranose glycosides that adds 10 mmoles is to stop neuroglial hypertrophy.Then neuroglia and one week of neuronic mixed culture feed twice.

In order to introduce the nerve injury that causes by NMDA or zine ion, in the HEPES-buffering regulation and control salts solutions of forming by following material (HCSS): (in mmole) 120NaCl, 5KCl, 1.6MgCl ₂, 2.3CaCl ₂, 15 glucose, 20HEPES and 10NaOH contact 10 minutes with the tegumental cell culture with the toxic dose of NMDA, or contact 30 minutes with the toxic dose of zine ion.After the contact, culture is cleaned three times and changes on the MEM that is adjusted to 25 mmole glucose and 26.2 moles of sodium bicarbonates, in the carbonic acid gas incubator, placed in ensuing 20-24 hour.In order to introduce the free radical neurotoxicity, the tegumental cell culture is placed on the MEM that is adjusted into 25 mmole glucose and 26.2 moles of sodium bicarbonates, with ferrous ion or BSO Continuous Contact 20-24 hour.

Differing microscopic evaluation general cell damage under the light, perhaps method (Koh and Choi as described above, JNeurosci Methods 20:83-90,1987), after 24 hours, come the damage of microscopic evaluation general cell at neurotoxic injury by the serum lactic dehydrogenase amount (LDH) that measurement is discharged in the substratum bath.The per-cent of neuronal death is standardized as the LDH mean value that contacts back or false contrast (=0) release continuously with 500 micromole NMDA (=100).

In order to detect antioxidant effect, a kind of stable free radical-DPPH (2,2-phenylbenzene-1-picryl-hydrazyl free radical) is dissolved in ethanol to form 100 micromolar solution.With compound and DPPH or ethanol synthesis.After cultivating 30 minutes, with the relative reduction of spectrophotometer measurement 517 nanometers DPPH absorption.

The antioxygenation of EXPERIMENTAL EXAMPLE 1:5-aminosallcylic acid

In the primary cortex cell culture, detect the neuroprotective of 5-aminosalicylic acid (AS).In 24 hours, containing the micromolar AS of 10-300 does not have to reduce and contacts the neuronal death (scheming .1.a) that causes after 10 minutes with 300 micromole NMDA.What is interesting is, increase 10-100 micromole AS, prevented to contact the free radical neurotoxicity (scheming .1b) that causes after 24 hours dose-dependently with ferrous ion.After zine ion treating processes neutralizing treatment, add the neuronal death after AS does not reduce by 24 hours constantly, these neurones have contacted 30 minutes with 300 micromole's zine ions.The neurovirulent neuroprotective of AS Green Tea Extract has reduced the level of DPPH (a kind of stabilized radical) owing to the direct anti-oxidant properties of compd A S.Compare with trolox (vitamin-E of the permeable type of a kind of film), AS is a kind of weak oxidant.

The antioxidant properties of table 1.AS

	Reactant	The concentration of Trolox or AS (micromole)
								0	3	10	30	100	300
		A 517nm	Has only DPPH	1.2±0.05
DPPH+Trolox								1.08±0.1	0.89±0.08 *	0.39±0.06 *	0.05±0.01 *	0.03±0.01 *
			DPPH+AS			1.03±0.05	0.9±0.06 *						0.45±0.07	0.16±0.00 *

AS or trolox and be dissolved in alcoholic acid 100 micromole DPPH reaction 30 minutes.After deducting the ethanol background value, by measuring its anti-oxidant properties of horizontal mutation analysis of 517 nanometers DPPH, mean value ± SEM (every kind of situation n=3 test tube). ^*ANOVA and Student-Neuman-Keuls test are used in expression, and p＜0.05 o'clock has significant difference when having only DPPH.

AS can make neurone avoid radical damage, but NMDA or zine ion neurotoxicity are not had good antagonism effect.Yet AS removes the effect of free radical not as trolox.

