WO2003086411A1

WO2003086411A1 - Compounds and preparation methods of carboxylate and monoester succinate derivative of diosgenin

Info

Publication number: WO2003086411A1
Application number: PCT/CN2003/000263
Authority: WO
Inventors: Zhongrong Liu; Xuechao Wang; Zhonglin Ye; Xiaoli Wang; Bogang Li; Liutang Wang; Shiyong Sun; Lingen Deng; Min He
Original assignee: Chengdu Di'ao Pharmaceutical Group Co., Ltd.
Priority date: 2002-04-15
Filing date: 2003-04-14
Publication date: 2003-10-23
Also published as: AU2003231427A1

Abstract

The present invention relates to carboxylate derivative of diosgenin represented by formula (I) and monoester succinate derivative of diosgenin represented by formula (III); wherein formula (I), R, is chosen from alkyl group or substituted alkyl group having 1 to 2 carbon atoms, M is H or Na; and in formula (III), AA is various of amino acid. The compounds of this invention have good water solubility, and can be used in treating and preventing cerebrovascular and cardiovascular diseases.

Description

Diosgenin carboxylic acid derivative and succinate monoester derivative and preparation method thereof

The present invention relates to a diosgenin derivative having water solubility, in particular to a diosgenin carboxylic acid and a succinic acid monoester derivative.

The present invention also relates to a method for preparing a diosgenin carboxylic acid derivative and a succinic acid monoester derivative.

The invention also relates to the use of diosgenin carboxylic acid derivatives and succinate monoester derivatives in the preparation and treatment of cardiovascular diseases. Background technique

Diosgenin has the effects of enhancing myocardial contractility, slowing heart rate, anti-arteriosclerosis, improving microcirculation, anti-aging and anti-arthritis. However, due to the very poor water solubility of diosgenin, its clinical application is greatly limited. So far, only the production method and clinical application of saponin, a diosgenin water-soluble derivative, have been reported, and no other types of water-soluble derivatives have been reported. In order to further understand the physiological activities of other water-soluble derivatives of diosgenin, the inventors designed and synthesized a series of new carboxylic acid derivatives and succinate monoester derivatives, and found that they all have physiological activities. Summary of the Invention

An object of the present invention is to provide a water-soluble diosgenin derivative. Another object of the present invention is to provide a method for preparing a diosgenin carboxylic acid derivative. Another object of the present invention is to provide the application of a diosgenin carboxylic acid derivative or a pharmaceutically acceptable salt thereof in the preparation of a cardio-cerebral-vascular disease.

Another object of the present invention is to provide a water-soluble diosgenin succinate monoester derivative. ·

Another object of the present invention is to provide a method for preparing a diosgenin succinate monoester derivative.

Yet another object of the present invention is to provide an application of a diosgenin succinic acid monoester derivative or a pharmaceutically acceptable salt thereof in the preparation of a cardio-cerebral vascular disease.

According to one aspect of the present invention, a diosgenin carboxylic acid derivative having the structure of formula (I) is provided:

In the formula (I), an alkyl group or a substituted alkyl group selected from 1-2 carbon atoms is preferably:

-C¾-, -CH ₂ CH _2- , -CH ₂ CH (OH) _-5 -CH (OH) CH (OH)-, most preferably -CH ₂ CH _2- ; M is H or Na.

According to another aspect of the present invention, a method for preparing a compound of formula (I) is provided. The method uses diosgenin as a raw material, reacts with an organic anhydride under reflux conditions, and then reacts with sodium bicarbonate to obtain diosgenin. Carboxylic acid derivatives. See the following reaction formula:

Most preferred is -CH ₂ CH _2- ; M is H or Na.

In the above method of the present invention, the reaction is preferably performed in an organic solvent under reflux conditions. In the step a, the preferred organic solvent is pyridine, and in the step b, the preferred solvent is ethanol.

In one embodiment of the present invention, diosgenin is reacted with succinate to obtain diosgenin succinate monoester, and then reacted with sodium bicarbonate to obtain diosgenin succinate monoester sodium salt, which Good water solubility has a good preventive effect on myocardial infarction in rats.

'' In another embodiment of the present invention, diosgenin ring deterrent and 2,2-dimethyl-dihydrofuran [3,4-d] [l, 3] -m-dioxoladiene- 4,6-dione reaction to obtain diosgenin 2,2 Difluorenyl- [1,3] dioxolane 4,5-dicarboxylic acid monoester, and then react with sodium bicarbonate to obtain diosgenin 2,2difluorenyl- [1,3] dioxolane 4,5-Dicarboxylic acid monoester sodium salt, which has good water solubility, has a good preventive effect on myocardial infarction in rats.

