CN107236013A - The alkane ester group pregnane compound of 20 hydroxyl pregnene 3 and its synthetic method and the application in antineoplastic is prepared - Google Patents

Abstract

The invention discloses a 20-hydroxy-pregnene-3-alkane ester-based pregnane compound, the steroid nucleus structural formula of the compound is as follows: Wherein, the R-group can be: R=-CCl ₃ ,-CH ₂ CH ₃ . The invention also discloses a synthesis method of the 20-hydroxyl-pregnene-3-alkane ester-based pregnene compound, comprising the following steps: taking the steroidal compound pregnenolone as a raw material, firstly preparing the 3-pregnenolone The hydroxyl group is esterified into an ester through alkane acid chloride, and the intermediate product is obtained through the reaction, and then the 17-carbonyl group of the intermediate product is reduced to obtain a 20-hydroxy-pregnene-3-alkane ester-pregnant compound. The 20-hydroxy-pregnene-3-alkane ester-based pregnant compound of the present invention has an inhibitory effect on the growth and proliferation of certain cancer cells, and can be used as a drug intermediate or drug for the manufacture and use of different anti-tumor drugs.

Description

20-Hydroxy-pregnene-3-alkane ester-based pregnan compound and its synthesis method and preparation Application in preparation of antineoplastic drugs

technical field

The invention relates to the technical field of compound synthesis, more specifically, the invention relates to a 20-hydroxy-pregnene-3-alkane ester group pregnant compound, a synthesis method thereof and an application in the preparation of antitumor drugs.

Background technique

Pregnenolone itself has weak anti-inflammatory activity. It has been reported in the literature that the C3-hydroxyl esterification of pregnenolone, salicylate and formate have anti-inflammatory effects equivalent to those of clinically used cortisone, while Trichloroacetate, chloroacetate, furan-2'-ester, p-toluenesulfonate, and phthalate have slightly stronger anti-inflammatory effects than anti-inflammatory pine (Li Zhensu, Wang Haiqing, etc. 5- Synthesis of Pregnene-3β,17α-Diol-20-keto-3-Acetate-17α-Fatty Acid Esters and Their Structure-Activity Relationship[J].Acta Pharmaceutica Sinica,1983,18(2):119-124 .).

The research group has carried out related research on the chemical modification of pregnenolone, "pregnenolone aromatic aldehyde azine steroid compound and its synthesis method and its application in the preparation of anti-tumor drugs" (Chinese invention patent, application number: 201310184358.X, application publication number CN103254264A), "3-substituted-B-Homo-steroid-B-cyclic lactam compound and its preparation method and application in the preparation of anti-tumor drugs" (Chinese invention patent, application number: 201110009124.2, application publication number: CN102146115A), "Preparation method of 3-hydroxy-5-ene steroid compound with ammonia derivative branched chain structure and its application in antitumor drugs" (Chinese invention patent, application number: 201210330275.2, Application publication number: CN102816198A).

However, there is no report about the 20-hydroxy-pregnene-3-alkane ester-pregnant compound, its synthesis method, and its application in the preparation of antitumor drugs.

Contents of the invention

The object of the present invention is to provide a 20-hydroxy-pregnene-3-alkane ester-pregnant compound.

Another object of the present invention is to provide a synthetic method for the above compound.

A further object of the present invention is to provide the application of the above compounds in the preparation of antitumor drugs.

The present invention realizes above-mentioned purpose through following technical scheme, and the steroid nuclear structural formula of 20-hydroxyl-pregnene-3-alkane ester prepregna compound of the present invention is as follows:

Wherein, the R-group may be: R=-CCl ₃ , -CH ₂ CH ₃ .

Preferably, the following steps are included: using the steroidal compound pregnenolone as a raw material, first esterifying the 3-hydroxyl group of the pregnenolone into an ester through alkane acid chloride, reacting to obtain an intermediate product, and then converting the 17-position of the intermediate product Carbonyl reduction, that is, to obtain 20-hydroxy-pregnene-3-alkane ester-pregnant compound.

