CN1793159A - Process for synthesizing cholester-3 bata, 5 alpha, 6 bata, 19-tetrahydroxy-24-one and derivative - Google Patents

Abstract

The invention discloses a method for synthesizing cholesteryl-3β, 5α, 6β, 19-tetrahydroxy-24-one and its derivatives. The method uses hyodeoxycholic acid as a raw material to prepare 3β-acetyloxy-cholesteryl- 5-ene-24-acid, prepared by reacting with thionyl chloride into cholesteryl chloride, reacting with isopropyl cadmium reagent under ultrasonic radiation to obtain 3β-acetoxy-cholest-5-en-24-one, and then React with N-bromosuccinimide/perchloric acid aqueous system to obtain 3β-acetoxy-cholesta-5α-bromo-6β-hydroxy-24-one, and then use diacetoxyiodobenzene to generate 3β -Acetoxy-cholesta-5α-bromo-6β,19-epoxy-24-one, reduced by zinc powder/glacial acetic acid system to generate 3β,19-diacetoxy-cholesta-5-ene-24- Ketone; then oxidize the 5 and 6 double bonds with an oxidizing agent, and hydrolyze to obtain cholesteryl-3β, 5α, 6β, 19-tetrahydroxy-24-ketone; finally react with methyl triphenylphosphine bromide to synthesize 24-methylene Cholesta-3β, 5α, 6β, 19-tetraol; react with hydrazine to synthesize cholesta-3β, 5α, 6β, 19-tetrahydroxy-24-dihydrazone. The synthesis method of the invention has the characteristics of cheap and easy-to-obtain raw materials, scientific and reasonable synthesis route, high synthesis yield of each step, etc., and the target compound has significant protective effect on neuron apoptosis cells induced by low potassium.

Description

Synthetic method of cholestan-3β, 5α, 6β, 19-tetrahydroxy-24-one and its derivatives

technical field

The invention relates to a synthesis method of cholesta-3β, 5α, 6β, 19-tetrahydroxyl-24-one (structure such as I) and derivatives.

Background technique

24-methylene-cholesta-3β, 5α, 6β, 19-tetraol (structure such as formula II) is a polyhydroxy steroid compound (Wang Guiyang, Li Fengying, Zeng Longmei, etc., Chemical Journal of Chinese Universities, 1992, 13, 623-627). The compound has strong killing ability to human rectal cancer cells, human lung cancer cells and mouse lymphoma cells (Chang-Yih Duh, Shang-Kwei, Mei-Jen Chu et al., J.Nat.Prod., 1998 , 61, 1022-1024), has significant anti-inflammatory effects on a variety of inflammations (Wang Zhenglian, Xu Shibo, Liu Qing et al., Tropical Ocean, 1999, 18, 27-32), the compound can inhibit the neuronal apoptosis induced by low potassium Cells have a significant protective effect.

The content of this compound in marine organisms is extremely low, and there are still many problems in the direct extraction of marine biological raw materials. The research and synthesis of this compound has high medicinal value. The intermediate cholestan-3β, 5α, 6β, 19-tetrahydroxyl-24-one and nitrogen-containing derivatives of the synthetic compound have significant protective effects on neuronal apoptosis cells induced by low potassium.

Contents of the invention

The object of the present invention is to provide a synthetic method of cholestan-3β, 5α, 6β, 19-tetrahydroxy-24-one and its derivatives.

