WO2007076655A1

WO2007076655A1 - METHOD FOR SYNTHESIS OF CHOLEST-3β,5α,6β,19-TETRAHYDROXY-24-ONE AND ITS DERIVATIVES

Info

Publication number: WO2007076655A1
Application number: PCT/CN2006/001627
Authority: WO
Inventors: Guangmei Yan; Jingxia Zhang; Pengxin Qiu; Yijun Huang; Qun Li
Original assignee: Sun Yat-Sen University
Priority date: 2005-12-31
Filing date: 2006-07-10
Publication date: 2007-07-12
Also published as: CN1793159A; CN100436474C

Abstract

A synthesis process of cholest-3β,5α,6β,19-tetrahydroxy-24-one and its derivatives includes the steps of: synthesis of 3β-acetoxy-cholest-5-en-24-acid with hyodesoxycholic acid as the starting material; synthesis of cholenic acid chloride by reaction with thionyl chloride; reaction with isopropylcadmium reagent under ultrasonic irradiation to produce 3β-acetoxy-cholest-5-en-24-one; then reaction with N-bromosuccinimide/water solution of perchloric acid system to obtain 3β-acetoxy-cholest-5α-bromo-6β-hydroxy-24-one; reaction with diacetoxy iodobenzene to give 3β-acetoxy-cholest-5α-bromo-6β,19-epoxy-24-one; reduction with Zn powder/glacial acetic acid system to give 3β,19-diacetoxy-cholest-5-en-24-one; oxidation of the double bond at position 5 and 6; then hydrolysis to produce cholest-3β,5α,6β,19-tetrahydroxy-24-one; finally reaction with methyltriphenylphosphonium bromide to produce 24-methylene-cholest-3β,5α,6β,19-tetraol; and reaction with hydrazine to give cholest-3β,5α,6β,19-tetrahydroxy-24-dihydrazone. The synthesis process according to the invention has the characteristic of lower price, material availability, scientific and reasonable synthetic routes, higher yield, and the like. The target compounds have significantly protective effect on neuronal apoptosis evoked by low K+.

Description

Method for synthesizing cholesteric-3β, 5α, 6β, 19-tetrahydroxy-24-ketone and derivatives

The present invention relates to a method for synthesizing cholestyl-3?,5?,6?,19-tetrahydroxy-24-ketone (structures such as I) and derivatives.

Background technique

24-methylene-cholesterol-35 a, 6 β, 19-tetraol (structure such as formula II) is a polyhydroxysteroid compound isolated from the soft corals Nephthea albida and N. tiexieralv erseveldt. Wang Guiyang, Li Fengying, Zeng Yongmei et al., Chemical Journal of Chinese Universities, 1992, 13, 623-627). The compound has strong killing ability on human rectal cancer cells, human lung cancer cells and murine lymphoid cancer cells (Chang- Yih Duh, Shang-Kwei, Mei-Jen Clu et al., J. Nat. Prod., 1998, 61, 1022-1024), has significant anti-inflammatory effects on a variety of inflammations (Wang Zhengyi, Xu Shibo, Liu Qing et al, Tropical Ocean, 1999, 18, 27-32), the compound for low potassium induced neuronal withering Dead cells have a significant protective effect.

Π

The content of this compound in marine organisms is extremely low. There are still many problems in the direct extraction of marine biological raw materials. The synthesis of this compound has high medicinal value. Intermediate cholesteric synthesis of the compound

3β, 5α, 6β, 19-tetrahydroxy- 24-ketone and nitrogen-containing derivatives have significant protective effects on hypokalemia-induced neuronal apoptotic cells.

Summary of the invention

SUMMARY OF THE INVENTION An object of the present invention is to provide a method for synthesizing cholestyl-3?,5?,6?,19-tetrahydroxy-24-one and derivatives.

