CN109761883A - A kind of 4- carbamate-cinnamamide -4- benzyl piperidine coumpound and its preparation method and application - Google Patents

Abstract

The invention discloses a kind of 4- carbamate-cinnamamide -4- benzyl piperidine coumpounds and its preparation method and application, the preparation method comprises the following steps: using 4- hydroxycinnamic acid or ferulic acid as starting material, it is reacted under the first solvent and condensing agent effect with 4- benzyl piepridine is replaced, obtains corresponding cinnamide compound；Under the second solvent and alkaline condition, with carbamyl chloride acylation reaction occurs for cinnamide compound, obtains product 4- carbamate-cinnamamide -4- benzyl piperidine coumpound.The compounds of this invention 4- carbamate-cinnamamide -4- benzyl piperidine coumpound chemical structure is novel, and thoroughly, product yield high is easy to operate for preparation process chemical reaction reaction, low in cost, can be widely used for the prevention or treatment of neurodegenerative disease.

Description

A kind of 4- carbamate-cinnamamide -4- benzyl piperidine coumpound and its preparation Method and purposes

Technical field

The invention belongs to field of medicinal chemistry, and in particular to a kind of 4- carbamate-cinnamamide -4- benzyl piepridine class Compound and its preparation method and application.

Background technique

Alzheimer's disease (Alzheimer ' s disease, AD) is disease incidence and the highest disease of lethality in the elderly One of disease." the 2015 of Alzheimer's disease international association (Alzheimer ' s disease International, ADI) publication Global Alzheimer's disease report " it points out, the whole world in 2015 has had more than 46,000,000 people and has suffered from dementia, it was predicted that 2050 Year, the whole world will have 1.315 hundred million populations by dull-witted puzzlement, wherein the disease incidence of Chinese Dementia patients has reached 6.61%. With the extension of existent age per capita, this disease has developed into the main burden of society and medical health system, and for society, suffer from Person and family members bring heavy spirit and economic pressures.Thus, it is significant to research and develop novel senile dementia therapeutic agent. From the point of view of the market demand, Alzheimer's disease international association prediction, to the global marketing of the year two thousand fifty curing senile dementia drug Volume will be up to 600,000,000,000 dollars；In China, with rising rapidly for senile dementia disease incidence, the market of this kind of drug is also quickly swollen It is swollen.

AD is that a kind of chronic, characterized by progressive memory and Cognitive function damage multi-pathogenesis, too many levels participate in Complicated neurodegenerative disease, key pathological feature are that beta amyloid peptide (β-amyloid peptide, A β) is largely deposited The neurofibrillary tangles that senile plaque (Senile plaque, SP), the Protein tau Hyperphosphorylationof of formation are formed (Neurofibrillary tangle, NFT), and the degeneration etc. of the apoptosis and nerve synapse with neuron.In recent years, many Researcher is dedicated to disclosing the pathogenesis of AD from molecule and cellular level, proposes a variety of hypothesis, such as: cholinergic neuron Damage, the imbalance of the deposition of amyloid protein, Protein tau Hyperphosphorylationof, inflammation, free-radical oxidation, metal ion etc., therefore, The novel therapeutic approach and means developed for these pathogenesis will be hopeful to alleviate and improve the state of an illness of AD patient.Mesh The drug of preceding clinically effective treatment AD is mainly based upon the choline that neurotransmitter acetylcholine deficiency causes cognitive function to be lacked of proper care Energy hypothesis, patient's intracerebral levels of acetylcholine is improved using acetylcholinesterase inhibitor, such as: Tacrine, Donepezil, Ravastigmine,Galantamine.But Long-term clinical use shows that these drugs can alleviate the symptom of AD in a short time, but not The course of disease fundamentally effectively can be prevented or be reversed, and also results in classical cholinergic toxicity, such as caused hallucinations, realized and mix Ignorant, dizzy, nauseous, hepatotoxicity, loss of appetite and stool frequency etc..Therefore, clinically there is an urgent need to research and develop with novel The AD therapeutic agent of mechanism of action.

The AD cause of disease is complicated, not yet illustrates its pathogenesis, but studies have shown that patient's intracerebral levels of acetylcholine completely so far It reduces, excessive generation and the deposition, metal ion metabolic disorder, Ca of beta-amyloid protein²⁺Dysequilibrium, the excessive phosphorus of tau- albumen Neurofibrillary tangles caused by being acidified, glutamate receptor activity are excessively high, oxidative stress generates a large amount of active oxygens (ROS) and free radical And many factors such as Neuroinflammation are played an important role in the pathogenic process of AD.For above-mentioned pathogenic factors, research Personnel use traditional " one target of a medicine " drug design strategies, it was found that largely have high activity and highly selective to a certain target spot Drug, such as: anticholinesterase and nmda receptor antagonist, these drugs there are action target spots single, clinical use The problems such as toxic side effect is more, not good enough to the long-term efficacy of AD patient.