In one embodiment of the present invention, the diosgenin 2,2 difluorenyl- [1,3] dioxolane 4,5-dicarboxylic acid monoester prepared above is reacted with concentrated sulfuric acid to obtain diosgenin This tartrate is reacted with sodium bicarbonate to obtain diosgenin tartaric acid ^ sodium salt, which has good water solubility and has a good preventive effect on myocardial infarction in rats.

According to another aspect of the present invention, the application of the compound of formula (I) of the present invention in the preparation of a medicament for treating or preventing cardiovascular and cerebrovascular diseases is provided by adding the compound of formula (I) of the present invention to a pharmaceutically acceptable carrier or excipient Agent or optional other ingredients to make a pharmaceutical composition suitable for clinical use, and to treat or prevent myocardial ischemia and myocardial infarction by administering to a human or an animal an effective amount of a compound of the formula (I) of the present invention.

According to still another aspect of the present invention, the diosgenin succinic acid monoester (compound of formula (II)) obtained by the above reaction can be subjected to a condensation reaction with various amino acids, or further prepared into a salt to obtain a compound of formula (III) Diosgenin succinic acid monoester ^ ^: Bio:

(III)

In formula (III), AA is various amino acids such as glycine, alanine, cystine, glutamic acid, lysine, proline, threonine, tyrosine, leucine, isoleucine Amino acid, phenylalanine, tryptophan, serine, threonine, cysteine, aspartic acid, arginine, histidine, and their corresponding amino or carboxylic acid salts, and other pharmaceutical Acceptable salt. The amino acid may be a (L) counterpart or a (D) counterpart; a pharmaceutically acceptable salt thereof is selected from a sodium salt, a potassium salt, a calcium salt, and a magnesium salt, or is selected from a hydrochloride salt, an acetate salt, a pyrene Sulfonates and citrates. Specifically, in this class of compounds, AA is glycine, or its sodium salt; or, AA is histidine, or its acetate salt; or, AA is glutamic acid, or its sodium salt; or, AA is asparagus Amino acid, or its sodium salt; or, AA is arginine, or its acetate. The water solubility of these compounds is obviously improved, and they have better anti-myocardial anemia, and can be used to prevent cardiac ischemia.

According to yet another aspect of the present invention, a method for preparing a compound of formula (III) includes combining diosgenin succinic acid monoester of formula (II) with N-hydroxysuccinimide, N, N-dicyclohexyl carbon The diimine is then reacted with various amino acids to obtain a compound of formula (III). The reaction is performed in a mixed solvent. The mixed solvent is preferably tetrahydrofuran and water. The reaction steps are as follows:

(II) (III)

The reaction step may further include a process of reacting with sodium bicarbonate or potassium bicarbonate under reflux conditions.

:

(II) (III) According to another aspect of the present invention, the application of the compound of formula (III) of the present invention in the preparation of a medicament for treating or preventing cardiovascular disease can be provided by adding the compound of formula (III) of the present invention to a pharmaceutically acceptable Carrier or excipient or optional other ingredients to make a pharmaceutical composition suitable for clinical use, by treating or preventing an effective amount of a compound of formula (III) of the present invention . DETAILED DESCRIPTION OF THE INVENTION

Hereinafter, the present invention is described in detail by describing the preferred embodiments of the present invention:

[Example 1] Preparation of Diosgenin Succinate Monoester

41.4 g (0.1 mol) of diosgenin and 20.1 g (0.2 mol) of succinic anhydride were dissolved in 80 ml of pyridine and stirred under reflux for 2 hours. Then a large amount of pyridine was distilled off, 80 ml of acetone was added thereto, and the mixture was stirred uniformly. After cooling, crystals were precipitated, filtered, washed with water, and dried. Ethanol recrystallization, white crystals 37.5g, 73% yield, Mp: l 96-201

MS m / z (%): 514, 470, 414;

toMR: 12.19 (s, 1H) ₅ 5.35 (d, J = 4Hz, 1H), 4.48 (dd, J = 4Hz, 8.4Hz, 1H), 4.29 (dd ₅ J = 4.1Hz, 8.4Hz, 1H), 3.39 (t, J = llHz, 1H), 3.20 (t, J = llHz, 1H), 2.47 (t, J = 8Hz, 2H), 2.25 (t, J = 8Hz, 2H), 0.95 (d, J = 6.8 Hz ₅ 3H), 0.89 (s, 3H), 0.76 (d, J = 6.7Hz, 3H), 0.73 (s, 3H).