Preferably, the reaction method for the 3-position hydroxyl group to be esterified through alkane acid chloride is: dissolve pregnenolone in pyridine, stir at 40°C, then slowly add alkane acid chloride, and follow the reaction by TLC until there is no raw material Stop the reaction after the point, add ice water to the obtained reaction product, then extract with ethyl acetate, combine the organic phases, wash with 1mol/L dilute hydrochloric acid, saturated NaHCO ₃ , distilled water, saturated NaCl successively, and then use anhydrous Na _{2 SO4} was dried, and finally the organic solvent was distilled off under reduced pressure, and separated by column chromatography to obtain white needle-like crystals, which were intermediate products;

Wherein, the alkanoyl chloride is trichloroacetyl chloride or propionyl chloride.

Preferably, the reaction method for reducing the 17-position carbonyl is: mix the intermediate product with CH ₂ Cl ₂ , stir at 40°C until the intermediate product is completely dissolved, add CeCl ₃ 7H ₂ O and CH ₃ OH, after CeCl ₃ 7H ₂ O is completely dissolved, weigh NaBH ₄ and add it slowly for reaction, react at room temperature, TLC detection, stop the reaction until there is no raw material, add 1mol/L dilute hydrochloric acid to quench the reaction, and distill under reduced pressure After removing the organic solvent, distilled water was added, extracted with ethyl acetate, the organic phases _were combined, washed with water and saturated NaCl solution in turn, then dried with anhydrous _Na2SO4 , and finally the organic solvent was distilled off under reduced pressure, separated by column chromatography, The final product obtained as a white solid is the 20-hydroxy-pregnene-3-alkane ester-pregnant compound.

The application of the 20-hydroxyl-pregnene-3-alkane ester group pregnoid compound in the preparation of antitumor drugs.

The application of the pharmaceutical composition in which the 20-hydroxy-pregnene-3-alkane ester-pregnant compound is an active ingredient in the preparation of antitumor drugs.

The reaction scheme for preparing the 20-hydroxyl-pregnene-3-alkane ester group pregnant compound is as follows:

Wherein, the R groups in compounds 1 to 4 are as follows: 1R=-CCl ₃ , 2R=-CH ₂ CH ₃ , 3R=-CCl ₃ , 4, R=-CH ₂ CH ₃ ; reagents and conditions: a , pyridine, CCl ₃ COCl, CH ₃ CH ₂ COCl; b: CH ₃ OH, CH ₂ Cl ₂ , CeCl ₃ ·7H ₂ O, NaBH ₄ .

Experimental studies on physiological activity show that the 20-hydroxy-pregnene-3-ester-pregnant compounds 3 and 4 of the present invention have an inhibitory effect on the growth and proliferation of human lung adenocarcinoma cell lines, and can be used as drug intermediates or drug applications For the manufacture and application of different anticancer drugs.

Other advantages, objectives and features of the present invention will partly be embodied through the following descriptions, and partly will be understood by those skilled in the art through the study and practice of the present invention.

detailed description

The present invention will be further described in detail below in conjunction with the embodiments, so that those skilled in the art can implement it with reference to the description.

It should be noted that the experimental methods described in the following embodiments, unless otherwise specified, are conventional methods, and the reagents and materials, unless otherwise specified, can be obtained from commercial sources.

<Example 1> Preparation of 20-pregnene acetylsalicylate-3-trichloroacetate (3)

Step 1: the preparation of pregnene-3-trichloroacetate (1)