In recent years, with constantly illustrating to AD pathogenesis, it is found that the occurrence and development of AD have multimachine system, multifactor The characteristics of effect, have between different mechanisms it is interrelated influence each other, constitute the network tune of AD occurrence and development process complexity Control system.Based on the above results, researcher proposes " multiple target point targeted drug (Multitarget-directed Ligands, MTDLs) " strategy researches and develops anti-neurodegenerative disease drug.So-called " multiple target point drug " refers to that single chemistry is real Body acts on multiple target spots in disease network simultaneously, can produce synergistic effect to the effect of each target spot, is greater than gross effect each Single-action the sum of is answered, and such medicine is also referred to as " Multifunctional " or " Multipotential " drug.Multiple target point drug and more Medicine use in conjunction and the main distinction of compound medicine are: can reduce dosage, improve therapeutic effect, avoid between drug Interaction and thus bring toxic side effect, uniform pharmacokinetic properties, be easy to use etc..Therefore, research and development tool There are novel chemical structure, novel mechanism of action, the anti-neurodegenerative disease medicine with multiple target effect, less toxic side effect Object not only conforms with the urgent need of social senilization's process, and has good market prospects.In report early period, it was found that Scutellarein carbamate derivates (CN10337956A, CN102603698A), talan or ethane amino first Acid esters compound (CN102816090A), isoflavones carbamate compound (CN102827131A), ferulic acid amino Formic ether compounds (CN105837497A, CA105601540A, CN105646289A) though these compounds have it is preferable Acetylcholinesterase (AChE) inhibitory activity and antioxidant activity, and have the inhibiting effect of any to A beta-aggregation, while to butyryl The inhibitory activity of cholinesterase (BuChE) is excessively poor, causes these compounds not good enough to the treatment curative effect of AD in animal model. Studies have shown that BuChE is the enzyme closely related with AChE, the auxiliary adjustment factor hydrolyzable acetyl as cholinergic nerve transmitting Choline (ACh), with the development of AD, AChE level is gradually decreased, and BuChE activity degree increases the 165% of normal level, BuChE inhibitor can make ACh level increase 5 times, and further animal experiments show that, the missing of BuChE is almost without health side The side effect in face.Therefore, it designs and finds double target spot inhibitor pair simultaneously with acetylcholinesterase and butyrylcholine esterase AD is treated great potential.

Vascular dementia (Vascular Dementia, VD) be by ischemic cerebrovascular disease, hemorrhagic cerebrovaseular disease, Intelligence caused by various types of cranial vascular diseases such as acute and chronic Hypoxic cranial vascular disease and cognition dysfunction face Bed syndrome, main clinical manifestation includes: the decline and emotion, personality of cognitive ability, memory and social-life ability Change, be a kind of chronic progressive disease.It is the first of senile dementia in the Asian countries such as China, Japan vascular dementia Position reason；As world population is to the continuous propulsion of aging, cerebrovascular disease is increasing, and Onset of Vascular Dementia rate has gradually The trend of rising seriously affects the work and life quality of the elderly, and brings heavy economy and spirit to society and family Burden.Therefore, VD has become an important research hotspot in current gerontology and psychologic medicine field.Vascular dementia by In pathogenesis complexity, the drug that disease can be blocked to develop there is no, clinical treatment is at present to improve brain blood circulation and brain Metabolism is reinforced based on brain nutrition.

In recent years, studies at home and abroad show that, cholinergic is also often accompanied by while VD patient shows cerebral damage The exception of system.VD patient's hippocampus ChAT positive neuron and fibre density reduce, under the ChAT activity of intracerebral different parts Drop, ACh concentration in VD Cerebrospinal Fluid in Patients are significantly lower than normal level, and the degree that reduces of its concentration and dementia is serious Degree is positively correlated；And cerebral ischemia can cause intracerebral acetylcholine esterase active to rise；Simultaneously it has also been found that acetylcholinesterase Inhibitor as: HuperzineA and Revastigmine can protect neure damage caused by ischemic, and can promote brain lack The recovery of neurotrosis and brain function after blood, this shows that acetylcholinesterase inhibitor can also be used for the treatment of vascular dementia.

Summary of the invention

To overcome drawbacks described above, the purpose of the present invention is to provide a kind of 4- carbamate-cinnamamide -4- benzyl piperazines Pyridine class compound.

Also residing in for the second object of the present invention provides a kind of 4- carbamate-cinnamamide -4- benzyl piepridine class Close the preparation method of object.

The third object of the present invention, which also resides in, provides a kind of 4- carbamate-cinnamamide -4- benzyl piepridine class chemical combination Purposes of the object in preparation treatment neurodegenerative disease drug.

The fourth object of the present invention, which also resides in, provides a kind of drug for treating neurodegenerative disease, including 4- carbamic acid The pharmaceutically acceptable salt of ester-cinnamamide -4- benzyl piperidine coumpound or itself and acid synthesis.