[Example 2] Preparation of diosgenin 2,2 difluorenyl- [1,3] dioxolane 4,5-dicarboxylic acid monoester

Diosgenin 41.4 g (0.1 mol) and 2,2-dimethyl-dihydrofuran [3,4-d] [l, 3] -m-dioxolane-4,6-dione It was dissolved in 80 ml of pyridine and stirred at reflux for 3 hours. Of course A large amount of pyridine was distilled off, and 80 ml of acetone was added thereto, followed by stirring. After cooling, crystals were precipitated, filtered, washed with water, and dried. Crystallization of ethanol to obtain 41.6 g of white crystals, yield 71%, Mp: 205-208 ° C

MS m / z (%): 586, 571,536, 542, 414;

¹ HNMR: 11.6 (s, IH), 5.37 (d, J = 4Hz, IH), 5.15 (d, J = 8.1Hz, IH), 5.04 (d, J = 8.1Hz, IH), 4.45 (dd, J = 4.2Hz, 8Hz, IH), 4.31 (dd ₅ J = 4.2Hz, 8 Hz, lH), 3.98 (m, IH), 3.41 (t, J = llHz ₅ IH), 3.26 (t, J = llHz, IH), 1.23 (s, 6H), 0.99 (d, J = 6.8Hz ₅ 3H), 0.92 (s, 3H), 0.83 (d, J = 6.7Hz, 3H), 0.79 (s, 3H).

[Example 3] Preparation of Diosgenin Tartrate Monoester

Diosgenin 2,2 diamidino- [1,3] dioxolane 4,5-dicarboxylic acid monoester 29.3 g (0.05 mol) was dissolved in 200 ml of ethanol, and 2 ml of concentrated sulfuric acid was added, and the mixture was stirred at room temperature until the reaction Completely and concentrated under reduced pressure. Ethanol recrystallization to obtain 19.2 g of white crystals, yield 70%, Mp: 219-224 ° C

MS m / z (%): 546, 528,543,414;

lHNMR 12.6 (s, IH), 5.35 (d, J = 4Hz, IH), 4.79 (d, J = 8Hz, IH), 4.71 (d, J = 8Hz, IH), 4.30 (dd, J = 4.2Hz, 8Hz, IH), 4.24 (dd, J = 4.2Hz, 8 Hz, lH), 4.01 (m, 1H), 3.38 (t, J = llHz, IH), 3.25 (t ₅ J = llHz, IH), 0.99 (d, J = 6.8Hz, 3H) ₅ 0.92 (s, 3H), 0.83 (d, J = 6.7Hz, 3H), 0.79 (s, 3H).

[Example 4] Preparation of diosgenin succinate monoester sodium salt (compound 1)

36 g (0.07 mol) of diosgenin succinate monoester was dissolved in 720 ml of absolute ethanol, and then 5.9 g (0.07 mol) of anhydrous sodium bicarbonate in 80 ml of an aqueous solution was added dropwise. After completion of the addition, reflux for 1 hour. Cool and filter to obtain white crystals 36.5g, yield 97%, Mp: 271-276 ° C

C ₃₁ H ₄₅ Na0 ₆ calculated C: 69.38 H: 8.45

Found C: 69.29 H: 8.50

[Example 5] Preparation of diosgenin 2,2 diamidino- [1,3] dioxolane 4,5-dicarboxylic acid monoester sodium salt (compound 2)

Diosgenin 2,2 dimethyl- [1,3] dioxolane 4,5-dicarboxylic acid monoester 29.3 g (0.05 mol) was dissolved in 500 ml of absolute ethanol, and then anhydrous hydrogen carbonate was added dropwise. Sodium 4.2 g (0.05 mol) in 80 ml of aqueous solution. After completion of the addition, reflux for 1 hour. Cool and filter to obtain white crystals 29.1g, yield 95%, Mp: 277-28rC

C ₃₄ H ₄₉ Na0 ₈ Calculate C: 67.08 H: 8.11

Found C: 67.19 H: 8.09

[Example 6] Preparation of diosgenin tartrate monoester sodium salt (compound 3) 27.3 g (0.05 mol) of diosgenin tartrate monoester was dissolved in 500 ml of absolute ethanol, and then 4.2 g (0.05 mol) of anhydrous sodium hydrogen carbonate in 60 ml of an aqueous solution was added dropwise. After completion of the addition, reflux for 1 hour. Cool and filter to obtain 26.7g of white crystals, yield 94%, Mp: 283-286 ° C

C ₃ iH ₄₅ Na0 ₆ Calculated C: 65.47 H: 7.98

Found C: 65.45 H: 8.03

[Example 7] Preparation of 4-L- (N-succinic acid-2-yl) amino-4-oxobutanoic acid diosgenin ester disodium salt (compound 4)

Diosgenin succinate monoester was prepared according to the method of Example 1. 30.8 g (60 mmol) of diosgenin succinic acid monoester and 9 g (78 mmol) of N-hydroxysuccinimide were dissolved in THF (350 ml) under stirring in a water-salt bath, and then Ν, Ν-bicyclohexyl 14.8 g (72 mmol) of carbodiimide. After 1 hour of reaction, remove the ice salt bath and react at room temperature overnight. The insoluble material was filtered off, and most of the THF was distilled off. The residue was poured into water (1000 ml) and stirred to precipitate a solid. The solid was washed with water. Then dissolved in THF (400ml) to prepare solution A.

