JPH0155277B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention relates to novel pomolic acid derivatives. The pomolic acid derivative of the present invention has the general formula [In the formula, R ¹ is a hydrogen atom or a group

[Formula], R ² is a hydrogen atom, a lower alkyl group, or a group

[Formula] is shown. However, when R ¹ represents a hydrogen atom, R ² represents a group.

[Formula] shall be shown. ] The present inventors have discovered that Aquifoliaceae
We have conducted intensive research on extracts from Chinese holly (Ilex cornuta). The present inventors recognized the presence of a compound in the extract that has thrombolytic activity, blood flow improving effect, and decholesterolizing effect, and succeeded in extracting and isolating this compound, thereby completing the present invention. In this specification, lower alkyl groups include, for example, methyl, ethyl, propyl, isopropyl,
Examples include butyl, tert-butyl, pentyl, hexyl, and the like. The compound of the present invention is produced, for example, according to the method shown below. Among the compounds of the present invention, the following formulas (1a) and (1b)
The compound represented by is extracted and isolated from Chinese holly in the following manner, for example. That is, first, Chinese holly Ilex cornuta is extracted using a common polar solvent such as methanol or ethanol, and the extract is concentrated under reduced pressure to obtain a primary extract. The method for collecting the target compound from the first extract is not particularly limited, and any of various known methods utilizing physical and chemical properties can be employed. For example, the difference in solubility with impurities, common adsorbents such as activated carbon, XAD-2, silica gel, ion exchange resin,
This can be carried out by a method that utilizes the difference in adsorption affinity for a cephadex, a difference in the distribution ratio between two liquid phases, or a combination of these methods. More specifically, the above-mentioned first extract is extracted using a solvent such as butanol by a solvent distribution method, the extract is concentrated under reduced pressure, and then subjected to silica gel column chromatography using an appropriate solvent such as chloroform, methanol, or It can be obtained by elution with a mixed solvent of water or the like. Among the compounds of the present invention, the formulas (1a) and (1b)
Compounds other than those represented by formula (1a) or (1b) can be produced by the method shown below using the compound of formula (1a) or (1b) as a starting material. <Reaction formula-1> [In the formula, R ² ' represents a lower alkyl group] According to reaction formula-1, among the compounds of the present invention, a compound in which R ² represents a lower alkyl group [compound of general formula (1c)] is a compound (1a) that is a lower alkyl group. It can be obtained by esterification using a conventional method. The esterification reaction is carried out by esterifying compound (1a) and a lower alcohol in the presence of a catalyst commonly used in esterification reactions. Typical catalysts used in this case include, for example, hydrochloric acid gas, concentrated sulfuric acid, phosphoric acid, polyphosphoric acid, boron trifluoride, inorganic acids such as perchloric acid, trifluoroacetic acid, trifluoromethanesulfonic acid, and naphthalenesulfonic acid. Examples include organic acids such as p-tosylic acid, benzenesulfonic acid, and ethanesulfonic acid, acid anhydrides such as trifluoromethanesulfonic anhydride, thionyl chloride, and acetone dimethyl acetal. Furthermore, cation exchange resins can also be used as the catalyst. The amount of these catalysts used may be within the range commonly used. The above esterification reaction proceeds either without a solvent or in a solvent. Examples of the solvent used include various solvents used in ordinary esterification reactions, such as aromatic hydrocarbons such as benzene, toluene, and xylene; halogenated hydrocarbons such as dichloromethane, dichloroethane, chloroform, and carbon tetrachloride;
Examples include ethers such as diethyl ether, tetrahydrofuran, dioxane, and ethylene glycol monomethyl ether. The ratio of compound (1a) and lower alcohol to be used may be appropriately selected over a wide range, but in order to improve the production rate of the target product, the latter is usually used in a solvent-free manner. When a solvent is used in excess, it is preferred to use the latter in an amount of equimolar to 5 times the former (especially preferably equimolar to 2 times). In addition,
In this esterification reaction, the production rate can be further increased by removing the produced water from the reaction system using a drying agent such as anhydrous calcium chloride, anhydrous copper sulfate, anhydrous calcium sulfate, or phosphorus pentoxide. be. The reaction temperature may be selected appropriately, and is usually in the range of about -20 to 200°C, especially about 0°C.
