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

Abstract

The invention discloses a new type of high isoflavone Mannich base compound (I) and its pharmaceutically acceptable salt, its preparation method, pharmaceutical composition and medicine for treating and/or preventing neurodegenerative related diseases Including but not limited to vascular dementia, Alzheimer's disease, Parkinson's disease, Huntington's disease, HIV-related dementia, multiple sclerosis, progressive lateral sclerosis, neuropathic pain, glaucoma, etc. neurodegenerative disease.

Description

High isoflavone Mannich base compound, its preparation method and use

technical field

The invention belongs to the field of medicinal chemistry, and relates to a new type of high-isoflavone Mannich base compound (I) and its pharmaceutically acceptable salt, its preparation method, pharmaceutical composition and preparation for treating and/or preventing neurodegeneration Sex-related diseases, including but not limited to vascular dementia, Alzheimer's disease, Parkinson's disease, Huntington's disease, HIV-related dementia, multiple sclerosis, progressive lateral sclerosis, neurological Pain, glaucoma and other neurodegenerative diseases.

Background technique

Alzheimer's disease (Alzheimer's disease, AD, senile dementia) is a degenerative disease of the central nervous system mainly characterized by progressive cognitive impairment and memory impairment. Vascular diseases and cancers are high-incidence diseases, which have risen to the fourth cause of death in developed countries such as Europe and the United States. According to the report of the World Health Organization, 10% of people over the age of 65 in the world have mental retardation, one-half of them develop dementia, and the incidence rate of people over the age of 85 is nearly 50%. The number of AD patients in my country is about 6-7 million, and the incidence rate exceeds 5%. With the acceleration of the aging process of the global population, its incidence has shown a clear upward trend. According to the report "Global Impact of Alzheimer's Disease: 2013-2050" published by Alzheimer's Disease International in December 2013, AD will become The biggest health challenge facing the world in the next few decades. By 2030, the number of patients will rise from 44 million in 2013 to 76 million. By 2050, this value will reach a staggering 135 million. The clinical manifestations of AD are decreased memory ability, orientation ability, thinking and judgment ability, reduced ability of daily life, and even abnormal mental behavior symptoms, which makes patient care more difficult and brings a heavy burden to society and families. Drugs currently approved for the treatment of mild/moderate AD include acetylcholinesterase (AChE) inhibitors, and N -methyl- D -aspartate (NMDA) receptor antagonists for the treatment of severe AD, but Clinical use shows that these drugs can relieve AD symptoms by increasing the level of acetylcholine in the patient's body or inhibiting the excitotoxicity of excitatory amino acids, but they cannot effectively prevent or reverse the course of the disease, and they can also cause hallucinations, confusion, dizziness, headache, nausea, liver Toxicity, loss of appetite and frequent defecation and other serious side effects, so the long-term efficacy is not ideal. Therefore, there is an urgent clinical need to develop AD therapeutic drugs with novel mechanisms of action.

AD is a disease caused by multiple factors, and its pathogenesis is complex. Its pathogenesis has not yet been fully elucidated. However, studies have shown that the decline in the level of acetylcholine in the brain of patients, the excessive production and deposition of β-amyloid protein, and the disorder of metal ion metabolism. Disorders of Ca ²⁺ balance, neurofibrillary tangles caused by hyperphosphorylation of tau -proteins, hyperactivity of glutamate receptors, oxidative stress producing a large amount of reactive oxygen species (ROS) and free radicals, and neuroinflammation play an important role in the pathogenesis of AD. In response to the above-mentioned pathogenic factors, researchers adopted the traditional "one drug, one target" drug design strategy, and discovered a large number of drugs with high activity and high selectivity for a certain target, such as: cholinesterase inhibitors and N -methyl - D -aspartate receptor antagonists, etc., but these drugs have problems such as a single target, more toxic and side effects in clinical use, and poor long-term curative effect on AD patients.