Under cooling and stirring in a water bath, 16 g (120 mmol) of aspartic acid and 22 g (264 mmol) of NaHC0 _{3 were} dissolved in ¾O (150 ml) and THF (50 ml), and then solution A was added. After stirring for 0.5 hours, the ice bath was removed, and then room temperature The reaction was completed for 4.5 hours, and the reaction was checked by TLC. The pH was adjusted to 3 with 6N hydrochloric acid, and the lower aqueous solution was separated. The upper reaction solution was added to water, the precipitate was filtered off, washed with water, and dried to obtain 35 g of solid. Silica gel column chromatography, using acetic acid: Ethyl acetate: Petroleum ether = 0.5: 1: 3 Eluted to obtain 17g of pure product. This product was dissolved in a mixed solvent of HF (50ml) and water (50ml), and then 4.55g of NaHC0 _{3 was} added. After stirring for 2 hours, the temperature was low. The solvent was evaporated and dried to obtain the product.

IH NMR (DMSO, ppm)): 5.38 (d, J = 4Hz, 1H), 4.72 (t, J = 6Hz, lH), 4.54 (m,

IH), 4.39 (dd, J = 7.6Hz ₅ IH), 3.21 (m, IH), 2.80 (dd, J = 6.4Hz, 2H),

2.58 (m, 3H), 2.32 (d, J = 6.8Hz, 2H), 1.06 (s, 3H), 0.95 (d, J = 7.2Hz, 3H),

0.81 (s, 3H), 0.78 (d, J = 6.4Hz, 3H).

MS m / z: 674 (M + l), 628, 397;

IR: 3415 (br), 2951, 2904, 1717, 1588, 1410.

[Example 8] Preparation of 4- (N-acetic acid-2-yl) amino-4-oxobutanoic acid diosgenin ester sodium salt (compound 5)

Diosgenin succinate monoester was prepared according to the method of Example 1. 30.8 g (60 mmol) of diosgenin succinic acid monoester and 9 g (78 mmol) of N-hydroxysuccinimide were dissolved in THF (350 ml) under stirring in a water and salt bath, and then N, N-dicyclohexyl 14.8 g (72 mmol) of carbodiimide, reacted for 1 hour and then reacted at room temperature overnight. The insolubles were filtered off and most of the THF was evaporated. The residue was poured into ice water (1000 ml) and stirred to precipitate a solid, and the solid was washed with water. Then dissolved in THF (400ml) to prepare solution A.

Glycine 9g (120mmol), NaHC0 ₃ 22g (144mmol) under cooling in a water bath N03 / 00263 was dissolved in a solution of H ₂ O (150 ml) and THF (150 ml), and then solution A was added. After 0.5 hours of reaction, the ice bath was removed and the reaction was stirred at room temperature. Adjust the pH to 3 with 6N hydrochloric acid. The lower aqueous solution was separated, and the upper solution was added to water (1500 ml) to precipitate a solid, which was filtered, washed with water, and dried. The residue was subjected to silica gel column chromatography using petroleum ether: ethyl acetate: HOAC = 3: 1:

Elution at 0.3 gave about 20.2 g of pure product. This product was dissolved in a mixed solvent of THF (60 ml) and water (60 ml), and then an equivalent amount of sodium bicarbonate was added, and the mixture was stirred at room temperature for 2 hours. The solvent was evaporated to dryness at low temperature, and the product was dried.

IH NMR (CDC1 ₃ , ppm)): 5.37 (d, J = 4Hz, IH), 4.61 (m, IH), 4.42 (dd,

J = 7.6Hz, IH), 4.07 (d, J = 3.6Hz, IH), 3.47 (m, IH), 3.37 (t, J = 6.8Hz ₅ IH),

2.65 (m, 2H), 2.58 (m, 2H). 2.3 l (m, 2H), 1.03 (s, 3H), 0.97 (d, J = 7.6Hz ₅

3H), 0.80 (d, J = 6Hz, 3H) 0.78 (s, 3H).

MS m / z: 594 (M + 1), 571, 397;

IR: 3500 ~ 3100 (br), 2950, 2907, 1731, 1650, 1600.

[Example 9] Preparation of 4-L- (N- (5-guanidino) -2-yl) amino-4-oxobutanoic acid diosgenin acetate (compound 6)

Diosgenin succinate monoester was prepared according to the method of Example 1. 30.8 g (60 mmol) of diosgenin succinic acid monoester and 9 g (78 mmol) of NH-hydroxysuccinimide were dissolved in THF (350 ml) under stirring in a water-salt bath, and then Ν, Ν-bicyclohexyl 14.8 g (72 mmol) of carbodiimide, reacted for 1 hour and then reacted at room temperature overnight. The insolubles were filtered off and most of the THF was evaporated. The residue was poured into ice water (1000 ml) and stirred to precipitate a solid, and the solid was washed with ice water. Then dissolved in THF (400ml) to prepare solution A.

