CN106632191B - Homoisoflavone Mannich alkaloid compound, preparation method and use - Google Patents

Abstract

The invention discloses a kind of novel homoisoflavone Mannich alkaloid compounds（I）And its pharmaceutically acceptable salt, preparation method, pharmaceutical composition and the purposes in preparing treatment and/or preventing nervus retrogression relevant disease drug, including but not limited to neurodegenerative diseases such as vascular dementia, Alzheimer's disease, parkinsonism, Huntingdon disease, HIV related dementia disorders, multiple sclerosis, progressive lateral sclerosis of spinal cord, neuropathic pain, glaucoma.

Description

Homoisoflavone Mannich alkaloid compound, preparation method and use

Technical field

The invention belongs to medicinal chemistry arts, are related to a kind of novel homoisoflavone Mannich alkaloid compound（I）And its medicine It acceptable salt on, preparation method, pharmaceutical composition and is preparing treatment and/or is preventing nervus retrogression relevant disease medicine Purposes in object, including but not limited to vascular dementia, Alzheimer's disease, parkinsonism, Huntingdon disease, HIV correlations are silly The neurodegenerative diseases such as slow-witted disease, multiple sclerosis, progressive lateral sclerosis of spinal cord, neuropathic pain, glaucoma.

Background technology

Alzheimer's disease（Alzheimer ' s disease, AD, senile dementia）Be one kind with progressive cognitive disorder With the central nervous system degenerative disease based on memory damage, incidence becomes in ascendant trend year by year and is only second to the heart The frequently-occurring disease of angiosis and cancer, it is the 4th of the cause of death to be had gone up in developed countries such as America and Europes.According to world health Organisation Report, global over-65s old man have 10% dysnoesia, wherein half to occur dull-witted, fall ill within 85 years old or more Rate nearly 50%.The AD patient numbers about 600-700 ten thousand in China, incidence are more than 5%.With adding for population in the world aging process Soon, incidence is in apparent ascendant trend, is announced in December, 2013 according to Alzheimer's Disease International 's《The global implication of Alzheimer's disease：2013-2050》It is pointed out in report, AD will face most as the coming few decades whole world Big Health challenges, to the year two thousand thirty, patient numbers will rise to 76,000,000 by 44,000,000 in 2013, to the year two thousand fifty, this numerical value It is up to surprising 1.35 hundred million.Due to AD clinical manifestations be memory capability, capacity of orientation, thinking and judgement decline, and Activity of daily living reduces, or even abnormal Behavioral and psychological symptom occur etc., keep patient care difficulty larger, to society and family's band Carry out heavy burden.The drug that approved is used to treat light/moderate AD at present has acetylcholinesterase（AChE）Inhibitor, Yi Jiyong In severe AD treatmentsNMethyl-DAspartic acid（NMDA）Receptor antagonist, but Clinical practice shows that these drugs can pass through It improves patient's body levels of acetylcholine or inhibits the exitotoxicity of excitatory amino acid to alleviate AD symptoms, but cannot be effective Prevent or reverse the course of disease, but also can cause hallucinations, misunderstanding, dizziness, headache, nausea, hepatotoxicity, loss of appetite and The serious toxic side effect such as stool frequency, thus long-term efficacy is not satisfactory.Therefore, clinically there is an urgent need to research and develop with novel work With the AD medicines of mechanism.

AD belongs to disease caused by many factors, and pathogenesis is complicated, does not illustrate its pathogenesis completely also so far, but study Show that the decline of patient's intracerebral levels of acetylcholine, the excessive generation of beta-amyloid protein and deposition, metal ion metabolism are disorderly Disorderly, Ca²⁺Dysequilibrium,tauNeurofibrillary tangles caused by protein hyperphosphorylation, glutamate receptor activity are excessively high, oxidation is answered Swash and generates a large amount of active oxygens（ROS）Play the part of weight in the pathogenic process of AD with many factors such as free radical and Neuroinflammations Want role.For above-mentioned pathogenic factors, researcher is using traditional " one target of a medicine " drug design strategies, it was found that largely to certain One target spot has high activity and highly selective drug, such as：Anticholinesterase andNMethyl-DAspartate receptor antagonism Agent etc., but that there are action target spots is single, Clinical practice toxic side effect is more, not good enough to the long-term efficacy of AD patient etc. for these drugs Problem.