In recent years, with the continuous elucidation of the pathogenic mechanism of AD, it has been found that the occurrence and development of AD have the characteristics of multi-mechanism and multi-factor effects. Network control system. Based on the above results, the researchers proposed a "Multitarget-directed Ligands, MTDLs" strategy to develop anti-neurodegenerative disease drugs. The so-called "multi-target drug" means that a single chemical entity acts on multiple targets in the disease network at the same time, and the effect on each target can produce a synergistic effect, making the total effect greater than the sum of the individual effects. It is a "Multifunctional" or "Multipotential" medicine. The main difference between multi-target drug, multi-drug combination and compound drug is that it can reduce the dosage, improve the therapeutic effect, avoid the interaction between drugs and the resulting toxic and side effects, uniform pharmacokinetic characteristics, Ease of use etc. Designing and discovering anti-acetylcholinesterase, anti-monoamine oxidase-B, anti-oxidative stress, metal ion complexation, inhibition of excessive production and deposition of β-amyloid protein, and a variety of active balanced multi-target AD therapeutic drugs are current research hotspot. Therefore, the research and development of anti-neurodegenerative disease therapeutic drugs with new chemical structures, new mechanisms of action, multi-target effects, and low side effects not only meets the urgent needs of the aging society, but also has good market prospects.

Contents of the invention

The purpose of the present invention is to disclose a novel high-isoflavone Mannich base compound (I) and pharmaceutically acceptable salts thereof.

Another object of the present invention is to disclose the preparation method of the high isoflavone Mannich base compound (I) and its pharmaceutically acceptable salt.

Another object of the present invention is to disclose a pharmaceutical composition comprising the high isoflavone Mannich base compound (I) and a pharmaceutically acceptable salt thereof.

Another object of the present invention is to disclose that the high isoflavone Mannich base compound (I) and its pharmaceutically acceptable salts have multi-target effects and can be used to prepare drugs for treating and/or preventing neurodegenerative related diseases Including but not limited to vascular dementia, Alzheimer's disease, Parkinson's disease, Huntington's disease, HIV-related dementia, multiple sclerosis, progressive lateral sclerosis, neuropathic pain, glaucoma, etc. neurodegenerative disease.

The general chemical structure formula of the high isoflavone Mannich base compound (I) provided by the present invention is:

In the formula: R ₁ , R ₂ and R ₃ each independently represent H, OH, CF ₃ O, C ₁ ~C ₁₂ alkoxy, NR ₆ R ₇ , R ₄ , R ₅ , R ₆ and R ₇ are each independently represent C ₁ ~ C ₁₂ alkyl, propargyl, benzyl, substituted benzyl, NR ₄ R ₅ and NR ₆ R ₇ can also represent tetrahydropyrrolyl, morpholinyl, piperidinyl, piperazinyl, Piperazinyl substituted by C ₁ ~C ₁₂ alkyl at the 4-position, piperazinyl substituted by benzyl or substituted benzyl at the 4-position; R ₁ , R ₂ , R ₃ , -CH ₂ NR ₄ R ₅ and OH can be in any possible position of the benzene ring;

The above-mentioned term "substituted benzyl" refers to a benzyl group substituted by 1-4 groups selected from the following group on the benzene ring: F, Cl, Br, I, C _1-4 alkyl, C _1-4 Alkoxy, trifluoromethyl, trifluoromethoxy, dimethylamino, nitro, cyano, these substituents can be in any possible position of the benzene ring.

The high isoflavone Mannich base compound (I) proposed by the present invention can be prepared by the following method:

In the formula: the definition of R ₁ to R ₅ is the same as the general chemical structure formula of the high isoflavone Mannich base compound (I).

Using the corresponding chromanone compound (1) and hydroxybenzaldehyde Mannich base compound (2) as starting materials, direct condensation under solvent and alkaline conditions to obtain the corresponding homoisoflavone Mannich base Compound (I). Wherein, the alkali used in the reaction is: alkali metal hydroxide, alkaline earth metal hydroxide, alkali metal carbonate, alkaline earth metal carbonate, alkali metal bicarbonate, alkaline earth metal bicarbonate, alkali of C _1-8 alcohol Metal salts, organic tertiary amines or quaternary ammonium bases (such as: triethylamine, tributylamine, trioctylamine, pyridine, N -methylmorpholine, N -methylpiperidine, triethylenediamine, tetrabutylhydrogen ammonium oxide), the preferred base is: potassium hydroxide, sodium hydroxide, potassium carbonate, triethylamine or pyridine; the solvent used for the reaction is: C _1-8 fatty alcohol, ether, tetrahydrofuran, 2-methyltetrahydrofuran, N,N - dimethylformamide, dimethylsulfoxide, dichloromethane, chloroform, 1,4-dioxane, benzene, toluene or acetonitrile, preferred solvents are: methanol, ethanol, isopropanol, N,N - Dimethylformamide, acetonitrile, tetrahydrofuran, dichloromethane or toluene; chromanone compound (1): hydroxybenzaldehyde Mannich base compound (2): the molar feed ratio of base is 1.0:1.0~3.0: 1.0~20.0, preferably the molar feed ratio is 1.0:1.0~2.0:1.0~10.0; the reaction temperature is 0~150°C, the preferable reaction temperature is room temperature~120°C; the reaction time is 1~120 hours, the preferable reaction time is 2~ 72 hours.