Add 20.9g (120mmol) of arginine and 12.1g of NaHC0 _{3 while} stirring in an ice bath.

(144mmol) was dissolved in a solution of H ₂ 0 (150ml) and THF (50ml), and then added to solution A, reacted for 0.5 hours and the water bath was removed, the reaction was completed at room temperature, the pH was adjusted to 3 with 6N hydrochloric acid, the aqueous layer was separated, and the upper layer The reaction solution was added to a NaCl solution (1500 ml), and the solid was filtered off and washed with water. The residue was subjected to silica gel column chromatography, and eluted with 11- (: ₄ ¾011: ¾0: 110); (== 8: 1: 1 to obtain 21 g of pure product. Yield: 52.3%

1H NMR (CD ₃ 0D), ppm): 5.38 (d, J = 4.8Hz, lH), 4.51 (m, IH), 4.40 (m, IH), 3.54 (m, 1H), 3.43 (m, IH), 3.20 (dd, J = 6.8Hz, IH), 2.62 (m, IH), 2.3 l (m, 4H), 1.95 (s, 3H), 1.06 (s, 3H), 0.95 (d, J = 6.8Hz, 3H), 0.81 (s, 3H) 0.78 (d, J = 6Hz, 3H).

MS m / z: 671, 397.

IR: 3396 (br), 2950, 2905, 1734, 1728, 1654, 1559.

[Example 10] Preparation of 4- (N- (3-imidazol-4-yl) propanoic acid-2-yl) amino-4-oxobutanoic acid diosgenin acetate (compound 7)

Diosgenin succinate monoester was prepared according to the method of Example 1. 30.8 g (60 mmol) of diosgenin succinic acid monoester and 9 g of N-hydroxysuccinimide under stirring in a water and salt bath (78 mmol) was dissolved in THF (350 ml), and then 14.8 g (72 mmol) of N, N-dicyclohexylcarbodiimide was added, and reacted at room temperature for 1 hour. The insolubles were filtered off and most of the THF was evaporated. The residue was poured into ice water (1000 ml) and stirred to precipitate a solid, and the solid was washed with water. Then dissolved in THF (400ml) to prepare solution A.

Under cooling and stirring in an ice bath, 18.6 g (120 mmol) of histidine hydrochloride, 22.2 g (264 mmol) of NaHC0 _{3 were} dissolved in H ₂ O (150 ml) and THF (50 ml), and then solution A. was added, and the reaction was stirred for 0.5 After an hour, remove the ice bath and react at room temperature for another 4.5 hours. The reaction was checked by TLC to completion. The pH value was adjusted to 3 with 6N hydrochloric acid, and the lower aqueous solution was separated. The upper reaction solution was added to the water to precipitate, and the precipitate was filtered off, washed with water, and dried to obtain 35 g of a solid. Silica gel column chromatography, eluting with nC ₄ H ₉ OH: H ₂ O: AcOH = 10: 1: 1, yielded 12 g of pure product, yield: 32.8%.

1H NMR (DMSO, ppm): 8.06 (s, 3H), 7.00 (s, 1H), 5.33 (d, J = 4.4HzHz, 1H), 4.43 (m, 2H), 4.43 (m, 2H), 4.28 ( dd, J = 7.6Hz, 1H), 3.40 (m, 1H), 3.21 (t, J = 11.6Hz, 1H), 2.98 (m, 1H), 2.70 (m, 1H), 2.38 (m, 4H), 2.24 (d, J = 7.6Hz, 2H), 1.96 (s, 3H), 0.98 (s, 3H), 0.90 (d, J = 6.8Hz, 3H), 0.74 (s, 3H), 0.73 (d, J = 5.6Hz, 3H).

MS m / z: 652;

IR: 3500-3100 (br), 2950, 2904, 1731, 1634.

[Example 11] Preparation of 4- (N-glutaric acid-2-yl) amino-4-oxobutanoic acid diosgenin disodium salt (Compound 8)

30.8 g (60 mmol) of diosgenin succinic acid monoester and 9 g (78 mmol) of N-hydroxysuccinimide were dissolved in THF (350 ml) under stirring in an ice-salt bath, and then Ν, Ν-bicyclo Hexylcarbodiimide 14.8 g (72 mmol), reacted at room temperature for 1 hour and then overnight. The insolubles were filtered off and most of the THF was evaporated. The residue was poured into water) (1000 ml) and stirred to precipitate a solid. The solid was washed with ice water, and then dissolved in THF (400 ml) to prepare solution A.

