CN106632191A - Homoisoflavonoid mannich base compound and preparation method and application thereof - Google Patents

Abstract

The invention discloses a novel homoisoflavonoid mannich base compound (I) and pharmaceutically acceptable salts, a preparation method, a pharmaceutical composition and application thereof in preparation of medicines for treatment and/or prevention of neurodegeneration related diseases. The neurodegeneration related diseases include but not limited to vascular dementia, Alzheimer's disease, Parkinson's disease, Huntington's disease, HIV (human immunodeficiency virus) related dementia, multiple sclerosis, progressive lateral sclerosis of spinal cord, neuropathic pain, glaucoma and the like.

Description

Homoisoflavone Mannich alkaloid compound, Preparation Method And The Use

Technical field

The invention belongs to medicinal chemistry art, is related to the new homoisoflavone Mannich alkaloid compound of a class（I）And its medicine Acceptable salt on, its preparation method, pharmaceutical composition and preparing treatment and/or preventing nervus retrogression relevant disease medicine Purposes in thing, including but not limited to vascular dementia, Alzheimer's disease, parkinsonism, Huntingdon disease, HIV are related silly The nerve degenerative diseases such as slow-witted disease, multiple sclerosis, progressive lateral sclerosis disease, 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 infringement, its incidence of disease becomes and is only second to the heart in ascendant trend year by year The frequently-occurring disease of angiosis and cancer, has gone up as the 4th of the cause of death in developed countries such as America and Europes.According to world health Organisation Report, global over-65s old man has 10% dysnoesia, wherein 1/2nd occur dull-witted, morbidity in more than 85 years old Rate nearly 50%.In China AD patient numbers about 600-700 ten thousand, the incidence of disease is more than 5%.With adding for population in the world aging process Hurry up, its incidence of disease is in obvious ascendant trend, is announced in December, 2013 according to Alzheimer's Disease International 's《The global implication of Alzheimer's disease：2013-2050》Point out in report, AD will become the coming few decades whole world and face most 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.Because AD clinical manifestations are gone down for memory capability, capacity of orientation, thinking and judgement, and Activity of daily living is reduced, or even abnormal Behavioral and psychological symptom etc. occurs, makes patient care difficulty larger, to society and family's band Carry out heavy burden.Current approved is used to treat the medicine of light/moderate AD acetylcholinesterase（AChE）Inhibitor, Yi Jiyong In the treatment of severe ADN- methyl-D- aspartic acid（NMDA）Receptor antagonist, but Clinical practice shows that these medicines can pass through Improve in patient's body levels of acetylcholine or suppress the exitotoxicity of excitatory amino acid alleviating AD symptoms, but can not be effective Prevent or reverse the course of disease, but also can cause hallucinations, misunderstanding, dizziness, headache, nausea, hepatotoxicity, poor appetite and The serious toxic and side effect such as stool frequency, thus long-term efficacy is not satisfactory.Therefore, clinically in the urgent need to research and development have new work With the AD medicines of mechanism.

The disease that AD category many factors cause, pathogenesis is complicated, does not also illustrate its pathogenesis completely so far, but studies Show, the decline of patient's intracerebral levels of acetylcholine, the excessive generation of amyloid-beta are disorderly with deposition, metal ion metabolism Disorderly, Ca²⁺Dysequilibrium,tauNeurofibrillary tangles, glutamate receptor activity are too high caused by-protein hyperphosphorylation, oxidation should Swash and produce 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 Neuroinflammation Want role.For above-mentioned pathogenic factors, researcher is using traditional " target of a medicine one " drug design strategies, it was found that a large amount of to certain One target spot has the medicine of high activity and high selectivity, such as：Anticholinesterase andN- methyl-D- aspartate receptor antagonism Agent etc., but these medicines are not present that action target spot is single, Clinical practice toxic and side effect is more, good enough to the long-term efficacy of AD patient etc. Problem.