In recent years, with constantly illustrating to AD pathogenesis, it is found that the occurrence and development of AD have multimachine system, multifactor It is the characteristics of effect, again interrelated between different mechanisms to influence each other, constitute complicated network during AD occurrence and development Regulator 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, and synergistic effect can be generated to the effect of each target spot, so that gross effect is more than each Single-action the sum of is answered, and such medicine is also referred to as " Multifunctional " or " Multipotential " drug.Multiple target point drug with it is more Medicine use in conjunction and the main distinction of compound medicine are：Dosage can be reduced, therapeutic effect is improved, avoids between drug Interaction and the toxic side effect thus brought, uniform pharmacokinetic properties, be easy to use etc..It designs and finds have simultaneously There are anti-acetylcholinesterase, anti-monoamine oxidase-B, anti-oxidation stress, complexing of metal ion, the mistake for inhibiting beta-amyloid protein Degree generates and deposition, and the multiple target point AD medicines of various active equilibrium are current research hotspots.Therefore, it researchs and develops With novel chemical structure, novel mechanism of action, and the anti-neurodegenerative disease with multiple target effect, less toxic side effect is controlled The active demand that drug not only conforms with social senilization's process is treated, and there are good market prospects.

Invention content

Present invention aims at open a kind of novel homoisoflavone Mannich alkaloid compounds（I）And its it can pharmaceutically connect The salt received.

Another object of the present invention is to disclose such homoisoflavone Mannich alkaloid compound（I）And its it is pharmaceutically acceptable Salt production process.

Another object of the present invention is to open comprising such homoisoflavone Mannich alkaloid compound（I）And its pharmaceutically The pharmaceutical composition of acceptable salt.

Still a further object of the present invention is to disclose such homoisoflavone Mannich alkaloid compound（I）And its it is pharmaceutically acceptable Salt have multiple target effect, can be used for prepare treatment and/or prevent nervus retrogression relevant disease drug in purposes, packet Include but be not limited to vascular dementia, Alzheimer's disease, parkinsonism, Huntingdon disease, HIV related dementia disorders, multiple hard Change the neurodegenerative diseases such as disease, progressive lateral sclerosis of spinal cord, neuropathic pain, glaucoma.

Homoisoflavone Mannich alkaloid compound provided by the present invention（I）Chemical structure of general formula be：

In formula：R₁、R₂And R₃Each independently represent H, OH, CF₃O、C₁~C₁₂Alkoxy, NR₆R₇, R₄、R₅、R₆And R₇Respectively Independently indicate C₁~C₁₂Alkyl, propargyl, benzyl, substituted benzyl, NR₄R₅And NR₆R₇It may also indicate that nafoxidine base, morpholine Base, piperidyl, piperazinyl, 4- by C₁~C₁₂Piperazinyl that alkyl is replaced, the 4- piperazines replaced by benzyl or substituted benzyl Piperazine base；R₁、R₂、R₃、-CH₂NR₄R₅It can be in the arbitrarily possible position of phenyl ring with OH；

Above-mentioned term " substituted benzyl " refers to the benzyl replaced by 1-4 groups selected from the group below on phenyl ring：F,Cl, Br、I、C_1-4Alkyl, C_1-4Alkoxy, trifluoromethyl, trifluoromethoxy, dimethylamino, nitro, cyano, these substituent groups can be The arbitrary possible position of phenyl ring.

Homoisoflavone Mannich alkaloid compound proposed by the invention（I）It can be prepared by the following method to obtain：

In formula：R₁~R₅Definition and homoisoflavone Mannich alkaloid compound（I）Chemical structure of general formula it is identical.