To achieve the above object, the present invention adopts the following technical scheme:

A kind of 4- carbamate-cinnamamide -4- benzyl piperidine coumpound, general formula of the chemical structure are as follows:

Wherein, R H, C₁~C₆Alkyl, hydroxyl or C₁~C₆Alkoxy；

C=X is CH₂, C=O, C=S or C-Ph；

Y is C or N；

Z is H ,-F ,-Cl ,-Br ,-I, C_1-4Alkyl, C_1-4Alkoxy, trifluoromethyl, trifluoromethoxy, nitro, cyano and Any one in phenoxy group, two kinds, three kinds or four kinds of groups；

R₁NR₂Indicate N- methyl-N ethyl, dimethyl, diethylamine, diisopropyl ammonia, dibutyl amine, two allylamines, morpholine ring, Piperidine ring, 4- benzyl piepridine ring, 4- substituted benzyl piperidine ring, 4- Phenylpiperidine ring, 4- substituted phenylpiperidines ring, benzyl diethylenediamine Ring, substituted benzyl piperazine ring, nafoxidine ring or 4- are by C₁~C₁₂Alkyl-substituted piperazine ring.

A kind of preparation method of 4- carbamate-cinnamamide -4- benzyl piperidine coumpound, comprising the following steps:

A. using 4- hydroxycinnamic acid or ferulic acid as starting material, the first solvent and condensing agent effect under with replace 4- benzyl Phenylpiperidines reaction, obtains corresponding cinnamide compound；

B. cinnamide compound occurs acylation reaction with carbamyl chloride, obtains under the conditions of the second solvent and alkali Product 4- carbamate-cinnamamide -4- benzyl piperidine coumpound.

It chemically reacts general formula are as follows:

Wherein, in the substitution of R, X, Y, Z and above-mentioned general formula of the chemical structure expression.

Preferably, the first solvent in the step a be tetrahydrofuran, n,N-Dimethylformamide, dimethyl sulfoxide, C_3-8Aliphatic ketone, benzene, toluene, acetonitrile, methylene chloride, chloroform, C_1-8Pure and mild C_5-8It is one or more of in alkane.

Preferably, the condensing agent in the step a is dicyclohexylcarbodiimide, 4-dimethylaminopyridine, 1- ethyl- It is one or more of in (3- dimethylaminopropyl) phosphinylidyne diimmonium salt hydrochlorate EDCI and I-hydroxybenzotriazole HOBT.

Preferably, 4- hydroxycinnamic acid or ferulic acid, the molar ratio for replacing 4- benzyl piepridine and condensing agent in the step a For 1:1~5:1~10, reaction temperature is 25~100 DEG C, and the reaction time is 12~48h.

Preferably, the second solvent in the step b is C_3-8Aliphatic ketone, N,N-dimethylformamide, ether, isopropyl ether, Methyl tertiary butyl ether(MTBE), tetrahydrofuran, glycol dimethyl ether, C_1-6Fatty acid and C_1-6The formed ester of fatty alcohol, methylene chloride, chlorine It is a kind of or several in imitative, 1,2- dichloroethanes, benzene,toluene,xylene, chlorobenzene, o-dichlorohenzene, acetonitrile, dimethyl sulfoxide and pyridine Kind.

Preferably, the alkali in the step b is alkali metal hydroxide, alkaline earth metal hydroxide, alkali metal or alkaline earth Metal carbonate, alkali or alkaline earth metal bicarbonate, C_1-6Fatty acid alkali metal salt, piperidines, nafoxidine, triethylamine, three It is a kind of or several in butylamine, trioctylamine, pyridine, N-methylmorpholine, N- methyl piperidine, triethylene diamine and tetrabutylammonium hydroxide Kind.

Preferably, cinnamide compound in the step b, carbamyl chloride, alkali molar ratio be than 1:1~10:1 ~20, reaction temperature be 25 DEG C~100 DEG C, the acylation reaction time be 10~for 24 hours.

A kind of 4- carbamate-cinnamamide -4- benzyl piperidine coumpound treats neurodegenerative disease in preparation Application in drug.

Preferably, the neurodegenerative disease is Alzheimer's disease, vascular dementia, parkinsonism, the prosperous court of a feudal ruler Disease, HIV related dementia disorders, multiple sclerosis, progressive lateral sclerosis of spinal cord, neuropathic pain or glaucoma.

It is a kind of to treat neurodegenerative disease drug, including above-mentioned 4- carbamate-ferulic amide -4- benzyl piepridine Class compound or the pharmaceutically acceptable salt synthesized by it with acid.

Preferably, it 4- carbamate-ferulic amide -4- benzyl piperidine coumpound or is closed by itself and acid At pharmaceutically acceptable salt weight percent 10%~99.5%.

Preferably, the acid is hydrochloric acid, hydrobromic acid, nitric acid, sulfuric acid, phosphoric acid, C_1-6Aliphatic carboxylic acid, oxalic acid, benzoic acid, Salicylic acid, maleic acid, fumaric acid, succinic acid, tartaric acid, citric acid, C_1-6Alkyl sulfonic acid, camphorsulfonic acid, benzene sulfonic acid or to first Benzene sulfonic acid.

