17beta-pregnenolone-amino-acid ester derivative as well as preparation method and application thereof
By preparing 17β-pregnenolone-amino acid ester derivatives, the problem of insufficient solubility and permeability of existing anti-tumor drugs has been solved, and effective inhibitory effect on a variety of cancer cells is achieved, and it is suitable for various forms of anti-tumor drugs.
Patent Information
- Application Number
- CN202510193470.2
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-02-21
- Publication Date
- 2025-08-05
AI Technical Summary
Existing anti-tumor drugs such as abiraterone have problems with poor solubility and low permeability, and existing amino acid ester derivatives have insufficient anti-tumor activity in some cancer cell lines.
By esterification, halogenation reaction and nucleophilic substitution of 17β-gesterone with different representative amino acid ester compounds, 17β-gesterone-amino acid ester derivatives were prepared to improve their biological activity.
The prepared 17β-pregnenolone-amino acid ester derivatives show significant inhibitory effects on human breast cancer, ovarian cancer, cervical cancer, liver cancer and prostate cancer cells, and are suitable for a variety of administration forms of anti-tumor drugs.
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Figure CN120424154A_ABST
Abstract
Description
Technical Field
[0001] The present invention relates to the field of anti-tumor drugs, in particular to a 17β-pregnenolone-amino acid ester derivative. Background Art
[0002] In recent years, steroidal compounds have garnered significant attention as anti-tumor drugs. The introduction of active functional groups at various positions within the steroid structure to create compounds with enhanced activity has been a research hotspot. Abiraterone, a clinically used anticancer steroidal drug, features a structural characteristic of pregnenolone with an active functional group at the 17-position. Its chemical structure is shown below.
[0003]
[0004] Amino acids are the building blocks of proteins, involved in energy metabolism and transport, and possess excellent biocompatibility and affinity. Studies have shown that amino acid modification can improve the poor solubility and low permeability of parent drugs, enhancing their bioactivity and leading to their widespread use in the design and development of anti-tumor drugs. Therefore, incorporating amino acids into drug molecules to create highly effective, low-toxic compounds is an important approach for the development of new anti-tumor drugs.
[0005] In recent years, steroid compounds have attracted great attention as anti-tumor drug precursors conjugated with amino acids. Zhou et al. designed and synthesized a series of 17β-estradiol-amino acid ester derivatives. These compounds can enhance the in vitro anti-tumor proliferation activity of the steroid nucleus by regulating the expression of apoptosis-related proteins in MDA-MB-231 (human breast cancer cells). The IC 50 The value can reach 4.89 µM. Yuan et al. modified the active ingredients of ginseng, 25-methoxyprotopanaxadiol (25-OCH3-PPD) and 25-hydroxyprotopanaxadiol (25-OH-PPD), with non-protein amino acids and synthesized a series of novel amino acid ester derivatives. Some of these compounds showed considerable antitumor activity against HCT-116 (human colon cancer cells) and BGC-823 (human gastric adenocarcinoma cells) cell lines, with IC 50 The values ranged from 4.76 to 9.03 μM. Lin et al. used ginsenoside (AD-2) as the lead compound and designed and synthesized a series of novel AD-2 amino acid ester derivatives, which showed good inhibitory effects on the proliferation of HepG2 (human liver cancer cells), with the best IC 50 The value is 8.25 µM. Summary of the Invention
[0006] An object of the present invention is to solve at least the above problems and to provide at least the advantages which will be described hereinafter.
[0007] Another object of the present invention is to provide a 17β-pregnenolone-amino acid ester derivative.
[0008] Another object of the present invention is to provide a method for preparing the above compound.
[0009] A further object of the present invention is to provide the use of the above compound in anti-tumor drugs.
[0010] In order to achieve these purposes and other advantages according to the present invention, a 17β-pregnenolone-amino acid ester derivative is provided, the chemical structure of which is as follows:
[0011] The preparation method of 17β-pregnenolone-amino acid ester derivatives comprises the following steps: selecting different representative amino acids constituting human proteins as raw materials, esterifying their carboxyl groups by dehydration condensation with ethanol to obtain various amino acid ester compounds 2 (a to e); using pregnenolone as a lead compound, protecting its 3-OH group with dihydropyran to obtain compound 4; reacting the 20-position carbonyl group with NaBrO to produce a 17-carboxyl group-containing compound 5; activating the 17-carboxyl group on the structure of compound 5 with thionyl chloride, and then reacting the 17-carboxyl group with compounds 2 (a to e) to produce corresponding amide compounds 6 (a to e); and finally deprotecting the 3-OH group in an acidic environment to obtain target compounds 7 (a to e), wherein: Compounds 2 (a~e) are glycine ethyl ester, L-valine ethyl ester, L-tyrosine ethyl ester, L-deoxyalliin ethyl ester, and L-alanine ethyl ester, respectively, and their structural formulas are as follows:
[0012] Preferably, the compounds 6(a-e) are dissolved in ethanol, and the 3-OH group is deprotected under acidic conditions to obtain target compounds 7(a-e). Their chemical names are: (3β-hydroxypregnane-17β-formyl)glycine ethyl ester, (3β-hydroxypregnane-17β-formyl)-L-valine ethyl ester, (3β-hydroxypregnane-17β-formyl)-L-tyrosine ethyl ester, (3β-hydroxypregnane-17β-formyl)-L-deoxyalliin ethyl ester, and (3β-hydroxypregnane-17β-formyl)-L-alanine ethyl ester.