With corresponding chromanone compound（1）With hydroxy benzaldehyde Mannich alkaloid compound（2）For starting material, Direct polycondensation under solvent and alkaline condition obtains corresponding homoisoflavone Mannich alkaloid compound（I）.Wherein, alkali used is reacted For：Alkali metal hydroxide, alkaline earth metal hydroxide, alkali carbonate, alkaline earth metal carbonate, alkali metal hydrogen carbonate Salt, alkali metal bicarbonates, C_1-8Alkali metal salt, trimethylamine class or the quaternary ammonium bases of alcohol（Such as：Triethylamine, tri-n-butylamine, three Octylame, pyridine,NMethyl morpholine,NMethyl piperidine, triethylene diamine, tetrabutylammonium hydroxide）, preferably alkali is：Potassium hydroxide, Sodium hydroxide, potassium carbonate, triethylamine or pyridine；Reacting solvent for use is：C_1-8Fatty alcohol, ether, tetrahydrofuran, 2- methyl four Hydrogen furans,N,NDimethylformamide, dimethyl sulfoxide (DMSO), dichloromethane, chloroform, Isosorbide-5-Nitrae-dioxane, benzene, toluene or acetonitrile, Preferred solvent is：Methanol, ethyl alcohol, isopropanol,N,NDimethylformamide, acetonitrile, tetrahydrofuran, dichloromethane or toluene；Color Full ketone compounds（1）：Hydroxy benzaldehyde Mannich alkaloid compound（2）：The molar feed ratio of alkali is 1.0：1.0~3.0：1.0 ~ 20.0, preferably molar feed ratio is 1.0：1.0~2.0：1.0~10.0；Reaction temperature is 0 ~ 150 DEG C, and preferable reaction temperature is room Temperature ~ 120 DEG C；Reaction time is 1 ~ 120 hour, and preferred reaction time is 2 ~ 72 hours.

Starting material --- the chromanone compound of the present invention（1）With hydroxy benzaldehyde Mannich alkaloid compound（2） The technology that this field can be used common is made, including but not limited to the method disclosed in following documents：1,Yoshihisa T.; Kenji M. European Journal of Organic Chemistry2001, (10), 1963-1966；2, Mahapatra T. et al. Tetrahedron: Asymmetry2008, 19(10), 1224-1232；3,Aissaoui H. et al.WO2014072903；4,Fattorusso C.et al.Journal of Medicinal Chemistry2008, 51(5), 1333-1343；5,Karki S.S.et al.Journal of Medicinal Chemistry2016, 59(2), 763-769；6,Torrente E.et al.Journal of Medicinal Chemistry2015, 58(15), 5900-5915。

The homoisoflavone Mannich alkaloid compound of gained according to the method described above（I）Contain amino in molecule, which is in Its pharmaceutically acceptable salt can be made by pharmaceutically conventional salifying method with any suitable acid in alkalinity, described Acid is：Hydrochloric acid, hydrobromic acid, nitric acid, sulfuric acid, phosphoric acid, C_1-6Aliphatic carboxylic acid（Such as：Formic acid, acetic acid, propionic acid etc.）, oxalic acid, benzoic acid, Salicylic acid, maleic acid, fumaric acid, succinic acid, tartaric acid, citric acid, malic acid, lipoic acid, C_1-6Alkyl sulfonic acid（Such as：Methyl sulphur Acid, ethylsulfonic acid etc.）, camphorsulfonic acid, benzene sulfonic acid or p-methyl benzenesulfonic acid.

Pharmaceutical composition disclosed in this invention includes one or more homoisoflavone Mannich bases of therapeutically effective amount Compound（I）Or its pharmaceutically acceptable salt, the pharmaceutical composition can further contain one or more pharmaceutically acceptable Carrier or excipient." therapeutically effective amount " refers to causing researcher or targeted tissue, system or the animal of doctor The amount of the drug or medicament of biology or medicine reaction；" composition " refers to by by more than one substances or component mixing At product；" pharmaceutically acceptable carrier " refers to pharmaceutically acceptable substance, composition or carrier, such as：Liquid Or solid-filling agent, diluent, excipient, solvent or packing substance, they carry or transport certain chemical substance.Institute of the present invention Its ideal ratio of the pharmaceutical composition of offer is homoisoflavone Mannich alkaloid compound（I）Or its is pharmaceutically acceptable Salt accounts for total weight than 5%~99.5% as active constituent, and rest part is to account for total weight than 95% or less.