Positive beneficial effect of the invention:

1. quoting carbamate groups in the compounds of this invention structure, it is based on multiple target point principle, to acetylcholinesterase With the IC of butyrylcholine esterase₅₀Respectively 0.18 μM~17.3 μM and 0.11 μM~7.6 μM, to A β under 25.0 μM of concentration_1-42 The inhibiting rate of self assemble is equal to acetylcholinesterase, butyrylcholine esterase and the A of auto-induction beta-aggregation 46.7% or more With effect is significantly inhibited, the treatment for the treatment of neurodegenerative disease can be widely used for.

2. the compounds of this invention is to AlCl₃The motor function and respond of the zebra fish AD model of induction have significant Improvement result all has statistical difference compared with model control group；The compounds of this invention causes mouse to be remembered hyoscine Obstacle is recalled with dose-dependent improvement result, and statistical difference is all had compared with model group.

Detailed description of the invention

Fig. 1 is the compounds of this invention to AlCl₃Induce the testing result figure of the motor function of zebra fish AD；

Fig. 2 is the compounds of this invention to AlCl₃Induce the testing result figure of the respond of zebra fish AD；

Fig. 3 is that the compounds of this invention causes mouse to obtain the testing result figure that memory disorders influence hyoscine.

Specific embodiment

Below with reference to some specific embodiments, the present invention is further described.

A kind of 4- carbamate-cinnamamide -4- benzyl piperidine coumpound, general formula of the chemical structure are as follows:

Wherein, in embodiment 1-29, R, X, Y, Z and R₁NR₂Substituent group is shown in Table 1 in detail.

By taking compound 1 as an example, a kind of preparation of 4- carbamate-cinnamamide -4- benzyl piperidine coumpound (I-1) Method, comprising the following steps:

A. in reaction flask be added 50mmol ferulic acid, 100mL dry THF, 75mmol EDCI, 75mmol HOBT, 50mmol 4- benzyl piepridine, 25 DEG C are stirred to react for 24 hours；After reaction, 250mL bis- is added in residue for evaporating solvent under reduced pressure Chloromethanes is successively washed with saturated aqueous sodium carbonate and saturated sodium-chloride water solution, after organic layer is dried over anhydrous sodium sulfate Filtering, evaporating solvent under reduced pressure, residue obtain accordingly through column chromatographic purifying (in eluent methylene chloride: methanol=30:1v/v) Cinnamide compound；

B. step a cinnamide compound is all dissolved in 100mL acetonitrile, 75mmol potassium carbonate, 55mmol is added Carbamyl chloride is warming up to 60 DEG C and is stirred to react 12h；After reaction, 200mL bis- is added in residue for evaporating solvent under reduced pressure Chloromethanes is successively washed with saturated aqueous sodium carbonate and saturated sodium-chloride water solution, after organic layer is dried over anhydrous sodium sulfate Filtering, evaporating solvent under reduced pressure, residue obtain product through column chromatographic purifying (in eluent methylene chloride: methanol=10:1v/v) I-1, yield 82.5%, chemical structure passes through¹H-NMR,¹³C-NMR and ESI-MS confirmation.

The 4- carbamate of above-described embodiment 2-10-cinnamamide-4- benzyl piperidine coumpound preparation method with Embodiment 1 is similar, and same section does not repeat, and difference is shown in Table 2 and table 3.

The 4- carbamate of 1 1-29 of the embodiment of the present invention of table-ferulic amide -4- benzyl piperidine coumpound

The compounds of this invention structural analysis data

4-N- methyl-N-ethylamino formic acid esters-(4- benzyl piepridine) asafoetide amide (I-1): Light yellow oil, 82.5%yield, 98.6%HPLC purity.¹H NMR 7.59 (d, J=15.2Hz, 1H, C=CH), 7.28 (t, J= 7.6Hz, 2H, 2 × Ar-H), 7.20 (t, J=7.2Hz, 1H, 1 × Ar-H), 7.14 (d, J=7.2Hz, 2H, 2 × Ar-H), 7.08 (d, J=7.2Hz, 2H, 2 × Ar-H), 7.04 (s, 1H, Ar-H), 6.81 (d, J=15.2Hz, 1H, C=CH), 4.69 (d, J=11.2Hz, 1H, 1/2phCH₂), 4.07 (d, J=11.6Hz, 1H, 1/2phCH₂),3.85(s,3H,OCH₃),3.48 (q, J=6.8Hz, 1H, 1/2NCH₂), 3.39 (q, J=6.8Hz, 1H, 1/2NCH₂),3.08(s,3/2H,1/2NCH₃),3.03 (t, J=9.6Hz, 1H, 1/2NCH₂), 2.61 (t, J=9.6Hz, 1H, 1/2NCH₂),2.98(s,3/2H,1/2NCH₃),2.56 (t, J=6.0Hz, 2H, NCH₂), 1.83-1.79 (m, 1H, CH), 1.73 (d, J=12.6Hz, 2H, CH₂),1.26-1.16(m, 5H,CH₃+CH₂).¹³C NMR 165.3,154.2,151.9,141.8,140.0,133.8,133.7,129.1(2C),128.3 (2C),126.1,123.6,120.3,117.5,111.6,56.0,46.2,44.2,43.0(2C),42.7,38.3(2C), 32.8,31.8.HR-ESI-MS:Calcd.for C₂₆H₃₂N₂O₄[M+H]⁺:437.2396,found:437.2430。

