CN101870692B

CN101870692B - Novel piperazinyl pyridazine compounds

Info

Publication number: CN101870692B
Application number: CN200910031106A
Authority: CN
Inventors: 胡文辉; 廖升荣; 杨玲; 徐宏江
Original assignee: Jiangsu Chia Tai Tianqing Pharmaceutical Co Ltd; Guangzhou Institute of Biomedicine and Health of CAS
Current assignee: Guangzhou Institute of Biomedicine and Health of CAS
Priority date: 2009-04-27
Filing date: 2009-04-27
Publication date: 2012-09-05
Anticipated expiration: 2029-04-27
Also published as: CN101870692A

Abstract

The invention relates to novel piperazinyl pyridazine compounds in a formula I or salts thereof, a preparation method thereof and use thereof as nervous inflammation inhibitors in treating or preventing neurodegenerative diseases such as senile dementia, Parkinson's disease, cognitive impairment and the like or treating diseases such as mental disorder, epilepsy, convulsions, stroke and the like.The compounds have the advantages of simple preparation process, readily available raw materials and suitability for industrial large-scale production. In addition, through the verification of in-vitro experiments, the compounds have the effect of inhibiting BV2 from secreting IL-1 beta efficiently, and the effect is superior to that of positive medicaments; and simultaneously, the compound have no inhibiting effect on the secretion of the IL-1 beta by C6, and thus the compounds have high safety.

Description

A kind of novel piperazinyl pyridazine compounds

Technical field

The present invention relates to a kind of novel piperazinyl pyridazine compounds, its preparation method, comprise the compsn of such material; And this compounds is as neural inflammation inhibitor for treating or prevention nerve degenerative diseases; Like the purposes of senile dementia, parkinson's disease, cognitive disorder etc., or the purposes of diseases such as treatment psychosis, epilepsy, convulsions or apoplexy.

Background technology

Senile dementia be also referred to as Alzheimer (Alzheimer ' s disease, AD), be a kind of be master's central nervous system degenerative disease with the infringement of carrying out property cognitive disorder and memory capability.The clinical manifestation of this disease is the recent memory dysfunction, is persistence hypophrenia, the forfeiture of judgement inferential capability, aphasia, dyspraxia etc. subsequently.Its pathological characters be a large amount of senile plaque (senile plaques, SPs) and neurofibrillary tangles (neurofibrilary tangles, NFTs).At present on the market medicine mainly contains two kinds: cholinesterase inhibitor (AChEI) and nmda receptor antagonist, the former is like tacrine (tacrine), E2020 (donepezil), sharp this bright (rivastigmine) and selagine (Huperzine A).Wherein, selagine is the original new drug of China Chinese Academy of Sciences institute of materia medica's independent research, is by a kind of new alkaloids that is separated in the plants of Huperzia Herba Lycopodii serrati; Nmda receptor antagonist such as DMAA (memantine, Namenda) be a kind ofly be used for treating, the medicine of severe senile dementia, gone on the market by drugs approved by FDA in 2003.

Find that at present the mechanism of action that is used for anti senile dementia drug mainly comprises: through the activity of acetylcholine esterase inhibition, reduce the degraded of vagusstoff, improve cholinergic level in the brain and the acetylcholinesterase depressant set up; Through suppressing each inner action site of nmda receptor surface or ionic channel, reduce the hormesis of nmda receptor, suppress its active and nmda receptor antagonist that set up.Occur a kind of popular and generally acknowledged mechanism of action in recent years and be " amyloid beta theory (amyloid-β hypothesis; A β) "; This mechanism thinks that the APP amyloid protein precursor is hydrolyzed the insoluble amyloid beta of fracture back formation and gathers formation senile plaque (amyloid plaques), has caused a series of pathological change thus, comprises neural inflammation; Neural cell loss and death etc. finally cause senile dementia.According to this machine-processed flow process, just can reach the purpose that alleviates or treat to change the process of disease through each step in the blocking-up circulation.At present according to the suppressor factor of the various new drugs of this thinking exploitation such as beta-hydrolase inhibitor, gama-hydrolase inhibitor, vaccine, tau phosphorylation, neuritis disease etc., these medicines overwhelming majority is based on the amyloid beta theory.