It is preferable to carry out the reaction at a temperature of about 150°C. The reaction time is generally about 10 minutes to 20 hours. The above esterification reaction can also be carried out using an activated derivative of the carboxy group of compound (1a), such as a carboxylic acid halide or carboxylic acid anhydride. Furthermore, the desired product can also be obtained by reacting compound (1a) with a diazoalkane such as diazomethane in the above esterification reaction. The reaction between compound (1a) and diazoalkane is carried out using the same solvent as used in the esterification reaction, usually at -5 to 20°C, preferably at about 0 to 5°C, for about 5 minutes to 1 hour. It can be done with In this case, the ratio of the compound (1a) and the diazoalkane (usually used as a solution in ether, dichloromethane, etc.) is usually about 1 to 2 times the mole of the former, preferably 1 to 1.5 times the mole of the latter. Good. <Reaction formula-2> According to Reaction Formula-2, compound (1a) can be obtained by hydrolyzing compound (1b). The reaction is usually carried out in the presence of a catalyst. As the catalyst, those used in ordinary hydrolysis reactions can be used, such as basic compounds such as sodium hydroxide, potassium hydroxide, barium hydroxide, sodium hydroxide, and potassium carbonate. The amount of these catalysts to be used is not particularly limited and can be appropriately selected from a wide range, but it is usually preferable to have them present in the reaction system in an amount of about 3 to 30 wt%.
Although the reaction may be carried out according to a conventional method, it generally proceeds advantageously in a solvent. The solvent used is
For example, water, lower alcohols such as methanol, ethanol, isopropanol, dioxane,
Examples include ethers such as THF, DMF, DMSO, or a mixture thereof. The reaction temperature is not particularly limited and may be selected appropriately from a wide range, but is usually room temperature to about 150°C, preferably 50 to 110°C.
It is better to do so. The reaction is generally completed in about 30 minutes to 10 hours. <Reaction formula-3> According to Reaction Formula-3, Compound (1d) can be obtained by hydrolyzing Compound (1b), and the same reaction conditions as the reaction in Reaction Formula-2 above can be adopted for the reaction. , preferably by enzymatic hydrolysis reaction. The enzyme to be used is not particularly limited, and a wide variety of so-called glycosidases and mixtures thereof can be used, and these are easily available, for example, as commercial products. The reaction is usually carried out under conditions customary for this type of enzymatic reaction, such as buffers such as acetate buffers and phosphate buffers, aqueous solutions of mineral acids such as hydrochloric acid, sulfuric acid, nitric acid, etc. in solvent, 35-40
The reaction is completed at 1 hour to 20 days at ℃. Among the pomol acid derivatives of the present invention thus obtained, the compound having an acidic group can easily form a salt by reacting with a basic compound. It also includes salts of derivatives. After completion of the reaction in each step described above, the target compound can be easily isolated and purified by commonly known separation means. As the separation means, for example, solvent distillation,
Solvent extraction, precipitation, recrystallization, column chromatography, thin layer preparative chromatography, etc. can be arbitrarily employed. When the pomolic acid derivatives and salts thereof of the present invention are used as pharmaceutical agents, they are usually put into the form of a pharmaceutical composition together with a pharmaceutical carrier. As carriers, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, surfactants, lubricants, etc., which are commonly used to prepare drugs according to the usage form, can be used. I can give an example. Various dosage unit forms of drugs can be selected depending on the therapeutic purpose, and representative examples include tablets, pills, powders, solutions, suspensions, emulsions, granules, capsules, suppositories, and injections. Examples include agents (liquid agents, suspension agents, etc.). When forming tablets, carriers include excipients such as lactose, sucrose, sodium chloride, glucose solution, urea, starch, calcium carbonate, kaolin, crystalline cellulose, and silicic acid, water, ethanol, propanol, and simple syrup. , glucose, starch solution, gelatin solution, carboxymethylcellulose, shellac, methylcellulose, potassium phosphate, binder such as polyvinylpyrrolidone, dry starch, sodium alginate, agar powder, laminaria powder, sodium bicarbonate, calcium carbonate, twin, lauryl sulfate Disintegrants such as sodium, stearic acid monoglyceride, starch, lactose, disintegration inhibitors such as sucrose, stearin, cocoa butter, hydrogenated oil, absorption enhancers such as quaternary ammonium bases, sodium lauryl sulfate, glycerin, starch, etc. moisturizer,
Adsorbents such as starch, lactose, kaolin, bentonite, and colloidal silicic acid, and lubricants such as purified talc, stearic acid groups, boric acid powder, macrogol, and solid polyethylene glycol can be used. When forming into a pill form, carriers include excipients such as glucose, lactose, starch, cocoa butter, hydrogenated vegetable oil, kaolin, and talc, binders such as gum arabic powder, tragacanth powder, gelatin, and ethanol, and laminaria. , agar, etc. can be used. Furthermore, the tablets can be made into conventionally coated tablets, such as sugar-coated tablets, gelatin-encapsulated tablets, enteric-coated tablets, film-coated tablets, double tablets, or multilayer tablets, if necessary. When forming into a suppository, for example, polyethylene glycol, cacao butter, higher alcohols, esters of higher alcohols, gelatin, semi-synthetic glycerides, etc. can be used as carriers. When prepared as injections, solutions and suspensions are preferably sterilized and isotonic with blood, and when formed into solutions, emulsions, and suspensions, diluents such as Water, ethyl alcohol, propylene glycol, ethoxylated isostearyl alcohol, polyoxylated isostearyl alcohol, polyoxyethylene sorbitol, sorbitan ester, etc. can be used. In this case, the drug may contain a sufficient amount of salt, glucose, or glycerin to prepare an isotonic solution, and may also contain conventional solubilizing agents, buffers, soothing agents, preservatives, etc. Furthermore, coloring agents, preservatives, fragrances, flavoring agents, sweeteners, etc., and other pharmaceuticals may be included in the drug, if necessary. The amount of the compound of the present invention to be contained in the drug is not particularly limited and can be appropriately selected within a wide range, but is usually 1 to 70% by weight, preferably 5 to 50% by weight of the total composition.