Homoisoflavone Mannich alkaloid compound disclosed in this invention（I）And its pharmaceutically acceptable salt carried out as Under bioactivity screening.

（1）Homoisoflavone Mannich alkaloid compound（I）To the inhibitory activity of acetylcholinesterase and butyrylcholine esterase

1.0 mmol/L acetylthiocholine iodides or iodine bisulfide are sequentially added into 96 orifice plates for BuCh（It is purchased from Sigma companies）30 μ L, 40 μ L of PBS buffer solution of pH7.4,20 μ L of testing compound solution（DMSO contents are less than 1%）With 10 μ L acetylcholinesterases（Rat brain cortex 5% is homogenized supernatant, and the phosphate buffer of pH7.4 makees homogenate medium）Or BuCh Esterase（25% supernatant of rat blood serum, pH7.4 phosphate buffers make homogenate medium）Solution, after finishing mixing, 37 DEG C of incubations 15min is added 0.2% 5,5 '-two thio-bis- (2- nitrobenzoic acids) into each hole（DTNB is purchased from Sigma companies）Solution 30 μ L colour developings, the optical density in each hole at 405nm is measured with microplate reader（OD values）, compared with the blank well for being not added with sample to be tested, meter Calculate inhibiting rate of the compound to enzyme（Enzyme inhibition rate (%)=(1- sample sets OD values/blank group OD values) × 100%）；Select compound Five to six concentration, measure its enzyme inhibition rate, and linearly return with the inhibiting rate of the negative logarithm of the compound molar concentration and enzyme Return, molar concentration when acquiring 50% inhibiting rate is the IC of the compound₅₀.Measurement result shows that institute is public in the embodiment of the present invention The homoisoflavone Mannich alkaloid compound opened（I）The effect of significantly inhibiting, IC are all had to acetylcholinesterase₅₀It is 2.49 nM~10.0 µM；And homoisoflavone Mannich alkaloid compound（I）The inhibitory activity of acetylcholinesterase is significantly higher than pair The inhibitory activity of butyrylcholine esterase（Selectivity is more than 10 times）, illustrate compound disclosed in this invention to acetylcholinesterase With selective inhibitory.Measurement result is also shown that homoisoflavone Mannich alkaloid compound（I）Parent nucleus --- 2 '-hydroxyls Base homoisoflavone（R₁、R₂、R₃With-CH₂NR₄R₅Indicate that H, OH are located at 2 ' positions）, 3 '-hydroxyl homoisoflavones（R₁、R₂、R₃With- CH₂NR₄R₅Indicate that H, OH are located at 3 ' positions）With 4 '-hydroxyl homoisoflavones（R₁、R₂、R₃With-CH₂NR₄R₅Indicate that H, OH are located at 4 ' Position）The IC that acetylcholinesterase is inhibited₅₀It is all higher than 500 μM.

（2）Homoisoflavone Mannich alkaloid compound（I）To the inhibitory activity of monoamine oxidase B