The block nerves inflammation has been proved the vicious cycle that can stop senile dementia effectively, and this experiment has obtained checking on cell and mouse model, and relevant compound gets in the clinical study.(with reference to Bioorganic & Medicinal Chemistry Letters17 (2007) 414-418) discloses the activity data of minozac and following formula II compound in the article like people such as WenhuiHu; The two has got into the clinical study stage at present because demonstrate good activity.The invention provides differently with these two kinds of compound structures, but effect is close or the compound of better anti-senile dementia.

(minozac)

(formula II)

Summary of the invention

One aspect of the present invention provides the compound or its salt of a kind of formula I:

formula I

Wherein, R ¹Be selected from low alkyl group or heteroaryl; R ³, R ⁴Be selected from hydrogen or low alkyl group respectively, or R ³And R ⁴Combine and form five yuan or six-membered carbon ring; R ²Be selected from replacement or unsubstituted heteroaryl, substituting group is selected from low alkyl group, lower alkoxy; N is selected from 0 or 1.

Wherein, R ¹Preferable methyl or pyrimidyl;

R ³, R ⁴Preferred hydrogen or methyl, perhaps R ³And R ⁴Five yuan or hexa-atomic aromatic carbocyclic combine; R ³, R ⁴Further preferred hydrogen or methyl or R ³And R ⁴Be combined together to form phenyl ring;

R ²Preferred replace or unsubstituted benzo is thick and heteroaryl or pyrimidine; Further preferred replacement or unsubstituted benzo nitrogenous or sulfur heteroatom thick and heteroaryl or pyrimidyl; Further be preferably replacement or unsubstituted thionaphthene, benzopyrrole, quinoline, pyrimidine or 2, the 4-dimethyl pyrimidine;

N preferred 0.

Formula I compound provided by the invention is preferably: R ¹Be selected from methyl or pyrimidyl, R ³, R ⁴Be selected from hydrogen or methyl respectively, perhaps R ³, R ⁴Be combined together to form phenyl ring, R ²Be selected from thionaphthene, benzopyrrole, quinoline, pyrimidine or 2,4-dimethyl pyrimidine, n are 0.

" low alkyl group " of the present invention is meant the saturated alkyl of the straight or branched of being made up of 1-4 carbon atom, and concrete example includes but not limited to methyl, ethyl, propyl group, sec.-propyl, butyl, sec.-butyl, isobutyl-, the tertiary butyl etc.

" heteroaryl " of the present invention is meant the monocycle or the bicyclic aromatic group of 5-10 atom, wherein contains 1 heteroatoms that is selected from N, O or S at least, and remaining atom is C, in addition, also has the πDian Zi system of total conjugated.The example of heteroaryl includes but not limited to thionaphthene, benzopyrrole, isoquinoline 99.9, quinoline, pyrimidine, pyridazine, indoles, quinazoline, pyrroles, thiophene, indazole, pyrazoles, pyridine, furans, imidazoles, pyrazine, thiazole, benzothiazole, naphthyridine etc.Heteroaryl can be substituted or unsubstituted.

Above-mentioned heterocycle hepyramine compounds provided by the invention can exist with the form of its salt, hydrate; They are converted into The compounds of this invention in vivo; For example, within the scope of the invention, according to technology well known in the art; The compounds of this invention is converted into the form of pharmacy acceptable salt, and uses them with salt form.

When The compounds of this invention possesses the form of free alkali; Make the free alkali form and the pharmaceutically acceptable inorganic or organic acid reaction of compound; The acid salt that can prepare The compounds of this invention, these salt include but not limited to: hydrochloride, hydrobromate, hydriodate, phosphoric acid salt, vitriol, nitrate salt, esilate, tosylate and benzene sulfonate, acetate, PHENRAMINE MALEATE, tartrate, SUMATRIPTAN SUCCINATE, Citrate trianion, benzoate, ascorbate salt and salicylate, malonate, adipate, hexanoate, arginic acid salt, fumarate, nicotinate, phthalate, oxalate etc.