The synthetic method of the present invention uses hyodeoxycholic acid as a raw material to prepare 3β-acetoxy-cholest-5-ene-24-acid by reacting with thionyl chloride to prepare cholesteryl chloride, and reacting with isopropyl under ultrasonic radiation The cadmium reagent is reacted to obtain 3β-acetoxy-cholest-5-en-24-one, and then reacted with N-bromosuccinimide/perchloric acid aqueous solution to obtain 3β-acetoxy-cholest-5- 5α-bromo-6β-hydroxy-24-one, oxidative ring closure of diacetoxyiodobenzene to 3β-acetoxy-cholesta-5α-bromo-6,19-epoxy-24-one, followed by zinc powder/glacial acetic acid system or zinc powder/alcohol system to generate 3β, 19-diacetoxy-cholest-5-en-24-one, and then get cholest-5-ene-3β, 19-hydroxyl by alkaline hydrolysis -24-ketone; Oxidation and hydrolysis with oxidant to obtain cholestan-3β, 5α, 6β, 19-tetrahydroxy-24-ketone; reaction with methyl triphenylphosphine bromide to synthesize 24-methylene-cholesteryl- 3β, 5α, 6β, 19-tetraol; react with hydrazine to synthesize cholesta-3β, 5α, 6β, 19-tetrahydroxy-24-dihydrazone. The reaction scheme is shown in the figure below:

I. Cholesta-3β, 5α, 6β, 19-tetrahydroxy-24-one;

II. 24-methylene-cholesta-3β, 5α, 6β, 19-tetrol;

III. Cholesta-3β, 5α, 6β, 19-tetrahydroxy-24-dihydrazone;

IV.3β-Acetoxy-cholest-5-en-24-oic acid;

V.3β-Acetoxy-cholest-5-en-24-one;

VI.3β-Acetoxy-cholesta-5α-bromo-6β-hydroxy-24-one;

VII.3β-Acetoxy-cholesta-5α-bromo-6β,19-epoxy-24-one;

VIII.3β,19-diacetoxy-cholest-5-en-24-one;

IXa. 3β,19-diacetoxy-cholesta-5,6β-epoxy-24-one;

IXb.3β,19-diacetoxy-cholesta-5,6α-epoxy-24-one;

X. 3β,19-Diacetoxy-cholesta-5,6-dihydroxy-24-one.

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

(1) IV was synthesized with reference to literature (Bharucha KR, Buckley GC, Cross CK, et al. Can J Chem., 1955, 34:982-990.);

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

(3) Weigh dry magnesium powder, add anhydrous diethyl ether, dropwise add isobromopropane/anhydrous diethyl ether solution, irradiate with ultrasound for 15-30 minutes after adding, add cadmium chloride in batches after cooling, at 25-40°C Stir under ultrasonic radiation, under ice bath cooling, slowly add 30-40% anhydrous benzene solution of the product in the previous step into the reaction system dropwise, continue ultrasonic radiation and reflux for 1-2 hours after dropping, after ice bath cooling, 1:1 hydrochloric acid aqueous solution was added dropwise, extracted with benzene, the benzene layer was washed with distilled water until neutral, dried, concentrated under reduced pressure, and subjected to silica gel column chromatography to obtain white solid V;

(4) Take the white solid V, add organic solvent to dissolve, then add HClO ₄ and water, then add N-succinimide in batches under the condition of avoiding light, control the temperature below 10°C, and then add it at 15-20°C The reaction was carried out for 1 to 2 hours, and 10% Na ₂ S ₂ O ₈ aqueous solution was added to the reaction liquid. After the layers were separated, the upper layer was concentrated, and the lower layer was extracted with CH ₂ Cl ₂ . The organic layers were combined, washed with saturated NaHCO ₃ solution, and then washed with Wash with distilled water until neutral, dry, concentrate under reduced pressure, and perform silica gel column chromatography to obtain white solid VI;

(5) Take compound VI, dissolve it into cyclohexane/benzene mixture, add an oxidizing agent, under stirring at 20-40°C, irradiate with ultraviolet light for 7-8 hours, and then use 10% sodium thiosulfate aqueous solution, saturated NaHCO ₃ The solution and distilled water were washed to neutrality, dried, and concentrated under reduced pressure to obtain light yellow compound VII;

(6) Take compound VIII and add glacial acetic acid to dissolve, add zinc powder, stir and reflux for 20 to 24 hours; suction filter while hot, extract the filtrate with dichloromethane, and wash the extract with saturated sodium bicarbonate solution and distilled water until neutral, Dry, concentrate under reduced pressure, and perform silica gel column chromatography to obtain light yellow viscous liquid VIII;