The starting materials of the present invention—chromanone compound (1) and hydroxybenzaldehyde Mannich base compound (2) can be prepared by common techniques in the art, including but not limited to the methods disclosed in the following documents : 1. Yoshihisa T.; Kenji M. European Journal of Organic Chemistry 2001, (10), 1963-1966; 2. Mahapatra T. et al . Tetrahedron: Asymmetry 2008, 19(10), 1224-1232; 3. AissaouiH . et al . WO 2014072903; 4. Fattorusso C. et al. Journal of Medicinal Chemistry 2008, 51(5), 1333-1343; 5. Karki SS et al. Journal of Medicinal Chemistry 2016, 59(2), 763- 769; 6. Torrente E. et al. Journal of Medicinal Chemistry 2015, 58(15), 5900-5915.

The homo-isoflavone Mannich base compound (I) obtained according to the above method contains an amino group in its molecule, which is basic and can be prepared with any suitable acid by a pharmaceutically conventional salt-forming method. Its pharmaceutically acceptable salt, the acid is: hydrochloric acid, hydrobromic acid, nitric acid, sulfuric acid, phosphoric acid, C _1-6 fatty carboxylic acid (such as: formic acid, acetic acid, propionic acid, etc.), oxalic acid, benzoic acid, salicylic acid, horse Toric acid, fumaric acid, succinic acid, tartaric acid, citric acid, malic acid, lipoic acid, C _1-6 alkyl sulfonic acid (such as: methylsulfonic acid, ethylsulfonic acid, etc.), camphorsulfonic acid, benzenesulfonic acid acid or p-toluenesulfonic acid.

The pharmaceutical composition disclosed in the present invention includes a therapeutically effective amount of one or more homoisoflavone Mannich base compounds (I) or pharmaceutically acceptable salts thereof, and the pharmaceutical composition may further contain one or more a pharmaceutically acceptable carrier or excipient. The "therapeutically effective amount" refers to the amount of the drug or agent that causes the biological or medical response of the tissue, system or animal targeted by the researcher or doctor; the "composition" refers to the combination of more than one substance or The mixed product; the "pharmaceutically acceptable carrier" refers to a pharmaceutically acceptable substance, composition or carrier, such as: liquid or solid filler, diluent, excipient, solvent or encapsulation Substances that carry or transport a chemical substance. The ideal proportion of the pharmaceutical composition provided by the present invention is that the high isoflavone Mannich base compound (I) or a pharmaceutically acceptable salt thereof is 5% to 99.5% of the total weight as the active ingredient, and the rest is Accounting for less than 95% of the total weight.

The high isoflavone Mannich base compound (I) disclosed in the present invention and its pharmaceutically acceptable salts were screened for their biological activity as follows.

(1) Inhibitory activity of high isoflavone Mannich base compounds (I) on acetylcholinesterase and butyrylcholinesterase

