CN103804309A

CN103804309A - Chloroxime compound, its preparation method and application in pharmacy

Info

Publication number: CN103804309A
Application number: CN201210447383.8A
Authority: CN
Inventors: 王玉强; 姚辉; 吴传斌; 陶亮; 孙业伟
Original assignee: GUANGZHOU XIQUE MEDICINE Co Ltd
Current assignee: GUANGZHOU XIQUE MEDICINE Co Ltd
Priority date: 2012-11-09
Filing date: 2012-11-09
Publication date: 2014-05-21
Anticipated expiration: 2032-11-09
Also published as: CN103804309B

Abstract

The invention relates to a chloroxime compound, its preparation method and application in pharmacy. The chloroxime compound has a structure shown as the general formula I in the specification. The compounds has a very strong effect in synergistic regulation of heat shock protein activity, can be used for treating neurodegenerative diseases caused by injection of Abeta1-42 to rats, and aims to treat human neurodegenerative diseases. The compound also has a significant stress resistant effect, and can be used for preparation of new drugs treating diseases caused by protein misfolding and/or aggregation, and oxidative stress.

Description

A kind of chlorine oxime compounds and preparation method thereof and the application in pharmacy

Technical field

The present invention relates to a kind of chlorine oxime compounds, and preparation method thereof and at the nerve degenerative diseases causing due to protein folding mistake or gathering for the preparation for the treatment of and prevention, metabolic system disease, the application in the medicine of the heart, cerebrovascular system disease and inflammation.

Background technology

Protein is all extremely important to human and animal's life, is one of three major nutrient of human body, is the basic substance of life, there is no life without protein.Therefore, it is the material together with being closely connected with life and with various forms of vital movements.In cell, the more stable natural structure of the spontaneous formation of most of native protein mass-energy, or stable by part and metabolic factor.But find that nearly 10%～20% new synthetic polypeptide chain need be combined with molecular chaperones, could be correct folding by molecular chaperones.Also have about 20% new synthetic polypeptide chain can not form correct three-dimensional structure and be degraded by proteolytic enzyme.Quality control after translation mainly guarantees the normal function of protein by molecular chaperones and this two cover system of proteolytic enzyme.The effect of molecular chaperones be help can not spontaneous folding protein folding and assembling, make it recover normal structure, the effect of protease system is the protein of removing false folding.

And during as infection, heat-shocked and oxidative damage, can cause protein stability to reduce in stress situation when cell, some protein can not be folding by correct mode.In oxidative stress situation, the amount of having a net increase of of Mitochondrial DNA Mutation or reactive oxygen species (reactive oxygen species, ROS) increases all can improve Parkinson's disease, the sickness rate of the nerve degenerative diseases such as senile dementia greatly.In animal and external model, Antioxidation Treatment to a certain extent can neuroprotective unit (Simpson, Lancet Neurol.4:266,2005; Neymotin et al., Free Radic Biol Med.51:88-96,2011).

When protein folding mistake, usually by exposed on surface containing amino acid one end of carbon rich, rather than it is wrapped in to the inside, and the similar group that can combine closely in other oroteins containing this type of group of carbon rich forms polymer macromolecule.These polymkeric substance are lethality for cell (neurone).In fact, nearly all nerve degenerative diseases all involves the abnormal process of neuronal protein.Abnormal process can cause a kind of false folding of protein, causes the rear new synthetic protein of translation normally not modified, and protein cleavage is abnormal, abnormal gene splicing, misrepresentation, or the ability reduction of removing paraprotein.The protein of false folding is often accumulated.Many nerve degenerative diseases, the most outstanding feature is the protein aggregate (Muchowski, Neuron.35:9-12,2002) that the filamentary structure of amyloid appears in the inside and outside cell of brain cell.Cell is dealt with improperly and is caused different neural tuples to break down these protein, just shows as clinically various nerve degenerative diseases (Prusiner, N Engl J Med.344:1516-1526,2001).

In senile dementia (AD), there are two kinds of protein aggregation precipitations.There is amyloid patch deposition in extracellular around brain essence and cerebrovascular wall, its main component is that the polypeptide of the composition of 40-42 amino-acid residue is called amyloid-beta (A β) (Glenner et al., Biochem Biophys Res Commun.120:885-890,1984).There is the gathering (Grundke-Iqbal et al., J Biol Chem.261:6084-6089,1986) of the Protein tau of neurofibrillary tangles and Hyperphosphorylationof in the tenuigenin of the neurocyte of degenerating.

Parkinson's disease (PD) patient in the endochylema of brain substantia nigra neuron, can observe Lewy body assembles, the main component of these aggregates is the protein fragments (Spillantini et al., Nature.388:839-840,1997) of α-synuclein.

Copper-zinc sudismase (copper-zinc superoxide dismutase of 20% typical Familial Occurrence lateral sclerosis of spinal cord (ALS), SOD1) genes encoding has sudden change, the mutant apparent error of SOD1 has folded, and causes highly gatheringization.And in sporadic ALS, be accompanied by all the time the aggregation of ubiquitin-dyeing inclusion body, (Kiernan et al., Lancet.377:942-955,2011) as shown in Figure 1A.Therefore protein Misfolding and be gathered in play an important role in the pathogenesis of ALS (Cudkowicz et al., Muscle Nerve.38:837-844,2008).

In huntington (HD) patient's nucleus, depositing a large amount of Huntington proteins (Huntingtin protein, Htt) poly glumine (polyglutamine, polyQ) aggregation, this is this patient's a typical brain feature (DiFiglia et al., Science.277:1990-1993,1997).This mutain, at thin intracellular accumulation, forms poisonous oligopolymer and polymkeric substance.No matter immunohistochemistry and electron microscope method, be in HD patient or experimental model, all shown that change has occurred the autophagy approach of cell.Observations (Davies et al., Cell.90:537-548,1997 that increase in a large number in a lot of HD models and patient's autophagic vacuole source of evidence the earliest; Sapp et al., Ann Neurol.42:604-612,1997; Kim et al., J Neurosci.19:964-973,1999; Kegel et al., J Neurosci.20:7268-7278,2000; Nagata et al., Neuroreport.15:1325-1328,2004; Heng et al., Hum MolGenet.19:3702-3720,2010).

When cell is when stress situation (as oxidative stress, ROS increases), cause protein folding mistake or gathering, finally cause the generation of nerve degenerative diseases, therefore repair the effective way that this protein Misfolding or degraded aggregation may stop the state of an illness to further develop exactly.

An importance for the treatment of protein Misfolding disease is exactly the quantity that increases molecular chaperones, the activity that improves heat shock protein(HSP), and the correct of accessory protein folds, and promotes the degraded of misfolded protein.Heat shock protein(HSP) (heat shock protein, HSP) stress protein (the stress proteins that is otherwise known as, SP) as a member in molecular chaperones, relevant with the active function performance of numerous albumen, albumen that can auxiliary adjustment false folding recovers normal configuration (Hartl et al., Nature.475:324-332,2011).In the time of cell generation stress reaction (heat shock response), heat shock protein(HSP) energy great expression, then protein and the aggregation thereof of the folding or unfolding of identification error, and mark they degrade for proteasome.For example, heat shock protein(HSP) HSP27 and HSP90 have been proved the activity that can improve Ubiquitin-proteasome system (ubiquitin-proteasome system, UPS) very early.Also have a kind of heat shock protein HSP70 to be combined with the protein surface hydrophobic region of carrying out autofolding mistake, then guide a kind of ligase enzyme of E3 ubiquitin (as CHIP, carboxy terminus of HSC70-interacting protein) at folding wrong protein and ubiquitin on mark above aggregation thereof, after this proteasome just can in conjunction with and degrade them.But high-intensity oxidative damage can be strengthened interconnective degree between protein fragments, proteasome is difficult to the protein polymer of the such high stability of degraded.Now may need more heat shock protein expression to improve the degradation capability of Ubiquitin-proteasome system.

Heat shock protein(HSP) activity also with a lot of disease-relateds, the diseases that almost stress cause with all close ties.Such as heat shock protein(HSP) can prophylactic treatment disease of immune system as molecular chaperones, prevent inflammation, such as Hsp60, Hsp70, Hsp90, with gp96 can be in conjunction with bacterial exotoxin LPS (Tsan et al., J Leukoc Biol.85:905-910,2009), with immunodeficient disease acquired immune deficiency syndrome (AIDS) closely bound up (Di Cesare et al., Immunology.76:341-343,1992), also (Pockley, Circulation.105:1012-1017,2002) are closely connected with cardiovascular and cerebrovascular diseases.

First chlorine 9 oxime derivate is the expression compound that a class energy selectivity of Biorex company of Hungary discovery is amplified HSPs, and clinical trial is for anti-glycosuria comprehensive sick (Biro et al., Brain Res Bull.44:259-263,1997; Nanasi et al., Cardiovasc Drug Rev.19:133-151,2001).Biorex company in 2004 is by CytRx corporate buyout of the U.S., and the chlorine 9 oxime derivate that exploitation makes new advances is as prodrug, is further used for treating the various disease nerve degenerative diseases relevant to heat shock protein(HSP) (molecular chaperones) activity.Studies confirm that in a large number chlorine oxime compounds has good chaperone activity regulating effect, can improve the activity of heat shock protein(HSP), can be used for the treatment of nerve degenerative diseases.In normal cell, chlorine 9 oxime derivate does not activate the expression of HSF1 or induction HSPs.Chlorine oxime compound only amplifies the chaperone activity (Soti et al., Br J Pharmacol.146:769-780,2005) in stress situation or diseased cells.So special molecular mechanism of action, particular target stress cell to acting on, have more potentiality, safer (Soti et al., Br J Pharmacol.146:769-780,2005) than those indiscriminate molecular chaperones conditioning agents that act on all cells.

Chlorine 9 oxime derivate activity is affirmed.Compd B imoclomol and arimoclomol (Figure 1B) are two representative chlorine 9 oxime derivates, have all entered the clinical II/III phase to study.Bimoclomol was just applied to the treatment of diabetes and diabetic nephropathy for many years in the past, and Abbott's infusion of financial resources in 1997 cooperates to carry out the second stage of clinical study (Kalman.1997) of bimoclomol diabetic syndrome with bimoclomol discoverer Biorex company.The activity that Bimoclomol energy selectivity is amplified multiple heat shock protein(HSP) and heat shock factor; for example Hsp60; Hsp70; Hsp90; Grp94(glucose regulated protein94kDa; grape regulate protein 94) etc. (Nat Med.3:1150-1154; 1997); and this quasi-molecule companion mechanism of action relates to various diseases; it is not only proved diabetes rat and multiple complications (the Biro et al. that can treat STZ modeling; Brain Res Bull.44:259-263,1997; Biro et al., Neuroreport.9:2029-2033,1998), and can treat cardiac dysfunction, the abnormal various diseases of the cerebrovascular (Erdo et al., Brain Res Bull.45:163-166,1998; Lubbers et al., Eur J Pharmacol.435:79-83,2002; Polakowski et al., Eur JPharmacol.435:73-77,2002).