In recent years, with constantly illustrating to AD pathogenesis, the occurrence and development for finding AD have many mechanism, multifactor It is the characteristics of effect, again interrelated between different mechanisms to influence each other, constitute network complicated during AD occurrence and development Regulator control system.Based on the above results, researcher proposes " Mutiple Targets targeted drug "（Multitarget-directed Ligands, MTDLs）Strategy is researching and developing anti-nerve degenerative diseases medicine.So-called " Mutiple Targets medicine " refers to that single chemistry is real Body acts on the multiple target spots in disease network simultaneously, and the effect to each target spot can produce cooperative effect, makes gross effect more than each Single-action answers sum, such medicine to be also referred to as " Multifunctional " or " Multipotential " medicine.Mutiple Targets medicine with it is many Medicine use in conjunction and compound medicine are differred primarily in that：Dosage can be reduced, therapeutic effect is improved, is avoided between medicine The toxic and side effect for interacting and thus bringing, homogeneous pharmacokinetic properties, be easy to use etc..Design and find tool simultaneously There are anti-acetylcholinesterase, anti-monoamine oxidase-B, anti-oxidation stress, complexing of metal ion, the mistake of suppression amyloid-beta Degree generate with deposition, and various active in a balanced way Mutiple Targets AD medicines are current study hotspots.Therefore, research and develop With novel chemical structure, new mechanism of action, and the anti-nerve degenerative diseases with multiple target effect, low toxicity side effect are controlled The active demand that medicine not only conforms with social senilization's process is treated, and with good market prospects.

The content of the invention

Present invention aim at disclosing the new homoisoflavone Mannich alkaloid compound of a class（I）And its pharmaceutically can 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.

A further object of the present invention is 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 there is multiple target effect, can be used for prepare treatment and/or prevent nervus retrogression relevant disease medicine in purposes, bag Include but be not limited to vascular dementia, Alzheimer's, parkinsonism, Huntingdon disease, HIV related dementia disorders, multiple hard Change the nerve degenerative diseases such as disease, progressive lateral sclerosis disease, neuropathic pain, glaucoma.

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

In formula：R₁、R₂And R₃H, OH, CF are represented independently of one another₃O、C₁~C₁₂Alkoxyl, NR₆R₇, R₄、R₅、R₆And R₇It is each independent Ground represents C₁~C₁₂Alkyl, propargyl, benzyl, substituted benzyl, NR₄R₅And NR₆R₇May also indicate that nafoxidine base, morpholinyl, piperazine Piperidinyl, piperazinyl, 4- positions are by C₁~C₁₂The piperazinyl that piperazinyl that alkyl is replaced, 4- positions are replaced by benzyl or substituted benzyl； R₁、R₂、R₃、-CH₂NR₄R₅Can be in phenyl ring arbitrarily possible position with OH；

Above-mentioned term " substituted benzyl " refers to the benzyl replaced by the group that 1-4 is selected from the group on phenyl ring：F、Cl、Br、 I、C_1-4Alkyl, C_1-4Alkoxyl, trifluoromethyl, trifluoromethoxy, dimethylamino, nitro, cyano group, these substituents can be in phenyl ring Any possible position.

Homoisoflavone Mannich alkaloid compound proposed by the invention（I）Can be prepared by the following method and 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 initiation material, Direct polycondensation under solvent and alkalescence 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_1-8The alkali metal salt of alcohol, trimethylamine class or quaternary ammonium bases（Such as：Triethylamine, tri-n-butylamine, three Octylame, pyridine,N- methyl morpholine,N- methyl piperidine, triethylene diamine, TBAH）, preferred alkali is：Potassium hydroxide, NaOH, potassium carbonate, triethylamine or pyridine；Reacting solvent for use is：C_1-8Fatty alcohol, ether, tetrahydrofuran, 2- methyl four Hydrogen furans,N,N- dimethylformamide, dimethyl sulfoxide (DMSO), dichloromethane, chloroform, Isosorbide-5-Nitrae-dioxane, benzene, toluene or acetonitrile, Preferred solvent is：Methyl alcohol, ethanol, isopropanol,N,N- dimethylformamide, 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, preferred 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.