When The compounds of this invention possesses the form of free acid; Make its free acid form and the reaction of pharmaceutically acceptable inorganic or organic bases can prepare the base addition salt of The compounds of this invention; This type salt includes but not limited to: lithium, sodium, potassium, barium, calcium, magnesium, aluminium, iron, ferrous, copper, zinc salt, or the salt of forming with morpholine, diethylamine, triethylamine, Isopropylamine, Trimethylamine 99, Methionin or histidine.

Compound or its salt provided by the invention specifically is exemplified below, but is not limited to following compounds or its salt:

Further aspect of the present invention provides formula I the preparation method of compound, comprises the steps:

formula I

Wherein, R ¹, R ², R ³, R ⁴, n is identical with the definition among the above-mentioned formula I.The compound of employed raw material formula e can adopt method well known in the art to prepare, such as, can obtain through following reactions step:

The compound of formula c can have been bought from market, perhaps adopts the method for knowing in this area to prepare, such as, can obtain through following reactions step:

Formula I compound provided by the invention can adopt following method to prepare particularly:

Wherein, R ¹, R ², R ³, R ⁴, n is identical with the definition among the above-mentioned formula I

The concrete compound method of compound provided by the invention is following:

1) is that 1: 1～1.5 formula a compound and Hydrazine Hydrate 80 are dissolved in the polar organic solvent with mol ratio, is heated to 80～120 ℃, reacted 2～6 hours; Slowly cooling then, concentrating under reduced pressure, resulting resistates add water and stir; Separate out white solid, filtration, washing, drying obtain formula b compound; This compound need not be further purified, and directly is used for next step reaction.

2) be that 1: 1.5～3 formula b compound and POCl3 are mixed with mol ratio, be heated to 100～150 ℃, reacted 2～4 hours; After reaction is accomplished, be chilled to room temperature, slowly pour in the trash ice and vigorous stirring 0.5～1 hour; Extraction is preferably used dichloromethane extraction 3 times then, merges organic phase; Dry, concentrated, resistates obtains formula c compound through column chromatography for separation.

3) be that 1: 1.5～4 formula c compound and Glacial acetic acid min. 99.5 are mixed with mol ratio, reflux 2～6 hours is after reaction is accomplished; Concentrating under reduced pressure, resistates is handled the compound that obtains formula d-1 and formula d-2, preferably adds water treatment; Ethyl acetate extraction 3 times; Merge organic layer, drying, filtration, filtrating concentrate, and get the simplification compound through column chromatography for separation.

4) be to splash in the dichloromethane solution of formula d-1 compound under 1.5～2: 1 the trifluoromethanesulfanhydride anhydride ice bath and insulation reaction 2～6 hours with mol ratio, logical nitrogen protection.After the reaction completion, handle obtaining formula e compound, preferably with washing 3 times, organic phase is dry, concentrated, and resistates is through column chromatography for separation.

5) be 1: 1.5～3 formula e compound, formula f compound and an amount of exsiccant N with mol ratio; Dinethylformamide is mixed, stirring at room 2～6 hours, and the compound that aftertreatment obtains formula g is accomplished in reaction; Preferably reactant directly is added to the water, leaches solid, the drying separated out.

6) be 1: 1: 4 with mol ratio: the heteroaryl boric acid of the compound of 0.03～0.08 formula g, formula h, Anhydrous potassium carbonate and triphenyl phosphatization palladium are woven in an amount of glycol dimethyl ether; Logical nitrogen protection, 100～150 ℃ were reacted 16～28 hours, and the compound that aftertreatment obtains formula I is accomplished in reaction; Preferably reactant is chilled to room temperature; Diatomite filtration, filtrating concentrates, and resistates is through column chromatography for separation.