The neuroprotective of EXPERIMENTAL EXAMPLE 2:5-phenmethyl aminosallcylic acid and derivative thereof

1.5-the neuroprotective of phenmethyl aminosallcylic acid

Synthetic BAS also detects the ability of its anti-nerve injury, and this nerve injury is induced generation in the tegumental cell culture.Adding 300 micromoles, 5 phenmethyl aminosallcylic acids (BAS) simultaneously approximately can make NMDA inductive neuronal death reduce by 50% (figure .2a).Under the situation that 1 micromole BAS exists, with the neuronal death that causes after (scheming .2c) that 50 micromole's ferrous ions (figure .2b) or 10 mmole BSO contacts sizable reduction is arranged, add 3 micromole BAS, neuronal death is blocked basically fully.Add 30-300 micromole BAS, can be in 24 hours, reduction of dose-dependently ground and caused neuronal death after 300 micromole's zine ions contact 30 minutes.As AS or trolox, BAS is a kind of direct antioxidant (table 2).The anti-oxidant properties of BAS is that 1 micromole just can observe in so low at dosage.

The antioxidant properties of table 2.BAS

[BAS], the micromole	0	1	3	10	30	100	300
								A 5l7nm	1.2±0.05	1.07±0.07	1.01±0.07	0.71±0.09 *	0.21±0.04 *	0.15±0.02 *	0.16±0.01 *

BAS be dissolved in alcoholic acid 100 micromole DPPH reaction 30 minutes.After deducting the ethanol background value, by measuring its anti-oxidant properties of horizontal mutation analysis of 517 nanometers DPPH, mean value ± SEM (every kind of situation n=3 test tube). ^*ANOVA and Student-Neuman-Keuls test are used in expression, p＜0.05 o'clock, and ([BAS]=O) has significant difference when having only DPPH.

2.BAS derivative

The BAS derivative is by the contraposition usefulness-NO at phenmethyl amino ₂[5-(4-oil of mirbane methyl amino) Whitfield's ointment, NBAS] ,-Cl[5-(4-chlorophenylmethyl amino) Whitfield's ointment, CBAS] ,-CF ₃[5-(4-trifluoromethylbenzene methylamino) Whitfield's ointment, TBAS] replacement-H comes synthetic.The replacement of this de-electronation group does not reduce BAS to NMDA, the neurovirulent resistance of zine ion or free radical (figure .3-5).These BAS derivatives more effective than BAS aspect the prevention free radical toxicity.

The antioxidant properties of table 3.NBAS

[NBAS], the micromole	0	10	30	100	300
						A 517nm	1.2±0.01	0.37±0.1 *	0.04±0.1 *	0.04±0.00 *	0.04±0.00 *

NBAS be dissolved in alcoholic acid 100 micromole DPPH reaction 30 minutes.After deducting the ethanol background value, by measuring its anti-oxidant properties of horizontal mutation analysis of 517 nanometers DPPH, mean value ± SEM (every kind of situation n=3 test tube). ^*ANOVA and Student-Neuman-Keuls test are used in expression, p＜0.05 o'clock, and ([reactant]=O) has significant difference when having only DPPH.

With-F or-OCH ₃Replacement-NO ₂As if the result causes the toxic neuroprotective of NMDA is reduced, but the neuroprotective potential of radical damage is risen.

The neuroprotective of EXPERIMENTAL EXAMPLE 3:5-(pentafluoro-benzyl) aminosallcylic acid (PBAS)

Synthetic 5-(pentafluoro-benzyl) aminosallcylic acid (PBAS) also detects the ability of its anti-nerve injury.Add 100-1000 micromole PBAS simultaneously, NMDA inductive neuronal death is reduced in dose-dependent mode.Approximately can make the NMDA neurotoxicity reduce by 65% (figure .8a) with 300 micromole PBAS.The dosage of PBAS is increased to 1 mmole, can blocks fully and the neuronal death that causes after 300 micromole NMDA contact.Add 1 micromolar PBAS and can obvious reduction continue to contact the caused neuronal death in back with 50 micromole's ferrous ions continuously.Under the situation that 3 micromole PBAS exist, neuronal death is blocked basically fully.By adding 1 micromolar PBAS, contact with 10 mmole BSO and the free radical neurotoxicity that causes is blocked.PBAS reduces the level (table 4) of DPPH, and hint PBAS is as a kind of direct oxidation agent blocking-up free radical neurotoxicity.The neurotoxicity (figure .8d) that adds the micromolar PBAS reduction of 100-300 zine ion simultaneously.