Recombined human MAO-B is made into 75 μ g/mL sample liquids with 7.4 kaliumphosphate buffers of pH of 100 mM.To black 96 20 μ L of testing compound solution are added in orifice plate, 80 μ L of monoamine oxidase, mixing, 37 °C are incubated 15 min in the place of being protected from light, and add Enter 200 μM of Amplex Red reagents, 2U/mL horseradish peroxidases, 2 mM benzene methanamine initiation reactions, 37 °C are incubated 20 Min, to fix 545 nm of excitation wavelength, is surveyed fluorescent emission intensity at 590 nm, is buffered with potassium phosphate on multi-function microplate reader It is blank that liquid, which replaces MAO-B,；Compound inhibit monoamine oxidase inhibiting rate calculation formula be：100-(IF_i)/(IF_c) * 100, In formula, IF_iAnd IF_cRespectively there is the difference of the fluorescence intensity and blank fluorescence intensity under inhibitor and no inhibitor.Each chemical combination Object measures 3 multiple holes every time, and every group of experiment is independent in triplicate.Five to six concentration for selecting compound measure the inhibition of its enzyme Rate, and with the inhibiting rate linear regression of the negative logarithm of the compound molar concentration and enzyme, acquire molar concentration when 50% inhibiting rate The as IC of the compound₅₀.Measurement result shows the homoisoflavone Mannich alkaloid compound disclosed in the embodiment of the present invention （I）The effect of significantly inhibiting, IC are all had to MAO-B₅₀It is 0.1 μM ~ 50.0 μM；And compound（I）Parent nucleus --- 2 '-hydroxyls Base homoisoflavone（R₁、R₂、R₃With-CH₂NR₄R₅Indicate that H, OH are located at 2 ' positions）, 3 '-hydroxyl homoisoflavones（R₁、R₂、R₃With- CH₂NR₄R₅Indicate that H, OH are located at 3 ' positions）With 4 '-hydroxyl homoisoflavones（R₁、R₂、R₃With-CH₂NR₄R₅Indicate that H, OH are located at 4 ' Position）The IC that MAO-B is inhibited₅₀>100 µM。

（3）Homoisoflavone Mannich alkaloid compound（I）Antioxidant activity（ORAC-FL methods）

Reference literature（Qiang, X.M.et al.Eur. J Med. Chem.2014, 76, 314-331）It is reported Method be measured, i.e.,：6- hydroxyl -2,5,7,8- tetramethyl primary colours alkane -2- carboxylic acids（Trolox）With the PBS buffer solution of pH7.4 It is made into the solution of 10-80 μm of ol/L, fluorescein（fluorescein）It is made into 250 nmol/L's with the PBS buffer solution of pH7.4 Solution, 2,2 '-azo diisobutyl amidine dihydrochlorides（AAPH）It is made into 40 mmol/L's with the PBS buffer solution of pH7.4 using preceding Solution.The compound solution and luciferin solution of 50-10 μm of ol/L, mixing are added into 96 orifice plates, 37 °C of incubation 15min add Enter AAPH solution, it is 200 μ L to make every hole total volume, and mixing is immediately placed on Varioskan Flash Multimode Reader In (Thermo Scientific) instrument, 90 min of METHOD FOR CONTINUOUS DETERMINATION under 485 nm excitation wavelengths and 535 nm launch wavelengths.It calculates Go out area AUC under fluorescence decay curve, wherein with 1-8 μm of ol/L'sTroloxAs standard, to be not added with sample to be tested as blank, The antioxidant activity results expression of compound isTroloxEquivalent, calculation formula is：[(AUC Sample-AUC blank)/(AUC Trolox-AUC blank)] ×[(concentration of Trolox/concentration of Sample)], each compound measures 3 multiple holes every time, and every group of experiment is independent in triplicate.Measurement result shows present invention reality Apply the homoisoflavone Mannich alkaloid compound disclosed in example（I）Antioxidant activity beTrolox1.0~3.0 times, say Such bright compound has strong anti-oxidative activity.

（4）Homoisoflavone Mannich alkaloid compound（I）To A β_1-42The inhibitory activity of self assemble