Weigh 0.320g (1.01mmol) of pregnenolone into a 100mL eggplant-shaped flask, add 15ml of pyridine under a fume hood, place in a 40°C oil bath and stir, and after the raw materials are dissolved, pipette 400uL (3.58mmol) three Chloroacetyl chloride was slowly added to the reaction, TLC tracked the reaction until the point of no raw material almost disappeared, then stopped the reaction, reacted for 12 hours, first added 15ml ice-distilled water to the reaction bottle, extracted 4 times with ethyl acetate (15mL×4), combined the extracts, followed by Wash with 1mol/L dilute hydrochloric acid, saturated NaHCO ₃ , distilled water, and saturated NaCl, then dry with anhydrous Na ₂ SO ₄ , then distill under reduced pressure, and separate by column chromatography (eluent: V petroleum ether: V ethyl acetate= 5:1) Obtained 0.396g of white needle-like crystals, mp174-177°C, yield: 85.0%, and the product structure was determined by IR, NMR and MS analysis;

Step 2: Preparation of 20-hydroxy-pregnene-3-trichloroacetate (3)

Weigh 0.289g (0.63mmol) of compound (1) into a 100mL eggplant-shaped flask, add 10mL CH ₂ Cl ₂ , place in a water bath at 40°C and stir, and after the solid dissolves, add 0.242g (0.65mmol) CeCl ₃ . 7H ₂ 0, 5mL CH ₃ OH, after completely dissolved, continue to stir for 15min, finally add 0.118g (3.12mmol) NaBH ₄ in batches, continue to stir for 5min after the addition, TLC traces the reaction to the point where there is no raw material, then stop the reaction. Quench the reaction with 1mol/L dilute hydrochloric acid, spin off most of the solvent, add 15mL of distilled water, extract 4 times with ethyl acetate (15mL×4), wash the organic phase with water and saturated NaCl successively, dry over anhydrous _Na2SO4 , and Pressure distillation, column chromatography separation (eluent: V petroleum ether: V ethyl acetate = 6:1) to obtain a white powdery solid 0.261g, mp177-179 ° C, yield: 90.0%, through IR, NMR and MS The analysis determined that the structure of the product was 20-pregnene acetylsalicylate-3-trichloroacetate.

<Example 2> Preparation of (4) of 20-pregnene acetylsalicylate-3-propionate

Step 1: Preparation of Pregnene-3-propionate (2)

Weigh 0.322g (1.02mmol) of pregnenolone into a 100mL eggplant-shaped flask, add 15ml of pyridine under a fume hood, place in a 40°C oil bath and stir, and after the raw materials are dissolved, pipette 400uL (4.59mmol) of acetone. Acyl chloride was slowly added to the reaction, TLC tracked the reaction until the point of no raw material almost disappeared, then stopped the reaction, reacted for 12 hours, first added 15ml ice-distilled water to the reaction bottle, extracted 4 times with ethyl acetate (15mL×4), combined the extracts, and sequentially used 1mol /L dilute hydrochloric acid, saturated NaHCO ₃ , distilled water, saturated NaCl, then dried with anhydrous Na ₂ SO ₄ , then distilled under reduced pressure, separated by column chromatography (eluent: V petroleum ether: V ethyl acetate = 5: 1) Obtained 0.368g of white needle-like crystals, mp124-126°C, yield: 97.1%, and the product structure was determined by IR, NMR and MS analysis;

Step 2: Preparation of 20-hydroxy-pregnene-3-propionate (4)

Weigh 0.290g (0.78mmol) of compound (2) into a 100mL eggplant-shaped flask, add 10mL CH ₂ Cl ₂ , place in a 40°C water bath and stir, and after the solid dissolves, add 0.302g (0.81mmol) CeCl3·7H ₂ 0, 5mL CH ₃ OH, after completely dissolving, continue to stir for 15min, finally add 0.141g (3.71mmol) NaBH ₄ in batches, continue to stir for 5min after the addition, TLC traces the reaction to the point where there is no raw material, stop the reaction, and use 1mol/ Quench the reaction with L dilute hydrochloric acid, spin off most of the solvent, add 15mL of distilled water, extract 4 times with ethyl acetate (15mL× ₄ ), wash the organic phase with water and saturated NaCl successively, dry over anhydrous _Na2SO4 , and distill under reduced pressure , separated by column chromatography (eluent: V petroleum ether: V ethyl acetate = 6:1) to obtain 0.274 g of white powdery solid, mp 153-155 ° C, yield: 90.0%. The structure of the product was determined to be 20-acetylsalicylate pregnene-3-propionate by IR, NMR and MS analysis.