Initiation material --- the chromanone compound of the present invention（1）With hydroxy benzaldehyde Mannich alkaloid compound（2） Can be obtained with the common technology in this area, including but not limited to the method disclosed in documents below：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, the amino is in Alkalescence, can be obtained its pharmaceutically acceptable salt with any suitable acid by pharmaceutically conventional salifying method, 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, butanedioic 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 be further pharmaceutically acceptable containing one or more Carrier or excipient." therapeutically effective amount " refers to the tissue, system or the animal that cause researcher or doctor targeted The amount of biological or medicine reaction medicine or medicament；" composition " is referred to by by more than one materials or component mixing Into product；" pharmaceutically acceptable carrier " refers to pharmaceutically acceptable material, composition or carrier, such as：Liquid Or solid-filling agent, diluent, excipient, solvent or packing material, they carry or transport certain chemical substance.Institute of the present invention The pharmaceutical composition of offer its preferable ratio is, homoisoflavone Mannich alkaloid compound（I）Or its is pharmaceutically acceptable Salt accounts for gross weight than 5%～99.5% as active component, and remainder is to account for gross weight than less than 95%.

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 acetylcholinesterase and the inhibitory activity of butyrylcholine esterase

1.0 mmol/L acetylthiocholine iodides or iodine bisulfide are sequentially added in 96 orifice plates for BuCh（It is purchased from Sigma companies）The μ L of PBS 40 of 30 μ L, pH7.4, the μ L of testing compound solution 20（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（The supernatant of rat blood serum 25%, pH7.4 phosphate buffers make homogenate medium）Solution, after finishing mixing, 37 DEG C of incubations 15min, adds 0.2% 5,5 '-two thio-bis- (2- nitrobenzoic acids) in each hole（DTNB, purchased from Sigma companies）Solution 30 μ L develop the color, and with ELIASA the optical density in each hole at 405nm is determined（OD values）, compare with the blank well for being not added with testing sample, count Calculate inhibiting rate of the compound to enzyme（Enzyme inhibition rate (%)=(1- sample sets OD value/blank group OD value) × 100%）；Select compound Five to six concentration, determine its enzyme inhibition rate, and linearly return with the negative logarithm of the compound molar concentration and the inhibiting rate of enzyme Return, molar concentration when trying to achieve 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 is respectively provided with to acetylcholinesterase, its IC₅₀For 2.49 nM~10.0 µM；And homoisoflavone Mannich alkaloid compound（I）It is right that the inhibitory activity of acetylcholinesterase is significantly higher than The inhibitory activity of butyrylcholine esterase（It is selective to be 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 '-hydroxyl Base homoisoflavone（R₁、R₂、R₃With-CH₂NR₄R₅Represent that H, OH are located at 2 ' positions）, 3 '-hydroxyl homoisoflavone（R₁、R₂、R₃With- CH₂NR₄R₅Represent that H, OH are located at 3 ' positions）With 4 '-hydroxyl homoisoflavone（R₁、R₂、R₃With-CH₂NR₄R₅Represent that H, OH are located at 4 ' Position）IC to acetylcholine ester enzyme level₅₀It is all higher than 500 μM.