(7) Take compound VIII and add dichloromethane to dissolve it, add an oxidizing agent, stir at room temperature, remove the dichloromethane under reduced pressure, dissolve the slag with ethyl acetate, then add 10% sodium sulfite solution to wash until colorless, and wash with water until neutral, Dry and concentrate under reduced pressure to obtain a white solid IXa, add acetone to dissolve it, add perchloric acid, stir at room temperature, adjust pH to neutral, evaporate acetone under reduced pressure, extract with dichloromethane, dry, and concentrate under reduced pressure, A white solid X was obtained;

(8) Dissolve compound VIII by adding 88% formic acid, heat in a boiling water bath for 5 minutes, cool to room temperature, add an oxidizing agent, place at room temperature for 24 hours, dilute with water, extract with ethyl acetate, then add 10% sodium sulfite solution to wash until colorless , washed with water to neutrality, dried, and concentrated under reduced pressure to obtain white solid IXb;

(9) Dissolve the product IXb or X of the previous step in methanol, add potassium hydroxide alcohol solution with a concentration of 3%, heat and reflux for 15 to 30 minutes, adjust the pH to neutral, remove methanol by distillation under reduced pressure, and extract with ethyl acetate , the organic layer was dried, and the solvent was evaporated under reduced pressure to give a light yellow solid, and silica gel column chromatography gave a white solid I;

(10) Add methyl triphenyl phosphine bromide into anhydrous tetrahydrofuran solution, inject n-butyllithium, the injected amount should be just dissolved in methyl triphenyl phosphine bromide, stir and react at room temperature for 4 to 6 hours, Add I in the previous step to the reaction system, continue to react for 2 to 3 hours, quickly pour the reaction solution into a saturated ammonium chloride solution, extract with ether, dry the ether layer, evaporate the solvent under reduced pressure, and obtain a light yellow solid. Silica gel column chromatography gave white solid II;

(11) Dissolve I in ethanol, add it to the aqueous solution of hydrazine, defrost and reflux, evaporate the solvent under reduced pressure, add water, adjust the pH to neutral, extract with ethyl acetate to obtain a white solid, and conduct silica gel column chromatography to obtain a white solid. Solid III.

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

The reaction route of the synthesis method of the present invention is scientific and reasonable, the yield of the compound is high, and the synthesized intermediates cholesta-3β, 5α, 6β, 19-tetrahydroxyl-24-ketone, 24-methylene-cholesta-3β , 5α, 6β, 19-tetrol; and cholesterin-3β, 5α, 6β, 19-tetrahydroxy-24-dihydrazone have significant protective effects on low potassium-induced neuronal apoptosis.

Detailed ways

The following examples are used to illustrate the present invention, but the protection scope of the present invention is not limited to the following examples. Unless otherwise specified, in the following examples: I represents cholesta-3β, 5α, 6β, 19-tetrahydroxy-24-one; II represents 24-methylene-cholesta-3β, 5α, 6β, 19-tetra Alcohol; III stands for cholest-3β, 5α, 6β, 19-tetrahydroxy-24-dihydrazone; IV stands for 3β-acetoxy-cholest-5-en-24-acid; V stands for 3β-acetoxy- Cholesta-5-en-24-one; VI represents 3β-acetoxy-cholesta-5α-bromo-6β-hydroxy-24-one; VII represents 3β-acetoxy-cholesta-5α-bromo-6β , 19-epoxy-24-one; VIII represents 3β, 19-diacetoxy-cholest-5-ene-24-one; IXa represents 3β, 19-diacetoxy-cholesta-5,6β- Epoxy-24-one; IXb represents 3β, 19-diacetoxy-cholesta-5,6α-epoxy-24-one; X represents 3β, 19-diacetoxy-cholesta-5,6- Dihydroxy-24-one.