Among the above-mentioned preparation method, the polar organic solvent described in the step 1) comprises alcoholic solvent etc., and said alcoholic solvent includes but not limited to the alcoholic solvent that this areas such as methyl alcohol, ethanol, propyl alcohol, Virahol, Ucar 35, butanols, the trimethyl carbinol are commonly used; Wherein, the mol ratio of formula a compound and Hydrazine Hydrate 80 is preferred 1: 1.2; Preferred 3 hours of reaction times; Preferred 80 ℃ of temperature of reaction;

Step 2) mol ratio of Chinese style b compound and POCl3 is preferred 1: 2.1; Preferred 4 hours of reaction times; Preferred 140 ℃ of temperature of reaction;

Preferred 1: 3 of the mol ratio of step 3) Chinese style c compound and Glacial acetic acid min. 99.5, preferred 3 hours of reaction times;

Preferred 1: 2 of the mol ratio of step 4) Chinese style d-1 compound and trifluoromethanesulfanhydride anhydride; Preferred 3 hours of reaction times; Preferred 0 ℃ of temperature of reaction;

Preferred 1: 2.5 of the mol ratio of step 5) Chinese style e compound and formula f compound; Preferred 5 hours of reaction times;

Preferred 1: 1: 3 of the compound of step 6) Chinese style g, the heteroaryl boric acid of formula h, Anhydrous potassium carbonate and triphenyl phosphatization palladium mol ratio: 0.05; Preferred 24 hours of reaction times; Preferred 120 ℃ of temperature of reaction.

Further aspect of the present invention provides the purposes of described formula I compounds for treating and/or prevention nervous system disorders; Described nervous system disorders comprises diseases such as nerve degenerative diseases, psychosis, epilepsy, convulsions and apoplexy, and described nerve degenerative diseases includes but not limited to diseases such as senile dementia, parkinson's disease or cognitive disorder.

Further aspect of the present invention provides a kind of pharmaceutical composition of treating nervous system disorders, comprises the described compound or its salt of the formula I that treats significant quantity, and suitable pharmaceutical excipient.

Compsn of the present invention can be liquid, semiliquid or solid form, prepares according to the mode that is suitable for used route of administration.Compsn of the present invention can be according to following administering mode administration: in oral, parenteral, intraperitoneal, intravenously, transdermal, hypogloeeis, intramuscular, rectum, oral cavity, the nose, mode such as liposome.

Oral compsns can be solid, gel or liquid.The instance of solid preparation includes but not limited to tablet, capsule, granule and pulvis in bulk.These preparations can selectively contain tackiness agent, thinner, disintegrating agent, lubricant, glidant, sweeting agent and correctives etc.The instance of tackiness agent includes but not limited to Microcrystalline Cellulose, glucose solution, mucialga of arabic gummy, gelatin solution, sucrose and starch paste; The instance of lubricant includes but not limited to talcum, starch, Magnesium Stearate, calcium stearate, Triple Pressed Stearic Acid; The instance of thinner includes but not limited to lactose, sucrose, starch, mannitol, Lin Suanergai; The instance of glidant includes but not limited to silicon-dioxide; The instance of disintegrating agent includes but not limited to Sodium Croscarmellose, primojel, alginic acid, W-Gum, yam starch, methylcellulose gum, agar and CMC 99.5.

Giving the present composition with parenteral, is main with injection generally, comprises subcutaneous, intramuscular or intravenous injection.Injection can be made into any conventionally form, like liquor or suspension, be suitable for before injection, being dissolved or suspended in solid form or the emulsion in the liquid.The instance that can be used for the pharmaceutically receivable carrier of injection of the present invention includes but not limited to aqueous carrier, non-aqueous carrier, biocide, isotonic agent, buffer reagent, oxidation inhibitor, suspension and dispersion agent, emulsifying agent, sequestrant and other pharmaceutically acceptable material.The instance of aqueous carrier comprise sodium chloride injection, Lin Geshi injection liquid, etc. ooze glucose injection, sterilized water injection liquid, glucose and lactic acid ringer's inj; The instance of non-aqueous carrier comprises fixed oil, Oleum Gossypii semen, Semen Maydis oil, til and the peanut oil of plant origin; The instance of biocide comprises meta-cresol, benzylalcohol, butylene-chlorohydrin, benzalkonium chloride etc.; The instance of isotonic agent comprises sodium-chlor and glucose; Buffer reagent comprises phosphoric acid salt and Citrate trianion.

