CN103450310A

Movatterモバイル変換

Info

Publication number: CN103450310A
Application number: CN2013103607948A
Authority: CN
Inventors: 陆豫; 肖志鹏; 余勃
Original assignee: Nanchang University
Current assignee: Nanchang University
Priority date: 2013-08-19
Filing date: 2013-08-19
Publication date: 2013-12-18

Abstract

一种豆甾醇衍生物及其在制备抗癌药物中的应用，为以下式Ⅰ、Ⅱ所示：

Figure 2013103607948100004DEST_PATH_IMAGE001

式Ⅰ中的R所代表的基团为：SO₂C₆H₅，COCH₃，COCH₂Cl，COC₆H₅，CH₂CH₃，NO₂。本发明的豆甾醇衍生物对多种肿瘤细胞诸如人肝癌上皮细胞、人胃癌细胞、人乳腺癌细胞、人卵巢癌细胞有显著的抑制作用，而对正常细胞毒性较低，可用于制备治疗癌症的药物。A stigmasterol derivative and its application in the preparation of anticancer drugs are represented by the following formulas I and II:

The groups represented by R in formula I are: SO₂ C₆ H₅ , COCH₃ , COCH₂ Cl, COC₆ H₅ , CH₂ CH₃ , NO₂ . The stigmasterol derivatives of the present invention have significant inhibitory effects on a variety of tumor cells such as human liver cancer epithelial cells, human gastric cancer cells, human breast cancer cells, and human ovarian cancer cells, but have low toxicity to normal cells, and can be used to prepare Drug.

Description

Translated fromChinese

豆甾醇衍生物及其在制备抗癌药物中的应用Stigmasterol derivatives and their application in the preparation of anticancer drugs

技术领域technical field

本发明涉及豆甾醇衍生物及其在制备抗癌药物中的应用。The present invention relates to stigmasterol derivatives and their application in the preparation of anticancer drugs.

背景技术Background technique

癌症目前已经成为威胁人类健康和生活素质的最大的疾病之一，利用我国丰富的自然资源，从中寻找前有效单体，并通过修饰改造获得高效、高选择性、低毒的抗癌药物是当前抗癌药物的主要研究方向。Cancer has become one of the biggest diseases that threaten human health and quality of life. It is currently the most important strategy to use our country's rich natural resources to find pre-effective monomers and modify them to obtain highly efficient, highly selective, and low-toxicity anticancer drugs. The main research directions of anticancer drugs.

肿瘤的化学预防是降低癌症发病率最直接的方法之一。为了战胜癌症等严重危害人体健康的疾病，人们不断地从植物中寻找新的治疗药物。近年来，倒捻子素的抗癌作用逐渐被人们得到证实。目前，针对高发癌症如肺癌、胃癌、肝癌、乳腺癌和结肠癌等进行研究，倒捻子素都体现很好的抗癌效果，并且逐步深入到倒捻子素的抗癌机制。Chemoprevention of tumors is one of the most direct ways to reduce the incidence of cancer. In order to overcome diseases that seriously endanger human health such as cancer, people are constantly looking for new therapeutic drugs from plants. In recent years, the anticancer effect of mangostin has been confirmed gradually. At present, researches on high-incidence cancers such as lung cancer, gastric cancer, liver cancer, breast cancer, and colon cancer have shown that mangostin has a good anti-cancer effect, and the anti-cancer mechanism of mangostin has gradually been penetrated.

以天然产物中的活性成分为母体化合物，根据药物分子学的原理进行结构修饰，设计出可能具有高活性、低毒副作用的半合成化合物，从而发现可能应用于临床的新分子实体是目前新药开发的一种重要手段。Taking the active ingredients in natural products as the parent compound, modifying the structure according to the principles of pharmaceutical molecular science, designing semi-synthetic compounds that may have high activity and low toxicity and side effects, so as to discover new molecular entities that may be applied clinically is the current development of new drugs. an important means.

发明内容Contents of the invention

本发明的目的是提供一种经过化学修饰的倒捻子素衍生物，及其在用于治疗癌症的药物的应用。The object of the present invention is to provide a chemically modified mangostin derivative and its application in medicine for treating cancer.