4- dimethylamino carbamate-(4- benzyl piepridine) asafoetide amide (I-2): Light yellow oil, 90.3%yield, 98.2%HPLC purity.¹H NMR 7.59 (d, J=15.2Hz, 1H, CH=CH), 7.29 (t, J= 7.6Hz, 2H, 2 × Ar-H), 7.21 (d, J=7.2Hz, 1H, Ar-H), 7.15-7.12 (m, 2H, 2 × Ar-H), 7.10 (s, 1H, ), Ar-H 7.07 (s, 1H, Ar-H), 7.05 (s, 1H, Ar-H), 6.80 (d, J=15.2Hz, 1H, CH=CH), 4.71-4.67 (m,1H,1/2phCH₂),4.08-4.05(m,1H,1/2phCH₂),3.86(s,3H,OCH₃),3.11(s,3H,NCH₃),3.07- 3.05(m,1H,1/2NCH₂),3.00(s,3H,NCH₃),2.65-2.61(m,1H,1/2NCH₂), 2.56 (t, J=5.6Hz, 2H, NCH₂), 1.83-1.77 (m, 1H, CH), 1.74 (d, J=13.6Hz, 2H, CH₂),1.25-1.19(m,2H,CH₂).¹³NMR 165.3,154.5,151.8,141.9,141.7,139.9,133.8,129.1(2C),128.3(2C),126.1,123.6, 120.2,117.6,111.6,56.1,46.3,43.0,42.7,38.3,36.8,36.6,32.8,31.8.HR-ESI-MS: Calcd.for C₂₅H₃₀N₂O₄[M+H]⁺:423.2239,found:423.2256。

4- diethylin carbamate-(4- benzyl piepridine) asafoetide amide (I-5): Light yellow oil, 84.7%yield, 98.8%HPLC purity.¹H NMR 7.59 (d, J=15.6Hz, 1H, CH=CH), 7.31-7.26 (m, 2H, 2 × Ar-H), 7.22-7.18 (m, 1H, Ar-H), 7.14 (d, J=7.2Hz, 2H, 2 × Ar-H), 7.10-7.08 (m, 2H, 2 × Ar-H), 7.04 (s, 1H, Ar-H), 6.80 (d, J=15.6Hz, 1H, CH=CH), 4.70 (d, J=7.6Hz, 1H, 1/ 2phCH₂), 4.08 (d, J=10.4Hz, 1H, 1/2phCH₂),3.85(s,3H,OCH₃),3.46-3.43(m,2H,CH₂), 3.39-3.36(m,2H,CH₂), 3.04 (t, J=10.0Hz, 1H, 1/2CH₂), 2.62 (t, J=11.8Hz, 1H, 1/2CH₂), 2.57 (t, J=7.2Hz, 2H, CH₂), 1.83-1.79 (m, 1H, CH), 1.74 (d, J=13.6Hz, 2H, CH₂),1.28-1.24 (m,6H,2×CH₃),1.21-1.19(m,2H,CH₂).¹³NMR 165.3,153.8,151.9,142.0,141.8,140.0, 133.7,129.1(2C),128.3(2C),126.1,123.6,120.2,117.4,111.7,56.0,43.0,42.7,42.3 (2C),42.1,38.3(2C),29.7,14.0,13.4.HR-ESI-MS:Calcd.for C₂₇H₃₄N₂O₄[M+H]⁺: 451.2552,found:451.2591。

4- benzhydryl carbamate-(4- benzyl piepridine) asafoetide amide (I-8): Light yellow oil, 73.9%yield, 98.9%HPLC purity.¹H NMR 7.57 (d, J=15.6Hz, 1H, CH=CH), 7.38-7.30 (m, 8H, 8 × Ar-H), 7.26 (t, J=7.2Hz, 2H, 2 × Ar-H), 7.19 (t, J=8.4Hz, 3H, 3 × Ar-H), 7.11 (d, J =7.2Hz, 2H, 2 × Ar-H), 7.06 (s, 2H, 2 × Ar-H), 7.03 (s, 1H, Ar-H), 6.79 (d, J=15.6Hz, 1H, CH =CH), 4.67 (d, J=10.0Hz, 1H, 1/2phCH₂), 4.03 (d, J=9.6Hz, 1H, 1/2phCH₂),3.86(s,3H, OCH₃), 2.98 (t, J=7.2Hz, 1H, 1/2CH₂), 2.57 (t, J=7.2Hz, 1H, 1/2CH₂), 2.52 (t, J=7.2Hz, 2H,CH₂), 1.77-1.75 (m, 1H, CH), 1.69 (d, J=13.6Hz, 2H, CH₂),1.19-1.16(m,2H,CH₂).¹³NMR 165.2,152.7,151.7,142.5,141.7,141.4,140.0,134.2,129.2,129.0,128.4,126.9, 126.4,126.1,123.3,120.3,117.9,111.7,56.2,46.2,43.0,38.3,32.9,31.9.HR-ESI-MS: Calcd.for C₃₅H₃₄N₂O₄[M+H]⁺:547.2552,found:547.2584。