The present composition can also be prepared into aseptic lyophilized injectable powder; Compound is dissolved in buffer solution of sodium phosphate; Wherein contain glucose or other vehicle that is fit to; Subsequently under standard conditions well known by persons skilled in the art with solution sterile filtration, succeeded by lyophilize, obtain required preparation.

Above-mentioned piperazinyl pyridazine compounds preparation technology provided by the invention is simple, and raw material is easy to get, and is fit to large-scale industrialization production, and through the experiment in vitro checking, The compounds of this invention has efficient inhibition BV ₂The effect of secretion IL-1 β, effect is better than positive drug, and The compounds of this invention does not have restraining effect to C6 secretion IL-1 β simultaneously, shows that this change thing is safe.

Embodiment

Compound provided by the invention can synthesize through multiple preparation method, and the exemplary process of synthetic these compounds only is provided among the embodiment.Here be noted that free acid and/or alkali form regardless of the The compounds of this invention of developing in which way, or the form of salt, scope of the present invention all belonged to.The purpose of specific embodiment is to further specify content of the present invention but do not mean that to limit the invention.The raw material that uses in the specific embodiment of the invention, reaction reagent etc. are if no specified otherwise is the commercially available prod.

Synthesizing of embodiment 13-(4-pyrimidylpiperazine)-5-methyl-6-(thionaphthene-2-yl) pyridazine (compound 1)

Synthetic route:

Step 1: compound 1-2's is synthetic:

With 50g 3-methyl furan-2,5-diketone, 33g 80% Hydrazine Hydrate 80 are dissolved in the 300ml absolute ethyl alcohol, reflux 4 hours; Reaction finishes, and is chilled to room temperature, removal of solvent under reduced pressure; Resistates adds 200ml water and stirs; Suction filtration, washing, the dry 37g white solid compound 1-2 that gets, yield 74%, MS:127 (M+H)+.

Step 2, compound 1-3's is synthetic:

It is mixed that 15g is gone up the compound 1-2 and the 56g POCl3 that make in the step, reflux 3 hours, and reaction finishes, and is chilled to room temperature; Slowly pour reaction solution in the trash ice and to stir 0.5 hour, ethyl acetate extraction (30ml * 3) merges organic layer, uses anhydrous magnesium sulfate drying; Filter, concentrate, oily matter obtains the 7.1g light yellow crystal through column chromatography for separation; Be compound 1-3, yield 44%, MS:163 (M+H)+.

Step 3, compound 1-4-1's is synthetic:

650mg is gone up the compound 1-3 that makes in the step mix reflux 3 hours with Glacial acetic acid min. 99.5.After reaction was accomplished, concentrating under reduced pressure, resistates add saturated sodium bicarbonate solution to be handled, and ethyl acetate extraction 3 times merges organic layer, dry, filter, filtrating concentrates, 0.3g compound 1-4-1, yield 52.08%, MS:145.0 (M+H) ⁺, ¹H NMR (MeOD, 400MHz): δ 7.37 (d, 1H, J=1.2Hz), 2.16 (d, 3H, J=1.2Hz); Get 0.2g compound 1-4-2, yield 34.72%, MS:145.2 (M+H) ⁺, ¹H NMR (MeOD, 400MHz): δ 6.87 (d, 1H, J=1.2Hz), 2.28 (d, 3H, J=0.8Hz).

Step 4, compound 1-5's is synthetic

1-4-1 is dissolved in the methylene dichloride with the 1g compound, logical nitrogen protection, and the ice bath cooling slowly splashes into the 2.92g trifluoromethanesulfanhydride anhydride, insulation reaction 3 hours.Reaction is washed 3 times after accomplishing, and organic phase is dry, concentrated, and resistates obtains 1.5g compound 1-5 through column chromatography for separation, yield 78.94%, MS:274.9 (M-H) ^-

Step 5, compound 1-6's is synthetic

22.6g compound 1-5 is dissolved among the exsiccant DMF, adds 33g pyrimidine piperazine, stirring at room 3 hours.Reaction is poured in the frozen water after accomplishing, stirs, and the deposition of separating out, suction filtration, drying obtains 17g compound 1-6, yield 85.99%, MS:291.1 (M+H) ⁺, ¹H NMR (CDCl ₃, 400MHz): δ 8.35～8.34 (d, 2H, J=4.8Hz), 7.21 (d, 1H, J=0.8Hz), 6.53 (t, 1H, J=4.8Hz), 4.00～3.97 (m, 4H), 3.37～3.35 (m, 4H), 2.36 (d, 3H, J=0.4Hz).