Arimoclomol shows outstanding activity on the multiple nerve degenerative diseases for the treatment of, and clinical trial (NCT00706147) result shows that it at least can delay the process of ALS ill 30%.Early-stage Study report has also confirmed that arimoclomol is HSPs inductor (amplifier); can obviously improve HSF1 in stress situation cell; HSP70; the expression amount of HSP90 etc.; and the expression of HSPs in standard state cell is affected to not obvious (Polakowski et al.; Eur JPharmacol.435:73-77,2002; Hargitai et al., Biochem Biophys Res Commun.307:689-695,2003; Kieran et al., Nat Med.10:402-405,2004).

Arimoclomol also as bimoclomol initial being developed be used for the treatment of diabetes, be found in afterwards that in animal model, to treat ALS effect remarkable.Arimoclomol is considered to the molecular chaperones conditioning agent that function is sound, conventionally can induce the heat shock protein(HSP) activity in all cells of human body, and to improve the natural repair ability of cell, treatment numerous disease, comprises that the impaired protein of ALS is abnormal.Another application, in the chlorine 9 oxime derivate Iroxanadine of clinical trial, is applied to the treatment of cardiovascular and cerebrovascular diseases.(http://www.cytrx.com/molecular_chaperone_regulation.html,2010)。

Ligustrazine (Tetramethylpyrazine, TMP), for extracting the main active ingredient obtaining in chuanxiong, has pharmacological action widely, for the treatment of multiple cardiovascular and cerebrovascular diseases.The calcium antagonist effect (Pang et al., Planta Med.62:431-435,1996) that Ligustrazine also shows in vascular tissue, this mechanism may be applied to nerve degenerative diseases.As ROS inhibitor, it can remove superoxide anion (O ₂ ^-), hydroxyl radical negative ion (OH ^-), lipid peroxy negative ion (lipid peroxyl, LOO ^-) etc. free radical (Zhang et al., Zhongguo Yao Li Xue Bao.15:229-231,1994; Zhang et al., Life Sci.72:2465-2472,2003).In central nervous system, TMP can significantly suppress the neurocyte oxidative damage of iron mediation and alleviate nerve cell death (Shih et al., Neuroreport.13:515-519,2002 that glutamate toxicity causes; Zhang et al., Eur J Pharmacol.467:41-47,2003; Liao et al., Neurosci Lett.372:40-45,2004).Ligustrazine systemic administration is protected ischemic to mouse and rabbit or the neurocyte of cerebral trauma or Spinal injury, can promote its functional rehabilitation (Fan et al., BMC Neurosci.7:48,2006; Kao et al., Neurochem Int.48:166-176,2006).What is interesting is, whole body administration Ligustrazine can alleviate the infringement of rodent learning and memory ability (Ni et al., Jpn J Pharmacol.67:137-141,1995 that D-semi-lactosi or ischemic brain injury cause; Zhang et al., Chin Med Sci J.19:180-184,2004).Significantly improve the amount (Tan, J Ocul Biol Dis Infor.2:57-64,2009) of cognitive function and the brain amyloid of dementia model mice.

But the antioxygenation of Ligustrazine is faint, bioavailability is low, needs clinically multiple dosing just can reach effective concentration.The treatment of nerve degenerative diseases at present does not also have specifics, and the medicine few in number having gone on the market is all because unsatisfactory curative effect or toxic side effect are difficult to greatly meet the demands.The protein that induction heat shock protein expression mis repair is folding or the effect of removing its aggregation neuroprotective cell are the emphasis of research both at home and abroad always.

Summary of the invention

The present invention aims to provide a kind of chlorine oxime compounds and pharmacy acceptable salt thereof, and this compounds can be induced the expression of heat shock protein(HSP), and antioxidant stress injury has very strong cytoprotective ability; And provide the preparation method of described chlorine oxime compounds; Also provide described chlorine oxime compounds and pharmacy acceptable salt thereof in treatment due to protein Misfolding or be extremely folded to form aggregation or the method for the disease that causes due to oxidative stress, and described chlorine oxime compounds and pharmacy acceptable salt thereof are in the application of preparing in relative medicine.

On the one hand, the invention provides a kind of chlorine oxime compounds and pharmacy acceptable salt thereof, described chlorine oxime compounds has the structure of following general formula I:

A kind of chlorine oxime compounds and pharmacy acceptable salt thereof, described chlorine oxime compounds has the structure of following general formula I:

Wherein:

R ₁, R ₂, R ₃, R ₄identical or different, be independently H separately, OH, NH ₂, COOH, straight or branched alkyl, replaces or unsubstituted monocycle or multi-ring alkyl aryl, heterocyclic base (O, S), nitrogenous heterocyclic base and N → O group thereof, acyl group, ester group, or amido; R ₂with R ₃also can form and replace or unsubstituted 4-8 ring with N atom;

Work as R ₄during for H, R ₁be not H, replace or unsubstituted phenyl naphthyl, pyridyl, or thienyl;

N is 0,1,2 or 3.

According to general formula I, preferred chlorine oxime compounds has the structure of following general formula I I:

Wherein: R ₂, R ₃, R ₄identical or different, be independently H separately, OH, NH ₂, COOH, straight or branched alkyl, replaces or unsubstituted monocycle or multi-ring alkyl aryl, heterocyclic base (O, S), nitrogenous heterocyclic base and N → O group thereof, acyl group, ester group, or amido; R ₂with R ₃also can form and replace or unsubstituted 4-8 ring with N atom;

R ₅, R ₆, R ₇identical or different, be independently H separately, F, Cl, Br, I, OH, NH ₂, COOH, straight or branched alkyl, aryl, heterocyclic base (O, N, S), acyl group, ester group, or amido;

X and Y have at least one to be N atom, and when X and Y only one be the N atomic time, R ₄be not H; On N atom, be connected with oxygen or do not connect oxygen;

N is 0,1,2 or 3.

According to general formula I I, preferred chlorine oxime compounds has the structure of general formula III:

According to general formula III, preferred chlorine oxime compounds is characterized in that R ₂be selected from methyl, ethyl, n-propyl, sec.-propyl, R ₃be selected from 3-methyl adamantane base, 3,5-dimethyladamantane base, sec.-propyl, or R ₂, R ₃connect into six-ring with N atom; Further preferred chlorine oxime compounds is characterized in that R ₅, R ₆, R ₇for methyl, R ₂, R ₃connect into six-ring with N atom, described chlorine oxime compounds has the structure of following general formula I V:

R ₄for H, OH, NH ₂, COOH, straight or branched alkyl, replaces or unsubstituted monocycle or multi-ring alkyl aryl, heterocyclic base (O, S), nitrogenous heterocyclic base and N → O group thereof, acyl group, ester group, amido.R ₈be selected from H, the straight or branched alkyl of C1-C4;

Above pyrazine ring N atom, be connected with oxygen or do not connect oxygen;

N is 0,1,2 or 3.

According to general formula I V, preferred chlorine oxime compounds has the structure of general formula V:

R ₄for H, OH, NH ₂, COOH, straight or branched alkyl, replaces or unsubstituted monocycle or multi-ring alkyl aryl, heterocyclic base (O, S), nitrogenous heterocyclic base and N → O group thereof, acyl group, ester group, or amido;

Above pyrazine ring N atom, be connected with oxygen or do not connect oxygen;

N is 0,1,2 or 3.

According to general formula V, preferred chlorine oxime compounds has the structure of general formula VI:

Wherein: on pyrazine ring N atom, be connected with oxygen or do not connect oxygen; N is 0,1,2 or 3.

According to general formula VI, preferred chlorine oxime compounds has the structure of following TCO-1 and TCO-2, but is not limited only to this two compounds

On the other hand, the present invention also provides the preparation method of described chlorine oxime compounds.Described method comprises: after the itrile group derivative of starting raw material is reacted with oxammonium hydrochloride, make the amino amino oxime compounds replacing again with the 2-hydroxyl-4-azepine-4-reactant salt replacing, last amino oxime and hydrochloric acid reaction make chlorine oxime compounds.

The present invention also provides and has comprised pharmaceutically above-mentioned arbitrary compound of effective dose or the pharmaceutical composition of its pharmaceutically-acceptable salts.Described effective dose is 1mg-10g.

The present invention further provides pharmaceutically above-mentioned arbitrary compound or its pharmaceutically-acceptable salts of effective dose, and comprise above-mentioned arbitrary compound of effective dose pharmaceutically or the pharmaceutical composition of its pharmaceutically-acceptable salts in treatment due to protein Misfolding or be extremely folded to form aggregation or the method for the disease that causes due to oxidative stress and preparing the application in relative medicine.

The present invention compared with prior art, has following novelty:

1) pyridine ring that adopts first active Chinese drug component molecule TMP ring and derivative thereof or pyrazine ring to replace clinical trial medicine bimoclomol and arimoclomol chlorine 9 oxime derivate has synthesized serial new chlorine 9 oxime derivate.

2) on replacing the basis of pyridine ring of clinical trial medicine bimoclomol and arimoclomol chlorine 9 oxime derivate, TMP ring adopt alkane or branched paraffin, diamantane and derivative thereof to replace the new chlorine 9 oxime derivate of piperidines synthesizing series on right side first; The activity of new synthetic compound TCO-1, TCO-2 is significantly higher than bimoclomol and arimoclomol.

3) on replacing the basis of pyridine ring of clinical trial medicine bimoclomol and arimoclomol chlorine 9 oxime derivate, TMP ring adopts active small molecular Salvianic acidA first, styracin, Thioctic Acid, vitamin H, TMP and derivative thereof are replaced the new chlorine 9 oxime derivate of middle hydroxyl hydrogen synthesizing series.

4) found that a set of practicable chemical synthesis route (method) synthesizes chlorine oxime-ligustrazine derivant.

Accompanying drawing explanation

Figure 1A. be bibliographical information diagram, wherein be described in the aggregation that is accompanied by all the time ubiquitin-dyeing inclusion body in sporadic ALS (referring to Cellular and molecular processes mediating neurodegeneration in ALS, Kiernan et al., Lancet.377:942-955,2011);

Figure 1B. show the chemical structure about chlorine oxime compounds;

Fig. 1 C. shows the chemical structure of TMP, TCO-1 and TCO-2;

Fig. 2. show a kind of synthetic route of compound TCO-1;

Fig. 3. show the another kind of synthetic route of compound TCO-1;

Fig. 4. show a kind of synthetic route of compound TCO-2;

Fig. 5. show the another kind of synthetic route of compound TCO-2;

Fig. 6. compound TCO-1 is described, TCO-2, the provide protection of PCO-1 and the BCO-1 cell to MG-132 induction damage;

Fig. 7. compound TCO-1 is described, TCO-2, PCO-1 and the BCO-1 restraining effect to ROS;

Fig. 8. compound TCO-1 is described, the provide protection of TCO-2 and the PCO-1 cell to MPP+ induction damage;

Fig. 9. for showing that compound TCO-1/TCO-2 removes the picture of the gathering of ubiquitin protein;

Figure 10. for showing that TCO-1 reduces the AD rat model data plot of arrival platform time;

Figure 11. for showing that TCO-1 increases the data plot of AD rat model through platform number of times.