The 4- carbamate of 2 1-10 of the embodiment of the present invention of table-ferulic amide -4- benzyl piperidine coumpound preparation side One of method parameter

The 4- carbamate of 3 1-10 of the embodiment of the present invention of table-ferulic amide -4- benzyl piperidine coumpound preparation side The two of method parameter

4- carbamate-ferulic amide -4- benzyl piperidine coumpound (I) and acid synthesis salt preparation

Gained 4- carbamate-cinnamamide -4- benzyl piepridine class in accordance with the above-mentioned embodiment 1 is added in reaction flask Object (I) 2.0mmol and acetone 50ml are closed, 6.0mmol tartaric acid is stirring evenly and then adding into, is warming up to return stirring reaction 20min, It is cooled to room temperature after reaction, evaporating solvent under reduced pressure, residue acetone recrystallization, filters the solid of precipitation to get 4- ammonia Carbamate-cinnamamide -4- benzyl piperidine coumpound (I) salt, chemical structure warp¹H-NMR and ESI-MS confirmation.

Biological activity test

1. 4- carbamate prepared by the present invention-cinnamamide -4- benzyl piperidine coumpound (I) is to acetylcholine ester Enzyme and butyrylcholine esterase inhibitory activity

1.0mmol/L acetylthiocholine iodide is sequentially added into 96 orifice plates or thio BuCh (is purchased from Sigma Company) 30 μ L, pH7.4 40 μ L of PBS buffer solution, 20 μ L of testing compound solution (DMSO content is less than 1%) and 10 μ L electric eels Acetylcholinesterase (EeAChE) or horse serum butyrylcholine esterase (eqBuChE), after finishing mixing, 37 DEG C of incubation 15min, to It is molten that thio-bis- (2- nitro) benzoic acid of 5,5'- bis- (DTNB is purchased from Sigma company) that mass fraction is 0.2% is added in each hole 30 μ L of liquid colour developing, the optical density (OD value) in each hole at 405nm is measured with microplate reader, compared with the blank well that sample to be tested is not added, Compound is calculated to the inhibiting rate [enzyme inhibition rate=(1- sample sets OD value/blank group OD value) × 100%] of enzyme；Select chemical combination Five to six concentration of object measure its enzyme inhibition rate, and linear with the inhibiting rate of the negative logarithm of the compound molar concentration and enzyme It returns, molar concentration when acquiring 50% inhibiting rate is the IC of the compound₅₀, using Rivastigmine as positive control eachization It closes object and measures 3 multiple holes every time, testing result is shown in Table 4.

4 the compounds of this invention of table is to cholinesterase inhibition and auto-induction A β_1-42Assemble inhibitory activity

Measurement result shows the compounds of this invention to the IC of acetylcholinesterase and butyrylcholine esterase₅₀Respectively 0.18 μ M~17.3 μM and 0.11 μM~7.6 μM, quite or more preferably with positive control medicine Rivastigmine testing result, this hair Bright compound all has the effect of significantly inhibiting to acetylcholinesterase and butyrylcholine esterase.

2. 4- carbamate prepared by the present invention-cinnamamide -4- benzyl piperidine coumpound (I) is to inhibition A beta-aggregation Determination of activity

Take the A β of 20 μ L_1-42(testing compound solution is that the application compound is molten to the testing compound solution of+20 μ L of solution Liquid or positive control curcumin solution), the A β of 20 μ L_1-42+ 20 μ L PBS buffer solution of solution (containing 2%DMSO), 20 μ L PBS Buffer (containing 2%DMSO)+20 μ L PBS buffer solution (containing 25%DMSO) are in 96 orifice plate of black, compound and A β_1-42Most Final concentration is 25 μM.37 DEG C are incubated for the glycine-NaOH buffering that the 50mM that 160 μ L contain 5 μM of thioflavine Ts is then added for 24 hours Liquid (pH=8.5) uses Varioskan Flash Multimode Reader (Thermo Scientific) immediately after shaking 5s Multi-function microplate reader measures fluorescent value under 446nm excitation wavelength and 490nm launch wavelength；Aβ_1-42+ testing compound solution Fluorescent value is recorded as IF_i, A β_1-42The fluorescent value of+PBS buffer solution is recorded as IF_c, the fluorescent value for containing only PBS buffer solution is recorded as IF₀, A β is inhibited by compound_1-42The inhibiting rate calculation formula of self assemble are as follows: 100- (IF_i-IF₀)/(IF_c-IF₀)*100.Often A each two multiple holes of concentration mensuration of compound, testing result are shown in Table 4.

Measurement result shows the compounds of this invention to A β_1-42The aggregation of auto-induction all has the effect of significantly inhibiting, To A β under 25.0 μM of concentration_1-42The inhibiting rate of self assemble is 46.7% or more；And inhibiting rate of the curcumin under same concentrations It is 43.1%, illustrates the compounds of this invention to A β_1-42The aggregation of auto-induction all has the effect of significantly inhibiting.