The anti-oxidant properties of table 4.PBAS

[PBAS], the micromole	0	1	3	10	30	100	300
								A 517nm	1.2±0.01	1.07±0.07	1.01±0.07	0.71±0.09 *	0.21±0.04 *	0.15±0.02 *	0.16±0.01 *

PBAS be dissolved in alcoholic acid 100 micromole DPPH reaction 30 minutes.After deducting the ethanol background value, by measuring its anti-oxidant properties of horizontal mutation analysis of 517 nanometers DPPH, mean value ± SEM (every kind of situation n=3 test tube). ^*ANOVA and Student-Neuman-Keuls test are used in expression, p＜0.05 o'clock, and ([PBAS]=O) has significant difference when having only DPPH.

EXPERIMENTAL EXAMPLE 4:NBAS derivative (X=CH ₂) neuroprotective

The derivative of several PBAS is for example: 5-(4-oil of mirbane methyl) amino-2-hydroxybenzoic acid ethyl ester (NAHE; R ₁=H, R ₂=CH ₂CH ₃, R ₃=H), 5-(4-oil of mirbane methyl)-N-acetamido-2 hydroxybenzoic acid ethyl ester (NNAHE; R ₁=COCH ₃, R ₂=CH ₂CH ₃, R ₃=H), and 5-(4-oil of mirbane methyl)-N-acetamido-2-acetoxyl group-ethyl benzoate (NNAAE; R ₁=COCH ₃, R ₂=CH ₂CH ₃, R ₃=COCH ₃) be synthesized, and in the tegumental cell culture, detect their neuroprotective.These derivatives have reduced the neurotoxicity (figure .9-11) of NMDA at 300 micromolar dosage.Contain 10 micromolar NAHE, NNAHE, or NNAAE can block ferrous ion inductive free radical neurotoxicity.

The neuroprotective of EXPERIMENTAL EXAMPLE 5:NBAS derivative (X=CO, SO ₂, CH ₂CH ₂Or CH ₂CH ₂CH ₂)

With CO[5-(4-oil of mirbane methyl) aminosallcylic acid; NBAA], SO ₂[5-(4-oil of mirbane alkylsulfonyl) aminosallcylic acid; NBSAA], CH ₂CH ₂[5-[2-(4-nitrophenyl)-ethyl] aminosallcylic acid; NPAA] or CH ₂CH ₂CH ₂[5-[3-(4-nitrophenyl) n-propyl] aminosallcylic acid; NPPAA] replace the X (CH of NBAS ₂) group.NBAA obviously reduces the neurotoxicity of NMDA when dosage is 30 micromoles.Yet, when dosage is micromolar up to 300, but do not have further to increase the provide protection (figure .12a) of its antagonism NMDA.What is interesting is that containing dosage is the micromolar NBSAA of 30-300, reduce NMDA neurotoxicity (figure .13a) in dose-dependent mode.When containing 300 micromole NBSA, obviously reduced neuronal death with the 90-100% that causes after 300 micromole NMDA contact.Still damage has neuroprotective to ferrous ion for NBAA and NBSAA, but they are weaker than NBAS (figure .12 and 13 on reduction free radical neurotoxic effect; Table 5).

CH with X ₂Use CH ₂CH ₂Or CH ₂CH ₂CH ₂Substitute, can reduce NBAS the neurovirulent protection potential of NMDA.Add 300 micromolar NPAA or NPPAA and reduce NMDA inductive neuronal death in the cortical neuron (figure .14 and 15) slightly.On the contrary, discover that NPAA or NPPAA are more effective than NBAS on blocking-up free radical neurotoxicity, when 1 micromole NPAA or NPPAA exist, block ferrous ion inductive neuronal death fully.