Under ice-cooling and stirring, dissolve 17.7 g (120 mmol) of glutamic acid, 22.2 g (264 mmol) of NaHC0 ₃ in H ₂ O (150 ml) and THF (50 ml), and then add solution A; remove the water bath after stirring for 0.5 hours The reaction was continued at room temperature for 4.5 hours. The reaction was checked by TLC to completion. 'The pH value was adjusted to 3 with 6N hydrochloric acid, and the lower aqueous solution was separated. The upper reaction solution was added to the water to precipitate, and the precipitate was filtered off, washed with water and dried to obtain 35 g of a solid. Silica gel column chromatography was performed with acetic acid: ethyl acetate: petroleum acid = 0.4: 1: 3 to obtain 12.8 g of pure product. This product was dissolved in a mixed solvent of THF Oml) and water (50ml), and then 3.35g of NaHC0 _{3 was} added. After stirring for 2 hours, the solvent was distilled off at low temperature and dried to obtain the product.

lH NMR (DMSO, ppm)): 12.3 (br), 5.34 (d, J = 4.4Hz, 1H), 4.44 (m, 1H) ₅ 4.28 (dd, J = 7.6Hz, 1H), 4.19 (m, 1H ), 3.42 (m, 1H), 3.21 (t, J = 6.8Hz, 1H), 2.49 (m, 4H), 2.27 (m, 2H), 0.99 (s, 3H), 0.91 (d ₅ J = 7.6Hz , 3H), 0.75 (s, 3H), 0.73 (d, J = 6.4Hz, 3H).

MS m / z: 642. PT / CN03 / 00263

IR: 3400 ~ 3300 (br), 2950, 1731, 1632.

[Example 12] Solubility experiment

Solubility in water was measured according to the regulations in the Pharmacopoeia of the People's Republic of China (2000 Edition). The results are shown in Table 2. Compounds 1 to 8 in Table 2 are the compounds prepared in Examples 4 to 11, respectively.

Table 2 Solubility test results

[Example 13] Effect of Diosgenin carboxylic acid derivative on myocardial infarction caused by myocardial ischemia in rats

In this experiment, a rat model of myocardial infarction caused by myocardial ischemia was used to observe the effect of diosgenin carboxylic acid derivatives on the degree of myocardial infarction. The test method is as follows:

The animals were randomly divided into 7 groups of 5 animals each: model group (M) (saline, 3ml), test drug group (T) 0.125, 0.063g / Kg group (respectively the maximum dose 1/20, 1 / 40), the tested drugs were: 1 diosgenin sodium succinate monoester sodium salt; 2 diosgenin 2,2 difluorenyl- [1,3] dioxolane 4,5-dicarboxylic acid monoester Ester sodium salt; 3 »Diosgenin tartrate monoester sodium salt. The test drugs were dissolved in physiological saline to a desired concentration, and the administration volume was 3 ml / kg, and the administration route was duodenum.

Animals were anesthetized with urethane (Urathan) intraperitoneally (1000mg / kg), fixed in the supine position, and ECG amplifier (Japan Optoelectronic ECG Amplifier, AC-601G) was connected to two recorders (Japan Optic Recticorder) monitoring standard II lead ECG; cut the trachea, insert the trachea, intubate, connect to a ventilator (SC-3, Shanghai Medical Equipment Factory) and perform artificial respiration (32 breaths / min, respiratory ratio 1: 3); open the chest, and break 3 ~ 5 ribs, Open the pericardium, expose the heart, and thread the root of the left anterior descending coronary artery (No. 0 suture) for ligation; isolate the duodenum for administration; stabilize for 10 minutes after threading, ligation (no ST segment and T wave Changes were eliminated), and the abdomen was closed after the test drug was administered; the chest wall was sutured and spontaneous breathing was restored.

The test was terminated 3 hours after the ligation. Five pieces were transected under the heart ligature line and stained with N-BT. The multimedia color pathology analysis system (MPIAS-500) was used to measure the area of normal myocardium and infarcted myocardium at a fixed image distance. Degree; results were statistically processed (t test).

Table 1 Effects of diosgenin carboxylic acid derivatives on myocardial infarction caused by myocardial ischemia in rats Normal myocardium infarct myocardial infarction cardiac infarct heart weight accounted for accounting infarction cardiac ventricle mm ^² mm ² g ^_0/0 Heart ⁰ / n ₀ doses.