3. 4- carbamate prepared by the present invention-ferulic amide -4- benzyl piperidine coumpound (I) is to AlCl₃It lures Lead the preventive and therapeutic effect (illustrating by taking I-28 as an example) of zebra fish Alzheimer's disease model.

3dpf wild type AB system zebra fish is randomly selected in six orifice plates, with alchlor (AlCl₃) induce zebra fish Ah Wurz sea is write from memory disease model (hereinafter referred to as AD zebra fish), water-soluble respectively to give 0.1 μ g/mL of I-28 compound, 0.3 μ g/mL and 0.9 μ g/mL concentration, positive controls are administered 8 μM of donepezil DPZ, while Normal group and model control group is arranged, every experiment 30 tail zebra fish of concentration group.After administration 3 days, each experimental group zebra fish is observed and recorded respectively 3 in 60min with behavioural analysis instrument The move distance and speed difference in light and shade period (that is: dark 10min, illumination 10min replace 3 periods), analyze zebra fish The move distance and speed difference of 60min, it is for statistical analysis with move distance and speed difference and model control group, with statistics Evaluation compound is learned to the preventive and therapeutic effect of zebra fish alzheimer's disease model, the result is shown in Figure 1 and Fig. 2.

As shown in Figure 1, the move distance (5513mm) of model control group zebra fish p compared with Normal group (7304mm) < 0.01, show model foundation success；P < 0.05 compared with model control group positive controls move distance (6566mm), fortune Dynamic improvement result is 59%, illustrates that DPZ has improvement result to the motor function of AD zebra fish.Concentration is 0.1 μ g/mL, 0.3 μ g/ The I-28 move distance of mL and 0.9 μ g/mL are respectively 6544,6741 and 7159mm, compared with model control group p>0.05, p< 0.05 and p < 0.01, illustrate that compound I-28 can significantly improve AlCl₃Induce the motor function of zebra fish AD.

As shown in Figure 2, the speed of model control group zebra fish changes (1.15mm/s) and Normal group (2.35mm/s) Compare p < 0.01, shows model foundation success；Positive controls speed change (1.97mm/s) compared with model control group p < 0.01, respond improvement result is 68%, illustrates that DPZ has improvement result to the respond of AD zebra fish.Concentration is 0.1 It is respectively 1.80,2.14 and 2.28mm/s that the I-28 speed of μ g/mL, 0.3 μ g/mL and 0.9 μ g/mL, which change, with model control group Compare p>0.05, p<0.05 and p<0.01, illustrates that compound I-28 can significantly improve AlCl₃Induce the reaction of zebra fish AD Ability.

4. 4- carbamate prepared by the present invention-cinnamamide -4- benzyl piperidine coumpound (I) is to hyoscine institute Cause the influence (illustrating by taking I-28 as an example) of mouse memory acquired disturbance.

SPF grades of ICR male mices, 25-30g are randomly divided into: normal group, model group, positive controls, test drug be high, In, low dose group, every group of 10 animals.Test medicine is given in disposable stomach-filling, and normal group and model group give solvent 0.5% CMC-Na, administered volume are 0.1ml/10g, positive controls stomach-filling Rivastigmine 6.0mg/kg, test drug is high, in, Low dose group feeds the compounds of this invention 20.0,10.0,5.0mg/kg respectively；45min after medicine is normal to organize mouse peritoneal injection life Salt water is managed, remaining groups of animals injects hyoscine 5mg/kg；After modeling 30min, by mouse be put into the labyrinth non-electro photoluminescence Y into Row Behavior test.Mouse is put in an arm end when test, it is allowed to travel freely 8min in labyrinth, records it into each arm Number and alternate frequency, calculate alternately rate according to following formula: alternately rate %=[alternate frequency/(always enter number -2)] × 100, it is as a result indicated with mean ± standard deviation, group difference uses one-way analysis of variance, as a result sees Fig. 3.

Measurement result shows that the alternating rate normally organized is 65.3%, and the alternating rate of model group is 40.8%, with model group phase Than the alternating rate of positive controls (Rivastigmine) is 60.2%, illustrates model running success.In the experiment condition Under, compound (I-28) disclosed in this invention, which causes mouse to obtain memory disorders hyoscine, has dose-dependent improvement Effect, wherein middle dose group (10.0mg/kg) shows highest alternating rate 62.2% (p < 0.01), compared with positive controls There is no statistical difference.Illustrate that disclosed compound I-28 can significantly improve hyoscine and mouse is caused to obtain memory disorders.

Finally, it is stated that the above examples are only used to illustrate the technical scheme of the present invention and are not limiting, this field is common Other modifications or equivalent replacement that technical staff makes technical solution of the present invention, without departing from technical solution of the present invention Spirit and scope, be intended to be within the scope of the claims of the invention.