The anti-oxidant properties of table 5.NBAS

Reactant	Has only DPPH	DPPH+NBAA	DPPH+NBSAA	DPPH+NPAA	DPPH+NPPAA
						A 517nm	1.2±0.01	0.42±0.22 *	0.15±0.05*	0.09±0.01 *	0.09±0.02 *

NBAS derivative (every kind 100 micromole) be dissolved in alcoholic acid 100 micromole DPPH reaction 30 minutes.After deducting the ethanol background value, by measuring its anti-oxidant properties of horizontal mutation analysis of 517 nanometers DPPH, mean value ± SEM (every kind of situation n=3 test tube). ^*ANOVA and Student-Neuman-Keuls test are used in expression, and p＜0.05 o'clock has significant difference when having only DPPH.

The above results shows, the compound of chemical formula (I) can be used to prevent and exitotoxicity, neurodegenerative disease that the zine ion neurotoxicity is relevant with the free radical neurotoxicity effectively.

In the chemical formula (I) compound to people's administration, can use any technology that compound can be introduced patient or Mammals blood flow, comprise oral administration, intravenous injection, intramuscular injection and subcutaneous injection.Mammals wherein includes, but are not limited to cat, dog, poultry, domestic animal or the like.

Dosage every day of prophylactic treatment required compound is preferably from 0.1 milligram-20 milligrams every kg body weight every days.More preferred dose is 0.2 milligram of-10 milligrams of every kg body weight.Most preferred dosage is 0.5 milligram-5 milligrams every kg body weight every days.The dosage that is fit to can be with a plurality of sub-doses administrations every day.These inferior agent can be with the unit dosage administration.Typically, dosage or sub-doses can contain 1 milligram of-100 milligrams of active compound per unit formulation.More preferred dose contains 2 milligrams-50 milligrams active compound per unit formulation.Most preferred dosage form contains 3 milligrams of-25 milligrams of active compound per unit dosage.

Active compound is usually with pharmaceutically acceptable preparation administration.These preparations can comprise active compound and one or more pharmaceutically acceptable carrier or thinners.Other therapeutical agent also can appear in the said preparation.Pharmaceutically acceptable carrier or thinner provide suitable launch vehicle for the transportation of active substance, and can not introduce and not wish the side reaction seen.The transmission of active compound in these preparations can realize that comprise oral administration, administration is gone in snuffing with following different approach, topical, oral cavity and sublingual administration, perhaps administered parenterally, for example: subcutaneous route, intramuscular approach, intravenous route and intradermal approach.

The preparation of oral administration can be capsular form, and this capsule contains active compound and one or more lubricants, sanitas, tensio-active agent or the dispersion agent that is scattered in tackiness agent (for example gelatin or HYDROXY PROPYL METHYLCELLULOSE).These capsules or tablet can contain controls-puts preparation, for example can be provided in the procatarxis of active compound in the HYDROXY PROPYL METHYLCELLULOSE.

The preparation of administered parenterally can be water-based or nonaqueous isotonic sterile injection solution or suspension.These solution or suspension can be by sterilized powder that contains one or more carriers or thinner or granules preparation, and carrier of wherein mentioning or thinner are employed carrier or thinners in the preparation of oral administration.

Although given description to specific embodiments of the present invention, the details of these invention schemes is not to be restriction.Not leaving under the spirit and scope of the present invention, can produce multiple Equivalent, change, and modification, should be appreciated that these equivalent embodiments are parts of the present invention.