The synthesis method of the invention uses porcine deoxycholic acid as a raw material to prepare 3 3-acetoxy-cholesteryl 5- The ene-24-acid is prepared by reacting with thionyl chloride to prepare choline chloride, which is reacted with isopropyl cadmium reagent under ultrasonic irradiation to obtain 3β-acetoxy-cholesteryl-5-ene-24-one, and then Reaction of bromosuccinimide/perchloric acid aqueous solution to obtain 3β-acetoxy-cholesteryl-5 α-bromo-6β-hydroxy-24-ketone, diacetoxyiodobenzene oxidation ring Generation of 3^-acetoxy-cholesteryl-5?-bromo-6,19-epoxy-24-ketone, followed by reduction with zinc powder/glacial acetic acid system or zinc powder/alcohol system to form 3β, 19-diacetyl Oxy-cholesteryl 5-alkenyl-2-ketone, and then choline-5-alkenyl-2-keto-ketone by alkali hydrolysis; oxidizing and hydrolyzing with oxidizing agent to obtain cholesteric _3β, 5α , 6β,19-tetrahydroxy-24-ketone; and methyltriphenylphosphonium bromide to synthesize 24-methylene-cholesteryl-3β, 5α, 6β,19-tetraol; Cholesterol - 3β, 5α, 6β, 19-tetrahydroxy-24-biguanide. The reaction route is shown below:

I. cholestyl-3 β,5 α, 6 19-tetrahydroxy-24-ketone; Π.24-methylene-cholesterol-3 β, 5 α, 6 β, 19-tetraol;

III. Cholesterol-3β,5α, 6β,19-tetrahydroxy-24-biguanide;

IV.3 β-acetoxy-cholesteryl 5-ene- 44-acid;

V.3 β-acetoxy-cholesteryl-5-ene-24-one;

VI.3 β-acetoxy-cholesteryl-5α-bromo-6β-hydroxy _24

W.3 β-acetoxy-cholesterol-5 α-bromo-6β, 19-epoxy

Shan.3β, 19-diacetoxycholine-5-ene-24-one;

Κ .3β 19-diacetoxy-cholesteryl-5,6β-epoxy-24-ketone;

Kb.3β 19-diacetoxy-cholesteryl-5, 6ci-epoxy

Β.3β, diacetoxy-cholesteryl-5,6-dihydroxy-24-one.

The specific steps of the synthetic method of the present invention are:

(1) IV reference (Bharuclia, Buckley GC, Cross CK, et al. Can J Chem., 1955, 34: 982-990.) Synthesis;

(2) Add IV to anhydrous benzene to thionyl chloride, add a catalytic amount of pyridine, stir at room temperature for 2 to 4 hours, concentrate to dryness under reduced pressure, and add benzene to continue concentration to bring out residual chlorination. Sulfoxide, a yellow viscous liquid;

(3) Weigh dry magnesium powder, add anhydrous ether, 'drop add isobromopropane / anhydrous ether solution, add ultrasonic radiation after 15~30 minutes, add cadmium chloride in batches after cooling, at 25~40° Stir under ultrasonic irradiation, and slowly add 30~40% anhydrous benzene solution of the above step product to the reaction system after cooling. After the completion of the dropping, continue ultrasonic wave reflux for 1~2 hours, cool in ice bath. After that, a 1:1 aqueous solution of hydrochloric acid is added dropwise, and extracted with benzene, and the benzene layer is washed with distilled water until neutral, dried, concentrated under reduced pressure, and purified by silica gel column chromatography

(4) Take a white solid V, add organic solvent to dissolve, add HC10 ₄ and water, then add N-succinimide in portions in the dark, control the temperature below 10 ° C, then at 15~20 The reaction was carried out at ° C for 1 to 2 hours, and a 10% aqueous solution of Na ₂ S ₂ O ₈ was added to the reaction mixture. After layering, the upper layer was concentrated, and the lower layer was extracted with C3⁄4C1 ₂ , and the organic layer was combined and washed with a saturated NaHC0 ₃ solution. Wash with distilled water until neutral, dry, concentrated under reduced pressure, silica gel column chromatography, to obtain white homo-VI;

(5) Take compound VI, dissolve in cyclohexane/benzene mixture, add oxidant, stir at 20~40 °C, irradiate with UV lamp for 7~8 hours, and then use 10% aqueous sodium thiosulfate solution, saturated NaHC0. ₃ solution and distilled water to wash to neutral, dry, concentrated under reduced pressure to give a pale yellow compound W; (6) Take the compound and add glacial acetic acid to dissolve, add zinc powder, stir and reflux for 20~24 hours; heat and filter, the filtrate is extracted with dichloromethane, and the extract is washed with saturated sodium bicarbonate solution and distilled water until neutral. , drying, concentration under reduced pressure, silica gel column chromatography, to obtain a pale yellow viscous liquid;