Example 1

Synthesis of Cholesta-3β, 5α, 6β, 19-tetrahydroxy-24-one

(1) Take 100 grams of compound (V), add 300 ml of benzene, 20 ml of thionyl chloride, and 1 ml of pyridine, stir at room temperature for 2 hours, concentrate under reduced pressure to dryness, remove residual thionyl chloride, and obtain 200 ml of acid chloride compound. Dissolve benzene and set aside. Take 20 grams of dry magnesium powder, add 60ml of anhydrous ether, drip 73ml of isobromopropane anhydrous ether solution under 60% ultrasonic radiation, continue ultrasonication for 15-30 minutes after adding, add 78 grams of chlorine in four batches after cooling Cadmium chloride, 60% ultrasonic vibration at 25-40°C for half an hour. Under ice-water cooling, the benzene solution of acid chloride was slowly added dropwise, and ultrasonic reflux was continued for 2 hours after the drop. After freezing with ice water for 30 minutes, add 150ml (1:1) hydrochloric acid aqueous solution dropwise, extract three times with benzene, wash the benzene layer with distilled water until neutral, dry over anhydrous sodium sulfate, concentrate under reduced pressure, and perform silica gel column chromatography to obtain white Solid (VI).

(2) Take 20 grams of white solid (VI), add 20 mL of tetrahydrofuran to dissolve, add appropriate amount of HClO ₄ and water, then add 20 grams of N-succinimide to the reaction solution in batches under dark conditions, and control the temperature. At 10°C, then react at 15-20°C for 2 hours. Add 10% Na ₂ S ₂ O ₈ aqueous solution to the reaction solution, separate the layers, extract with CH ₂ Cl ₂ , combine the organic layers, wash with saturated NaHCO ₃ solution, and then wash with distilled water until neutral, dry over anhydrous sodium sulfate , concentrated under reduced pressure, and subjected to silica gel column chromatography to obtain a white solid (VII).

(3) Take 5 grams of compound (VII), dissolve it into 1000 mL of cyclohexane/benzene mixed solution, add 4 grams of diacetoxyiodobenzene, 2.8 grams of iodine, stir, and irradiate with ultraviolet light at 40 ° C for 2 hours to react The solution was successively washed with 10% aqueous sodium thiosulfate solution, saturated NaHCO ₃ solution and distilled water until neutral, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain light yellow viscous compound (VIII).

(4) Take 2 g of compound (VIII) and add 40 g of glacial acetic acid to dissolve it, add 6 g of zinc powder, stir and reflux for 20 to 24 hours. Suction filtration while hot, pour the filtrate into 5 times the amount of water, extract with dichloromethane, wash the extract with saturated sodium bicarbonate solution and distilled water to neutrality, dry over anhydrous sodium sulfate, concentrate under reduced pressure, and perform silica gel column chromatography. A light yellow viscous liquid (III) was obtained.

(5) Dissolve compound (VIII) in dichloromethane (concentration is about 2.5-10%), add crystalline copper sulfate and potassium permanganate mixture powder, add appropriate amount of tert-butanol and water, and stir at room temperature for 2 hours. The dichloromethane was removed under reduced pressure, and the residue was dissolved in ethyl acetate, then washed with 10% sodium sulfite solution until it was colorless, washed with water until neutral, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a white solid (IXa). Dissolve it in acetone, add an appropriate amount of perchloric acid, stir at room temperature for 24 hours, adjust the pH to neutral with saturated sodium bicarbonate solution, evaporate the acetone under reduced pressure, extract with dichloromethane, dry over anhydrous sodium sulfate, and depressurize Concentration gave white solid (X).

(6) Add 1 g of the product (X) from the previous step 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, remove methanol by distillation under reduced pressure, and extract three times with ethyl acetate , the organic layers were combined and dried with anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure to obtain a light yellow solid, which was subjected to silica gel column chromatography to obtain a white solid (I).