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Claims (43)

1. with the compound of following chemical formula (I) expression:

Wherein,

X is CO, SO ₂Or (CH ₂) n (wherein n is the integer of 1-5, comprises 1 and 5);

R1 is a hydrogen, alkyl or alkyloyl;

R2 is a hydrogen or alkyl;

R3 is hydrogen or a kind of acetoxyl; With

R4 is a phenyl, and this phenyl is not substituted or is selected from one or more following groups and replaces: nitro, halogen, alkylhalide group and C ₁-C ₅Alkoxyl group; Or a kind of its pharmacy acceptable salt.

2. the compound of claim 1, wherein X is CO, SO ₂Or (CH ₂) n (wherein n is the integer of 1-5, comprises 1 and 5); R ₁Be hydrogen, C ₁-C ₅Alkyl or C ₂-C ₅Alkyloyl; R ₂Be hydrogen or C ₁-C ₅Alkyl; R ₃Be hydrogen or a kind of acetoxyl; And R ₄Be phenyl, this phenyl is not substituted or is selected from one or more following groups and replaces: nitro, halogen, alkylhalide group and C ₁-C ₅Alkoxyl group; Or a kind of its pharmacy acceptable salt.

3. the compound of claim 1, wherein X is CO, SO ₂Or (CH ₂) n (wherein n is the integer of 1-5, comprises 1 and 5); R ₁Be hydrogen, C ₁-C ₃Alkyl or C ₂-C ₃Alkyloyl; R ₂Be hydrogen or C ₁-C ₃Alkyl; R ₃Be hydrogen or a kind of acetoxyl; And R ₄Be phenyl, this phenyl is not substituted or is selected from one or more following groups and replaces: nitro, halogen, halogen (C ₁-C ₃) alkyl and C ₁-C ₃Alkoxyl group; Or a kind of its pharmacy acceptable salt.

4. the compound of claim 1, this compound is selected from following material:

5-phenmethyl aminosallcylic acid (BAS)

5-(4-oil of mirbane methyl) aminosallcylic acid (NBAS)

5-(4-chlorophenylmethyl) aminosallcylic acid (CBAS)

5-(4-phenylfluoroform methyl) aminosallcylic acid (TBAS)

5-(4-fluorobenzene methyl) aminosallcylic acid (FBAS)

5-(4-methoxybenzyl) aminosallcylic acid (MBAS)

5-(4-pentafluoro-benzyl) aminosallcylic acid (PBAS)

5-(4-oil of mirbane methyl) amino-2-hydroxybenzoic acid ethyl ester

5-(4-oil of mirbane methyl)-N-acetamido-2-acetoxy-benzoic acid ethyl ester

5-(4-oil of mirbane methyl)-N-acetamido-2-acetoxyl group-ethyl benzoate

5-(4-nitro benzoyl) aminosallcylic acid

5-(4-oil of mirbane alkylsulfonyl) aminosallcylic acid

5-(4-oil of mirbane ethyl) aminosallcylic acid

5-[3-(4-nitrophenyl) n-propyl] aminosallcylic acid, or a kind of its pharmacy acceptable salt.

5. the purposes of the compound of any one on medicine is made among the claim 1-4, this medicine are used for protecting relay cell to avoid acute and chronic injury to central nervous system (CNS).

6. the purposes of claim 5, wherein said CNS damage be from ischemic, anoxic, hypoglycemia, traumatic brain injury, traumatic Spinal injury, epilepsy, Huntington, Parkinson's disease, Alzheimer, or amyotrophic lateral sclerosis.

7. the purposes in the claim 5, wherein said CNS damage are the activation by N-methyl-D-aspartate (NMDA) glutamate receptor, the entering and accumulating of zine ion, or free radical caused.

8. any one described purposes among the claim 5-7, wherein said compound reduces the NMDA neurotoxicity, zine ion neurotoxicity and blocking-up free radical neurotoxicity as direct antioxidant.

9. the purposes of the compound in any on medicine is made among the claim 1-4, this medicine are used for treatment or prevention and NMDA neurotoxicity, the nervous system disorders that zine ion neurotoxicity or oxidative stress are correlated with.