[0013] The chemical structures are shown below:
[0014] Preferably, 1.00-2.00 g of one of glycine, L-valine, L-tyrosine, L-deoxyalliin or L-alanine is dissolved in 20-40 mL of ultra-dry thionyl chloride, the nitrogen is replaced, and 600-1200 μL of anhydrous ethanol is slowly added under ice bath conditions, and the mixture is stirred for 10 h under ice bath conditions. The reaction end point is monitored by thin layer chromatography. After the reaction is completed, the mixture is evaporated under reduced pressure, the solvent is dried, and the mixture is extracted with ethyl acetate three times, washed with water three times, and the organic phases are combined, dried over anhydrous sodium sulfate, evaporated under reduced pressure, and purified by column chromatography to obtain a white solid 2 (a~e).
[0015] Preferably, pregnenolone is used as the lead compound, and its 3-OH is protected by dihydropyran, specifically comprising: taking 2.00-4.00 g of pregnenolone and dissolving it in 20-40 mL of ultra-dry DMF, adding 30-60 mg of p-toluenesulfonamide, replacing argon, adding 200-400 μL of DHP at room temperature, continuing to stir for 12 hours, and monitoring the reaction end point by thin layer chromatography. After the reaction is completed, vacuum rotary evaporation is performed to dry the solvent, and the mixture is extracted with dichloromethane three times, washed with water three times, and the organic phases are combined, dried over anhydrous sodium sulfate, vacuum rotary evaporation, and purified by column chromatography to obtain a white solid as compound 4.
[0016] Preferably, NaBrO is used to perform a haloform reaction on the 20-carbonyl group of compound 4, specifically comprising: taking 2.00 g-3.00 g of compound 4, dissolving it in 30-50 mL of ultra-dry 1,4-dioxane, replacing nitrogen, slowly adding 20-30 mL of NaBrO dropwise under stirring at -20 ° C, controlling the addition to be completed within 1 h, maintaining stirring at -20 ° C for 12 h, monitoring the reaction end point by thin layer chromatography, after the reaction is completed, vacuum rotary evaporation, spin-drying the solvent, extracting with dichloromethane 3 times, washing with water 3 times, combining the organic phases, drying over anhydrous sodium sulfate, vacuum rotary evaporation, and column chromatography analysis and purification to obtain a white solid as compound 5.
[0017] Preferably, thionyl chloride is used to activate the 17-carboxyl group on the structure of compound 5, and then a nucleophilic substitution reaction is carried out with compound 2 (a~e), specifically comprising: taking 400-800 mg of compound 5, dissolving it in 10-20 mL of ultra-dry thionyl chloride, adding 10-20 μL of DMF, replacing nitrogen, refluxing at 80 ° C for 6 h, removing the solvent under reduced pressure, dissolving it with 10-20 mL of dry dichloromethane, placing it at -10 ° C, adding 200-300 mg of one of compounds 2 (a~e), then adding 50-100 μL of triethylamine, maintaining stirring at -10 ° C for 10 min, transferring it to room temperature and stirring for 3 h, monitoring the reaction end point by thin layer chromatography, after the reaction is completed, vacuum rotary evaporation, after the solvent is dried, dichloromethane extraction 3 times, washing 3 times, combining the organic phases, drying over anhydrous sodium sulfate, vacuum rotary evaporation, column chromatography analysis and purification, to obtain a white solid as compound 6 (a~e).
[0018] Preferably, the 3-OH of compound 6 (a~e) is deprotected under an acidic environment, specifically comprising: taking 300-400 mg of compound 6 (a~e), dissolving it in 20-30 mL of anhydrous ethanol, replacing nitrogen, adding 10% dilute hydrochloric acid at -10 ° C, controlling stirring at -10 ° C for 12 h, monitoring the reaction end point by thin layer chromatography, and after the reaction is completed, vacuum rotary evaporation, after the solvent is dried, extracting with dichloromethane 3 times, washing with water 3 times, combining the organic phases, drying over anhydrous sodium sulfate, vacuum rotary evaporation, and column chromatography analysis and purification to obtain a white solid as compound 7 (a~e).