Claims (10)

1. a kind of 4- carbamate-cinnamamide -4- benzyl piperidine coumpound, general formula of the chemical structure are as follows:

Wherein, R H, C₁~C₆Alkyl, hydroxyl or C₁~C₆Alkoxy；

C=X is CH₂, C=O, C=S or C-Ph；

Y is C or N；

Z is H ,-F ,-Cl ,-Br ,-I, C_1-4Alkyl, C_1-4Alkoxy, trifluoromethyl, trifluoromethoxy, nitro, cyano and benzene oxygen Any one in base, two kinds, three kinds or four kinds of groups；

R₁NR₂Indicate N- methyl-N ethyl, dimethyl, diethylamine, diisopropyl ammonia, dibutyl amine, two allylamines, morpholine ring, piperidines Ring, 4- substituted benzyl piperidine ring, 4- Phenylpiperidine ring, 4- substituted phenylpiperidines ring, benzyl diethylenediamine ring, takes 4- benzyl piepridine ring For benzyl diethylenediamine ring, nafoxidine ring or 4- by C₁~C₁₂Alkyl-substituted piperazine ring.

2. 4- carbamate according to claim 1-cinnamamide-4- benzyl piperidine coumpound preparation method, Characterized by comprising the following steps:

A. using 4- hydroxycinnamic acid or ferulic acid as starting material, the first solvent and condensing agent effect under with replace 4- benzyl piperazine Pyridine reaction, obtains corresponding cinnamide compound；

B. cinnamide compound occurs acylation reaction with carbamyl chloride, is produced under the second solvent and alkaline condition Object 4- carbamate-cinnamamide -4- benzyl piperidine coumpound.

3. 4- carbamate according to claim 2-cinnamamide-4- benzyl piperidine coumpound preparation method, It is characterized in that, the first solvent in the step a is tetrahydrofuran, n,N-Dimethylformamide, dimethyl sulfoxide, C_3-8Rouge Fat ketone, benzene, toluene, acetonitrile, methylene chloride, chloroform, C_1-8Pure and mild C_5-8It is one or more of in alkane；

Condensing agent in the step a is dicyclohexylcarbodiimide, 4-dimethylaminopyridine, 1- ethyl-(3- dimethylamino Propyl) it is one or more of in phosphinylidyne diimmonium salt hydrochlorate and I-hydroxybenzotriazole.

4. 4- carbamate according to claim 2-cinnamamide-4- benzyl piperidine coumpound preparation method, It is characterized in that, the molar ratio of 4- hydroxycinnamic acid or ferulic acid, substitution 4- benzyl piepridine and condensing agent is 1 in the step a: 1~5:1~10, reaction temperature are 25~100 DEG C, and the reaction time is 12~48h.

5. 4- carbamate according to claim 2-cinnamamide-4- benzyl piperidine coumpound preparation method, It is characterized in that the second solvent in the step b is C_3-8Aliphatic ketone, N,N-dimethylformamide, ether, isopropyl ether, methyl Tertbutyl ether, tetrahydrofuran, glycol dimethyl ether, C_1-6Fatty acid and C_1-6The formed ester of fatty alcohol, methylene chloride, chloroform, 1, It is one or more of in 2- dichloroethanes, benzene,toluene,xylene, chlorobenzene, o-dichlorohenzene, acetonitrile, dimethyl sulfoxide and pyridine；

Alkaline condition in the step b is alkali metal hydroxide, alkaline earth metal hydroxide, alkali metal or alkaline earth gold with alkali Belong to carbonate, alkali or alkaline earth metal bicarbonate, C_1-6Fatty acid alkali metal salt, piperidines, nafoxidine, triethylamine, three fourths It is one or more of in amine, trioctylamine, pyridine, N-methylmorpholine, N- methyl piperidine, triethylene diamine and tetrabutylammonium hydroxide.

6. 4- carbamate according to claim 2-cinnamamide-4- benzyl piperidine coumpound preparation method, It is characterized in that, the molar ratio of cinnamide compound in the step b, carbamyl chloride, alkali be than 1:1~10:1~ 20, reaction temperature be 25 DEG C~100 DEG C, the acylation reaction time be 10~for 24 hours.

7. a kind of 4- carbamate-cinnamamide -4- benzyl piperidine coumpound treats neurodegenerative disease medicine in preparation Application in object.

8. application according to claim 7, which is characterized in that the neurodegenerative disease is Alzheimers Disease, vascular dementia, parkinsonism, Huntingdon disease, HIV related dementia disorders, multiple sclerosis, progressive lateral spinal sclerosis Disease, neuropathic pain or glaucoma.

9. a kind of treat neurodegenerative disease drug, including 4- carbamate-ferulic amide -4- benzyl piepridine class chemical combination Object or the pharmaceutically acceptable salt synthesized by it with acid.

10. treatment neurodegenerative disease drug according to claim 9, which is characterized in that the acid is hydrochloric acid, hydrogen Bromic acid, nitric acid, sulfuric acid, phosphoric acid, C_1-6Aliphatic carboxylic acid, oxalic acid, benzoic acid, salicylic acid, maleic acid, fumaric acid, succinic acid, winestone Acid, citric acid, C_1-6Alkyl sulfonic acid, camphorsulfonic acid, benzene sulfonic acid or p-methyl benzenesulfonic acid.