(7) The compound is dissolved in dichloromethane, added with an oxidizing agent, stirred at room temperature, and the dichloromethane is removed under reduced pressure. The residue is dissolved in ethyl acetate, and then washed with a 10% sodium sulfite solution until colorless, washed with water until neutral. Dry, concentrate under reduced pressure to give a white solid Ka, dissolve it in acetone, add perchloric acid, stir at room temperature, adjust pH to neutral, distill off acetone under reduced pressure, 'extract with dichloromethane, dry, decompress Concentrated to give a white solid X;

(8) Compound I was dissolved in 88% formic acid, heated in a boiling water bath for 5 minutes, cooled to room temperature, added with oxidizing agent, allowed to stand at room temperature for 24 hours, diluted with water, extracted with ethyl acetate, and then washed with 10% sodium sulfite solution until colorless. , washed to neutral, dried, concentrated under reduced pressure to give a white solid Kb;

(9) The upper step product Kb or X is dissolved in methanol, and a 3% potassium hydroxide alcohol solution is added thereto, and the mixture is heated under reflux for 15 to 30 minutes to adjust the pH to neutrality, and methanol is distilled off under reduced pressure, and ethyl acetate is extracted. , the organic layer is dried, the solvent is evaporated under reduced pressure to give a pale yellow solid.

(10) Add methyltriphenylphosphonium bromide to an anhydrous tetrahydrofuran solution, and inject n-butyllithium. The amount of the injection is preferably dissolved in methyltriphenylphosphonium bromide. The reaction is stirred at room temperature for 4 to 6 hours. The I in the above step is added to the reaction system, and the reaction is continued for 2 to 3 hours. The reaction solution is poured into a saturated ammonium chloride solution, extracted with diethyl ether, the ether layer is dried, and the solvent is evaporated to give a pale yellow solid. Silica gel column chromatography gave white solid II;

(11) I was dissolved in ethanol, added to an aqueous solution of hydrazine, refluxed under reduced pressure, and the solvent was evaporated under reduced pressure. Water was added, and the mixture was adjusted to neutral, and ethyl acetate was evaporated to give a white solid. White solid III.

In the above reaction step, the concentration of the benzene solution in the step (2) is 25 to 45%, the amount of the thionyl chloride is 1.0 to 2.0 times the number of moles of II; the amount of the magnesium powder in the step (3) is II. The molar amount of the organic solvent is 1.2 to 3.0 times; the organic solvent in the step (4) is tetrahydrofuran or dioxane; the oxidizing agent in the step (5) is diacetoxyiodobenzene or lead tetraacetate; in the step (7) The oxidizing agent is potassium permanganate/copper sulfate or m-chloroperbenzoic acid; the oxidizing agent in the step (8) is 30% hydrogen peroxide.

The synthetic method of the invention has scientific and reasonable reaction route and high yield of the synthetic compound, and the synthesized intermediates are cholesteric-3β, 5α, 6β, 19-tetrahydroxy-24-one, 24-methylene-cholesteryl-3 β , 5 α, 6 β , 19—four Alcohol; and cholesteric-3β, 5α, 6β, 19-tetrahydroxy-; 24-biguanide have significant protective effects on low potassium-induced neuronal apoptosis cells.

detailed description

The following examples are intended to illustrate the invention, but the scope of protection of the invention is not limited to the following examples. Unless otherwise stated, in the following examples: I represents cholesteryl-3β,5α,60,19-tetrahydroxy-24-one; II represents 24-methylene-cholesteryl-3β,5α', 6β,19 Tetrahydrin; III represents cholestyl-3 β, 5 α, 6β, 19-tetrahydroxy-24-biguanide; IV represents 3 β-acetoxy-cholesteryl 5-alkenyl 24-acid; V represents 3 β -Acetoxy-cholesteryl-5-ene-24-one; VI represents 3?-acetoxy-cholesteryl-5?-bromo-6?-hydroxy-24-one; VII represents 3 acetoxy-cholesterium-5? - bromo-6β, 19-epoxy-24-ketone; VDI stands for 3β, 19-diacetoxy-cholester-5-ene-24'-ketone; Ka stands for 3β, 19-diacetoxy-chol甾5, 6β-epoxy-24-ketone; Kb stands for 3β, 19-diacetoxy-cholesteryl-5,6α-epoxy-24-one; X stands for 3β, 19-diacetoxy-chol甾5,6-dihydroxy-24-ketone.