Step 6, compound 1 synthetic:

With 450mg compound 1-6,670mg Anhydrous potassium carbonate, 93mg Pd (PPh3) ₄, 390mg thionaphthene-2-boric acid and 40ml glycol dimethyl ether mix, and nitrogen protection is heated to 120 ℃ of reaction 24h, and solution colour gradually becomes black.Reaction finishes, and is chilled to room temperature, and is overanxious through zeyssatite, and filtrating concentrates, and residuum obtains the 240mg white solid through column chromatography purification, is compound 1, yield 45%, MS:389.1 (M+H) ⁺ ¹H NMR (CDCl ₃, 400MHz): δ 8.35 (d, 2H, J=4.8Hz), 7.88～7.86 (m, 1H), 7.80～7.77 (m; 1H), 7.75 (s, 1H), 7.63 (d, 1H, J=0.8Hz); 7.76～7.34 (m, 2H), 6.53 (t, 1H, J=4.8Hz), 4.03～4.00 (m; 4H), 3.46～3.43 (m, 4H), 2.42 (d, 3H, J=0.4Hz).

(indoles-5-) pyridazine (compound 2) is synthetic for embodiment 23-(4-pyrimidylpiperazine)-5-methyl-6-

Adopting the same compound method of embodiment 1, is raw material with 5-indoles boric acid, prepares target compound 2, is white solid, yield 42%, MS:372.1 (M+H) ⁺ ¹H NMR (CDCl ₃, 400MHz): δ 9.13 (s, 1H), 8.35～6.51 (9H), 4.00～3.79 (t, 8H, J=10.4,10.4Hz), 2.34 (d, 3H, J=9.2Hz).

(quinolyl-4-) pyridazine (compound 3) is synthetic for embodiment 33-(4-pyrimidylpiperazine)-5-methyl-6-

Adopting the same compound method of embodiment 1, is raw material with quinoline 4-boric acid, prepares target compound 3, is faint yellow solid, yield 28%, MS:384.1 (M+H) ⁺, ¹H NMR (CDCl3,400MHz): δ 9.31 (s, 1H), 8.51 (s, 1H), 8.34 (d, 2H, J=8.8Hz); 8.04 (q, 1H, J=8.8Hz), 7.60 (m, 2H, J=3.2Hz), 7.55 (d, 1H; J=1.6Hz), 6.91 (s, 1H), 6.54 (t, 1H, J=4.8Hz), 4.28 (t, 4H; J=5.2Hz), 8.60 (t, 4H, J=5.2Hz), 2.04 (d, 3H, J=14.8Hz).

Synthesizing of embodiment 43-(4-pyrimidylpiperazine)-5-methyl-6-[(2, the 4-dimethoxy) pyrimidine-5-yl] pyridazine (compound 4)

Adopt the same compound method of embodiment 1, with 2,4-dimethoxypyridin-5-boric acid is raw material, prepares 280mg target compound 4, yield 33%, MS:395.1 (M+H) ⁺, ¹H NMR (CDCl ₃, 400MHz): δ 9.34 (t, 2H, J=3.6Hz), 6.80 (s, 1H), 6.54 (t, 2H, J=4.8Hz), 4.05 (s, 3H), 3.99 (t, 4H, J=6Hz), 3.97 (s, 3H), 3.80 (t, 4H, J=5.2Hz), 2.14 (s, 3H).