Add 30 μL of 1.0 mmol/L thioacetylcholine iodide or thiobutyrylcholine iodide (both purchased from Sigma), 40 μL of PBS buffer solution with pH 7.4, and 20 μL of the test compound solution to the 96-well plate in sequence. μL (DMSO content is less than 1%) and 10 μL acetylcholinesterase (5% homogenate supernatant of rat cerebral cortex, pH 7.4 phosphate buffer as homogenization medium) or butyrylcholinesterase (rat serum 25 % supernatant, pH 7.4 phosphate buffer as homogenization medium) solution, after adding and mixing, incubate at 37°C for 15min, add 0.2% 5,5'-dithio-bis(2- Nitrobenzoic acid) (DTNB, purchased from Sigma Company) solution 30 μL for color development, measured the optical density (OD value) of each well at 405 nm with a microplate reader, compared with the blank well without the sample to be tested, and calculated the compound pair Enzyme inhibition rate (enzyme inhibition rate (%)=(1-sample group OD value/blank group OD value)×100%); select five to six concentrations of the compound, determine the enzyme inhibition rate, and use the compound mole The negative logarithm of the concentration and the inhibition rate of the enzyme are linearly regressed, and the molar concentration at which the 50% inhibition rate is obtained is the IC ₅₀ of the compound. The measurement results show that the high-isoflavone Mannich base compounds (I) disclosed in the examples of the present invention all have a significant inhibitory effect on acetylcholinesterase, and its _IC50 is 2.49nM~10.0 μM; and the high-isoflavone Mannich base The inhibitory activity of the alkaloid compound (I) to acetylcholinesterase is significantly higher than the inhibitory activity to butyrylcholinesterase (selectivity greater than 10 times), indicating that the compounds disclosed in the present invention have selective inhibition of acetylcholinesterase effect. The measurement results also showed that the core of the homoisoflavone Mannich base compound (I) - 2'-hydroxy homoisoflavone (R ₁ , R ₂ , R ₃ and -CH ₂ NR ₄ R ₅ all represent H, OH is at the 2' position), 3'-hydroxy homoisoflavones (R ₁ , R ₂ , R ₃ and -CH ₂ NR ₄ R ₅ all represent H, and OH is at the 3' position) and 4'-hydroxy homoisoflavones ( R ₁ , R ₂ , R ₃ and -CH ₂ NR ₄ R ₅ all represent H, and OH is at the 4' position), and the IC ₅₀ for the inhibition of acetylcholinesterase is greater than 500 μM.

(2) Inhibitory activity of high isoflavone Mannich base compound (I) on monoamine oxidase B

Recombinant human MAO-B was prepared into a 75 μg/mL sample solution with 100 mM pH 7.4 potassium phosphate buffer. Add 20 μL of the test compound solution and 80 μL of monoamine oxidase to a black 96-well plate, mix well, incubate at 37°C in a dark place for 15 min, add 200 μM Amplex Red reagent, 2U/mL horseradish peroxidase, 2 Initiate the reaction with mM benzylamine, incubate at 37°C for 20 min, measure the fluorescence emission intensity at 590 nm on a multi-functional microplate reader with a fixed excitation wavelength of 545 nm, and use potassium phosphate buffer instead of MAO-B as a blank; compound inhibition The formula for calculating the inhibition rate of monoamine oxidase is: 100-(IF _i )/(IF _c )*100, where IF _i and IF _c are the difference between the fluorescence intensity with and without the inhibitor and the blank fluorescence intensity, respectively. Each compound was measured in triplicate wells, and each group of experiments was repeated three times independently. Select five to six concentrations of the compound, measure its enzyme inhibition rate, and linearly regress the negative logarithm of the molar concentration of the compound and the enzyme inhibition rate to obtain the molar concentration when the 50% inhibition rate is the _IC50 of the compound . The measurement results show that the high-isoflavone Mannich base compounds (I) disclosed in the examples of the present invention have significant inhibitory effects on MAO-B, with an _IC50 of 0.1 μM to 50.0 μM; while the compound (I) Mother nucleus - 2'-Hydroxy isoflavone (R ₁ , R ₂ , R ₃ and -CH ₂ NR ₄ R ₅ all represent H, OH is at the 2' position), 3'-Hydroxy isoflavone (R ₁ , R ₂ , R ₃ and -CH ₂ NR ₄ R ₅ all represent H, OH is at the 3' position) and 4'-hydroxyhomoisoflavones (R ₁ , R ₂ , R ₃ and -CH ₂ NR ₄ R ₅ all represent H, OH at the 4' position) IC ₅₀ >100 µM for MAO-B inhibition.

(3) Antioxidant activity of high isoflavone Mannich base compound (I) (ORAC-FL method)

Determination with reference to the method reported in the literature (Qiang, XM et al. Eur. J Med. Chem. 2014, 76, 314-331), namely: 6-hydroxy-2,5,7,8-tetramethylchromane -2-Carboxylic acid ( Trolox ) was made into a 10-80 μmol/L solution with pH 7.4 PBS buffer, and fluorescein was made into a 250 nmol/L solution with pH 7.4 PBS buffer, 2 , 2'-Azobisisobutylamidine dihydrochloride (AAPH) was made into a 40 mmol/L solution with PBS buffer at pH 7.4 before use. Add 50-10 μmol/L compound solution and fluorescein solution to the 96-well plate, mix well, incubate at 37°C for 15 minutes, add AAPH solution to make the total volume of each well 200 μL, mix well, and immediately place in Varioskan Flash In a Multimode Reader (Thermo Scientific) instrument, continuous measurement was performed for 90 min at an excitation wavelength of 485 nm and an emission wavelength of 535 nm. Calculate the AUC of the area under the fluorescence decay curve, wherein the Trolox of 1-8 μmol/L is used as the standard, and the sample to be tested is not added as a blank, and the antioxidant activity of the compound is expressed as the equivalent of Trolox , and its calculation formula is: [(AUC Sample-AUCblank)/(AUC Trolox -AUC blank)] ×[(concentration of Trolox /concentration of sample)], each compound was measured in 3 replicate wells, and each group of experiments was repeated three times independently. The measurement results show that the antioxidant activity of the high-isoflavone Mannich base compound (I) disclosed in the examples of the present invention is 1.0-3.0 times that of Trolox , indicating that this type of compound has strong antioxidant activity.