The 20-hydroxy-pregnene-3-alkane ester-pregnant compounds of the present invention are used to carry out cytotoxicity tests on some tumor cells, and it is found that some of the compounds have a significant effect of inhibiting the growth and proliferation of tumor cells. Such as the inhibitory effect of compounds 3 and 4 in the table on human lung adenocarcinoma cell line (A549).

_IC50 values (μmol/L) of compounds 3 and 4 in Table 1 on human lung adenocarcinoma cell lines

compound 3 4 _IC50 7.9 10.3

Although the embodiment of the present invention has been disclosed as above, it is not limited to the use listed in the specification and implementation, it can be applied to various fields suitable for the present invention, and it can be easily understood by those skilled in the art Therefore, the invention is not limited to the specific details without departing from the general concept defined by the claims and their equivalents.

Claims (6)

1. a 20-hydroxyl-pregnene-3-alkane ester group pregnant compound, is characterized in that, the steroid nucleus structural formula of this compound is as follows: Wherein, the R-group may be: R=-CCl ₃ , -CH ₂ CH ₃ . 2. the synthetic method of 20-hydroxyl-pregnene-3-alkane ester base pregnant compound as claimed in claim 1, is characterized in that, comprises the following steps: take steroidal compound pregnenolone as raw material, at first pregnenolone The 3-hydroxyl group of the enolone is converted into an ester through alkane acid chloride, and the intermediate product is obtained through the reaction, and the 17-position carbonyl group of the intermediate product is reduced to obtain the 20-hydroxy-pregnene-3-alkane ester-pregnant compound. 3. the synthetic method of 20-hydroxyl-pregnene-3-alkane ester group pregnant compound as claimed in claim 2, is characterized in that, the reaction method that 3-position hydroxyl is converted into ester through acid chloride esterification is: Dissolve pregnenolone in pyridine, stir at 40°C, then slowly add alkanyl chloride, follow the reaction by TLC, stop the reaction at the point where there is no raw material, add ice water to the obtained reaction product, and then extract with ethyl acetate , combined the organic phases, washed with 1mol/L dilute hydrochloric acid, saturated NaHCO ₃ , distilled water, and saturated NaCl successively, dried with anhydrous Na ₂ SO ₄ , and finally distilled off the organic solvent under reduced pressure, and separated by column chromatography to obtain white needles. crystal, which is the intermediate product; Wherein, the alkanoyl chloride is trichloroacetyl chloride or propionyl chloride. 4. the synthetic method of 20-hydroxyl-pregnene-3-alkane ester group pregnant compound as claimed in claim 3, is characterized in that, the reaction method of 17-position carbonyl reduction is: Mix the intermediate product with CH ₂ Cl ₂ , stir at 40°C until the intermediate product is completely dissolved, add CeCl ₃ 7H ₂ O and CH ₃ OH, after CeCl ₃ 7H ₂ O is completely dissolved, weigh Add _NaBH4 slowly to react, react at room temperature, detect by TLC, stop the reaction until there is no raw material, add 1mol/L dilute hydrochloric acid to quench the reaction, distill off the organic solvent under reduced pressure, add distilled water, extract with ethyl acetate, The organic phases were combined, washed with water and saturated NaCl solution in turn, then dried with anhydrous Na ₂ SO ₄ , and finally the organic solvent was distilled off under reduced pressure, and separated by column chromatography to obtain the final product of white solid, which was the 20-hydroxyl - Pregnene-3-alkane ester prepregnant compounds. 5. The application of the 20-hydroxyl-pregnene-3-alkane ester group pregnant compound as claimed in claim 1 in the preparation of antitumor drugs. 6. The application of the 20-hydroxyl-pregnene-3-alkane ester-based pregnant compound as claimed in claim 1 in the preparation of antitumor drugs in the pharmaceutical composition containing the active ingredient.