（2）Homoisoflavone Mannich alkaloid compound（I）Inhibitory activity to MAO-B

Recombined human MAO-B is made into 75 μ g/mL sample liquids with the kaliumphosphate buffers of pH 7.4 of 100 mM.To the orifice plate of black 96 Middle addition the μ L of testing compound solution 20, the μ L of monoamine oxidase 80, mix, and 37 °C are incubated 15 min at lucifuge, add 200 μM Amplex Red reagents, 2U/mL horseradish peroxidases, 2 mM benzene methanamine initiation reactions, 37 °C of 20 min of incubation, many On function ELIASA, to fix the nm of excitation wavelength 545, fluorescent emission intensity at 590 nm is surveyed, with kaliumphosphate buffer replacement MAO-B is blank；Compound suppress monoamine oxidase inhibiting rate computing formula be：100-(IF_i)/(IF_c) * 100, in formula, IF_iAnd IF_cRespectively there is the difference of inhibitor and the fluorescence intensity under no inhibitor and blank fluorescence intensity.Each compound is every 3 multiple holes of secondary measure, per group of experiment is independent in triplicate.Five to six concentration of compound are selected, its enzyme inhibition rate is determined, and With the inhibiting rate linear regression of the negative logarithm of the compound molar concentration and enzyme, molar concentration when trying to achieve 50% inhibiting rate is The 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 is respectively provided with to MAO-B, its IC₅₀For 0.1 μM ~ 50.0 μM；And compound（I）Parent nucleus --- 2 '-hydroxyl Homoisoflavone（R₁、R₂、R₃With-CH₂NR₄R₅Represent that H, OH are located at 2 ' positions）, 3 '-hydroxyl homoisoflavone（R₁、R₂、R₃With- CH₂NR₄R₅Represent that H, OH are located at 3 ' positions）With 4 '-hydroxyl homoisoflavone（R₁、R₂、R₃With-CH₂NR₄R₅Represent that H, OH are located at 4 ' Position）The IC that MAO-B is suppressed₅₀>100 µM。

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

Reference literature（Qiang, X.M.et al.Eur. J Med. Chem.2014, 76, 314-331）The side for being reported Method is measured, i.e.,：6- hydroxyl -2,5,7,8- tetramethyl primary colours alkane -2- carboxylic acids（Trolox）It is made into the PBS of pH7.4 The solution of 10-80 μm of ol/L, fluorescein（fluorescein）The solution of 250 nmol/L is made into the PBS of pH7.4, 2,2 '-azo diisobutyl amidine dihydrochloride（AAPH）Using the front solution that 40 mmol/L are made into the PBS of pH7.4. The compound solution and luciferin solution of 50-10 μm of ol/L are added in 96 orifice plates, is mixed, 37 °C of incubation 15min are added AAPH solution, makes every hole cumulative volume be 200 μ L, mixes, and is immediately placed on Varioskan Flash Multimode Reader In (Thermo Scientific) instrument, the min of METHOD FOR CONTINUOUS DETERMINATION 90 under 485 nm excitation wavelengths and 535 nm launch wavelengths.Calculate Go out area AUC under fluorescence decay curve, wherein with 1-8 μm of ol/L'sTroloxAs standard, to be not added with testing sample as blank, The antioxidation activity results expression of compound isTroloxEquivalent, its computing formula is：[(AUC Sample-AUC blank)/(AUC Trolox-AUC blank)] ×[(concentration of Trolox/concentration of Sample)], each compound determines every time 3 multiple holes, and per group of experiment is independent in triplicate.Measurement result shows that the present invention is real Apply the homoisoflavone Mannich alkaloid compound disclosed in example（I）Antioxidation activity beTrolox1.0～3.0 times, say Bright such 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）The side for being reported Method is measured, i.e.,：Pretreated Aβ _1-42Storing solution is made into DMSO, is diluted to the PBS of pH7.4 using front 50μM；Testing compound DMSO is made into 2.5 mM storing solutions, and respective concentration is diluted to the PBS of pH7.4 using front, Take the A of 20 μ Lβ _1-42The A of the testing compound solution of the μ L of solution+20,20 μ Lβ _1-42The PBS of the μ L of solution+20（Containing 2% DMSO）In 96 orifice plates, 37 °C of incubation 24h, the glycine-NaOH for being subsequently adding the 50mM that 160 μ L contain 5 μM of thioflavine Ts delays Rush liquid（pH=8.5）, determined under 446 nm excitation wavelengths and 490 nm launch wavelengths with multi-function microplate reader immediately after shaking 5s Fluorescent value；Aβ _1-42The fluorescent value of+testing compound is designated as IF_i, Aβ _1-42The fluorescent value of+PBS is designated as IF_c, comprise only The fluorescent value of PBS is designated as IF₀, compound suppression Aβ _1-42The inhibiting rate of self assemble is：100-(IF_i-IF₀)/(IF_c- IF₀)*100；Five to six concentration of compound are selected, its inhibiting rate is determined, each compound each concentration repetition measurement 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 is respectively provided with 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 medicines：Many how piperazines Together, Rivastigmine, memantine hydrochloride and compound（I）Parent nucleus --- 2 '-hydroxyl homoisoflavone（R₁、R₂、R₃With- CH₂NR₄R₅Represent that H, OH are located at 2 ' positions）, 3 '-hydroxyl homoisoflavone（R₁、R₂、R₃With-CH₂NR₄R₅Represent that H, OH are located at 3 ' Position）With 4 '-hydroxyl homoisoflavone（R₁、R₂、R₃With-CH₂NR₄R₅Represent 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 measure of complexing of metal ion effect