Group ■ / kg

M 5 323.41 ± 21.79 103.06 ± 10.51 0.278 + 0.027 31.8 + 2.1 27.1 ± 1.8

1 5 0.125 327.47 ± 21.98 82.60 ± 6.38 ** 0.230 ± 0.032 * 25.3 ± 1.8 ** 21.6 ± 1.6 *

1 5 0.063 341.09 ± 11.43 93.81 ± 7.34 0.272 + 0.022 28.1 ± 1.8 * 24.0 + 1.6 *

2 5 0.125 325.41 ± 21.79 88.06 ± 10.51 0.256 ± 0.027 27.0 ± 2.1 22 · 5 ± 1 · 8

2 5 0.063 322.86 + 21.98 94.60 ± 6.38 ** 0.268 + 0.032 * 29.3 ± 1.8 ** 25.6 ± 1.6 *

3 5 0.125 330.11 ± 11.43 87.81 ± 7.34 0.249 + 0.022 26.6 ± 1.8 * 22.0 + 1.6 *

3 5 0.063 321.88 ± 11.43 92.70 ± 6.38 ** 0.258 ± 0.032 * 28.8 + 1.8 * 25.1 ± 1.6 * Compared with the model group P <0.05, P <0.01

The test results confirmed that the percentage of infarcted area in the model group to the ventricle and heart were

31.8 and 27.1%; Diosgenin carboxylic acid derivative sodium salt 0.125g / kg group significantly reduced myocardial infarction, infarct area decreased, infarct area weight reduced, infarct area occupied ventricle and heart percentage decreased, 0.063g / Kg group infarction The percentage of ventricular area and heart decreased, and there were significant differences compared with the model group (P <0.05 ~ P <0.01). ''

[Example 14] Effect of diosgenin succinate monoester derivative on myocardial infarction caused by myocardial ischemia in rats

1. Test method: Healthy Wistar rats, male and female, weighing 250-300. Randomly grouped according to body weight, 8 animals in each group. Distilled water (sham operation control, model control group), test substance 125, 250 mg / kg '(test drug group), and heartache 10mg / kg

(Tool Drug Group). The administration volume was 10ml / kg, once a day for 3 days. One hour after the last dose, anesthesia was injected intraperitoneally with chloral hydrate (350 mg / kg), and the normal ECG was measured. The left chest was depilated, the intersection of the midline of the clavicle and the fourth rib was taken as the center, and the skin was cut 2 cm along the midline of the clavicle. The muscle layer was separated sacrifice, and the circumlunar / L layer was used for purse suture. Cut the fourth rib, open the thorax, pull the heart out of the thorax with a circular hook, and ligate the left anterior descending coronary artery with a 6/0 silk thread at the coronary vein between the arterial cone and the midpoint of the left atrial appendage. The heart was placed in the chest cavity, the air in the chest cavity was extracted, and the L-layer purse was ligated, and the skin was sutured. Blood was taken from the abdominal aorta 24 hours later, and the serum was prepared by centrifugation at 4rpm, 3000 rpm and lOmin for 10min, and stored at -20Ό, and the serum biochemical indicators (AST, LDH, CK-MB) were measured within one month. After the blood was taken, 1% Evans Blue 0.3ml / head was injected through the external jugular vein. After 1 min, the heart was taken out and washed several times with normal saline. The heart was sliced along the vertical direction of the long axis of the heart to a thickness of 1.5 mm. After staining in 1% TTC solution for 15 minutes, the heart sections were removed, and the pathological image analysis system was used to determine and calculate the percentage of myocardial infarction area to the left ventricle area.

2. Test results

1) Effect of myocardial infarction range in myocardial ischemia rats

The results showed that the infarct size of the rats in the model control group before the ligation was 17.1%. Compared with the sham operation group, the myocardial infarction area was obvious. The rats infused the compound 7 and the infarct size was significantly reduced. The results are shown in Table 3. N03 / 00263 Table 3 Diosgenin succinate monoester derivative on myocardium induced by coronary ligation in rats

Effect of infarct size

Note: 1, compared with the sham operation group: "^ ΊΡΟ.ΟΟΙ;

2, compared with the model control group: * P <0.05, ** P <0.01

2) Effects on myocardial enzymes AST, LDH, CK-MB in myocardial ischemia rats

The results showed that the myocardial enzymes AST, LDH, and CK-MB in the model control group of the descending branch before ligation were significantly increased, and the difference was very significant compared with the sham operation group (P <0.01). Rats ingested compounds 4 and 7, and their AST was significantly reduced. Compound 7 was administered and its LDE [significantly decreased.

The CK-MB of compound 7 was significantly reduced after administering the compound. The results are shown in Table 4.

The tool drug Xintongding significantly reduced the ST segment elevation of the ischemic ECG, narrowed the scope of myocardial infarction, and decreased serum myocardial enzymes (LDH, CK-MB,).