10. one kind makes relay cell avoid composition to central nervous system (CNS) acute and chronic injury, and said composition contains the compound in any among the claim 1-4 an amount of on the therapeutics.

11. compound or its pharmacy acceptable salt by following formula (I) expression:

Wherein,

X is CO, SO ₂Or (CH ₂) n (wherein n is the integer of 2-5, comprises 2 and 5);

R ₁Be hydrogen, C ₁-C ₅Alkyl or C ₂-C ₅Alkyloyl;

R ₂Be hydrogen or C ₁-C ₅Alkyl;

R ₃Be hydrogen or ethanoyl; With

R ₄Be phenyl, this phenyl is not substituted or is replaced by one or more groups that are selected from down group: nitro, halogen, halo (C ₁-C ₃) alkyl, and C ₁-C ₅Alkoxyl group.

12. according to the compound of claim 11, wherein R ₁Be hydrogen; R ₂Be hydrogen; R ₃Be hydrogen; And R ₄By halo (C ₁-C ₃) phenyl that replaces of alkyl.

13. according to the compound of claim 11, wherein X is (CH ₂) _n(wherein n is the integer of 2-5, comprises 2 and 5).

14. according to the compound of claim 13, wherein R ₁Be hydrogen; R ₂Be hydrogen; R ₃Be hydrogen; And R ₄By halo (C ₁-C ₃) phenyl that replaces of alkyl.

15. according to the compound of claim 11, wherein X is (CH ₂) _n(wherein n is 2 or 3).

16. according to the compound of claim 15, wherein R ₁Be hydrogen; R ₂Be hydrogen; R ₃Be hydrogen; And R ₄By halo (C ₁-C ₃) phenyl that replaces of alkyl.

17. according to the compound of claim 16, wherein n is 2.

18. one kind is used for protecting axoneuron to avoid pharmaceutical composition to the acute and chronic injury of central nervous system (CNS), said composition contains the compound of useful following formula (I) expression or its pharmacy acceptable salt as active principle:

Wherein,

X is CO, SO ₂Or (CH ₂) _n(wherein n is the integer of 1-5, comprises 1 and 5);

R ₁Be hydrogen, C ₁-C ₅Alkyl or C ₂-C ₅Alkyloyl;

R ₂Be hydrogen or C ₁-C ₅Alkyl;

R ₃Be hydrogen or ethanoyl; With

R ₄Be phenyl, this phenyl is not substituted or is replaced by one or more groups that are selected from down group: nitro, halogen, halo (C ₁-C ₃) alkyl, and C ₁-C ₅Alkoxyl group.

19. according to the pharmaceutical composition of claim 18, wherein R ₁Be hydrogen; R ₂Be hydrogen; R ₃Be hydrogen; And R ₄The phenyl that replaced by one or more groups that are selected from down group: halogen and halo (C ₁-C ₃) alkyl.

20. according to the pharmaceutical composition of claim 19, wherein R ₄By halo (C ₁-C ₃) phenyl that replaces of alkyl.

21. according to the pharmaceutical composition of claim 18, wherein X is (CH ₂) _n(wherein n is the integer of 1-5, comprises 1 and 5).

22. according to the pharmaceutical composition of claim 21, wherein R ₁Be hydrogen; R ₂Be hydrogen; R ₃Be hydrogen; And R ₄The phenyl that replaced by one or more groups that are selected from down group: halogen and halo (C ₁-C ₃) alkyl.

23. according to the pharmaceutical composition of claim 18, wherein X is (CH ₂) _n(wherein n is the integer of 2-5, comprises 2 and 5).

24. according to the pharmaceutical composition of claim 23, wherein R ₁Be hydrogen; R ₂Be hydrogen; R ₃Be hydrogen; And R ₄By halo (C ₁-C ₃) phenyl that replaces of alkyl.

25. according to the pharmaceutical composition of claim 18, wherein X is (CH ₂) _n(wherein n is 2 or 3).