Example 2

Synthesis of Cholesta-3β, 5α, 6β, 19-tetrahydroxy-24-one

(1)-(4) are the same as steps (1)-(4) of Example 1.

(5) Take compound (VIII) and add dichloromethane to dissolve (concentration is about 2.5-10%), add m-chloroperoxybenzoic acid, and stir at room temperature for 2 hours. The dichloromethane was removed under reduced pressure, and the residue was dissolved in ethyl acetate, then washed with 10% sodium sulfite solution until it was colorless, washed with water until neutral, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a white solid (IXa). Dissolve it in acetone, add an appropriate amount of perchloric acid, stir at room temperature for 24 hours, adjust the pH to neutral with saturated sodium bicarbonate solution, evaporate the acetone under reduced pressure, extract with dichloromethane, dry over anhydrous sodium sulfate, and depressurize Concentration gave white solid (X).

(6) is the same as step (6) of Implementation 1.

Example 3

Synthesis of Cholesta-3β, 5α, 6β, 19-tetrahydroxy-24-one

(1)-(4) are the same as steps (1)-(4) of Example 1.

(5) Take compound (VIII) and add 88% formic acid to dissolve (concentration is about 5%), heat in a boiling water bath for 5 minutes, cool to room temperature, add 30% hydrogen peroxide solution, leave it at room temperature for 24 hours, dilute with water, and wash with acetic acid Extract with ethyl ester, then add 10% sodium sulfite solution to wash until colorless, wash with water until neutral, dry over anhydrous sodium sulfate, and concentrate under reduced pressure to obtain a white solid (IXb).

(6) Add 1 g of the previous step product (IXb) to 500 L of 3% potassium hydroxide methanol solution, heat to reflux for 30 minutes, adjust the pH of the solution to be neutral with 5% hydrochloric acid, remove methanol by distillation under reduced pressure, and extract three times with ethyl acetate , the organic layers were combined and dried with anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure to obtain a light yellow solid, which was subjected to silica gel column chromatography to obtain a white solid (I).

Example 4

Synthesis of 24-methylene-cholesta-3β, 5α, 6β, 19-tetrol

(1)-(6) are the same as steps (1)-(6) of Example 1.

(7) Add 2 grams of methyltriphenylphosphine bromide to 50 mL of anhydrous tetrahydrofuran solution, ventilate nitrogen, inject an appropriate amount of n-butyllithium, stir and react at room temperature for 6 hours, inject 1 gram of tetrahydrofuran solution of (I) The reaction system was continued to react for 2 hours, the reaction solution was quickly poured into a saturated ammonium chloride solution, extracted with ether, the ether layer was dried with anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure to obtain a light yellow solid, which was subjected to silica gel column chromatography. , to give white solid (II).

Example 5

Synthesis of Cholesta-3β, 5α, 6β, 19-tetrahydroxy-24-dihydrazone

(1)-(6) are the same as steps (1)-(6) of Example 1.

(7) Dissolve 1 gram of (I) in 10 mL of ethanol, add 1 mL of 50% hydrazine aqueous solution, heat to reflux for 4 hours, evaporate the solvent under reduced pressure, add an appropriate amount of water, adjust the pH to neutral, extract with ethyl acetate to obtain a white solid , silica gel column chromatography to give white solid (III).

Example 6

Synthesis of Cholesta-3β, 5α, 6β, 19-tetrahydroxy-24-dihydrazone

(1)-(6) are the same as steps (1)-(6) of Example 1.

(7) Dissolve 1 gram of (I) in 20 mL of ethanol, add 1 mL of 80% hydrazine aqueous solution, heat to reflux for 2 hours, evaporate the solvent under reduced pressure, add an appropriate amount of water, adjust the pH to neutral, extract with ethyl acetate to obtain a white solid , silica gel column chromatography to give white solid (III).