[0019] The above 17β-pregnenolone-amino acid ester derivative is used for preparing anti-tumor drugs.
[0020] The reaction route for preparing the above-mentioned 17β-pregnenolone-amino acid ester derivative is as follows:
[0021] The present invention can achieve at least the following beneficial effects: In vitro anti-tumor proliferation activity tests have shown that the present invention has inhibitory effects on human breast cancer cells, human ovarian cancer cells, human cervical cancer cells, human liver cancer cells, and prostate cancer cells. Therefore, the 17β-pregnenolone-amino acid ester derivatives of the present invention can be used to prepare medicaments for treating tumors. These medicaments can be formulated into injections, tablets, pills, capsules, suspensions, or emulsions, and can be administered orally, or by subcutaneous, intravenous, or intramuscular injection. Other advantages, objectives, and features of the present invention will be apparent in part from the following description and will also be understood by those skilled in the art through study and practice of the present invention.
Claims
1. A 17β-pregnenolone-amino acid ester derivative, characterized in that: Its chemical structure is as follows: in, 2. A method for preparing the 17β-pregnenolone-amino acid ester derivative according to claim 1, characterized in that: The preparation method comprises the following steps: selecting different representative amino acids constituting human proteins as raw materials, esterifying their carboxyl groups by dehydration condensation with ethanol to obtain various amino acid ester compounds 2 (a to e); using pregnenolone as a lead compound, protecting its 3-OH group with dihydropyran to obtain compound 4; and using NaBrO to react with the 20-position carbonyl group to produce a 17-carboxyl group-containing compound 5; further using thionyl chloride to activate the 17-carboxyl group on the structure of compound 5, and then reacting with compounds 2 (a to e) for nucleophilic substitution to obtain corresponding amide compounds 6 (a to e); and finally deprotecting the 3-OH group in an acidic environment to obtain target compounds 7 (a to e); The amino acid ester compounds 2 (a to e) are prepared by esterifying the carboxyl groups of the amino acids glycine, L-valine, L-tyrosine, L-deoxyalliin, and L-alanine, respectively, using them as raw materials. The amino acids are dehydration condensation with ethanol. The chemical names of the amino acid ester compounds 2 (a to e) are glycine ethyl ester, L-valine ethyl ester, L-tyrosine ethyl ester, L-deoxyalliin ethyl ester, and L-alanine ethyl ester, respectively. The chemical structures thereof are as follows: The compound 4 is 3β-(tetrahydro-2H-pyran-2-yl)oxypregnesterone, and its chemical structure is as follows: The compound 5 is 3β-(tetrahydro-2H-pyran-2-yl)oxypregnane-17β-carboxylic acid, and its structural formula is as follows:
3. The 17β-pregnenolone-amino acid ester derivative according to claim 2, characterized in that: The compounds 6 (a to e) are obtained by activating the 17-carboxyl group on the structure of compound 5 with thionyl chloride, and then undergoing nucleophilic substitution reactions with compounds 2 (a to e). The chemical names of the obtained compounds 6 (a to e) are (3β-(tetrahydro-2H-pyran-2-yl)oxypregnane-17β-formyl)glycine ethyl ester, (3β-(tetrahydro-2H-pyran-2-yl)oxypregnane-17β-formyl)glycine ethyl ester, -formyl)-L-valine ethyl ester, (3β-(tetrahydro-2H-pyran-2-yl)oxypregnan-17β-formyl)-L-tyrosine ethyl ester, (3β-(tetrahydro-2H-pyran-2-yl)oxypregnan-17β-formyl)-L-deoxyalliin ethyl ester, (3β-(tetrahydro-2H-pyran-2-yl)oxypregnan-17β-formyl)-L-alanine ethyl ester; their chemical structural formulas are as follows:
4. The 17β-pregnenolone-amino acid ester derivative according to claim 2, characterized in that: The compounds 6 (a-e) were dissolved in ethanol, and the 3-OH group was deprotected under acidic conditions to obtain target compounds 7 (a-e); their chemical names are: (3β-hydroxypregnane-17β-formyl)glycine ethyl ester, (3β-hydroxypregnane-17β-formyl)-L-valine ethyl ester, (3β-hydroxypregnane-17β-formyl)-L-tyrosine ethyl ester, (3β-hydroxypregnane-17β-formyl)-L-deoxyalliin ethyl ester, (3β-hydroxypregnane-17β-formyl)-L-alanine ethyl ester; their chemical structures are as follows:
5. The 17β-pregnenolone-amino acid ester derivative according to claim 2, characterized in that: Different representative amino acids constituting human proteins are selected as raw materials, and their carboxyl groups are subjected to an esterification reaction through dehydration condensation with ethanol. Specifically, the method comprises: dissolving 1.00-2.00 g of one of glycine, L-valine, L-tyrosine, L-deoxyalliin or L-alanine in 20-40 mL of ultra-dry thionyl chloride, replacing nitrogen, slowly adding 600-1200 μL of anhydrous ethanol in an ice bath, stirring for 10 hours in an ice bath, monitoring the reaction endpoint by thin-layer chromatography, and after the reaction is completed, performing vacuum rotary evaporation to dry the solvent, extracting with ethyl acetate three times, washing with water three times, combining the organic phases, drying over anhydrous sodium sulfate, vacuum rotary evaporation, and purifying by column chromatography to obtain a white solid 2(a-e).