26. according to the pharmaceutical composition of claim 25, wherein R ₁Be hydrogen; R ₂Be hydrogen; R ₃Be hydrogen; And R ₄By halo (C ₁-C ₃) phenyl that replaces of alkyl.

27. according to the pharmaceutical composition of claim 26, wherein n is 2.

28. according to any one pharmaceutical composition among the claim 18-27, wherein said CNS damage is from ischemic, anoxic, hypoglycemia, traumatic brain injury, traumatic Spinal injury, epilepsy, Huntington Chorea, Parkinson's disease, Alzheimer, or amyotrophic lateral sclerosis.

29. according to the pharmaceutical composition of claim 28, wherein said composition reduces N-methyl-D-aspartate (NMDA) neurotoxicity, Zn as direct antioxidant ^{+ 2}Neurotoxicity, or blocking-up free radical neurotoxicity.

30. according to any one pharmaceutical composition among the claim 18-27, wherein said CNS damage is the activation by N-methyl-D-aspartate (NMDA) glutamate receptor, Zn ^{+ 2}Enter and accumulate, or free radical caused.

31. according to the pharmaceutical composition of claim 30, wherein said composition reduces N-methyl-D-aspartate salt (NMDA) neurotoxicity, Zn as direct antioxidant ^{+ 2}Neurotoxicity, or blocking-up free radical neurotoxicity.

32. the application in the preparation medicine of the compound of following formula (I) expression or its pharmacy acceptable salt, described medicine are used to protect axoneuron to avoid acute and chronic injury to central nervous system (CNS):

Wherein,

X is (CH ₂) _n(wherein n is the integer of 1-5, comprises 1 and 5);

R ₁Be hydrogen, C ₁-C ₅Alkyl or C ₂-C ₅Alkyloyl;

R ₂Be hydrogen or C ₁-C ₅Alkyl;

R ₃Be hydrogen or ethanoyl; With

R ₄Be phenyl, this phenyl is not substituted or is replaced by one or more groups that are selected from down group: nitro, halogen, halo (C ₁-C ₃) alkyl, and C ₁-C ₅Alkoxyl group.

33. according to the application of claim 32, wherein R ₁Be hydrogen; R ₂Be hydrogen; R ₃Be hydrogen; And R ₄The phenyl that replaced by one or more groups that are selected from down group: halogen and halo (C ₁-C ₃) alkyl.

34. according to the application of claim 33, wherein R ₄By halo (C ₁-C ₃) phenyl that replaces of alkyl.

35. according to the application of claim 32, wherein n is 2-5.

36. according to the application of claim 35, wherein R ₁Be hydrogen; R ₂Be hydrogen; R ₃Be hydrogen; And R ₄By halo (C ₁-C ₃) phenyl that replaces of alkyl.

37. according to the application of claim 32, wherein n is 2 or 3.

38. according to the application of claim 37, wherein R ₁Be hydrogen; R ₂Be hydrogen; R ₃Be hydrogen; And R ₄By halo (C ₁-C ₃) phenyl that replaces of alkyl.

39. according to the application of claim 38, wherein n is 2.

40. according to any one application among the claim 32-39, wherein said CNS damage is from ischemic, anoxic, hypoglycemia, traumatic brain injury, traumatic Spinal injury, epilepsy, Huntington Chorea, Parkinson's disease, Alzheimer, or amyotrophic lateral sclerosis.

41. according to the application of claim 40, wherein said compound reduces N-methyl-D-aspartate salt (NMDA) neurotoxicity, Zn as direct antioxidant ^{+ 2}Neurotoxicity, or blocking-up free radical neurotoxicity.

42. according to any one application among the claim 32-39, wherein said CNS damage is activation, the Zn by N-methyl-D-aspartate (NMDA) glutamate receptor ^{+ 2}Enter and accumulate or free radical caused.

43. according to the application of claim 42, wherein said compound reduces N-methyl-D-aspartate salt (NMDA) neurotoxicity, Zn as direct antioxidant ^{+ 2}Neurotoxicity or blocking-up free radical neurotoxicity.

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