Table 4.Effects of Diosgenin 4 Succinate Monoester 4 on Blood Serum Biochemistry in Rats with Myocardial Ischemia Induced by Coronary Ligation

2, compared with the model control group: PO.05, P <0.01, *** P <0.001

[Example 14] Effect of Diosgenus Sodium or Yuanyuan Succinate Monoderivative on Cerebrovascular Resistance in Rats with Constant Perfusion of Cone Artery

In this experiment, the literature method (Chen Qi, Research Methodology of Pharmacology of Traditional Chinese Medicine, People's Medical Publishing House, 1996: 538-540) was used to observe the injection of drugs (50 mg / kg) and phentolamine (0.5 mg / kg) into the cone artery. Effects on blood pressure and cerebrovascular resistance in rats. The experimental results show that after the test substance, phentolamine is injected into the cone arteries, compared with before injection, it has a significant effect on reducing vascular resistance and blood pressure. The results are shown in Table 5. The groups in the table are the corresponding compound groups. It can be known from technical common sense that the present invention may be implemented by other embodiments without departing from the spirit or essential characteristics thereof. Therefore, the above-disclosed implementations are, for all aspects, merely examples, and are not the only ones. All changes that are within the scope of the invention or within the scope equivalent to the invention are encompassed by the invention. Table 5 Effects of diosgenin succinate monoester derivatives on blood pressure and cerebrovascular resistance in rats

Note: Compared with phentolamine group: P <0.05, P <0.01, *** P <0.001

Claims

Rights request

1. Diosgenin carboxylic acid derivatives of the formula (I):

In the formula (I), an alkyl group having 1 to 2 carbon atoms or a substituted alkyl group, and M is H or Na.

2, according to claim 2, diosgenin monocarboxylic acid derivative, wherein the selected _{_{_{-CH r, -CH 2 CH 2 -}}} 5 -CH 2 CH (OH) - 3 -CH (OH) CH ( OH) -and

3. The diosgenin carboxylic acid derivative according to claim 2, wherein the -CH ₂ C¾- and M is Na.

4. The diosgenin carboxylic acid derivative according to claim 2, wherein said is -CH (OH) CH (OH)-, and M is Na.

5. The diosgenin carboxylic acid derivative according to claim 2, wherein the

M is Na.

6. The diosgenin carboxylic acid derivative according to claim 2, wherein the -CH ₂ CH _2- , and M is an amino acid.

7. A method for preparing a diosgenin carboxylic acid derivative, comprising the following steps:

1) Diosgenin is reacted with acid anhydride under reflux conditions to obtain a diosgenin derivative;

2) Diosgenin saponin carboxylic acid 4-fouling organism is prepared by reacting with sodium bicarbonate or potassium bicarbonate under reflux conditions; the reaction formula is as follows:

8. The method according to claim 7, wherein the reaction is performed in an organic solvent under reflux conditions. .

9. The method according to claim 8, wherein the organic solvent in the step a reaction is pyridine, and the organic solvent in the b step reaction is ethanol.

10. Application of diosgenin carboxylic acid derivatives and pharmaceutically acceptable salts thereof in the preparation of medicines for treating cardiovascular and cerebrovascular diseases.

11. A diosgenin succinic acid monoester having the structure of formula (III) ^ · Biological:

(III)

In formula (III), AA is selected from glycine, alanine, cystine, glutamic acid, lysine, proline, threonine, tyrosine, leucine, isoleucine, benzene Alanine, tryptophan, serine, threonine, cysteine, aspartic acid, arginine, histidine, and their corresponding amino or carboxylic acid salts, and other pharmaceutically acceptable salt.

The diosgenin succinate monoester derivative according to claim 11, wherein the amino acid is selected from the (L) counterpart.

13. The diosgenin succinate monoester derivative according to claim 11, wherein the amino acid is selected from the (D) counterpart.

The diosgenin succinate monoester derivative according to claim 11, wherein the pharmaceutically acceptable salt is selected from a sodium salt, a potassium salt, a calcium salt and a magnesium salt. '

15. The diosgenin succinate monoester derivative according to claim 11, wherein the pharmaceutically acceptable salt is selected from the group consisting of a hydrochloride, an acetate, a mesylate, and a citric acid salt.

16. The diosgenin succinate monoester derivative according to claim 11, wherein AA is glycine or a sodium salt thereof.

17. The diosgenin succinate monoester derivative according to claim 11, wherein AA Is histidine or its acetate.

The diosgenin succinate monoester derivative according to claim 11, wherein AA is glutamic acid or a sodium salt thereof.

19. The diosgenin succinate monoester derivative according to claim 11, wherein AA is aspartic acid or a sodium salt thereof.

20. The diosgenin succinate monoester derivative according to claim 11, wherein AA is arginine or an acetate thereof.

21. A method for preparing a diosgenin succinate monoester derivative, comprising reacting a compound of formula (II) with N-hydroxysuccinimide, N, N-dicyclohexylcarbodiimide, and then reacting with Various amino acid reactions:

(Π)

22. The method according to claim 21, wherein the reaction is performed in a mixed solvent of tetrahydrofuran and water.

23. The method according to claim 22, further comprising the step of reacting a compound of formula (III) with sodium bicarbonate or potassium bicarbonate under reflux conditions.

24. The method according to claim 22, further comprising the step of salting the compound of formula (III) with acetic acid.

25. Application of a diosgenin succinate monoester derivative and a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating cardiovascular and cerebrovascular diseases.