Embodiment 53-(N methyl piperazine base)-5-methyl-6-[(2, the 4-dimethoxy) pyrimidine-5-yl)] pyridazine (compound 5) synthetic

Adopt the same compound method of embodiment 1, change the raw material pyrimidine piperazine of step 3 wherein into N methyl piperazine, and change the thiophthene of the benzene feedstock in the step 4-2-boric acid into 2; 4-dimethoxypyridin-5-boric acid; Prepare 270mg target compound 5, yield 51%, MS:331.1 (M+H) ⁺, ¹H NMR (CDCl ₃, 400MHz): δ 8.31 (s, 1H), 6.73 (s, 1H), 4.02 (s, 3H), 3.94 (s, 3H), 3.68 (t, 4H, J=5.6Hz), 2.52 (t, 4H, J=4.8Hz), 2.30 (s, 3H), 2.03 (s, 3H).

Synthesizing of embodiment 63-(N methyl piperazine base)-5-methyl-6-(quinolyl-4) pyridazine (compound 6)

Adopt the same compound method of embodiment 5, use quinoline-4-boric acid to be raw material, prepare 280mg target compound 6, yield 54%, MS:320.1 (M+H) ⁺

Embodiment 71,4-two (4-pyrimidine piperazine) phenol piperazine (compound 7) synthetic

Synthetic route:

Step 1 compound 7-2's is synthetic

Adopt the compound method of step 2 among the embodiment 1, prepare compound 7-2 by the raw material Tetra hydro Phthalic anhydride, MS:200.1 (M+H) ⁺

Synthesizing of step 2 compound 7

Adopt the similar compound method of step 2 among the embodiment 1, the normal pyrimidylpiperazine reaction of compound 7-2 and twice obtains 0.3g target compound 7, yield 66%, MS:455.2 (M+H) ⁺, ¹H NMR (DMSO, 400MHz): δ 8.39 (t, 6H, J=4.8Hz), 8.18 (m, 2H), 7.95 (m; 2H), 6.72 (t, 1H, J=4.8Hz), 6.66 (t, 2H, J=4.8Hz); 4.00 (t, 8H, J=1.6Hz), 3.91 (t, 4H, J=5.2Hz); 3.35 (t, 8H, J=5.2Hz), 3.12 (t, 4H, J=5.2Hz).

The embodiment of biological activity determination method

The active testing of The compounds of this invention adopts methods known in the art to measure; Employed positive control is following formula II compound; Its preparation method adopts disclosed method among the WO2008109437, and employed compound 1, compound 4, compound 6 adopt disclosed method preparation in the foregoing descriptions.

formula II

The embodiment of embodiment 8 active determination in vitro methods

Microglia is an a group cell of carrying out immunologic function in the cns; In the AD pathogenic process; The susceptor of their not only former as causing a disease (comprising bacterium composition, unusual endogenous protein product, some antibody component etc.) transmits the maincenter damage signal sensitively, and can play the immunity decision through changing active state.Microglia secrete a variety of pro-inflammatory cytokines (like IL-1 β, TNF α, IFN γ etc.) and anti-inflammatory factors (like IL-4, IL-10, IL-13, TGF _βDeng), balance is between the two determining " selection mode " of microglia, has only the microglia that is activated fully to carry out and removes the pathology function of neurons.The microglia overactivity is to cause one of major reason that dopaminergic neuron is lost in a large number in AD patient's brain.The level of cns IL-1 β is played an important role by the transformation of selection mode to complete active state for microglia.Therefore we assess its potential using value according to the efficient of the single-minded inhibition of candidate compound microglia secretion IL-1 β.

Experimental technique:

1. mouse source microglia is BV ₂Cultivate respectively in containing the DMEM substratum of 10%FBS with glioma cell line C6, the cell of cultivating 15 generations of the 6th generation to the is used for the candidate compound screening active ingredients.Culturing cell is inoculated in 24 porocyte culture plates by 50000 cells/well, after cultivating 1 day, is replaced by low blood serum medium (adding 2%FBS) and continues to cultivate 16 hours.In substratum, add 300ng/ml and (be used to induce BV ₂Cell) or 1mg/ml (being used to induce the C6 cell) LPS (Salmonella typhimutium) inducing culture emiocytosis IL-1 β.And pressing final concentration 100nM simultaneously, 3 μ M and 100 μ M add testing sample (DMSO≤0.1%); Add 0.1%DMSO as solvent control group to blank.