Example 1

Synthesis of cholesteric 3β,5α, 6β, 19-tetrahydroxy-24-ketone

(1) Take 100 g of the compound (V), add 300 ml of benzene, 20 ml of thionyl chloride, 1 ml of pyridine, stir at room temperature for 2 hours, and concentrate to dryness under reduced pressure to remove residual thionyl chloride to obtain an acid chloride compound. 200 ml of benzene was dissolved and used. Take 20 grams of dry magnesium powder, add 60 ml of anhydrous ether, instill 73 ml of isobromopropene in anhydrous ether solution under 60% ultrasonic irradiation, continue to ultrasonic for 15 to 30 minutes, and then add in four batches after cooling. 78 g of cadmium chloride was shaken at 60 to 40 ° C for 60% ultrasonic wave for half an hour. While the ice water was cold, the benzene solution of the acid chloride was slowly added dropwise, and the ultrasonic wave was further refluxed for 2 hours. After chilling with ice water for 30 minutes, 150 ml (1:1) aqueous hydrochloric acid solution was added dropwise, and extracted with benzene three times. The benzene layer was washed with distilled water until neutral, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. White solid (VI).

(2) Take 20g of white solid (VI), add 20mL of tetrahydrofuran to dissolve, add appropriate amount of HC10 ₄ and water, then add 20g of N-succinimide in the dark to the reaction solution in batches, the temperature is low. The reaction was carried out at 10 ° C for 2 hours at 15 to 20 ° C. A 10% aqueous solution of Na ₂ S ₂ O ₈ was added to the reaction mixture, and the layers were separated, and then extracted with CH ₂ C _{2 2} , and the organic layer was combined, washed with a saturated NaHCO ₃ solution, and washed with distilled water to neutral, anhydrous sulfuric acid The organic layer was dried (MgSO4).

(3) Take 5 g of compound (VII), dissolve it into 1000 mL of cyclohexane/benzene mixture, and add 4 g of diacetyl. Oxyiodobenzene, 2.8 g of iodine, irradiated with a UV lamp at 40 ° C for 2 hours while stirring, and the reaction solution was washed with a 10% aqueous sodium thiosulfate solution, a saturated NaHC0 ₃ solution and distilled water to neutral, anhydrous sodium sulfate. Dry and concentrate under reduced pressure to give a pale yellow viscous compound ().

(4) Take 2 gram of compound (Shan), add 40 glacial acetic acid to dissolve, add 6 zinc powder, and stir to reflux for 20 to 24 hours. The mixture was filtered while hot, and the filtrate was poured into 5 times of water and extracted with dichloromethane. The extract was washed with saturated sodium bicarbonate and distilled water to dryness, dried over anhydrous sodium sulfate, A pale yellow viscous liquid (111).

(5) Take the compound (Shan) dissolved in dichloromethane (concentration is about 2.5~10%), add the mixture of crystalline copper sulfate and potassium permanganate mixture, add appropriate amount of t-butanol and water, stir at room temperature for 2 hours. . The methylene chloride was removed under reduced pressure, and the residue was crystallised from ethyl acetate (EtOAc). Adding to acetone, adding appropriate amount of perchloric acid, stirring at room temperature for 24 hours, adjusting the pH to neutral with saturated sodium bicarbonate solution, distilling off acetone under reduced pressure, extracting with dichloromethane, drying over anhydrous sodium sulfate, decompression Concentration gave a white solid (X).

(6) Add the above product (X) lg to 500 mL of 3% potassium hydroxide methanol solution, heat to reflux for 30 minutes, adjust the pH of the solution to neutral with 5% hydrochloric acid, distill off methanol under reduced pressure, and extract with ethyl acetate three times. The organic layer was dried over anhydrous sodium sulfate (MgSO4).

Example 2

Synthesis of cholestyl-3e,5a, 6β,19-tetrahydroxy-4-one

(1) to (4) The same as steps (1) to (4) of the first embodiment.