Embodiment

Definition:

To give a definition to illustrate and define implication and the scope of various terms used in this invention.

Term used herein " alkyl " refers to the alkyl carbon chain of 15 carbon atoms of as many as of unsubstituted or substituted straight chain, side chain or annular.Straight chained alkyl comprises as methyl, ethyl, n-propyl, normal-butyl, n-pentyl, n-hexyl, n-heptyl and n-octyl.Branched-chain alkyl comprises as sec.-propyl, sec-butyl, isobutyl-, the tertiary butyl, neo-pentyl.Monocycle alkyl comprises as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and suberyl.Alkyl can be replaced by one or more substituting groups.Above-mentioned substituent indefiniteness example comprises OH, F, Cl, Br, I, NH ₂, NO ₂, ONO ₂.Term " multi-ring alkyl " also refers to two rings that do not replace or replace, triatomic ring, and substituent non-limiting example comprises OH, F, Cl, Br, I, NH ₂, NO ₂, ONO ₂.

Term used herein " aryl " refers to unsubstituted or substituted aromatics, carbon ring group and heteroaryl.Aryl or monocycle or fused polycyclic compounds.For example, phenyl is monocyclic aryl.Naphthyl is to have the examples that encircle the aryl condensing more.Aryl can be replaced by one or more substituting groups, and substituent nonrestrictive example comprises OH, F, Cl, Br, I, NH ₂, NO ₂, ONO ₂.Term " heteroaryl " relates to the group of monocycle replacement or non-substituted or many rings, at least comprises a heteroatoms in ring, such as nitrogen, oxygen and sulphur.For instance, typical heterocyclic group comprises that one or more nitrogen-atoms such as tetrazyl, pyrryl, pyridyl are (as 4-pyridyl, 3-pyridyl, 2-pyridyl etc.), pyridazinyl, indyl, quinolyl be (as 2-quinolyl, 3-quinolyl etc.), imidazolyl, isoquinolyl, pyrazolyl, pyrazinyl, pyrimidyl, pyriconyl or pyridazinyl; The typical heterocyclic group containing a Sauerstoffatom comprises 2-furyl, 3-furyl or benzofuryl; Typical sulfur heteroatom group comprises thienyl, benzothienyl; The typical heteroatom group that mixes comprises furan a word used for translation Ji, oxazolyl, isoxazolyl, thiazolyl and phenothioxin base.Heterocyclic group can be replaced by one or more substituting groups.These substituting groups comprise NH ₂, NO ₂, O-alkyl, NH-alkyl, N (alkyl) ₂, NHC (O)-alkyl, ONO ₂, F, Cl, Br, I, OH, OCF ₃, OSO ₂cH ₃, CO ₂h, CO ₂-alkyl, CN and low alkyl group, aryl and polyaryl.These situations comprise that in ring, heteroatoms is oxidized simultaneously, for example form N-oxide compound, ketone or sulfone.

Nitrogen-containing hetero aryl N → O compound relates to the N → O compound that contains 1 or 2 N atom aromatic heterocycle.

Term used herein " pharmaceutically acceptable " refers at compound does not have unacceptable toxicity in as salt.Pharmacy acceptable salt comprises inorganic anion, such as chlorion, sulfate radical, inferior sulfate radical, nitrate radical, nitrite anions, phosphate radical, hydrogen phosphate etc.Organic anion comprises acetate moiety, propionate, cinnamate, benzene methanesulfonic acid root, citrate, lactate, glucose acid group etc.

The new compound with general formula (I) structure providing in the specific embodiment of the present invention comprises following preferred compound: (TCO-1), (TCO-2), (PCO-1), (PCO-2), (ACO-1a), (ACO-1b), (ACO-2a), (ACO--2b), (GCO-1c-7c), (GCO-1d-7d), (FCO-1a-6a), (FCO-1b-6b), (LCO-1), (LCO-2).

There is the compound (TCO-1) of general formula (I) structure, (TCO-2), (PCO-1), (PCO-2), (ACO-1a), (ACO-1b), (ACO-2a), (ACO--2b), (GCO-1c-7c), (GCO-1d-7d), (FCO-1a-6a), (FCO-1b-6b), (LCO-1), (LCO-2) related chlorine oxime and oxime TMP or its N → Oization derivative are all the abilities with induction heat shock protein expression, they can induce the heat shock protein expression under stress situation on the one hand, can repair on the other hand and stress cause folding wrong protein or remove its aggregation, therefore, they can be used for prevention and treatment protein folding mistake or abnormal gathering and form the disease causing.These diseases include but not limited to nervous system disorders, as ischemic-hypoxic brain injury, apoplexy, cerebral trauma, senile dementia, epilepsy, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis, aids dementia disease, multiple sclerosis, chronic pain, priapism, cystic fibrosis, schizophrenia, dysthymia disorders, premenstrual tension syndrome, anxiety, habituation and migraine etc.; Also comprise cardiovascular disorder, as heart effluent, ischemia reperfusion injury, ischemia-reperfusion, toxic shock syndrome, adult respiratory distress are waited group, emaciation, myocarditis, atherosclerosis, coronary heart disease heart disease and heart attack etc.; Also comprise inflammation infection disease, as inflammatory intestinal tract disease, diabetes, rheumatoid arthritis, asthma, liver cirrhosis, allosome rejection, encephalomyelitis, meningitis, pancreatitis, peritonitis, vasculitis, lymphocytic choriomeningitis, glomerulonephritis, systemic lupus erythematous, gastrointestinal motility dysfunction, obesity, hyperalimentation, hepatitis and renal failure; Also comprise ophthalmic diseases, as diabetic retinopathy, uveitis, glaucoma, blepharitis, tarsal cyst, allergic ophthalmopathy, keratohelcosis, keratitis, cataract, age-related macular degeneration and optic neuritis.

The chlorine oxime compounds the present invention relates to can a kind of pharmacologically acceptable salts or the form of pharmaceutical composition to patient's administration.Thereby certain mixture need form pharmaceutical composition with suitable carrier or mixed with excipients and guarantee to reach dose therapeutically effective." dose therapeutically effective " refers to that chlorine oxime compounds reaches result for the treatment of and (repairs protein folding mistake, remove the abnormal protein of assembling, reduce because the cell injury that nerve degenerative diseases, metabolic disease or cardiovascular and cerebrovascular diseases cause etc.) necessary dosage.

Chlorine oxime compounds and the composition that contains this compounds thereof can be made multiple formulation, comprise solid dosage, semisolid dosage form, liquid preparation and aerosol (Remington ' s Pharmaceutical Sciences, Mack Publishing Company (1995), Philadelphia, PA, 19 ^thed).Concrete formulation in these a few class formulations comprises tablet, pill, dragee, granule, gelifying agent, paste, solution, suppository, injection, inhalation and sprays.These formulations can be used for part or whole body administration can be used for again quick-release or slow continuous administration, the administering mode of this type of medicine has a variety of, except aforesaid way, also has the interior administration of orally administering, cheek administration, rectal administration, peritoneal administration, intraperitoneal administration, epidermis administration, subcutaneous administration and tracheae etc.

When chlorine oxime compounds and contain these compounds composition drug administration by injection time, can with water-soluble or fat-soluble solvent, this compounds be mixed with to solution, clouding agent and emulsion.Fat-soluble solvent specifically comprises vegetables oil and similar oils, synthetic fat acid glyceride, high-grade aliphatic ester and glycol ester (proylene glycol).This compounds is more soluble in ethanolic soln, micro-DMSO solution.

When chlorine oxime compounds and contain these compounds composition oral administration time, can adopt common technology that itself and the acceptable vehicle of pharmacy are made to mixture.These vehicle these compounds can be made multiple can be by patient's formulation, as tablet, pill, suspensoid, gelifying agent etc.The preparation of oral preparations has several different methods, and as first compound and solid excipient mixed, fully grinding mixture, adds suitable auxiliary material, and processing treatment becomes particle.Can comprise for the auxiliary material of making oral dosage form: carbohydrate is as lactose, sucrose, N.F,USP MANNITOL or sorbyl alcohol; Cellulose family is as W-Gum, wheat starch, yam starch, gelatin, western twelve month yam glue, methylcellulose gum, Walocel MT 20.000PV (hydroxyproylmethyl-cellulose), cellulose sodium carboxymethyl, Povidone etc.

The chlorine oxime compounds the present invention relates to and the composition that contains these compounds thereof also can be made sprays, and this kind of formulation realizes by a pressurizer and an atomizer or a powder inhaler.Can be as propellant suitable in injector as Refrigerant 12, fluoro trichloromethane, dichloro tetrafluoro ethane, carbonic acid gas and dme etc.The dosage of aerosol drug delivery can regulate by the valve of injector.

The dose therapeutically effective of the composition that the various formulations that the present invention relates to are all related to chlorine oxime compounds and contain these compounds.The dose therapeutically effective of this compounds depends on the patient who receives treatment.In the time determining suitable dosage, patient's body weight, the state of an illness, medicining mode and prescriber's subjective judgement factor all will be included consideration in.The treatment significant quantity of chlorine oxime compounds and the composition that contains these compounds thereof should be by having the ability and the prescriber of rich experiences determine.

Although the dose therapeutically effective of chlorine oxime compounds and the composition that contains these compounds thereof can change according to patient's situation, suitable dosage scope is 10mg-10g conventionally.

The present invention compared with prior art; tool has the following advantages: the compound that the invention provides a kind of brand new; there is dual function mechanism (repair protein folding mistake & remove its abnormal aggregation or/and cytoprotective) simultaneously, can be by hemato encephalic barrier and compound safely and effectively.These compounds are to be worth the treatment of exploitation and prevention due to protein folding mistake or the nerve degenerative diseases of extremely assembling or causing due to oxidative stress, metabolic system disease, the heart, cerebrovascular system disease, the medicine of infectious diseases and ageing disorders etc.

The following examples are used for describing for enforcement of the present invention, but should not be interpreted as the present invention and can be subject to the restriction of these embodiment.