CuCl is dissolved with methyl alcohol₂·2H₂O、ZnCl₂、FeSO₄·7H₂O、AlCl₃And testing compound, it is made into the molten of 75 μm of ol/L Liquid, 100 μ L testing compound solutions and 100 μ L metal ion solutions are added in 96 orifice plates, are mixed, and are stored at room temperature 30 min, Mixture is recorded on Varioskan Flash Multimode Reader (Thermo Scientific) instrument in 200-600 Ultraviolet absorption curve in the range of nm, and with 100 μ L testing compound solutions and 100 μ L methyl alcohol mixed liquors as control, observation gold The Red Shift Phenomena and the intensity of maximum absorption band of the maximum absorption band of category ion and testing compound mixed liquor.Measurement result table Bright, disclosed in the embodiment of the present invention homoisoflavone Mannich alkaloid compound（I）Show selective to copper ion 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, and 20 μ L Cu are taken respectively²⁺The μ L A of solution+20β _1-42The μ L test compounds of solution+20 Thing solution, 20 μ L Cu²⁺The μ L A of solution+20β _1-42The μ L HEPES buffer solutions of solution+20 and 60 μ L HEPES buffer solutions are in 96 holes In plate, mix, 37 °C of 24 h of incubation are subsequently adding the glycine-NaOH buffer of the 50mM that 190 μ L contain 5 μM of thioflavine Ts （pH=8.5）, fluorescence is determined under 446nm excitation wavelengths and 490nm launch wavelengths with multi-function microplate reader immediately after shaking 5s Value；Cu²⁺+Aβ _1-42The fluorescent value of+testing 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 comprising 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, with 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 homoisoflavone（R₁、R₂、R₃With-CH₂NR₄R₅Represent that H, OH are located at 2 ' positions）, 3 '-hydroxyl Base homoisoflavone（R₁、R₂、R₃With-CH₂NR₄R₅Represent that H, OH are located at 3 ' positions）With 4 '-hydroxyl homoisoflavone（R₁、R₂、R₃With- CH₂NR₄R₅Represent that H, OH are located at 4 ' positions）Inhibiting rate under same concentrations is less than 20.0%.

Specific embodiment

The present invention can be conducted further description 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 on the premise of without departing substantially from the spirit and scope of the present invention The present invention carries out various change and modification.

The homoisoflavone Mannich alkaloid compound of embodiment 1（I）The logical method of preparation