(5) The compound (Shan) was dissolved in dichloromethane (concentration: about 2.5 to 10%), and m-chloroperoxybenzoic acid was added thereto, and the mixture was stirred at room temperature for 2 hours. The methylene chloride was removed under reduced pressure, and the residue was crystallised from ethyl acetate (EtOAc). Add it to acetone, add appropriate amount of perchloric acid, stir at room temperature for 24 hours, adjust the pH to neutral with saturated sodium bicarbonate solution, distill off acetone under reduced pressure, extract with dichloromethane, dry over anhydrous sodium sulfate, decompress Concentration gave a white solid (X).

(6) Same as step (6) of implementation 1.

Example 3

Synthesis of cholestyl-3β,5α, 6β,19-tetrahydroxy-i24-one (1) to (4) The same as steps (1) to (4) of the first embodiment.

(5) Take compound (VI) dissolved in 88% formic acid (concentration is about 5%), heat in boiling water bath for 5 minutes, cool to room temperature, add 30% hydrogen peroxide solution, let stand at room temperature for 24 hours, dilute with water, use acetic acid The ethyl ester was extracted and washed with aq. EtOAc (EtOAc) (EtOAc)

(6) Add the above product (Kb) lg to 500 L of 3% potassium hydroxide in methanol, heat to reflux for 30 minutes, adjust the pH of the solution to neutral with 5% hydrochloric acid, distill off methanol under reduced pressure, and extract with ethyl acetate three times. The organic layer was dried over anhydrous sodium sulfate (MgSO4). '

Example 4

Synthesis of 24-methylene-cholesteryl-3β,5α,6]β,19-tetraol

(1) to (6) The same as steps (1) to (6) of the first embodiment.

(7) Add 2 g of methyltriphenylphosphonium bromide in 50 mL of anhydrous tetrahydrofuran solution, pass nitrogen gas, inject an appropriate amount of n-butyllithium, and stir the reaction at room temperature for 6 hours to give 1 g of '(I) tetrahydrofuran solution. The reaction mixture was poured into the reaction system, and the reaction was continued for 2 hours. The reaction solution was poured into a saturated ammonium chloride solution and extracted with diethyl ether. The ether layer was dried over anhydrous sodium sulfate. Analysis gave a white solid (11).

Example 5

Synthesis of cholesteric 3β,5α, 6β, 19-tetrahydroxy-24-biguanide

(1) to (6) are the same as steps (1) to (6) of the first embodiment.

(7) 1 g (I) was dissolved in 10 mL of ethanol, and 1 mL of a 50% hydrazine aqueous solution was added thereto, and the mixture was heated under reflux for 4 hours. The solvent was evaporated under reduced pressure, and the mixture was adjusted to pH. Silica gel column chromatography gave white solid (111). - Example 6

Cholesterol - Synthesis of 3β, 5α, 6β, 19-tetrahydroxy _24-biguanide

(1) to (6) are the same as steps (1) to (6) of the embodiment 1.

(7) 1 g (I) was dissolved in 20 mL of ethanol, and 1 mL of an 80% aqueous solution of hydrazine was added thereto, and the mixture was heated under reflux for 2 hours. The solvent was evaporated under reduced pressure, and the mixture was adjusted to pH. Silica gel column chromatography gave white solid (m). Structural confirmation of the target product h cholestyr-3β,5α, 6β, 19-tetrahydroxy-24-ketone

Melting point: 213-214 ° C

Elemental analysis: C ₂₇ H «0 ₅ Measured: C, 71.60; H, 20.50; Theory: C, 71.92; H, 10.29, MS: [M + Na] ⁺ = 473

IR: 3324, 2941, 2889, 1708, 1464, 1380, 1342, 1048, 975.

1H NMR (DMSO):

3.99 (IH, d, 19-CH), 3.86 (ra, IH, 3 β-CH), 3.52 (IH, in, 19-CH), 3.23 (m, IH, 60-CH), 2.59 (IH, m , 25 - CH) , 1.18 (3H, d, 26-C3⁄4, 27-CH ₃ ) , 1.01 (3H, s, 19-CH ₃ ) , 0.87 (3H, d , 21- C3⁄4), 0.65 (3H, s , 18-C3⁄4).