Target product 1: Confirmation of the structure of cholestan-3β, 5α, 6β, 19-tetrahydroxy-24-one

Melting point: 213-214°C

Elemental analysis: C ₂₇ H ₄₆ O ₅ Found: C, 71.60; H, 20.50; Theoretical: C, 71.92; H, 10.29,

MS: [M+Na] ⁺ = 473

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

^1HNMR (DMSO):

3.99 (1H, d, 19-CH), 3.86 (m, 1H, 3β-CH), 3.52 (1H, m, 19-CH), 3.23 (m, 1H, 6β-CH), 2.59 (1H, m, 25-CH), 1.18 (3H, d, 26-CH ₃ , 27-CH ₃ ), 1.01 (3H, s, 19-CH ₃ ), 0.87 (3H, d, 21-CH ₃ ), 0.65 (3H, s, 18-CH ₃ ).

¹³ CNMR (DMSO):

214.2(C), 74.5(C), 73.8(CH), 65.8(CH), 62.0( _CH2 ), 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(CH ₃ ), 18.1(CH ₃ ), 18.0(CH ₃ ), 13.1(CH ₃ ).

Target product 2: Confirmation of the structure of 24-methylene-cholesta-3β, 5α, 6β, 19-tetrol

Melting point: 223-225°C

Elemental analysis: C ₂₈ H ₄₈ O ₄ Found: C, 75.20; H, 10.36 Theoretical: C, 74.95; H, 10.78,

MS: [MH ₂ O] ⁺ = 432,

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

^1HNMR (DMSO):

5.25 (1H, d, 6β-OH), 4.71 (1H, d, J = 1.5Hz, 28-CH), 4.65 (1H, d, J = 1.5Hz, 28-CH), 4.51 (1H, d, 19 -OH), 4.16 (1H, d, 3-CH), 4.01 (1H, d, 19-CH), 3.84 (1H, m, 3-CH), 3.64 (1H, s, 5-OH), 3.23 ( 1H, m, 6-CH), 2.25 (1H, m, 25-CH), 1.13 (6H, d, 26-CH ₃ , 27-CH ₃ ), 1.01 (3H, s, 19-CH ₃ ), 0.87 (3H, d, 21-CH ₃ ), 0.67 (3H, s, 18-CH ₃ ).

¹³ CNMR (DMSO):

156.6(C), 107.3( _CH2 ), 75.4(C), 74.7(CH), 66.7(CH), 63.0( _CH2 ), 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(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 ), 22.64(CH ₃ ), 22.57(CH ₃ ), 19.4(CH ₃ ), 13.1(CH ₃ ).

Target product 3: Confirmation of the structure of cholestan-3β, 5α, 6β, 19-tetrahydroxy-24-dihydrazone

Melting point: 270-272°C

Elemental analysis: Found for C ₅₄ H ₉₂ O ₄ N ₂ : C, 70.92; H, 10.44; N, 2.98 Theoretical: C, 72.27; H, 10.33; N, 3.12

MS: [M+1] ⁺ = 897

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

^1HNMR (DMSO):

5.38 (1H, t, 6β-H), 4.71 (1H, d, J=1.5Hz, 28-CH), 4.65 (1H, d, J=1.5Hz, 28-CH), 3.52 (1H, m, 3 -CH), 2.28 (1H, m, 25-CH), 1.13 (6H, d, 26-CH ₃ , 27-CH ₃ ), 1.01 (3H, s, 19-CH ₃ ), 0.91 (3H, d, 21-CH ₃ ), 0.69 (3H, s, 18-CH ₃ ).