Synthetic (Fig. 2) of embodiment mono-, compound TCO-1

Take Tetramethylpyrazine 10g (73.5mmol) and be dissolved in 20ml acetic acid, add 30% hydrogen peroxide 8ml.Under 70 ° of C conditions, oil bath reaction was added 8.3ml30% (73.5mmol) hydrogen peroxide after 5 hours, continued reaction 5 hours, and TLC monitors level of response, stopped reaction.Be chilled to room temperature, sodium hydroxide saturated solution regulates pH to 10.Dichloromethane extraction (50ml*5 time), gets the underpressure distillation of organic layer Rotary Evaporators and does to obtain white solid (1a-1).Add wherein 125 ° of C oil bath reactions of 20ml diacetyl oxide (212mmol) 2.5 hours, TLC detection reaction is complete.Diacetyl oxide is removed in underpressure distillation, obtains acetylize crude product (1a-2).Then, sodium hydroxide saturated solution is adjusted to pH to 12, stirring at room temperature, and hydrolysis reaction spends the night.Ethyl acetate extractive reaction liquid (50ml*5 time), underpressure distillation is dry, wet method loading column chromatography, moving phase EA:PE (1:1), TLC detects cut, collects TMP-OH cut.Merge required identical cut, underpressure distillation is dry, obtains Off-white solid powder 7.5g (1a-3, TMP-OH, 49.3mmol), [M+Na] ⁺=175, productive rate 67%.

Gained 7.5g TMP-OH (49.3mmol) is dissolved in to 50ml dehydrated alcohol and adds activated manganese dioxide (6.4g, 73.6mmol) oxidation, 84 ° of C oil bath reaction 2h, after TLC detection reaction is complete, adopt while hot the careful suction filtration reaction solution of five metafiltration paper, remove manganese dioxide powder.After filter, obtain clear filtrate, Rotary Evaporators underpressure distillation is dry.Wet method loading column chromatography: moving phase EA:PE (1:5) separation and purification, TLC monitors cut.Collect compound 1a-4 cut, merge identical cut distillation and do to obtain lenticular white solid 5.5g (36.7mmol), [M+H] ⁺=151 is 1a-4 (TMP-CHO), productive rate 75%.

5.5g (36.7mmol) compound 1a-4 sample is dissolved with 30ml formic acid, taking 3.2g (45.9mmol) oxammonium hydrochloride adds, add subsequently 3.13g (45.9mmol) sodium formiate, miscible, 120 ° of C oil bath reactions reflux and spend the night.Saturated sodium hydroxide solution regulates reaction solution pH to 10, and dichloromethane extraction (30ml*5 time) is got organic layer underpressure distillation and done.100-200 order silica gel mixed sample, dry method loading column chromatography: moving phase EA:PE(1:5), fraction compositions is flowed out in TLC monitoring, collects compound 1a-5 cut.Merge identical cut distillation dry, obtaining white solid is compound 1a-5, productive rate 65%.

Get compound 1a-5 (3.5g, 23.9mmol) be dissolved in 30ml dehydrated alcohol, add oxammonium hydrochloride 2.1g (29.9mmol), get sodium bicarbonate 2.6g(30.9mmol) in 20ml distilled water heating for dissolving, miscible, 105 ° of C oil baths are reacted 3 hours.TLC monitors level of response, stopped reaction.Underpressure distillation is dry, dry method loading column chromatography, moving phase EA:PE:Et ₃n (60:40:0.5), collects cut, and TLC monitors fraction compositions.Merge identical cut underpressure distillation dry, obtaining white solid is compound 1a-6, productive rate 95%.

Get compound 1a-6 (4.1g, 22.7mmol) and be dissolved in 30ml ethanol, get sodium hydroxide (1.4g, 35mmol) be dissolved in 30ml distilled water, after add quaternary ammonium salt 1a-75.1g (28.8mmol) miscible, 80 ° of C oil baths reaction 3 hours, TLC detection reaction degree.Ethyl acetate extractive reaction liquid (30ml*5 time), collects organic layer underpressure distillation dry, wet method loading column chromatography, moving phase EA:PE:Et ₃n (70:30:0.5) wash-out, collects cut, and TLC detects fraction compositions.Merge the underpressure distillation of compound cut and do to obtain transparent oily matter 1a-8 (4.4g, 13.7mmol), productive rate 60%.

Take Sodium Nitrite (1.2g, 17.4mmol) and be dissolved in 4ml distilled water, salt ice bath is down to-5 ° of C, makes sub-acid sodium solution for subsequent use.The solution that is 1M by concentrated hydrochloric acid dilution, is chilled to-5 ° of C for subsequent use.Get compound 1a-8 (4.4g, 13.7mmol) and be dissolved in the cold hydrochloric acid soln of 20ml, salt ice bath stirs, and then slowly drips wherein sodium nitrite solution reaction 1h, then under condition of ice bath, regulates pH to 10 with chilled NaOH solution (7M).Finally, adopt dichloromethane extraction reaction solution (15ml*5 time), collect organic layer.The sample liquid of underpressure distillation extraction gained, wet method loading column chromatography, moving phase EA:PE:Et ₃n (17:83:0.5), TLC monitors fraction compositions, merges identical cut underpressure distillation dry, and oil pump decompression extracts residual solvent, and obtaining water white transparency oily thing is TCO-1 (2.2g, 6.8mol), productive rate 50%. ¹H-NMR(CDCl ₃,300MHz)(ppm)δ:1.37-1.60(m,6H,3CH ₂),2.30-2.40(m,6H,3NCH ₂),2.52(s,6H,2CH ₃),2.60(s,3H,CH ₃),4.04(m,1H,CH),4.28(m,2H,NOCH ₂). ¹³C-NMR(CDCl ₃,75.47MHz)(ppm)δ:21.57(CH ₃);21.97(CH ₃);22.40(CH ₃);24.29,26.16(3C,piperidine,3CH ₂);55.78,55.41(2C,piperidine,2NCH ₂);61.09(CHOH);65.34(CH ₂N);78.31(NOCH ₂);135.89(2-pyrazine);141.98(C(Cl)=N);148.96(5-pyrazine);149.24(3-pyrazine);152.20(6-pyrazine).MS(ESI)[M+H] ⁺m/z341;Analysis?calculated?for(C ₁₆H ₂₅ClN ₄O ₂):C,56.38;H,7.39;N,16.44.Found:C,56.14;H,7.58;N,16.19.

The another kind of synthetic method (Fig. 3) of embodiment bis-, compound TCO-1

Get the 1a-6 (2.1g, 11.6mmol) having prepared and be dissolved in 20mlNaOH solution, get epoxy chloropropane (2.05ml, 21.5mmol) and be dissolved in a small amount of DMSO, miscible, ice bath stirs and spends the night, and TLC monitors level of response, stopped reaction.Dichloromethane extraction reaction solution (50ml*5 time), column chromatography purification obtains compound 1a-9 (1.64g, 6.96mmol), productive rate 60%.Gained 1a-9 is dissolved in 10ml DMF, adds 80 ° of C of piperidines (1.04ml, 10.3mmol), reaction 3h, TLC monitors level of response, stopped reaction.Ethyl acetate extraction (30ml*5 time), column chromatography purification obtains compound 1a-8(2.12g, 6.6mmol), productive rate 95%.Take Sodium Nitrite (0.57g, 8.3mmol) and be dissolved in 2ml distilled water, salt ice bath is down to-5 ° of C, makes sub-acid sodium solution for subsequent use.The solution that is 1M by concentrated hydrochloric acid dilution, is chilled to-5 ° of C for subsequent use.Get compound 1a-8 (2.12g, 6.6mmol) and be dissolved in the cold hydrochloric acid soln of 13ml, salt ice bath stirs, and then slowly drips wherein sodium nitrite solution reaction 1h, then under condition of ice bath, regulates pH to 10 with chilled NaOH solution (7M).Finally, adopt dichloromethane extraction reaction solution (10ml*5 time), collect organic layer.The sample liquid of underpressure distillation extraction gained, wet method loading column chromatography, moving phase EA:PE:Et ₃n (17:83:0.5), TLC monitors fraction compositions, merges identical cut underpressure distillation dry, and oil pump decompression extracts residual solvent, and obtaining water white transparency oily thing is TCO-1 (1.1g, 3.4mol), productive rate 51.5%.Detect data identical with a upper method.

Synthetic (Fig. 4) of embodiment tri-, compound TCO-2

Get the intermediate 1a-5 (3g, 20.4mmol) having prepared, add 30ml methylene dichloride to dissolve, then add metachloroperbenzoic acid (MCPBA, 3.5g, 20.4mmol) room temperature (20-24 ° of C) to stir 24h, TLC monitors level of response, stopped reaction.Methylene dichloride is removed in underpressure distillation, dry method loading column chromatography.Moving phase EA:PE (1:2), collects cut, and TLC monitors fraction compositions, merges identical cut.Underpressure distillation is dry, obtains white solid 1b-1 (2.8g, 17.2mmol).Productive rate 84%.

By the 1b-1 (2.8g of gained, 17.2mmol) be dissolved in 30ml dehydrated alcohol, add oxammonium hydrochloride 1.5g (21.7mmol), get sodium bicarbonate 1.9g (22.6mmol) and dissolve in 20ml distilled water, miscible, stirring at room temperature reaction 24 hours.TLC monitors level of response, stopped reaction.Underpressure distillation is dry, dry method loading column chromatography, moving phase EA:PE:Et ₃n (80:20:0.5), collects cut, and TLC monitors fraction compositions.Merge identical cut underpressure distillation dry, obtaining white solid is compound 1b-2, productive rate 95%.

Get compound 1b-2 (3.2g, 16.3mmol) and be dissolved in 30ml ethanol, get sodium hydroxide (0.8g, 20mmol) be dissolved in 20ml distilled water, after add quaternary ammonium salt 1a-7 (3.5g, 19.7mmol) miscible, 80 ° of C oil baths are reacted 3 hours, TLC detection reaction degree.Ethyl acetate extractive reaction liquid (30ml*5 time), collects organic layer underpressure distillation dry, wet method loading column chromatography, moving phase EA:PE:Et ₃n (85:15:0.5) wash-out, collects cut, and TLC detects fraction compositions.Merge the underpressure distillation of compound 1b-2 cut and do to obtain transparent oily matter (3.3g, 9.8mmol), productive rate 60%.

Take Sodium Nitrite (1.06g, 12.25mmol) and be dissolved in 3.5ml distilled water, salt ice bath is down to-5 ° of C, makes sub-acid sodium solution for subsequent use.The solution that is 1M by concentrated hydrochloric acid dilution, is chilled to-5 ° of C for subsequent use.Get compound 1b-3 (3.3g, 9.8mmol) and be dissolved in the cold hydrochloric acid soln of 20ml, salt ice bath stirs, and then slowly drips wherein sodium nitrite solution reaction 1h, then under condition of ice bath, regulates pH to 10 with chilled NaOH solution (7M).Finally, adopt dichloromethane extraction reaction solution (15ml*5 time), collect organic layer.The sample liquid of underpressure distillation extraction gained, wet method loading column chromatography, moving phase EA:PE:Et ₃n (20:80:0.5), TLC monitors fraction compositions, merges identical cut underpressure distillation dry, oil pump decompression extracts residual solvent, to obtain white solid after water white transparency oily thing multigelation be TCO-2 (1.8g, 4.9mmol), productive rate 50%. ¹H-NMR(CDCl ₃,300MHz)(ppm)δ:1.23-1.60(m,6H,3CH ₂),2.04-2.41(m,6H,3NCH ₂),2.51(s,3H,CH ₃),2.56(s,3H,CH ₃),2.59(s,3H,CH ₃),4.01-4.35(m,3H,CH?and?NOCH ₂). ¹³C-NMR(CDCl ₃,75.47MHz)(ppm)δ:13.81(CH ₃);14.23(CH ₃);22.57(CH ₃);24.33,26.22(3C,piperidine,3CH ₂);54.78,54.81,60.97(3C,3NCH ₂);65.29(CHOH);78.58(NOCH ₂);134.22(C(Cl)=N);141.48(2-pyrazine);143.11(6-pyrazine);?144.96(3-pyrazine);152.27(5-pyrazine).ESI-MS[M+H] ⁺m/z357;Analysis?calculated?for?(C ₁₆H ₂₅ClN ₄O ₃):C,53.85;H,7.06;N,15.70.Found:C,53.91;H,6.91;N,16.04.