(4) Inhibitory activity of high isoflavone Mannich base compound (I) on Aβ _1-42 self-aggregation

Refer to the method reported in the literature (Qiang, XM et al. Eur. J Med. Chem. 2014, 76, 314-331), that is, the pretreated A β _1-42 is made into a stock solution with DMSO, using Dilute to 50 μM with PBS buffer at pH 7.4 before use; make 2.5 mM stock solution of the compound to be tested with DMSO, dilute with PBS buffer at pH 7.4 to the corresponding concentration before use, take 20 μL of A β _1-42 solution+ 20 μL of the test compound solution, 20 μL of A β _1-42 solution + 20 μL of PBS buffer (containing 2% DMSO) were placed in a 96-well plate, incubated at 37°C for 24 hours, and then 160 μL of 50 mM thioflavin T containing 5 μM was added. Glycine-NaOH buffer solution (pH = 8.5), after shaking for 5 s, measure the fluorescence value with a multi-functional microplate _reader at the excitation wavelength of 446 nm and the emission wavelength of 490 nm; IF _i , the fluorescence value of A β _1-42 + PBS buffer is recorded as IF _c , the fluorescence value of only PBS buffer is recorded as IF ₀ , and the inhibition rate of the compound to inhibit A β _1-42 self-aggregation is: 100- (IF _i -IF ₀ )/(IF _c -IF ₀ )*100; five to six concentrations of the compound were selected, and the inhibition rate was determined. Each compound was tested three times at each concentration, and curcumin was used as a positive control. The measurement results show that the high isoflavone Mannich base compound (I) disclosed in the examples of the present invention has significant inhibitory activity on A β _1-42 self - _aggregation . The inhibition rate of aggregation was greater than 50.0%; while the inhibition rate of curcumin at the same concentration was 41.3%. Anti-AD drugs widely used in clinical practice: donepezil, rivastigmine, memantine hydrochloride, and the parent Core—2'-Hydroxyhomoisoflavones (R ₁ , R ₂ , R ₃ and -CH ₂ NR ₄ R ₅ all represent H, OH is at the 2' position), 3'-Hydroxyhomoisoflavones (R ₁ , R ₂ , R ₃ and -CH ₂ NR ₄ R ₅ all represent H, OH is at the 3' position) and 4'-hydroxyl homoisoflavones (R ₁ , R ₂ , R ₃ and -CH ₂ NR ₄ R ₅ all represent H , OH is located at the 4' position), the inhibition rate of A β _1-42 self-aggregation was less than 10% at the concentration of 25.0 μM.

(5) Determination of complexation between high isoflavone Mannich base compounds (I) and metal ions

Dissolve CuCl ₂ 2H ₂ O, ZnCl ₂ , FeSO ₄ 7H ₂ O, AlCl ₃ and the test compound in methanol to make a 75 μmol/L solution, add 100 μL test compound solution and 100 μL metal Ionic solution, mix well, stand at room temperature for 30min, record the UV absorption curve of the mixture in the range of 200-600 nm on Varioskan Flash Multimode Reader (Thermo Scientific) instrument, and use 100 μL test compound solution and 100 μL methanol mixture as In contrast, observe the red shift of the maximum absorption peak and the intensity of the maximum absorption peak of the mixture of metal ions and the compound to be tested. The measurement results show that the high isoflavone Mannich base compounds (I) disclosed in the examples of the present invention all exhibit selective complexation to copper ions.