1 ³ C NMR (DMSO):

214.2(C), 74.5(0, 73.8 (CH), 65.8 (CH), 62.0 (CH ₂ ), 56.5 (CH), 55.5 (CH), 44.9 (CH), 42.53(C), 42.47 (CH), 41.0(C), 40.3(CH ₂ ), 39.8 (CH) , 36.4 (CH), 34.9 (CH ₂ ) , 33.9 (CH ₂ ) , 31.1 (CH ₂ ), 30.8 (CH ₂ ), 29.3 (CH ₂ ) , 27.7 (CH ₂ ), 26.8 (CH ₂ ), 23.8 (CH ₂ ), 21.7 (CH ₃ ), 18.3 (C3⁄4), 18.1 (CH ₃ ), 18.0 (CH ₃ ), 13.1 (CH ₃ ). 2: 24-Methylene-cholesteryl- 3 5 (i, 6β, 19-tetraol structure confirmation

Melting point: 223-225 °C '

Elemental analysis: C ₂₈ H ₄₈ 0 ₄ Measured: C, 75.20; H, 10.36 Theory: C, 74.95; H, 10.78, MS: [MH ₂ 0] ⁺ = 432,

IR: 3416, 3028, 2936, 2868, 1642, 1464, 1377, 1057

^NMRiDMSO):

5.25 (IH, d, 6 β - OH), 4.71 (IH, d, - 5Hz, 28- CH), 4.65 (IH, d, J=l.5Hz, 28- CH) ,

4.51(lH,d, 19-OH), 4.16(1H, d, 3—CH), 4.01 (IH, d, 19-CH), 3.8 (IH, ra, 3-CH), 3.64 (IH, s, 5-0H), 3.23 (lH,m, 6-CH), 2.25 (IH, m, 25- CH) ,

1.13 (6H, d, 26-C, 27-CH ₃ ) , 1.01 (3H, s, 19-CH ₃ ), 0.87 (3H, d, 21-CH ₃ ) , 0.67 (3H, s, 18-C3⁄4) .

1 ³ C NMR (DMSO):

156.6(C), 107.3 (CH ₂ ), 75.4(C), 74.7 (CH), 66.7 (CH), 63.0(CH ₂ ), 57.4(CH), 56.7(CH), 45.9(CH), 43.5(C ), 43.3(CH), 42.8(C), 41.3(CH ₂ ), 40.9(CH), 37.4 (CH ₂ ), 36.1 (CH), 33.9 (C3⁄4), 31.8 (C3⁄4), 31.4 (CH ₂ ), 30.5 (CH ₂ ), 28.7 (CH ₂ ), 27.9 (C3⁄4), 24.70 (CH ₂ ), 22.64(CH ₃ ), 22.57 (CH ₃ ) , 19.4(CH ₃ ), 13.1 (CH ₃ ) . The target product 3: the structure of cholestyl-3β,5α, 6 e, 19-tetra-light-one-bi-indole Confirmation

Melting point: 270-272 °C

Elemental analysis: C ₅₄ H _∞ 0 Measured: C, 70.92; H, 10.44; N, 2.98 Theory: C, 72.27;

H, 10.33; N, 3.12 - MS: [M+l] ⁺ =897

IR: 3318, 2943, 2870, 1628, 1464, 1379, 1344, 1060, 976c

1H NMR (DMSO):

5.38 (1H, ί,6-H), 4.71 (1H, d, J l- 5Hz, 28-CH), 4.65 (1H, d, J=l.5Hz, 28-CH), 3. 52 (IH, m, 3-CH), 2.28 (1H, m, 25-CH) , 1.13'(6H, d, 26-C3⁄4, 27 - CH ₃ ) , 1.01 (3H, s, 19- CH ₃ ), 0.91 (3H , d, 21-CH ₃ ), 0.69 (3H, s, 18- CH ₃ ) .