The another kind of synthetic method (Fig. 5) of embodiment tetra-, compound TCO-2

Get the 1b-2 (1.6g, 8.2mmol) having prepared and be dissolved in 10ml NaOH solution, get epoxy chloropropane (1.2ml, 12.6mmol) and be dissolved in a small amount of DMSO, miscible, ice bath stirs and spends the night, and TLC monitors level of response, stopped reaction.Dichloromethane extraction reaction solution (10ml*5 time), column chromatography purification obtains compound 1b-4 (1.23g, 4.9mmol), productive rate 60%.Gained 1b-4 is dissolved in 10ml DMF, adds 80 ° of C of piperidines (0.74ml, 7.3mmol), reaction 3h, TLC monitors level of response, stopped reaction.Ethyl acetate extraction (10ml*5 time), column chromatography purification obtains compound 1b-3(1.57g, 4.65mmol), productive rate 95%.Take Sodium Nitrite (0.4g, 5.8mmol) and be dissolved in 4ml distilled water, salt ice bath is down to-5 ° of C, makes sub-acid sodium solution for subsequent use.The solution that is 1M by concentrated hydrochloric acid dilution, is chilled to-5 ° of C for subsequent use.Get compound 1b-3(1.57g, 4.66mmol) be dissolved in the cold hydrochloric acid soln of 10ml, salt ice bath stirs, and then slowly drips wherein sodium nitrite solution reaction 1h, then under condition of ice bath, regulates pH to 10 with chilled NaOH solution (7M).Finally, adopt dichloromethane extraction reaction solution (10ml*5 time), collect organic layer.The sample liquid of underpressure distillation extraction gained, wet method loading column chromatography, moving phase EA:PE:Et ₃n (20:80:0.5), TLC monitors fraction compositions, merges identical cut underpressure distillation dry, and oil pump decompression extracts residual solvent, and obtaining water white transparency oily thing is TCO-2 (0.86g, 2.3mol), productive rate 51.7%.Detect data identical with a upper method.

A kind of synthetic method of embodiment five, compd B CO-1

Get synthetic midbody compound 1a-3 (TMP-OH, 4g, 26.3mmol), add wherein 79ml ammoniacal liquor (30%) to dissolve, then add elemental iodine 0.67g (2.63mmol), tertbutyl peroxide (TBHP) 8.3ml (57.86mmol) to be miscible in oil bath reaction under 60 ° of C conditions, TLC monitoring level of response after 15h, stopped reaction.Ethyl acetate extraction (30ml*5 time), collected organic layer underpressure distillation is dry.100-200 order silica gel mixed sample, dry method loading column chromatography.Moving phase: Acetone:PE:Et ₃n (20:80:0.5) wash-out, fraction compositions is flowed out in TLC monitoring, merges identical cut underpressure distillation dry, obtains faint yellow solid powder 2a-1 (0.45g, 3.06mmol), productive rate 11.6%.

Get synthetic 2a-1 (0.45g, 3.06mmol) be dissolved in 10ml dehydrated alcohol, then add oxammonium hydrochloride (0.27g, 3.88mmol), take in addition sodium bicarbonate (0.33g, 3.93mmol) be dissolved in 5ml distilled water miscible, 105 ° of C oil baths reaction 3 hours, TLC detection reaction degree, stopped reaction.Underpressure distillation is dry, silica gel mixed sample dry method loading column chromatography, moving phase EA:PE:Et ₃n (80:20:0.5) wash-out, fraction compositions is flowed out in TLC monitoring, merges identical component, and white solid 2a-2 (0.53g, 2.93mmol), productive rate 95.7% are done to obtain in underpressure distillation.

Get compound 2a-2 (0.53g, 2.93mmol) be dissolved in 15ml ethanol, get sodium hydroxide (0.18g, 4.5mmol) be dissolved in 5ml distilled water, after add quaternary ammonium salt 1a-70.65g (3.66mmol) miscible, 80 ° of C oil baths are reacted 3 hours, TLC detection reaction degree.Ethyl acetate extractive reaction liquid (10ml*5 time), collects organic layer underpressure distillation dry, wet method loading column chromatography, moving phase EA:PE:Et ₃n (80:20:0.5) wash-out, collects cut, and TLC detects fraction compositions.Merge the underpressure distillation of compound 2a-3 cut and do to obtain transparent oily matter 2a-3 (0.28g, 0.87mmol), productive rate 30%.

Take Sodium Nitrite (0.075g, 1.09mmol) and be dissolved in 0.5ml distilled water, salt ice bath is down to-5 ° of C, makes sub-acid sodium solution for subsequent use.The solution that is 1M by concentrated hydrochloric acid dilution, is chilled to-5 ° of C for subsequent use.Get compound 2a-3 (0.28g, 0.87mmol) and be dissolved in the cold hydrochloric acid soln of 1.3ml, salt ice bath stirs, and then slowly drips wherein sodium nitrite solution, reaction 1h.Then under condition of ice bath, regulate pH to 10 with chilled NaOH solution (7M).Finally, adopt dichloromethane extraction reaction solution (2ml*5 time), collect organic layer.The sample liquid of underpressure distillation extraction gained, wet method loading column chromatography, moving phase EA:PE:Et ₃n (15:85:0.5), TLC monitors fraction compositions, merges identical cut underpressure distillation dry, and oil pump decompression extracts residual solvent, and obtaining water white transparency oily thing is 2a, BCO-1 (0.063g, 0.0017mol), productive rate 20%. ¹H-NMR(CDCl ₃,300MHz)(ppm)δ:1.42-1.62(m,6H,3CH ₂),2.24-2.66(m,12H,3NCH ₂,2CH ₃),4.02-4.37(m,3H,CH,NOCH ₂). ¹³C-NMR(CDCl ₃,75.47MHz)(ppm)δ:21.75(CH ₃);22.52(CH ₃);24.32,26.20(3C,piperidine,3CH ₂);54.79,54.86(2C,piperidine,2NCH ₂);60.96(CHOH);65.26(CH ₂N);78.60(NOCH ₂);135.17(2-pyrazine,CCl);142.85(5-pyrazine);147.81(C(Cl)=N);149.61(3-pyrazine);150.45(6-pyrazine).ESI-MS[M+H] ⁺m/z361;Analysis?calculated?for(C ₁₅H ₂₂Cl ₂N ₄O ₂):C,49.87;H,6.14;N,15.51.Found:C,50.27;H,6.18;N,15.40.

A kind of synthetic method of embodiment six, compound PCO-1

Get compound 2-itrile group pyrazine (3g, 28.6mmol) be dissolved in 35ml dehydrated alcohol, add oxammonium hydrochloride 2.5g (36.2mmol), get sodium bicarbonate 3.1g (36.9mmol) in 25ml distilled water heating for dissolving, miscible, 105 ° of C oil baths are reacted 3 hours.TLC monitors level of response, stopped reaction.Underpressure distillation is dry, dry method loading column chromatography, moving phase EA:PE:Et ₃n (70:30:0.5), collects cut, and TLC monitors fraction compositions.Merge identical cut underpressure distillation dry, obtaining white solid is compound 3a-1 (3.75g, 27.2mmol, productive rate 95%).

Get compound 3a-1 (3.75g, 27.2mmol) and be dissolved in 30ml ethanol, get sodium hydroxide (1.4g, 35mmol) be dissolved in 30ml distilled water, after add quaternary ammonium salt 1a-75.1g (28.8mmol) miscible, 80 ° of C oil baths reaction 3 hours, TLC detection reaction degree.Ethyl acetate extractive reaction liquid (30ml*5 time), collects organic layer underpressure distillation dry, wet method loading column chromatography, moving phase EA:PE:Et ₃n (85:15:0.5) wash-out, collects cut, and TLC detects fraction compositions.Merge the underpressure distillation of compound 3a-2 cut and do to obtain transparent oily matter (4.5g, 16.1mmol), productive rate 59%.

Take Sodium Nitrite (1.4g, 20.3mmol) and be dissolved in 4ml distilled water, salt ice bath is down to-5 ° of C, makes sub-acid sodium solution for subsequent use.The solution that is 1M by concentrated hydrochloric acid dilution, is chilled to-5 ° of C for subsequent use.Get compound 3a-2 (4.5g, 16.1mmol 13.7mmol) be dissolved in the cold hydrochloric acid soln of 25ml, salt ice bath stirs, and then slowly drips wherein sodium nitrite solution reaction 1h, then under condition of ice bath, regulates pH to 10 with chilled NaOH solution (7M).Finally, adopt dichloromethane extraction reaction solution (15ml*5 time), collect organic layer.The sample liquid of underpressure distillation extraction gained, wet method loading column chromatography, moving phase EA:PE:Et ₃n (20:80:0.5), TLC monitors fraction compositions, merges identical cut underpressure distillation dry, and oil pump decompression extracts residual solvent, and obtaining water white transparency oily thing is 3a, PCO-1 (2.2g, 6.8mol), productive rate 50%. ¹H-NMR(CDCl ₃,300MHz)(ppm)δ:1.35-1.51(m,6H,3CH ₂),2.25-2.38(m,6H,3NCH ₂),3.96-4.40(m,3H,CH,NH ₂). ¹³C-NMR(CDCl ₃,75.47MHz)(ppm)δ:23.99,25.67(3C,piperidine,3CH ₂);58.40(2C,piperidine,2NCH ₂);61.71(CH ₂N),66.09(CHOH);79.36(NOCH ₂);135.09(5-pyrazine);143.75(3-pyrazine);144.30(6-pyrazine);145.11(2-pyrazine);145.76(C(Cl)=N).ESI-MS[M+H] ⁺m/z299;Analysis?calculated?for?(C ₁₃H ₁₉ClN ₄O ₂):C,52.26;H,6.41;N,18.75.Found:C,52.19;H,6.53;N,18.62.

A kind of synthetic method of embodiment seven, compound F 17-hydroxy-corticosterone CO-1a

Get 2,6-lupetidine (10ml, about 8.23g) is dissolved in 25ml methyl alcohol, adds epoxy chloropropane (9ml, about 10g), 80 ° of C oil baths are reacted 10 hours, and underpressure distillation goes out methyl alcohol, adds 25ml water, remove surplus stock by washed with dichloromethane (15ml*3 time), collect water evaporated under reduced pressure and obtain incarnadine solid 5a-1(8.6g, 41.9nmol), productive rate 60%.