Reference literature（Qiang, X.M.et al.Eur. J Med. Chem.2014, 76, 314-331）It is reported Method be measured, i.e.,：Pretreated Aβ _1-42It is made into storing solution with DMSO, using preceding dilute with the PBS buffer solution of pH7.4 It releases to 50 μM；Untested compound is made into 2.5 mM storing solutions with DMSO, is diluted to accordingly with the PBS buffer solution of pH7.4 using preceding Concentration takes the A of 20 μ Lβ _1-42The testing compound solution of+20 μ L of solution, the A of 20 μ Lβ _1-42The PBS buffer solution of+20 μ L of solution（Contain 2%DMSO）In 96 orifice plates, 37 °C are incubated for 24 hours, and the glycine-NaOH for the 50mM that 160 μ L contain 5 μM of thioflavine Ts is then added Buffer solution（pH=8.5）, surveyed under 446 nm excitation wavelengths and 490 nm launch wavelengths with multi-function microplate reader immediately after shaking 5s Determine fluorescent value；Aβ _1-42The fluorescent value of+untested compound is denoted as IF_i, Aβ _1-42The fluorescent value of+PBS buffer solution is denoted as IF_c, contain only The fluorescent value of PBS buffer solution is denoted as IF₀, compound inhibition Aβ _1-42The inhibiting rate of self assemble is：100-(IF_i-IF₀)/(IF_c- IF₀)*100；Five to six concentration for selecting compound, measure its inhibiting rate, each each concentration repetition measurement of compound three times, with Curcumin is positive control.Measurement result shows the homoisoflavone Mannich alkaloid compound disclosed in the embodiment of the present invention （I）To Aβ _1-42Self assemble all has remarkable inhibiting activity, to A under 25.0 μM of concentrationβ _1-42The inhibiting rate of self assemble is equal More than 50.0%；And inhibiting rate of the curcumin under same concentrations is 41.3%, clinically widely used anti-AD drugs：It is more how piperazine Together, Rivastigmine, memantine hydrochloride and compound（I）Parent nucleus --- 2 '-hydroxyl homoisoflavones（R₁、R₂、R₃With- CH₂NR₄R₅Indicate that H, OH are located at 2 ' positions）, 3 '-hydroxyl homoisoflavones（R₁、R₂、R₃With-CH₂NR₄R₅Indicate that H, OH are located at 3 ' Position）With 4 '-hydroxyl homoisoflavones（R₁、R₂、R₃With-CH₂NR₄R₅Indicate that H, OH are located at 4 ' positions）To A under 25.0 μM of concentrationβ _1-42The inhibiting rate of self assemble is respectively less than 10%.

（5）Homoisoflavone Mannich alkaloid compound（I）With the measurement of complexing of metal ion effect

CuCl is dissolved with methanol₂·2H₂O、ZnCl₂、FeSO₄·7H₂O、AlCl₃And untested compound, it is made into 75 μm of ol/L Solution, 100 μ L testing compound solutions and 100 μ L metal ion solutions are added into 96 orifice plates, mixing is stored at room temperature 30 Min, record mixture exists on Varioskan Flash Multimode Reader (Thermo Scientific) instrument Ultraviolet absorption curve within the scope of 200-600 nm, and be pair with 100 μ L testing compound solutions and 100 μ L methyl alcohol mixed liquors According to observation metal ion and the Red Shift Phenomena of the maximum absorption band of untested compound mixed liquor and the intensity of maximum absorption band.It surveys It is fixed the result shows that, the homoisoflavone Mannich alkaloid compound disclosed in the embodiment of the present invention（I）It shows to copper ion Selective complexing.

（6）Homoisoflavone Mannich alkaloid compound（I）To Cu²⁺The A β of induction_1-42The inhibitory activity of aggregation

By CuCl₂75 μM of solution are made into HEPES buffer solution, with HEPES buffer solution by compound stock solution（2.5 mM） With 200 μM of Aβ _1-42Storing solution is diluted to 75 μM, takes 20 μ L Cu respectively²⁺+ 20 μ L A of solutionβ _1-42+ 20 μ L to be measuredization of solution Polymer solution, 20 μ L Cu²⁺+ 20 μ L A of solutionβ _1-42+ 20 μ L HEPES buffer solutions of solution and 60 μ L HEPES buffer solutions are in 96 In orifice plate, then the glycine-NaOH buffer for the 50mM that 190 μ L contain 5 μM of thioflavine Ts is added in mixing, 37 °C of 24 h of incubation （pH=8.5）, after shaking 5s fluorescence is measured under 446nm excitation wavelengths and 490nm launch wavelengths with multi-function microplate reader immediately Value；Cu²⁺+Aβ _1-42The fluorescent value of+untested compound is recorded as IF_i, Cu²⁺+Aβ _1-42The fluorescent value of+HEPES buffer solution is recorded as IF_c, the fluorescent value for containing only HEPES buffer solution is recorded as IF₀, compound is to Cu²⁺The A of inductionβ _1-42The inhibiting rate of aggregation is： 100-(IF_i-IF₀)/(IF_c-IF₀)*100.Each compound three multiple holes of each concentration mensuration, using curcumin as positive control. Measurement result shows the homoisoflavone Mannich alkaloid compound disclosed in the embodiment of the present invention（I）Under 25.0 μM of concentration To Cu²⁺The A of inductionβ _1-42The inhibiting rate of aggregation is all higher than 60.0%；And inhibiting rate of the curcumin under same concentrations is 52.1%, Compound（I）Parent nucleus --- 2 '-hydroxyl homoisoflavones（R₁、R₂、R₃With-CH₂NR₄R₅Indicate that H, OH are located at 2 ' positions）, 3 '-hydroxyls Base homoisoflavone（R₁、R₂、R₃With-CH₂NR₄R₅Indicate that H, OH are located at 3 ' positions）With 4 '-hydroxyl homoisoflavones（R₁、R₂、R₃With- CH₂NR₄R₅Indicate that H, OH are located at 4 ' positions）Inhibiting rate under same concentrations is less than 20.0%.