6. The 17β-pregnenolone-amino acid ester derivative according to claim 2, characterized in that: Using pregnenolone as the lead compound, its 3-OH group was protected by dihydropyran, specifically comprising: dissolving 2.00-4.00 g of pregnenolone in 20-40 mL of ultra-dry DMF, adding 30-60 mg of p-toluenesulfonamide, replacing argon, adding 200-400 μL of DHP at room temperature, continuing stirring for 12 hours, and monitoring the reaction end point by thin-layer chromatography. After the reaction is completed, vacuum rotary evaporation is performed to dry the solvent, and the compound is extracted with dichloromethane three times, washed with water three times, and the organic phases are combined, dried over anhydrous sodium sulfate, vacuum rotary evaporation is performed, and purification is performed by column chromatography to obtain a white solid as compound 4.
7. The 17β-pregnenolone-amino acid ester derivative according to claim 2, characterized in that: The 20-carbonyl group of compound 4 was subjected to a haloform reaction using NaBrO, specifically comprising: taking 2.00 g-3.00 g of compound 4, dissolving it in 30-50 mL of ultra-dry 1,4-dioxane, replacing nitrogen, slowly adding 20-30 mL of NaBrO dropwise under stirring at -20°C, and controlling the addition to be completed within 1 hour, maintaining stirring at -20°C for 12 hours, and monitoring the reaction endpoint by thin-layer chromatography. After the reaction is completed, the mixture is evaporated under reduced pressure, the solvent is dried, extracted with dichloromethane three times, washed with water three times, the organic phases are combined, dried over anhydrous sodium sulfate, evaporated under reduced pressure, and purified by column chromatography to obtain a white solid as compound 5.
8. The 17β-pregnenolone-amino acid ester derivative according to claim 2, characterized in that: The 17-carboxyl group on the structure of compound 5 was activated by thionyl chloride, and then a nucleophilic substitution reaction was carried out with compound 2 (a~e), specifically comprising: taking 400-800 mg of compound 5, dissolving it in 10-20 mL of ultra-dry thionyl chloride, adding 10-20 μL of DMF, replacing nitrogen, refluxing at 80° C. and stirring for 6 hours, removing the solvent under reduced pressure, dissolving it with 10-20 mL of dry dichloromethane, placing it at -10° C. and adding 200-300 mg of one of compounds 2 (a~e), then adding 50-100 μL of triethylamine, maintaining stirring at -10° C. for 10 minutes, transferring it to room temperature and stirring for 3 hours, monitoring the reaction end point with thin layer chromatography, and after the reaction is completed, vacuum rotary evaporation, after the solvent is dried, extracting with dichloromethane 3 times, washing with water 3 times, combining the organic phases, drying over anhydrous sodium sulfate, vacuum rotary evaporation, and column chromatography analysis and purification to obtain a white solid as compound 6 (a~e).
9. The 17β-pregnenolone-amino acid ester derivative according to claim 2, characterized in that: The 3-OH of compound 6 (a ~ e) was deprotected under an acidic environment, specifically comprising: taking 300-400 mg of compound 6 (a ~ e), dissolving it in 20-30 mL of anhydrous ethanol, replacing nitrogen, adding 10% dilute hydrochloric acid at -10 ° C, controlling stirring at -10 ° C for 12 hours, and monitoring the reaction end point by thin layer chromatography. After the reaction is completed, vacuum rotary evaporation is performed, the solvent is dried, extracted with dichloromethane 3 times, washed with water 3 times, and the organic phases are combined, dried over anhydrous sodium sulfate, vacuum rotary evaporation, and purified by column chromatography to obtain a white solid as compound 7 (a ~ e).
10. Use of the 17β-pregnenolone-amino acid ester derivative according to claim 1 in the preparation of anti-tumor drugs.