Get compound 1a-6 (1.0g, 5.6mmol) and be dissolved in 20ml ethanol, get sodium hydroxide (0.6g, 15mmol) be dissolved in 10ml distilled water, after add quaternary ammonium salt 5a-12.28g (11.1mmol) miscible, 80 ° of C oil baths reaction 3 hours, TLC detection reaction degree.Ethyl acetate extractive reaction liquid (20ml*5 time), collects organic layer underpressure distillation dry, wet method loading column chromatography, moving phase EA:PE:Et ₃n (80:20:0.5) wash-out, collects cut, and TLC detects fraction compositions.Merge the underpressure distillation of compound cut and do to obtain transparent oily matter 5a-2 (1.1g, 3.2mmol), productive rate 59%.

Take Sodium Nitrite (0.43g, 6.3mmol) and be dissolved in 2ml distilled water, salt ice bath is down to-5 ° of C, makes sub-acid sodium solution for subsequent use.The solution that is 1M by concentrated hydrochloric acid dilution, is chilled to-5 ° of C for subsequent use.Get compound 5a-2 (1.1g, 3.2mmol) and be dissolved in the cold hydrochloric acid soln of 10ml, salt ice bath stirs, and then slowly drips wherein sodium nitrite solution reaction 1h, then under condition of ice bath, regulates pH to 10 with chilled NaOH solution (7M).Finally, adopt dichloromethane extraction reaction solution (10ml*5 time), collect organic layer.The sample liquid of underpressure distillation extraction gained, wet method loading column chromatography, moving phase EA:PE:Et ₃n (20:80:0.5), TLC monitors fraction compositions, merges identical cut underpressure distillation dry, and oil pump decompression extracts residual solvent, and obtaining water white transparency oily thing is FCO-1 (0.57g, 1.5mmol), productive rate 50%.ESI-MS[M+H] ⁺m/z369,ESI-MS[M+Na] ⁺m/z391。

A kind of synthetic method of embodiment eight, compound F 17-hydroxy-corticosterone CO-1b

Get compound 1b-2 (1.0g, 5.1mmol) be dissolved in 20ml ethanol, get sodium hydroxide (0.43g, 10.7mmol) be dissolved in 10ml distilled water, after add quaternary ammonium salt 5a-1 (2.00g, 9.75mmol) miscible, 80 ° of C oil baths are reacted 3 hours, TLC detection reaction degree.Ethyl acetate extractive reaction liquid (20ml*5 time), collects organic layer underpressure distillation dry, wet method loading column chromatography, moving phase EA:PE:Et ₃n (80:20:0.5) wash-out, collects cut, and TLC detects fraction compositions.Merge the underpressure distillation of compound cut and do to obtain transparent oily matter 5b-1 (1.12g, 3.07mmol), productive rate 60%.

Take Sodium Nitrite (0.42g, 6.1mmol) and be dissolved in 2ml distilled water, salt ice bath is down to-5 ° of C, makes sub-acid sodium solution for subsequent use.The solution that is 1M by concentrated hydrochloric acid dilution, is chilled to-5 ° of C for subsequent use.Get compound 5b-1 (1.12g, 3.07mmol) and be dissolved in the cold hydrochloric acid soln of 9.2ml, salt ice bath stirs, and then slowly drips wherein sodium nitrite solution reaction 1h, then under condition of ice bath, regulates pH to 10 with chilled NaOH solution (7M).Finally, adopt dichloromethane extraction reaction solution (10ml*5 time), collect organic layer.The sample liquid of underpressure distillation extraction gained, wet method loading column chromatography, moving phase EA:PE:Et ₃n (20:80:0.5), TLC monitors fraction compositions, merges identical cut underpressure distillation dry, and oil pump decompression extracts residual solvent, obtaining water white transparency oily thing is FCO-2 (0.58g, 1.5mmol), productive rate 50%, ESI-MS[M+H] ⁺m/z385, ESI-MS[M+Na] ⁺m/z407.

A kind of synthetic method of embodiment nine, compound F 17-hydroxy-corticosterone CO-2a

Get 3,5-lupetidine (10ml, about 8.23g) is dissolved in 25ml methyl alcohol, adds epoxy chloropropane (9ml, about 10g), 80 ° of C oil baths are reacted 10 hours, and underpressure distillation goes out methyl alcohol, adds 30ml water, remove surplus stock by washed with dichloromethane (15ml*3 time), collect water evaporated under reduced pressure and obtain white solid 6a-1(9.7g, 47.3nmol), productive rate 65%.

Get compound 1a-6 (1.0g, 5.6mmol) and be dissolved in 20ml ethanol, get sodium hydroxide (0.6g, 15mmol) be dissolved in 10ml distilled water, after add quaternary ammonium salt 6a-12.3g (11.2mmol) miscible, 80 ° of C oil baths reaction 3 hours, TLC detection reaction degree.Ethyl acetate extractive reaction liquid (20ml*5 time), collects organic layer underpressure distillation dry, wet method loading column chromatography, moving phase EA:PE:Et ₃n (80:20:0.5) wash-out, collects cut, and TLC detects fraction compositions.Merge the underpressure distillation of compound cut and do to obtain transparent oily matter 6a-2 (1.04g, 2.97mmol), productive rate 54%.

Take Sodium Nitrite (0.41g, 6.0mmol) and be dissolved in 2ml distilled water, salt ice bath is down to-5 ° of C, makes sub-acid sodium solution for subsequent use.The solution that is 1M by concentrated hydrochloric acid dilution, is chilled to-5 ° of C for subsequent use.Get compound 6a-2 (1.04g, 2.97mmol) and be dissolved in the cold hydrochloric acid soln of 8.9ml, salt ice bath stirs, and then slowly drips wherein sodium nitrite solution reaction 1h, then under condition of ice bath, regulates pH to 10 with chilled NaOH solution (7M).Finally, adopt dichloromethane extraction reaction solution (10ml*5 time), collect organic layer.The sample liquid of underpressure distillation extraction gained, wet method loading column chromatography, moving phase EA:PE:Et ₃n (20:80:0.5), TLC monitors fraction compositions, merges identical cut underpressure distillation dry, and oil pump decompression extracts residual solvent, obtaining water white transparency oily thing is FCO-2a (0.58g, 1.5mmol), productive rate 51%, ESI-MS[M+H] ⁺m/z369, ESI-MS[M+Na] ⁺m/z391.

A kind of synthetic method of embodiment ten, compound F 17-hydroxy-corticosterone CO-2b

Get compound 1b-2 (1.0g, 5.1mmol) be dissolved in 20ml ethanol, get sodium hydroxide (0.49g, 12.3mmol) be dissolved in 10ml distilled water, after add quaternary ammonium salt 6a-1 (2.1g, 10.0mmol) miscible, 80 ° of C oil baths are reacted 3 hours, TLC detection reaction degree.Ethyl acetate extractive reaction liquid (20ml*5 time), collects organic layer underpressure distillation dry, wet method loading column chromatography, moving phase EA:PE:Et ₃n (80:20:0.5) wash-out, collects cut, and TLC detects fraction compositions.Merge the underpressure distillation of compound cut and do to obtain transparent oily matter 6b-1 (0.93g, 2.5mmol), productive rate 50%.

Take Sodium Nitrite (0.35g, 5.1mmol) and be dissolved in 2ml distilled water, salt ice bath is down to-5 ° of C, makes sub-acid sodium solution for subsequent use.The solution that is 1M by concentrated hydrochloric acid dilution, is chilled to-5 ° of C for subsequent use.Get compound 6b-1 (0.93g, 2.5mmol) and be dissolved in the cold hydrochloric acid soln of 7.6ml, salt ice bath stirs, and then slowly drips wherein sodium nitrite solution reaction 1h, then under condition of ice bath, regulates pH to 10 with chilled NaOH solution (7M).Finally, adopt dichloromethane extraction reaction solution (10ml*5 time), collect organic layer.The sample liquid of underpressure distillation extraction gained, wet method loading column chromatography, moving phase EA:PE:Et ₃n (20:80:0.5), TLC monitors fraction compositions, merges identical cut underpressure distillation dry, and oil pump decompression extracts residual solvent, obtaining water white transparency oily thing is FCO-2b (0.58g, 1.5mmol), productive rate 50%, ESI-MS[M+H] ⁺m/z385, ESI-MS[M+Na] ⁺m/z407.

A kind of synthetic method of embodiment 11, compound F 17-hydroxy-corticosterone CO-3a

Get Diisopropylamine (10ml, about 7g) be dissolved in 25ml methyl alcohol, add epoxy chloropropane (10ml, about 11.7g), 80 ° of C oil baths are reacted 10 hours, underpressure distillation goes out methyl alcohol, add 30ml water, remove surplus stock by washed with dichloromethane (20ml*3 time), collect water evaporated under reduced pressure and obtain white solid 7a-1(7.4g, 38.3nmol), productive rate 56%.

Get compound 1a-6 (1.0g, 5.6mmol) and be dissolved in 20ml ethanol, get sodium hydroxide (0.6g, 15mmol) be dissolved in 10ml distilled water, after add quaternary ammonium salt 5a-12.14g (11.1mmol) miscible, 80 ° of C oil baths reaction 3 hours, TLC detection reaction degree.Ethyl acetate extractive reaction liquid (20ml*5 time), collects organic layer underpressure distillation dry, wet method loading column chromatography, moving phase EA:PE:Et ₃n (80:20:0.5) wash-out, collects cut, and TLC detects fraction compositions.Merge the underpressure distillation of compound cut and do to obtain transparent oily matter 7a-2 (1.04g, 3.08mmol), productive rate 60%.

Take Sodium Nitrite (0.43g, 6.3mmol) and be dissolved in 2ml distilled water, salt ice bath is down to-5 ° of C, makes sub-acid sodium solution for subsequent use.The solution that is 1M by concentrated hydrochloric acid dilution, is chilled to-5 ° of C for subsequent use.Get compound 7a-2 (1.04g, 3.08mmol) and be dissolved in the cold hydrochloric acid soln of 9.2ml, salt ice bath stirs, and then slowly drips wherein sodium nitrite solution reaction 1h, then under condition of ice bath, regulates pH to 10 with chilled NaOH solution (7M).Finally, adopt dichloromethane extraction reaction solution (10ml*5 time), collect organic layer.The sample liquid of underpressure distillation extraction gained, wet method loading column chromatography, moving phase EA:PE:Et ₃n (20:80:0.5), TLC monitors fraction compositions, merges identical cut underpressure distillation dry, and oil pump decompression extracts residual solvent, and obtaining water white transparency oily thing is FCO-3a (0.42g, 1.2mmol), productive rate 38%.ESI-MS[M+H] ⁺m/z357,ESI-MS[M+Na] ⁺m/z379。

A kind of synthetic method of embodiment 12, compound F 17-hydroxy-corticosterone CO-3b

Get compound 1b-2 (1.0g, 5.1mmol) be dissolved in 20ml ethanol, get sodium hydroxide (0.43g, 10.7mmol) be dissolved in 10ml distilled water, after add quaternary ammonium salt 7a-1 (1.97g, 10.2mmol) miscible, 80 ° of C oil baths are reacted 3 hours, TLC detection reaction degree.Ethyl acetate extractive reaction liquid (20ml*5 time), collects organic layer underpressure distillation dry, wet method loading column chromatography, moving phase EA:PE:Et ₃n (80:20:0.5) wash-out, collects cut, and TLC detects fraction compositions.Merge the underpressure distillation of compound cut and do to obtain transparent oily matter 7b-1 (0.83g, 2.35mmol), productive rate 46%.