1 ³ C NMR (DMSO):

169.3 (0,75.4 (0, 74.7 (CH), 66.8 (CH), 63.6 (CH ₂ ), 57.5 (CH), 56.4 (CH), 45.9 (CH), 43.5 (C), 42.8 (C), 41.3 (CH ₂ ), 40.8 (CH) , 37.4 (CH ₂ ), 35.8 (CH), 35.6 (CH ₂ ) , 33.9 (CH ₂ ) , 31.8 (CH ₂ ) , 31.4 (CH ₂ ) , 30.5 (CH ₂ ) , 28.7 (CH ₂ ) , 27.9 (CH ₂ ) , 24.70 (CH ₂ ), 22.5 (CH ₃ ) , 21.4 ( CH ₃ ) , 19.2 ( CH ₃ ) , 18.9 ( CH ₃ ) , 13.1 ( CH ₃ ) .

Claims

Rights request

A method for synthesizing cholesteric-3β, 5α, 6β, 19-tetrahydroxy-24-ketone and a derivative thereof, characterized in that porphyoxycholic acid is used as a raw material to prepare 3 β-acetoxy-cholesteryl 5-ene-24-acid, prepared by reacting with thionyl chloride to prepare choline chloride, and reacting with isopropyl cadmium reagent under ultrasonic irradiation to obtain 3β-acetoxy-cholester-5-ene-24-one. Reaction with hydrazine-bromosuccinimide/perchloric acid aqueous solution system to obtain 3β-acetoxy-cholesteryl-5 α-bromo-6β-hydroxy- ₂₄ -ketone, followed by diacetoxyiodo Benzene produces 3 β-acetoxy-cholesterol_5 α-bromo-6β, 19-epoxy-24-ketone, which is reduced by zinc powder/glacial acetic acid system to form 3β,19-diacetoxy-cholesteryl a 5-alkenyl-2-ketone; oxidizing 5, 6 double bonds with an oxidizing agent, hydrolyzing to give 3β, 5α, 6β, 19-tetrahydroxy-24-one; and finally methyltriphenylphosphonium bromide The reaction synthesizes 24-methylene-cholesterol-3β, 5α, 6β, 19-tetraol; and hydrazine reacts to synthesize cholesteric-3β, 5α, 6β, 19-3⁄4 hydroxy-24-biguanide.

2. The method of synthesis according to claim 1, wherein the specific reaction steps are:

(1) Add IV to anhydrous benzene to thionyl chloride, add a catalytic amount of pyridine, stir at room temperature for 2 to 4 hours, concentrate to dryness under reduced pressure, and add benzene to continue concentration to bring out residual chlorination. Sulfoxide, a yellow viscous liquid;

(2) Weigh dry magnesium powder, add anhydrous ether, add isobromopropane / anhydrous ether solution, add ultrasonic radiation for 15~30 minutes, add cadmium chloride in batches after cooling, at 25~40 °C Under the ultrasonic irradiation, the 30~40% anhydrous benzene solution of the above step product is slowly added dropwise to the reaction system after the ice bath is cooled. After the completion of the dropping, the ultrasonic irradiation is continued for 1 to 2 hours, and the ice bath is cooled. , adding 1 : 1 hydrochloric acid aqueous solution, extracted with benzene, the benzene layer was washed with distilled water until neutral, dried, concentrated under reduced pressure, silica gel column chromatography to give a white solid V;

(3) Take a white solid V, add organic solvent to dissolve, add HC10 ₄ and water, then add N-succinimide in portions in the dark, control the temperature below 10 ° C, then at 15~20 after the reaction at ° C 1~2 hours, was added 10% Na _₂ S ₂ O ₈ was added to the reaction solution, the layers were separated, the upper layer was concentrated, the lower layer was extracted with CH _₂ C1 _2, the organic layers were combined, washed with saturated NaHC0 ₃ solution, Washing with distilled water until neutral, drying, concentration under reduced pressure, silica gel column chromatography to give a white solid VI;

(4) Take compound VI, dissolve in cyclohexane/benzene mixture, add oxidant, stir at 20~40 °C, irradiate with UV lamp for 7~8 hours, and then use 10% aqueous sodium thiosulfate solution, saturated NaHC0. ₃ solution and distilled water to wash to neutral, dry, concentrated under reduced pressure to give a pale yellow compound W;

(5) Take the compound and add it to the glacial acetic acid to dissolve, add the zinc powder, and stir to reflux for 20 to 24 hours; After suction filtration, the filtrate was extracted with dichloromethane, and the extract was washed with a saturated aqueous sodium hydrogen carbonate solution and distilled water to neutral, dried, concentrated under reduced pressure, and purified by silica gel column chromatography to obtain a pale yellow viscous liquid.