¹³ CNMR (DMSO):

169.3(C), 75.4(C), 74.7(CH), 66.8(CH), 63.6( _CH2 ), 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 (8)

1. courage steroid-3 β, 5 α, 6 β, the synthetic method of 19-tetrahydroxy-24-ketone and derivative, it is characterized in that with the Hyodeoxycholic Acid being raw material, prepare 3 β-acetoxyl group-courage steroid-5-alkene-24-acid, by become courage steroid acyl chlorides with the sulfur oxychloride prepared in reaction, the ultrasonic wave radiation down and the sec.-propyl cadion react 3 β-acetoxyl group-courage steroid-5-alkene-24-ketone, obtain 3 β-acetoxyl group-courage steroid-5 α-bromo-6 beta-hydroxies-24-ketone with N-bromo-succinimide/high chloro acid solution's system reaction again, generate 3 β-acetoxyl group-courage steroid-5 α-bromo-6 β with the diacetoxy phenyl-iodide afterwards, 19-epoxy-24-ketone, generate 3 β with zinc powder/Glacial acetic acid reduction system, 19-diacetoxy-courage steroid-5-alkene-24-ketone; Use 5,6 two keys of oxygenant oxidation again, hydrolysis reaction gets courage steroid-3 β, 5 α, 6 β, 19-tetrahydroxy-24-ketone; Last and methyltriphenylphosphonium bromide reacts Synthetic 2 4-methylene radical-courage steroid-3 β, 5 α, 6 β, 19-tetrol; With synthetic courage steroid-3 β of hydrazine reaction, 5 α, 6 β, the two hydrazones of 19-tetrahydroxy-24-.

2. synthetic method according to claim 1 is characterized in that concrete reactions steps is:

(1) IV is dissolved in dry-out benzene and adds sulfur oxychloride, add the pyridine of catalytic amount, at room temperature stirred 2～4 hours, be evaporated to driedly, add benzene again and continue to concentrate, take residual sulfur oxychloride out of, yellow heavy-gravity liquid;

(2) take by weighing dry magnesium powder, add anhydrous diethyl ether, drip different N-PROPYLE BROMIDE/anhydrous ether solution, add back ultrasonic radiation 15～30 minutes, add Cadmium chloride fine powder after the cooling in batches, stirring under the ultrasonic wave radiation under 25～40 ℃, the ice bath cooling down, slowly be added drop-wise to 30～40% dry-out benzene solution of product of last step in the reaction system, drip off the back and continue ultrasonic wave radiation backflow 1～2 hour, after the ice bath cooling, drip 1: 1 aqueous hydrochloric acid, extract with benzene, the benzene layer is washed till neutrality with distilled water, drying, concentrating under reduced pressure, silica gel column chromatography gets white solid V;

(3) extracting waste solid V adds organic solvent dissolution, adds HClO again ₄And water, under the lucifuge condition, adding the N-succimide then in batches, controlled temperature is lower than 10 ℃, reacts 1～2 hour down at 15～20 ℃ afterwards, adds 10%Na in reaction solution ₂S ₂O ₈The aqueous solution, after the layering, the upper strata concentrates, the CH of lower floor ₂Cl ₂Extraction merges organic layer, uses saturated NaHCO ₃Solution washing is washed with distilled water to neutrality again, drying, and concentrating under reduced pressure, silica gel column chromatography gets white solid VI;

(4) get compound VI, dissolve in hexanaphthene/benzene mixed solution, add oxygenant, under 20～40 ℃ of stirrings, ultra violet lamp 7～8 hours, reaction solution is used 10% sodium thiosulfate solution successively, saturated NaHCO ₃Solution and distilled water wash are extremely neutral, drying, and concentrating under reduced pressure gets light yellow compound VI I;

(5) get compound VIII and add the Glacial acetic acid dissolving, add zinc powder, stirring and refluxing 20～24 hours; Suction filtration while hot, filtrate is used dichloromethane extraction, extraction liquid is washed till neutrality with saturated sodium bicarbonate solution and distilled water respectively, drying, concentrating under reduced pressure, silica gel column chromatography, light yellow thick liquid VIII;