Specific implementation mode

The present invention can be further described by the following examples, however, the scope of the present invention is not limited to Following embodiments.One of skill in the art, can be right it is understood that under the premise of without departing substantially from the spirit and scope of the present invention The present invention carries out various change and modification.

1 homoisoflavone Mannich alkaloid compound of embodiment（I）Preparation lead to method

The corresponding chromanone compounds of 2.0 mmol are added in reaction bulb（1）, the corresponding hydroxy benzenes first of 3.0 mmol Aldehyde Mannich alkaloid compound（2）With 30 ml methanol, after stirring evenly, it is added dropwise to 30% KOH aqueous solutions, 12.0 mmol, is heated up Return stirring reacts 3.0~24.0 hours（Reaction process is tracked with TLC）；After reaction, it is cooled to room temperature, with 10% hydrochloric acid Aqueous solution adjusts reaction solution pH to highly acid, then adjusts reaction solution pH to alkalescent with saturated sodium bicarbonate aqueous solution, and decompression is steamed Except methanol, 80 mL deionized waters are added in residual solution, are extracted in three times with 240 mL dichloromethane, organic layer is used full after merging It is washed with sodium-chloride water solution, is filtered after being dried over anhydrous sodium sulfate, remove solvent under reduced pressure, residue is purified through column chromatography（It washes De- liquid：Dichloromethane:Acetone=10:1 v/v）, obtain corresponding homoisoflavone Mannich alkaloid compound（I）, yield 20.0%- 75.0%, chemical constitution passes through¹H-NMR、¹³C-NMR and ESI-MS confirmations；The purity of gained object measures big through HPLC In 97.0%.The object structure being prepared using above-mentioned logical method is as follows：

；

。

2 homoisoflavone Mannich alkaloid compound of embodiment（I）It is prepared at salt with acid and leads to method

The homoisoflavone Mannich alkaloid compound of gained in accordance with the above-mentioned embodiment 1 is added in reaction bulb（I）2.0 50 ml of mmol and acetone, is stirring evenly and then adding into that 8.0 mmol are sour accordingly, and temperature rising reflux is stirred to react 20 minutes, reaction knot Beam postcooling removes solvent under reduced pressure to room temperature, residue acetone recrystallization, and the solid for filtering precipitation is graceful to get homoisoflavone Buddhist nun wishes alkaloid compound（I）Salt, chemical constitution warp¹H NMR and ESI-MS confirmations.

Claims (7)

1. a kind of homoisoflavone Mannich alkaloid compound or its pharmaceutically acceptable salt, it is characterised in that such compound Chemical structure of general formula is such as（I）It is shown：

In formula：R₁、R₂And R₃Each independently represent H, OH, CF₃O、C₁~C₁₂Alkoxy, NR₆R₇, R₄、R₅、R₆And R₇It is respectively independent Ground indicates C₁~C₁₂Alkyl, propargyl, benzyl, substituted benzyl, NR₄R₅And NR₆R₇It may also indicate that nafoxidine base, morpholinyl, piperazine Piperidinyl, piperazinyl, 4- by C₁~C₁₂Piperazinyl that alkyl is replaced, the 4- piperazinyls replaced by benzyl or substituted benzyl, R₁、R₂、R₃、-CH₂NR₄R₅It can be in the arbitrarily possible position of phenyl ring with OH；" substituted benzyl " refers to by a by 1-4 on phenyl ring The benzyl that group selected from the group below is replaced：F,Cl,Br,I,C_1-4Alkyl, C_1-4Alkoxy, trifluoromethyl, trifluoromethoxy, two Methylamino, nitro, cyano, these substituent groups can be in the arbitrary possible positions of phenyl ring.