Take Sodium Nitrite (0.32g, 4.7mmol) and be dissolved in 2ml distilled water, salt ice bath is down to-5 ° of C, makes sub-acid sodium solution for subsequent use.The solution that is 1M by concentrated hydrochloric acid dilution, is chilled to-5 ° of C for subsequent use.Get compound 7b-1 (0.83g, 2.35mmol) and be dissolved in the cold hydrochloric acid soln of 7.1ml, salt ice bath stirs, and then slowly drips wherein sodium nitrite solution reaction 1h, then under condition of ice bath, regulates pH to 10 with chilled NaOH solution (7M).Finally, adopt dichloromethane extraction reaction solution (10ml*5 time), collect organic layer.The sample liquid of underpressure distillation extraction gained, wet method loading column chromatography, moving phase EA:PE:Et ₃n (20:80:0.5), TLC monitors fraction compositions, merges identical cut underpressure distillation dry, and oil pump decompression extracts residual solvent, obtaining water white transparency oily thing is FCO-6 (0.304g, 0.8mmol), productive rate 34.7%, ESI-MS[M+H] ⁺m/z373, ESI-MS[M+Na] ⁺m/z395.

A kind of synthetic method of embodiment 13, compd A CO-1a

Get diamantane 10g (73.5mmol), drip bromine 23.5g (147.0mmol), within 1 hour, be slowly warmed up to 70 ° of C, react 6 hours, hold over night, dripping saturated bisulfite receives, until solution becomes colorless, ethyl acetate extraction (30ml*3), get organic phase concentrating under reduced pressure, the sample liquid of underpressure distillation extraction gained, wet method loading column chromatography, moving phase is sherwood oil, TLC monitors fraction compositions, merge identical cut underpressure distillation dry, oil pump decompression extracts residual solvent, obtaining colourless transparent oil liquid is 8a-1(11.0g, 51.4mmol), productive rate 70%.

Get compound 8a-1(11.0g, 51.4mmol), add N-METHYLFORMAMIDE (10g, 169.4mmol), 170 ° of C reflux 2 hours, are cooled to 100 ° of C, add 10% hydrochloric acid 30ml, 100 ° of C stir 2 hours, cooling, toluene wash (20ml*3), in water, add gac 1g, 70 ° of C stir 30 minutes, cooling, filter, saturated sodium hydroxide regulates pH to 12, anhydrous diethyl ether extraction (30ml*3), underpressure distillation evaporate to dryness obtains white solid 8a-2(6.6g, 40mmol), productive rate 78%.

Get 8a-2(6.6g, 40mmol) be dissolved in 50ml methyl alcohol, add epoxy chloropropane (4.6g, 80mmol), 80 ° of C oil baths are reacted 10 hours, underpressure distillation goes out methyl alcohol, add 50ml water, remove surplus stock by washed with dichloromethane (25ml*3 time), collect water evaporated under reduced pressure and obtain white solid 8a-3(4.4g, 17.2mmol), productive rate 43%.

Get compound 1a-6 (1.0g, 5.6mmol) and be dissolved in 20ml ethanol, get sodium hydroxide (0.6g, 15mmol) be dissolved in 10ml distilled water, after add quaternary ammonium salt 8a-3 (2.85g, 11.1mmol) miscible, 80 ° of C oil baths are reacted 3 hours, TLC detection reaction degree.Ethyl acetate extractive reaction liquid (20ml*5 time), collects organic layer underpressure distillation dry, wet method loading column chromatography, moving phase EA:PE:Et ₃n (67:33:0.5) wash-out, collects cut, and TLC detects fraction compositions.Merge the underpressure distillation of compound cut and do to obtain transparent oily matter 8a-4 (1.1g, 2.7mmol), productive rate 50%.

Take Sodium Nitrite (0.37g, 5.4mmol) and be dissolved in 2ml distilled water, salt ice bath is down to-5 ° of C, makes sub-acid sodium solution for subsequent use.The solution that is 1M by concentrated hydrochloric acid dilution, is chilled to-5 ° of C for subsequent use.Get compound 8a-4 (1.1g, 2.7mmol) and be dissolved in the cold hydrochloric acid soln of 8.5ml, salt ice bath stirs, and then slowly drips wherein sodium nitrite solution reaction 1h, then under condition of ice bath, regulates pH to 10 with chilled NaOH solution (7M).Finally, adopt dichloromethane extraction reaction solution (10ml*5 time), collect organic layer.The sample liquid of underpressure distillation extraction gained, wet method loading column chromatography, moving phase EA:PE:Et ₃n (20:80:0.5), TLC monitors fraction compositions, merges identical cut underpressure distillation dry, and oil pump decompression extracts residual solvent, obtaining water white transparency oily thing is ACO-1a (0.44g, 1.1mmol), productive rate 38%, ESI-MS[M+H] ⁺m/z421, ESI-MS[M+Na] ⁺m/z443. ¹H-NMR(CDCl ₃,300MHz)(ppm)δ:1.49-1.68(m,12H,CH ₂),1.98(s,3H,CH),2.27(s,3H,NCH ₃),2.47(s,6H,2CH ₃),2.56(s,3H,CH ₃),2.14-2.18,2.63-2.73(m,2H,OCH ₂),3.84-3.92(m,1H,OCH),4.21-4.26(m,2H,OCH ₂). ¹C-NMR(CDCl ₃,300MHz)(ppm)δ:21.47(CH ₃),21.87(CH ₃),22.30(CH ₃),29.69?(3CH),34.25(3CH ₂),36.68(3NCH ₂),38.96(NCH ₃),51.46(OCH ₂),56.44(NCH ₂),65.44(HOCH),78.26(NOCH ₂),135.67(2-pyrazine);141.91(C(Cl)=N);148.86(5-pyrazine);149.11(3-pyrazine);152.35(6-pyrazine).

A kind of synthetic method of embodiment 14, compd A CO-1b

Get compound 1b-2 (1.0g, 5.1mmol) be dissolved in 20ml ethanol, get sodium hydroxide (0.43g, 10.7mmol) be dissolved in 10ml distilled water, after add quaternary ammonium salt 8a-3 (2.6g, 10.2mmol) miscible, 80 ° of C oil baths are reacted 3 hours, TLC detection reaction degree.Ethyl acetate extractive reaction liquid (20ml*5 time), collects organic layer underpressure distillation dry, wet method loading column chromatography, moving phase EA:PE:Et ₃n (80:20:0.5) wash-out, collects cut, and TLC detects fraction compositions.Merge the underpressure distillation of compound cut and do to obtain transparent oily matter 8b-1 (0.95g, 2.29mmol), productive rate 45%.

Take Sodium Nitrite (0.31g, 4.58mmol) and be dissolved in 2ml distilled water, salt ice bath is down to-5 ° of C, makes sub-acid sodium solution for subsequent use.The solution that is 1M by concentrated hydrochloric acid dilution, is chilled to-5 ° of C for subsequent use.Get compound 8b-1 (0.95g, 2.29mmol) and be dissolved in the cold hydrochloric acid soln of 7.0ml, salt ice bath stirs, and then slowly drips wherein sodium nitrite solution reaction 1h, then under condition of ice bath, regulates pH to 10 with chilled NaOH solution (7M).Finally, adopt dichloromethane extraction reaction solution (10ml*5 time), collect organic layer.The sample liquid of underpressure distillation extraction gained, wet method loading column chromatography, moving phase EA:PE:Et ₃n (20:80:0.5), TLC monitors fraction compositions, merges identical cut underpressure distillation dry, and oil pump decompression extracts residual solvent, obtaining water white transparency oily thing is ACO-1b (0.8g, 1.8mmol), productive rate 36%, ESI-MS[M+H] ⁺m/z437, ESI-MS[M+Na] ⁺m/z459. ¹H-NMR(CDCl ₃,300MHz)(ppm)δ:1.61-1.76(m,12H,6CH ₂),2.09(s,3H,3CH),2.23(s,3H,CH ₃),2.26-2.34,2.73-2.81(m,2H,CH ₂),2.52(s,3H,CH ₃),2.57(s,3H,CH ₃),2.70(s,3H,CH ₃),3.09-3.98(m,1H,CHOH),4.33-4.34(d,2H,OCH ₂). ¹C-NMR(CDCl ₃,300MHz)(ppm)δ:13.64(CH ₃),14.04(CH ₃),22.39(CH ₃),29.48(3CH),33.57(3CH ₂),36.66(3NCH ₂),38.95(NCH ₃),50.93(OCH ₂),54.10(NCH ₂),65.40(HOCH),82.17(NOCH ₂),133.83(2-pyrazine);141.66(C(Cl)=N);142.91(5-pyrazine);144.79(3-pyrazine);152.08(6-pyrazine).

Embodiment 15, compound TCO-1, TCO-2, PCO-1, the provide protection (Fig. 6) of the cell of BCO-1 to MG-132 induction damage

10000, every hole SY5Y cell, first protects 1h with compound, and then adding final concentration is that the MG-132 of 15 μ Μ is hatched 24h.Adopt bimoclomol, arimoclomol, and TMP is as positive control, and DMSO contrasts as solvent, and mtt assay detects cytoactive.

Result shows the compound TCO-1 that we are synthetic, and TCO-2 can rely on ground neuroprotective cellular stress damage by dosage, and the biological activity of TCO-1 and the bimoclomol of CytRx company and the activity of arimoclomol are suitable; And the activity of TCO-2 is obviously better than bimoclomol and arimoclomol.#P<0.01versus?control?group.*P<0.05versus?MG-132group.**P<0.01versus?MG-132group.Ctrl:control,Ari:arimoclomol,Bim:bimoclomol.

Embodiment 16, compound TCO-1, TCO-2, PCO-1, the restraining effect (Fig. 7) of BCO-1 to ROS

Oxidation stimulation can cause cell in stress situation, and protein expression is abnormal, may produce wrong folding or gathering, causes the generation of nerve degenerative diseases.First we damage by tertbutyl peroxide induced oxidation, then detect each compound with oxidation resistance ability with the compound protection of different concns gradient.10000, every hole SY5Y cell, first protects 1h with compound, and then adding final concentration is that the TBHP of 150 μ Μ is hatched 4h.Adopt bimoclomol, arimoclomol, and TMP is as positive control, and DMSO contrasts as solvent, and mtt assay detects cytoactive.Experimental result shows: positive reference substance bimoclomol, and arimoclomol, TMP is all without antioxygenation, and new synthetic compound TCO-1, TCO-2 has obvious antioxygenation.#P<0.01versus?control?group.*P<0.05versus?t-BHP?group.