The corresponding chromanone compounds of 2.0 mmol are added in reaction bulb（1）, the corresponding hydroxy benzaldehydes of 3.0 mmol it is graceful Buddhist nun wishes alkaloid compound（2）With 30 ml methyl alcohol, after stirring, the mmol of the 30% KOH aqueous solution 12.0, temperature rising reflux are added dropwise to Stirring reaction 3.0～24.0 hours（Reaction process is tracked with TLC）；After reaction terminates, room temperature is cooled to, it is water-soluble with 10% hydrochloric acid Liquid adjusts reactant liquor pH to highly acid, then adjusts reactant liquor pH to alkalescent with saturated sodium bicarbonate aqueous solution, removes first under reduced pressure Alcohol, adds 80 mL deionized waters in residual solution, extracted in three times with 240 mL dichloromethane, and organic layer uses saturation chlorine after merging Change sodium water solution washing, filter Jing after anhydrous sodium sulfate drying, remove solvent, residue Jing column chromatographies purifying under reduced pressure（Eluent： Dichloromethane:Acetone=10:1 v/v）, obtain corresponding homoisoflavone Mannich alkaloid compound（I）, yield 20.0%-75.0%, The equal Jing of its chemical constitution¹H-NMR、¹³C-NMR and ESI-MS is confirmed；The purity Jing HPLC of gained object is determined and is all higher than 97.0%.The object structure prepared using above-mentioned logical method is as follows：

；

。

The homoisoflavone Mannich alkaloid compound of embodiment 2（I）Logical method is prepared with acid into salt

Add in reaction bulb according to the homoisoflavone Mannich alkaloid compound of the gained of above-described embodiment 1（I）2.0 mmol and The ml of acetone 50, it is sour accordingly to be stirring evenly and then adding into 8.0 mmol, temperature rising reflux stirring reaction 20 minutes, and reaction is cold after terminating But to room temperature, solvent is removed under reduced pressure, residue acetone recrystallization filters the solid for separating out, obtains final product homoisoflavone Mannich base Class compound（I）Salt, its chemical constitution Jing¹H NMR and ESI-MS are confirmed.

Claims (7)

1. a class 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 R₃H, OH, CF are represented independently of one another₃O、C₁~C₁₂Alkoxyl, NR₆R₇, R₄、R₅、R₆And R₇It is each independent Ground represents C₁~C₁₂Alkyl, propargyl, benzyl, substituted benzyl, NR₄R₅And NR₆R₇May also indicate that nafoxidine base, morpholinyl, piperazine Piperidinyl, piperazinyl, 4- positions are by C₁~C₁₂The piperazinyl that piperazinyl that alkyl is replaced, 4- positions are replaced by benzyl or substituted benzyl, R₁、R₂、R₃、-CH₂NR₄R₅Can be in phenyl ring arbitrarily possible position with OH；" substituted benzyl " is referred to by individual by 1-4 on phenyl ring The benzyl that the group being selected from the group is replaced：F、Cl、Br、I、C_1-4Alkyl, C_1-4Alkoxyl, trifluoromethyl, trifluoromethoxy, two Methylamino, nitro, cyano group, these substituents can be in any possible position of phenyl ring.

2. homoisoflavone Mannich alkaloid compound as claimed in claim 1 or its pharmaceutically acceptable salt, its feature exists In described 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, butanedioic 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. as described in any one of claim 1-2 homoisoflavone Mannich alkaloid compound or its pharmaceutically acceptable salt system Preparation Method, it is characterised in that the compound can be prepared by the following method and 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 initiation material, in solvent With direct polycondensation under alkalescence condition, corresponding homoisoflavone Mannich alkaloid compound is obtained（I）；Using the height of said method gained Contain amino in isoflavones Mannich bases compound molecule, the amino, can be with any suitable acid by pharmaceutically in alkalescence Conventional salifying method is obtained its pharmaceutically acceptable salt.

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,N- methyl morpholine,N- methyl piperidine, triethylene diamine or TBAH；Reacting solvent for use is：C_1-8Fat Alcohol, ether, tetrahydrofuran, 2- methyltetrahydrofurans,N,N- dimethylformamide, 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 class pharmaceutical composition, it is characterised in that comprising the homoisoflavone Mannich base as described in any one of claim 1-2 Class compound or its pharmaceutically acceptable salt and one or more pharmaceutically acceptable carrier or excipient.

7. the homoisoflavone Mannich alkaloid compound or its pharmaceutically acceptable salt as described in any one of claim 1-2 exists Treatment and/or the purposes in prevention nervus retrogression relevant disease medicine are prepared, 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.