2. homoisoflavone Mannich alkaloid compound as described in claim 1 or its pharmaceutically acceptable salt, feature exist In the pharmaceutically acceptable salt be such homoisoflavone Mannich alkaloid compound and hydrochloric acid, hydrobromic acid, nitric acid, sulphur Acid, phosphoric acid, C_1-6Aliphatic carboxylic acid, oxalic acid, benzoic acid, salicylic acid, maleic acid, fumaric acid, succinic acid, tartaric acid, citric acid, apple Tartaric acid, lipoic acid, C_1-6The salt of alkyl sulfonic acid, camphorsulfonic acid, benzene sulfonic acid or p-methyl benzenesulfonic acid.

3. the system of homoisoflavone Mannich alkaloid compound or its pharmaceutically acceptable salt as described in claim any one of 1-2 Preparation Method, it is characterised in that the compound can be prepared by the following method to obtain：

In formula：R₁~R₅Definition and homoisoflavone Mannich alkaloid compound（I）Chemical structure of general formula it is identical；

With corresponding chromanone compound（1）With hydroxy benzaldehyde Mannich alkaloid compound（2）For starting material, in solvent With direct polycondensation under alkaline condition, corresponding homoisoflavone Mannich alkaloid compound is obtained（I）；Utilize the height of above method gained Contain amino in isoflavones Mannich bases compound molecule, which, can be with any suitable acid by pharmaceutically in alkalinity Its pharmaceutically acceptable salt is made in conventional salifying method.

4. the preparation method of homoisoflavone Mannich alkaloid compound as claimed in claim 3 or its pharmaceutically acceptable salt, It is characterized in that reaction alkali used is：Alkali metal hydroxide, alkaline earth metal hydroxide, alkali carbonate, alkaline-earth metal Carbonate, alkali metal hydrogencarbonate, alkali metal bicarbonates, C_1-8The alkali metal salt of alcohol, triethylamine, tri-n-butylamine, trioctylamine, Pyridine,NMethyl morpholine,NMethyl piperidine, triethylene diamine or tetrabutylammonium hydroxide；Reacting solvent for use is：C_1-8Fat Alcohol, ether, tetrahydrofuran, 2- methyltetrahydrofurans,N,NDimethylformamide, dimethyl sulfoxide (DMSO), dichloromethane, chloroform, 1, 4- dioxane, benzene, toluene or acetonitrile.

5. the preparation method of homoisoflavone Mannich alkaloid compound as claimed in claim 3 or its pharmaceutically acceptable salt, It is characterized in that chromanone compound（1）：Hydroxy benzaldehyde Mannich alkaloid compound（2）：The molar feed ratio of alkali is 1.0：1.0~3.0：1.0~20.0；Reaction temperature is 0 ~ 150 DEG C；Reaction time is 1 ~ 120 hour.

6. a kind of pharmaceutical composition, it is characterised in that comprising such as claim 1-2 any one of them homoisoflavone Mannich bases Class compound or its pharmaceutically acceptable salt and one or more pharmaceutically acceptable carriers or excipient.

7. as claim 1-2 any one of them homoisoflavone Mannich alkaloid compound or its pharmaceutically acceptable salt exist It prepares treatment and/or prevents the purposes in nervus retrogression relevant disease drug, this kind of nervus retrogression relevant disease is：Blood vessel Property dementia, Alzheimer's disease, parkinsonism, Huntingdon disease, HIV related dementia disorders, multiple sclerosis, progressive spinal cord Lateral schlerosis, neuropathic pain or glaucoma.