Embodiment 17, compound TCO-1, TCO-2, PCO-1 is to MPP ⁺the provide protection (Fig. 8) of the cell of induction damage

10000, every hole SY5Y cell, first protects 1h with compound, then adds the MPP of final concentration 2m Μ ⁺hatch 24h.Adopt bimoclomol, arimoclomol, and clinical antiparkinsonism drug selegiline (MAO-B) inhibitor is as positive control, and DMSO contrasts as solvent, and mtt assay detects cytoactive.Preliminary experimental results show: positive reference substance bimoclomol, arimoclomol, selegiline except the arimoclomol of 10 μ M all without nonreactive MPP ⁺the cells apoptosis of induction, and new synthetic compound TCO-1, TCO-2 is at 1 μ M, and 10 μ M all have obvious anti-apoptotic effect.#P<0.01versus?control?group.*P<0.05versus?MPP ⁺group.Sel:Selegiline.

Embodiment 18, compound TCO-1, TCO-2 suppresses the gathering (Fig. 9) of ubiquitin protein

TCO-1, TCO-2 can regulate HSPs protein expression under heat-shocked state; And in MG-132 stress cell model, the TCO-1 of 100 μ M and TCO-2 can obviously remove the gathering of ubiquitin protein.

Embodiment 19, compound TCO-1 are to A β _1-42the provide protection of the AD rat model of induction

Injection A β _1-42manufacture rat Senlie dementia model, then, with synthetic chlorine 9 oxime derivate TCO-1 treatment, investigate the provide protection of compound to rat.Adopt clinical anti senile dementia drug E2020 (donepezil, DPH) as positive control, physiological saline group is Normal group.Morris water maze system detects study of behaviour index, the provide protection size of observing new synthetic compound.First, rat intracerebroventricular injection A β _1-42after (10 μ g/ only), observing each group of rats'swimming speed does not have obvious significant difference, illustrates that the state of rat is identical.

Rat intracerebroventricular injection A β _1-42after (10 μ g/ only), arrive for the first time time (latent period) significant prolongation of platform, * P<0.01versus A β group.Positive control drug E2020 (3mg/kg) can resist A β _1-42effect, obviously shortening arrives the time of platform, * P<0.01versus A β group for the first time.The TCO-1 testing can significantly shorten the time (latent period) that rat arrives platform for the first time, * P<0.01versus A β group (Figure 10).

Rat intracerebroventricular injection A β _1-42after (10 μ g/ only), the number of times of spanning platform obviously reduces, * P<0.01versusA β group.DPH (3mg/kg) can resist A β _1-42effect, obviously increase the number of times of spanning platform, * P<0.01versus A β group.New synthetic chlorine 9 oxime derivate TCO-1 can significantly increase the number of times of spanning platform, * P<0.01versus A β group (Figure 11).

Above-mentioned to specific descriptions of the present invention clear illustrated this new compound to protein folding mistake or the abnormal disease forming of assembling as nerve degenerative diseases, the heart, cerebrovascular disease, the treatment of metabolic trouble and inflammation or prevention provide unique method.The above-mentioned detailed description about specific embodiment, only conduct illustrates the example of technical solution of the present invention, and does not limit the scope of the claims in the present invention.Particularly point out, inventor is through scrupulous consideration, and replacement, change and the modification that the present invention is different do not depart from the defined the connotation and extension of the claims in the present invention.

Claims

1. chlorine oxime compounds and a pharmacy acceptable salt thereof, described chlorine oxime compounds has the structure of following general formula I:

Wherein:

R ₁, R ₂, R ₃, R ₄identical or different, be independently H separately, OH, NH ₂, COOH, straight or branched alkyl, replaces or unsubstituted monocycle or multi-ring alkyl, and aryl, containing the heterocyclic base of O or S, nitrogenous heterocyclic base and N → O group thereof, acyl group, ester group, or amido, R ₂with R ₃also can form and replace or unsubstituted 4-8 ring with N atom;

Work as R ₄during for H, R ₁be not H, replace or unsubstituted naphthyl pyridyl, or thienyl;

N is 0,1,2 or 3.

2. chlorine oxime compounds and pharmacy acceptable salt thereof according to claim 1, described chlorine oxime compounds has the structure of following general formula I I:

Wherein:

R ₂, R ₃, R ₄identical or different, be independently H separately, OH, NH ₂, COOH, straight or branched alkyl, replaces or unsubstituted monocycle or multi-ring alkyl, and aryl, containing the heterocyclic base of O or S, nitrogenous heterocyclic base and N → O group thereof, acyl group, ester group, or amido; R ₂with R ₃also can form and replace or unsubstituted 4-8 ring with N atom;

R ₅, R ₆, R ₇identical or different, be independently H separately, F, Cl, Br, I, OH, NH ₂, COOH, straight or branched alkyl, aryl, containing the heterocyclic base of O, N or S, acyl group, ester group, or amido;

At least one is N atom for X and Y, and when X and Y only one be the N atomic time, R ₄be not H; Above N atom, be connected with oxygen or do not connect oxygen;

N is 0,1,2 or 3.

3. chlorine oxime compounds and pharmacy acceptable salt thereof according to claim 2, described chlorine oxime compounds has the structure of following general formula III:

Wherein:

R ₂, R ₃, R ₄identical or different, be independently H separately, OH, NH ₂, COOH, straight or branched alkyl, replaces or unsubstituted monocycle or multi-ring alkyl, and aryl, containing the heterocyclic base of O or S, nitrogenous heterocyclic base and N → O group thereof, acyl group, ester group, or amido, R ₂with R ₃also can form and replace or unsubstituted 4-8 ring with N atom;

Above pyrazine ring N atom, be connected with oxygen or do not connect oxygen;

N is 0,1,2 or 3.

4. chlorine oxime compounds and pharmacy acceptable salt thereof according to claim 3, is characterized in that in general formula III: R ₂be selected from methyl, ethyl, n-propyl, sec.-propyl, R ₃be selected from 3-methyl adamantane base, 3,5-dimethyladamantane base, sec.-propyl, or R ₂, R ₃connect into and replace or unsubstituted six-ring with N atom;

R ₄be selected from H, OH, NH ₂, COOH, straight or branched alkyl, replaces or unsubstituted monocycle or multi-ring alkyl, and aryl, containing the heterocyclic base of O or S, nitrogenous heterocyclic base and N → O group thereof, acyl group, ester group, amido;

Above pyrazine ring N atom, be connected with oxygen or do not connect oxygen;

N is 0,1,2 or 3.

5. chlorine oxime compounds and pharmacy acceptable salt thereof according to claim 4, described chlorine oxime compounds has the structure of following general formula I V:

Wherein:

R ₄for H, OH, NH ₂, COOH, straight or branched alkyl, replaces or unsubstituted monocycle or multi-ring alkyl, and aryl, containing the heterocyclic base of O or S, nitrogenous heterocyclic base and N → O group thereof, acyl group, ester group, or amido;

R ₈be selected from H, the straight or branched alkyl of C1-C4;

Above pyrazine ring N atom, be connected with oxygen or do not connect oxygen;

N is 0,1,2 or 3.

6. chlorine oxime compounds and pharmacy acceptable salt thereof according to claim 5, described chlorine oxime compounds has the structure of following general formula V:

Wherein:

Above pyrazine ring N atom, be connected with oxygen or do not connect oxygen.

N is 0,1,2 or 3.

7. chlorine oxime compounds and pharmacy acceptable salt thereof according to claim 6, described chlorine oxime compounds has the structure of following general formula VI:

Wherein:

Above pyrazine ring N atom, be connected with oxygen or do not connect oxygen;

N is 0,1,2 or 3.

8. chlorine oxime compounds and pharmacy acceptable salt thereof according to claim 7, is characterized in that in general formula VI, n is 1, on pyrazine ring N atom, is not connected with O, and described chlorine oxime compounds has the structure as shown in the formula TCO-1:

。

9. chlorine oxime compounds and pharmacy acceptable salt thereof according to claim 7, is characterized in that in general formula VI, n is 1, on pyrazine ring N atom, is connected with O, and described chlorine oxime compounds has the structure as shown in the formula TCO-2:

。

10. according to the preparation method of the chlorine oxime compounds described in claim 1-9 any one, it comprises: will after the reaction of the itrile group derivative of starting raw material and oxammonium hydrochloride, make the amino amino oxime compounds replacing again with corresponding reactant salt, more described amino oxime compounds is carried out to diazotization chlorination make described chlorine oxime compounds.

11. according to the chlorine oxime compounds described in claim 1-9 any one and the pharmaceutical composition that contains described compound thereof in the purposes of preparing in medicine, described medicine is used for the disease of prevention or treatment and comprises cranial nerve or cerebrovascular disease, cardiovascular disorder, inflammation infection disease and ophthalmic diseases.

12. purposes of preparing in medicine according to claim 11, wherein said cranial nerve or cerebrovascular disease comprise alzheimer (family name) disease, parkinsonism, Huntington's disease, prion disease, encephalomalacia disease, lethal familial insomnia, the mankind's striatum spinal cord degeneration disease, scrapie, mad cow disease, amyotrophic lateral sclerosis (spinal cord) lateral sclerosis, dementia, multiple sclerosis, chronic pain class encephalopathic, cerebral trauma, epilepsy, apoplexy, brain of neonatal rat on ischemia hypoxia, acquired immune deficiency syndrome (AIDS).

13. purposes of preparing in medicine according to claim 11, wherein said cardiovascular disorder comprise cardiopulmonary effluent, pulmonary emphysema, respiratory distress syndrome, heart ischemia or pour into again, toxic shock syndrome, adult respiratory distress syndrome, emaciation, myocarditis, atherosclerosis, coronary heart disease and sudden heart disease class heart and vascular conditions.

14. purposes of preparing in medicine according to claim 11, wherein said inflammation infection disease comprises inflammatory bowel, diabetes, rheumatoid arthritis, asthma, liver cirrhosis, allosome repulsion, encephalomyelitis, brain (ridge) film inflammation, pancreatitis, peritonitis, vasculitis (disease), lymphocytic choriomeningitis, choriomeningitis, glomerulonephritis, systemic lupus erythematosus, gastrointestinal peristalsis disorder, obesity, hungry disease, hepatitis and renal failure.

15. purposes of preparing in medicine according to claim 11, wherein said ophthalmic diseases comprises diabetic retinopathy, uveitis, glaucoma, blepharitis, chalazion, allergic ophthalmopathy, keratohelcosis, keratitis, cataract, senile macula lutea degenerative change and optic neuritis.