It is better to There are no particular restrictions on the use of the above-mentioned drugs, and they can be administered in a manner appropriate for various forms. For example, tablets
In the case of pills, solutions, suspensions, emulsions, granules and capsules, they are administered orally. In the case of an injection, it is administered intravenously alone or mixed with a normal replacement fluid such as glucose or an amino acid, and if necessary, it is administered alone intramuscularly, intradermally, subcutaneously, or intraperitoneally. Suppositories are administered rectally. Although the dosage of the drug is appropriately selected depending on the purpose of use, symptoms, etc., it is usually preferable that the dosage of the compound of the present invention is about 0.5 to 3 mg/Kg per day. Thus, a drug containing the compound of the present invention as an active ingredient is useful as a prophylactic or therapeutic agent for thrombosis or a lipid metabolism improving agent. The present invention will be explained in more detail with reference to Examples below. Example 1 9 kg of air-dried leaves of Chinese holly were ground and mixed with methanol (80%) at room temperature for 20 days.
Extracted with. The solvent was distilled off under reduced pressure to obtain 806 g of syrup. 628g of n-butanol
It was distributed in a water (1:1 V/V) system. The organic layer was separated and the solvent was distilled off under reduced pressure to obtain 510 g of a residue. After dissolving 478g of it in the minimum amount of methanol,
Poured into a large volume of stirred ethyl acetate. The deposited precipitate was separated to obtain 69 g. 40 g of the precipitate obtained was applied to Kieselgel 60 (Merck), and the portion eluted with chloroform/methanol/water (11:3:1, lower layer) was repeatedly purified by precipitation from methanol-ether and methanol. 3β-O-[β-D-
1.5 g of glucopyranosyl-(1→2)-α-L-arabinopyranosyl]-pomolic acid was obtained. Amorphous IRν ^KBr _nax ; 3380 (OH), 1690 (COOH), 1636 (sh,
C=C), 1070 (ether) cm ^-1 FD-MS m/z (%); 789 [(M+Na) ⁺ ] (100), and further added to chloroform/methanol/water (7:3:1, lower layer). The eluted portion was recrystallized from methanol to give 3β-O-[β
-D-glucopyranosyl-(1→2)-α-L-
Arabinopyranosyl]-pomolic acid-(28→1)
9.4 g of -β-D-glucopyranosyl ester was obtained. White powder crystal Melting point 268-270℃ [α] ²⁰ _D , -15.0 (methanol; c = 0.71) IRν ^KBr _nax ; 3380 (OH), 1725 (ester C=O),
1630 (C=0) 1060 (ether) cm ^-1 FD-MS m/z (%); 951 [(M+Na) ⁺ ] (33),
789 [(M+Na) ⁺ −162] (100), 657
[(M+Na) ⁺ −294] (5.9), 627 [(M+
Na) ⁺ −324] (11.8) C H Calculated value C ₄₇ H ₇₆ O ₁₈・2H ₂ O 58.49 8.36 (%) Analytical value 58.69 8.37 (%) Example 2 3β-O- [obtained in Example 1-] Dissolve 300 mg of β-D-glucopyranosyl-(1→2)-α-L-arabinopyranosyl]-pomolic acid in 20 ml of methanol, add about 2 g of Dowex 50W x 8 (Hani Chemical Co., Ltd.), and leave at room temperature. After stirring for 30 minutes, it was filtered. 24 to liquid
An ether solution containing mg of diazomethane was added and reacted at room temperature for 1 hour. After purification by preparative thin layer chromatography (silica gel; Merck & Co., Ltd., PF254, solvent: chloroform/methanol/water = 7:1:1, lower layer eluent: methanol), it was recrystallized from methanol to obtain 3β-O-[ β-D-glucopyranosyl-