(6) Inhibitory activity of high isoflavone Mannich base compound (I) on Cu2 ⁺ -induced aggregation of _Aβ1-42

Make a 75 μM solution of CuCl ₂ with HEPES buffer, dilute the compound stock solution (2.5 mM) and 200 μM Aβ _1-42 stock solution to 75 μM with HEPES buffer, take 20 μL Cu ²⁺ solution + 20 μL respectively A β _1-42 solution + 20 μL test compound solution, 20 μL Cu ²⁺ solution + 20 μL A β _1-42 solution + 20 μL HEPES buffer and 60 μL HEPES buffer in a 96-well plate, mix well, and incubate at 37°C for 24 h, then add 190 μL of 50 mM glycine-NaOH buffer solution (pH=8.5) containing 5 μM Thioflavin T, shake for 5 s, and immediately measure the fluorescence value with a multifunctional microplate reader at an excitation wavelength of 446 nm and an emission wavelength of 490 nm; Cu ^{2 +} +A β _1-42 +The fluorescence value of the compound to be tested is recorded as IF _i , the fluorescence value of Cu ²⁺ +A β _1-42 +HEPES buffer is recorded as IF _c , and the fluorescence value of the buffer containing only HEPES is recorded as IF ₀ , the inhibition rate of the compound on Cu ²⁺ -induced A β _1-42 aggregation is: 100-(IF _i -IF ₀ )/(IF _c -IF ₀ )*100. Three replicate wells were measured for each concentration of each compound, and curcumin was used as a positive control. The measurement results show that the high-isoflavone Mannich base compound (I) disclosed in the examples of the present invention has an inhibitory rate of more than 60.0% on Cu ²⁺ -induced aggregation of Aβ _1-42 at a concentration of 25.0 μM; and The inhibition rate of curcumin at the same concentration was 52.1%. The core of compound (I) - 2'-hydroxyhomoisoflavone (R ₁ , R ₂ , R ₃ and -CH ₂ NR ₄ R ₅ all represent H, OH is at the 2' position), 3'-hydroxy homoisoflavones (R ₁ , R ₂ , R ₃ and -CH ₂ NR ₄ R ₅ all represent H, and OH is at the 3' position) and 4'-hydroxy homoisoflavones ( R ₁ , R ₂ , R ₃ and -CH ₂ NR ₄ R ₅ all represent H, and OH is at the 4' position), and the inhibition rate at the same concentration is less than 20.0%.

Detailed ways

The present invention can be further described by the following examples, however, the scope of the present invention is not limited to the following examples. Those skilled in the art can understand that various changes and modifications can be made in the present invention without departing from the spirit and scope of the present invention.

Example 1 General method for the preparation of high isoflavone Mannich base compounds (I)

Add 2.0 mmol of the corresponding chromanone compound (1), 3.0 mmol of the corresponding hydroxybenzaldehyde Mannich base compound (2) and 30 ml of methanol into the reaction flask. After stirring evenly, add 30% KOH aqueous solution 12.0 mmol, heated and refluxed and stirred for 3.0 to 24.0 hours (the reaction process was tracked by TLC); after the reaction, cooled to room temperature, adjusted the pH of the reaction solution to strong acidity with 10% aqueous hydrochloric acid solution, and then adjusted the pH of the reaction solution with saturated aqueous sodium bicarbonate solution To weak alkalinity, methanol was evaporated under reduced pressure, 80 mL of deionized water was added to the residual liquid, extracted three times with 240 mL of dichloromethane, the organic layers were combined, washed with saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate Filtrate, evaporate the solvent under reduced pressure, and purify the residue by column chromatography (eluent: dichloromethane: acetone = 10:1 v/v) to obtain the corresponding high isoflavone Mannich base compound (I), The yields ranged from 20.0% to 75.0%, and the chemical structures were confirmed by ¹ H-NMR, ¹³ C-NMR and ESI-MS; the purity of the obtained target compounds were all greater than 97.0% as determined by HPLC. The structure of the target object prepared by the above-mentioned general method is as follows:

;

.

Example 2 General method for the preparation of high isoflavone Mannich base compound (I) and acid salt formation

Add 2.0 mmol of the high isoflavone Mannich base compound (I) obtained according to the above-mentioned Example 1 and 50 ml of acetone into the reaction flask, stir evenly, add 8.0 mmol of the corresponding acid, heat up and reflux and stir for 20 minutes, and the reaction is over After cooling to room temperature, the solvent was distilled off under reduced pressure, the residue was recrystallized with acetone, and the precipitated solid was filtered to obtain the salt of the high isoflavone Mannich base compound (I), whose chemical structure was verified by ¹ H NMR and ESI- MS confirmed.

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.