2.LPS induce/drug-treated is after 24 hours, and collection nutrient solution detection by quantitative is IL-1 β level wherein.The nutrient solution sample is removed the suspended particle impurity in the nutrient solution in 4 ℃ centrifugal (8000G) 10 minutes.With 1 times of supernatant dilution, get 150 μ l samples and be used for ELISA (Bioso μ rce) detection.

3. culturing cell processing and ELISA detect and adopt double-blind method to carry out.

4. 1. candidate compound inhibition efficient calculated according to formula, and calculate the IC of every kind of candidate compound ₅₀

Formula is 1.: inhibiting rate (%)=([IL-1 β] _{LPS induces}-[IL-1 β] _Drug-treated)/[IL-1 β] _{LPS induces}* 100%.

5. candidate compound therapeutic evaluation: with compound to BV ₂The inhibiting rate of secretion IL-1 β is positive in judging the effective standard of compound; And with negative as the index of judging compound safety to the inhibiting rate of C6 secretion IL-1 β.

Experimental result:

Table 1 The compounds of this invention is to BV ₂IL-1 β excretory IC ₅₀

Group	IC ₅₀
		Positive control drug	0.69μM
Compound 1	49.037nM
		Compound 4	33.94nM
Compound 6	0.0124nM

When table 2 The compounds of this invention concentration is 20nM to BV ₂The inhibiting rate of secretion IL-1 β

Group	The inhibiting rate of 20nM (%)
		Positive control drug	39.6635±0.0914
Compound 1	44.5375±0.0219

Compound 4	47.3142 ± 0.0753
		Compound 6	55.5237 ± 0.0384

Table 1,2 result show that The compounds of this invention can effectively suppress BV ₂Secretion IL-1 β, and restraining effect is better than the positive drug of being selected for use.Another experiment in vitro confirms that The compounds of this invention does not have restraining effect to C6 secretion IL-1 β, shows that this compound has good security simultaneously.So the cumulated volume test-results, those skilled in the art can understand compound provided by the invention and have the prospect that is developed to the treatment medicine for senile dementia.

Claims

1. the compound or its salt shown in the formula I:

Formula I

Wherein, R ¹Be selected from methyl or pyrimidyl;

R ³, R ⁴Be selected from hydrogen or C respectively _1-4Alkyl, or R ³And R ⁴Combine and form five yuan or six-membered carbon ring;

R ²Be selected from replacement or unsubstituted thionaphthene, benzopyrrole, quinoline, pyrimidine or 2, the 4-dimethoxypyridin; Substituting group is selected from C _1-4Alkyl, C _1-4Alkoxyl group;

N is selected from 0 or 1.

2. the described compound or its salt of claim 1, wherein:

R ¹Be selected from methyl or pyrimidyl;

R ³, R ⁴Be selected from hydrogen or methyl respectively, perhaps R ³And R ⁴Be combined together to form phenyl ring;

R ²Be selected from thionaphthene, benzopyrrole, quinoline, pyrimidine or 2, the 4-dimethoxypyridin;

N is 0 or 1.

3. claim 1 or 2 described compound or its salts are selected from following compounds or its salt:

4. the preparation method of any described compound among the claim 1-3, it comprises following reactions step:

formula I

Wherein, R ¹, R ², R ³, R ⁴, in n and the claim 1 definition identical.

5. claim 1,2 or 3 described compounds treat and/or prevent the purposes in the nervous system disorders medicine in preparation.

6. the said nervous system disorders of claim 5 comprises nerve degenerative diseases or diseases such as psychosis, epilepsy, convulsions or apoplexy.

7. the described nerve degenerative diseases of claim 6 comprises senile dementia, parkinson's disease or cognitive disorder.

8. the described nerve degenerative diseases of claim 7 comprises senile dementia.

9. a pharmaceutical composition of treating nervous system disorders comprises claim 1, the 2 or 3 described compound or its salts of treating significant quantity, and suitable pharmaceutical excipient.