(6) Compound VI is added to methylene chloride to dissolve, oxidizing agent is added, stirring at room temperature, dichloromethane is removed under reduced pressure, the residue is dissolved in ethyl acetate, and then washed with 10% sodium sulfite solution until colorless, washed with water until neutral. , dry, concentrated under reduced pressure to give white solid IXa, which is dissolved in acetone, added with perchloric acid, stirred at room temperature, adjusted to pH neutral, acetone evaporated under reduced pressure, extracted with dichloromethane, dried, decompressed Concentrated to give a white solid X;

(7) The compound was dissolved in 88% formic acid, heated in a boiling water bath for 5 minutes, cooled to room temperature, added with an oxidizing agent, allowed to stand at room temperature for 24 hours, diluted with water, extracted with ethyl acetate, and then washed with 10% sodium sulfite solution until colorless. Washed to neutral, dried, concentrated under reduced pressure to give a white solid Kb;

(8) Dissolve the above-mentioned product Kb or X in methanol, add a 3% potassium hydroxide alcohol solution, heat to reflux for 15 to 30 minutes, adjust the pH to neutral, distill off methanol under reduced pressure, and extract with ethyl acetate. , the organic layer is dried, the solvent is evaporated under reduced pressure to give a pale yellow solid.

(9) Add methyltriphenylphosphonium bromide to an anhydrous tetrahydrofuran solution, and inject n-butyllithium. The amount of the injection is preferably dissolved in methyltriphenylphosphonium bromide, and the reaction is stirred at room temperature for 4 to 6 hours. Will be in the previous step

I, the reaction system is added to the reaction system, and the reaction is continued for 2 to 3 hours. The reaction solution is poured into a saturated ammonium chloride solution, and extracted with diethyl ether. The ether layer is dried, and the solvent is evaporated to give a pale yellow solid. Obtained a white solid II;

(10) I was dissolved in ethanol, added to an aqueous solution of hydrazine, and refluxed under reduced pressure. The solvent was evaporated under reduced pressure. Water was added, and the mixture was adjusted to neutral, and ethyl acetate was evaporated to give a white solid. solid

III;

In the above steps, I represents cholestyl-3 P,5 ci, 6 β,19-tetrahydroxy-24-one; Π represents 24-methylene-cholesteryl-3β, 5α, 6β,19-14 Alcohol; III represents cholesteryl-3β, 5α, 6β, 19-tetrahydroxy-24-biguanide; IV represents 3β-acetylindolyl-cholester-5-ene-24-acid; V represents 3 β -Acetoxy-cholesteryl-5-ene-24-one; "VI stands for 3β-acetoxy-cholesterium-5α-bromo-6β-hydroxy-24-one; W stands for 3β-acetoxy Base one cholesteryl-5 α-bromo-6 epoxide-epoxy-24-ketone; Shan represents 3 β, 19-diacetoxy-cholesteryl 5-alkenyl 24-ketone; IXa stands for 3 β , 19 — Diacetoxy-cholesteryl-5,6β-epoxy-24-one; Kb stands for 3β,19-diacetoxy-cholesteryl-5,6α-epoxy-24-one; 'X stands for 3 β, 19-diacetoxy-cholesteryl 5,6-dihydroxy-24-ketone.

The synthesis method according to claim 2, wherein the concentration of the benzene solution of II in the step (1) is 25 to 45%, and the amount of the thionyl chloride is 1.0 to 2.0 times the number of moles of II.

The synthetic side according to claim 2, wherein the amount of the magnesium powder in the step (2) is 1.2 to 3.0 times the number of moles of II.

The method according to claim 2, wherein the organic solvent in the step (3) is tetrahydrofuran or dioxane.

6. The method according to claim 2, wherein the oxidizing agent in the step (4) is diacetoxyiodobenzene or lead tetraacetate. -

7. The method according to claim 2, wherein the oxidizing agent in the step (6) is potassium permanganate / copper sulfate or m-chloroperbenzoic acid.

8. The method of synthesis according to claim 2, wherein the oxidizing agent in the step (7) is 30% hydrogen peroxide.