220 mg of (1→2)-α-L-arabinopyranosyl]-pomolic acid methyl ester was obtained. Colorless crystal Melting point 240-243℃ [α] ²⁰ _D , +15.6 (methanol; c=0.80) IRν ^KBr _nax ; 3310 (OH), 1718 (COOCH ₃ ), 1635
(C=C), 1065 (ester) cm ^-1 FD-MS m/z (%); 803 [(M+Na) ⁺ ]
(100), 641 [(M+Na) ⁺ −162]
(13.7), 509 [(M+Na) ⁺ -294] (5.9) C H Calculated value C ₄₂ H ₆₃ O ₁₃・2H ₂ O 61.74 8.88 (%) Analytical value 61.75 8.80 (%) Example 3 In Example 1- 1 g of the obtained 3β-O-[β-D-glucopyranosyl-(1→2)-α-L-arabinopyranosyl]-pomoric acid (28→1)-β-D-glucopyracyl ester was added to 20% The mixture was added to 60 ml of a methanol solution of potassium hydroxide and refluxed for 3 hours. The reaction solution was poured into a large amount of hydrochloric acid-ice water and extracted with n-butanol/ethyl acetate (1:1). After washing the organic layer with water, it was distilled off under reduced pressure and the residue was dissolved in methanol.
Mix to 100 ml, add Dowex 50 x 8 and mix at room temperature.
After stirring for an hour and desalting, the mixture was filtered. After distilling off the solvent of the liquid, the resulting residue was precipitated with a methanol-ether solution to obtain amorphous 3β-O-[β-D-glucopyranosyl-(1→2)-α-L-arabinopyranosyl. ]-Pomolic acid was obtained, and it was confirmed that the physical properties were the same as those of Example 1-. This was dissolved in methanol and methyl esterified in the same manner as in Example 2 above. The 3β-
O-[β-D-glucopyranosyl-(1→2)-α
802 mg of -L-arabinopyranosyl]-pomolic acid methyl ester was obtained. This compound had the same physical properties as the compound obtained in Example 2 above. The melting point of the mixture with the compound obtained in Example 2 is 240
˜243° C., showing no melting point depression. Example 4 3β-O-[β-D-glucopyranosyl-(1→2)-α-L-arabinopyranosyl]-pomolic acid (28→1)-β-D obtained in Example 1- - Mixed glycidase obtained from 'Tvrbo cornutus' (Seikagaku Corporation, lot no.
ET9401) was added and stirred at 37°C for 17 days. 100 ml of cold water and n-butanol/ethyl acetate (2:
1) After adding 100 ml and filtering, the liquid was separated into an aqueous layer and an organic layer. The aqueous layer was further extracted with n-butanol/ethyl acetate (2:1). The organic layers were combined, washed with water, the solvent was distilled off under reduced pressure, and the resulting residue was dried in vacuo. This was performed using preparative thin layer chromatography (silica gel; Merck & Co., Ltd.,
PF254, solvent; chloroform/methanol/water,
12:3:1 Lower layer Eluent: Purified with methanol) and then recrystallized from methanol to obtain pomol acid (28→1)-β-D-glucopyranosyl ester
Obtained 111 mg. Colorless crystal Melting point 279-280℃ [α] ²⁰ _D , +17.8 (methanol, c=0.11) IRν ^KBr _nax ; 3400 (OH), 1723 (ester, C=
0), 1630 (C=C), 1070 FD-MS m/z (%); 657 [(M+Na) ⁺ ]
(100), 639 [(M+Na) ⁺ −H ₂ O]
(9.8), 495 [(M+Na) ⁺ −162] (11.8),
472 (M ⁺ −162) (2.9) MS m/z (%); 472.355 (M ⁺ −C ₆ H ₁₀ O ₅ ,
The calculated value for C ₃₀ H ₄₈ O ₄ is 472.355)
(2.2), 454 (4.9), 429 (8), 264
(46.4), 246 (76.5), 201 (100) C H Calculated value C ₃₆ H ₅₈ O ₉ 68.11 9.21 (%) Analyzed value 67.92 9.42 (%)

Claims (1)

[Claims] 1. General formula [In the formula, R ¹ represents a hydrogen atom or a group [Formula], and R ² represents a hydrogen atom, a lower alkyl group, or a group [Formula]]. However, when R ¹ represents a hydrogen atom, R ² represents a group [formula]. ] A pomolic acid derivative represented by these and its salt.