(6) get compound VIII and add the methylene dichloride dissolving, add oxygenant, stir under the room temperature, methylene dichloride is removed in decompression, the clinker acetic acid ethyl dissolution adds the washing of 10% sodium sulfite solution then to colourless, is washed to neutrality, drying, concentrating under reduced pressure gets white solid IXa, it is added acetone solution, add perchloric acid, stir under the room temperature, transfer pH to neutral, pressure reducing and steaming acetone is used dichloromethane extraction, drying, concentrating under reduced pressure gets white solid X;

(7) get compound VIII and add the dissolving of 88% formic acid, boiling water bath heating 5 minutes is cooled to room temperature, adds oxygenant, room temperature was placed 24 hours, behind the dilute with water, use ethyl acetate extraction, add the washing of 10% sodium sulfite solution then to colourless, be washed to neutrality, drying, concentrating under reduced pressure gets white solid IXb;

(8) will go up step product IXb or X is dissolved in the methyl alcohol, adding concentration is 3% potassium hydroxide alcoholic solution, reflux 15～30 minutes, it is neutral transferring pH, methyl alcohol is removed in underpressure distillation, ethyl acetate extraction, the organic layer drying, the pressure reducing and steaming solvent gets light yellow solid, silica gel column chromatography gets white solid I;

(9) methyltriphenylphosphonium bromide is added in the anhydrous tetrahydrofuran solution, inject n-Butyl Lithium, injection rate just is dissolved as suitable with methyltriphenylphosphonium bromide, stirring reaction is 4～6 hours under the room temperature, I in the last step is added reaction system, continue reaction 2～3 hours, reaction solution is poured into rapidly in the saturated ammonium chloride solution, use extracted with diethyl ether, ether layer drying, the pressure reducing and steaming solvent gets light yellow solid, silica gel column chromatography gets white solid II;

(10) I is added dissolve with ethanol, add in the aqueous solution of hydrazine, separate thermal backflow, the pressure reducing and steaming solvent adds entry, and it is neutral transferring pH, and ethyl acetate extraction gets white solid, and silica gel column chromatography gets white solid III;

In the above-mentioned steps, I represents courage steroid-3 β, 5 α, 6 β, 19-tetrahydroxy-24-ketone; II represents 24-methylene radical-courage steroid-3 β, 5 α, 6 β, 19-tetrol; III represents courage steroid-3 β, 5 α, 6 β, the two hydrazones of 19-tetrahydroxy-24-; IV represents 3 β-acetoxyl group-courage steroid-5-alkene-24-acid; V represents 3 β-acetoxyl group-courage steroid-5-alkene-24-ketone; VI represents 3 β-acetoxyl group-courage steroid-5 α-bromo-6 beta-hydroxies-24-ketone; VII represents 3 β-acetoxyl group-courage steroid-5 α-bromo-6 β, 19-epoxy-24-ketone; VIII represents 3 β, 19-diacetoxy-courage steroid-5-alkene-24-ketone; IXa represents 3 β, 19-diacetoxy-courage steroid-5,6 beta epoxides-24-ketone; IXb represents 3 β, 19-diacetoxy-courage steroid-5,6 α-epoxy-24-ketone; X represents 3 β, 19-diacetoxy-courage steroid-5,6-dihydroxyl-24-ketone.

3. synthetic method according to claim 2 is characterized in that the concentration of the benzole soln of II in the step (1) is 25～45%, and the amount of sulfur oxychloride is 1.0～2.0 times of mole number of II.

4. synthetic method according to claim 2, the amount of magnesium powder in the step (2) of it is characterized in that are 1.2～3.0 times of mole number of II.

5. synthetic method according to claim 2 is characterized in that the organic solvent in the step (3) is tetrahydrofuran (THF) or dioxane.

6. synthetic method according to claim 2 is characterized in that the oxygenant in the step (4) is diacetoxy phenyl-iodide or lead tetra-acetate.

7. synthetic method according to claim 2 is characterized in that the oxygenant in the step (6) is potassium permanganate/copper sulfate or metachloroperbenzoic acid.

8. synthetic method according to claim 2 is characterized in that the oxygenant in the step (7) is 30% hydrogen peroxide.