本发明的豆甾醇衍生物为以下式Ⅰ或II所示：The stigmasterol derivatives of the present invention are shown in the following formula I or II:

式ⅠFormula Ⅰ

式Ⅰ中的R所代表的基团为： SO₂C₆H₅、COCH₃、COCH₂Cl、COC₆H₅、CH₂CH₃或NO₂。The groups represented by R in formula I are: SO₂ C₆ H₅ , COCH₃ , COCH₂ Cl, COC₆ H₅ , CH₂ CH₃ or NO₂ .

式II。Formula II.

本发明所述的豆甾醇衍生物，是从大豆植物中提取，经分离纯化得到的豆甾醇经过化学修饰得到的化合物。The stigmasterol derivatives of the present invention are compounds obtained by extracting from soybean plants, separating and purifying the stigmasterol and chemically modifying it.

制备本发明的豆甾醇衍生物的反应如以下反应式所示：The reaction of preparing stigmasterol derivatives of the present invention is shown in the following reaction formula:

3-苯磺酸基-5,22-二烯-24-乙基-胆甾（AB-1）的合成路线图Synthetic route of 3-benzenesulfonate-5,22-diene-24-ethyl-cholesteryl (AB-1)

3-乙酰氧基-5,22-二烯-24-乙基-胆甾（AB-2）的合成路线图Synthetic route of 3-acetoxy-5,22-diene-24-ethyl-cholesteric (AB-2)

3-氯乙酰氧基-5,22-二烯-24-乙基-胆甾（AB-3）的合成路线图Synthetic route of 3-chloroacetoxy-5,22-diene-24-ethyl-cholesteric (AB-3)

3-苯甲酰基-5,22-二烯-24-乙基-胆甾（AB-4）的合成路线图Synthetic route of 3-benzoyl-5,22-diene-24-ethyl-cholesteric (AB-4)

3-羟基-5,6,22,23-四溴-24-乙基-胆甾（AB-5）的合成路线图Synthetic route of 3-hydroxy-5,6,22,23-tetrabromo-24-ethyl-cholesteric (AB-5)

3-乙氧基-5,22-二烯-24-乙基-胆甾（AB-6）的合成路线图Synthetic route of 3-ethoxy-5,22-diene-24-ethyl-cholesteric (AB-6)

3-硝基-5,22-二烯-24-乙基-胆甾（AB-7）的合成路线图Synthetic route of 3-nitro-5,22-diene-24-ethyl-cholesteryl (AB-7)

通过体外癌细胞抑制试验表明，本发明的豆甾醇衍生物对多种肿瘤细胞诸如人肝癌上皮细胞（HEPG-2）、人胃癌细胞（MKN45）、人乳腺癌细胞（MDA-MB-231）、人卵巢癌细胞（SKOV3）有显著的抑制作用，而对正常细胞毒性较低。部分衍生物对HEPG-2、MDA-MB-231和SKOV3肿瘤细胞的抑制作用比相应的豆甾醇和ADM都强，因此发明的部分倒捻子素衍生物可用于制备治疗癌症的药物。In vitro cancer cell inhibition tests show that the stigmasterol derivatives of the present invention can inhibit various tumor cells such as human liver cancer epithelial cells (HEPG-2), human gastric cancer cells (MKN45), human breast cancer cells (MDA-MB-231), Human ovarian cancer cells (SKOV3) have a significant inhibitory effect, while less toxic to normal cells. Part of the derivatives have stronger inhibitory effects on HEPG-2, MDA-MB-231 and SKOV3 tumor cells than the corresponding stigmasterol and ADM, so the invented part of mangostin derivatives can be used to prepare medicines for treating cancer.

具体实施方式Detailed ways

本发明将通过以下实施例作进一步说明。The invention will be further illustrated by the following examples.

实施例1。3-苯磺酸基-5,22-二烯-24-乙基-胆甾（AB-1）的合成。Example 1. Synthesis of 3-benzenesulfono-5,22-diene-24-ethyl-cholesteric (AB-1).

取豆甾醇0.5g（1.21mmol）于氮气保护的50mL三口瓶中，加入干燥的氯仿20mL，加入干燥的三乙胺0.6 mL（约4.3mmol），控制温度在60℃，缓慢滴加1ml 苯磺酰氯（6.0mmol）与3 mL 氯仿的混合液，半小时滴加完毕，再继续搅拌，薄层色谱跟踪反应进程，7小时后终止反应。过滤，滤液用饱和氯化钠溶液洗3次，有机相用无水硫酸钠干燥过夜，再次过滤，蒸干滤液，硅胶柱层析（硅胶，200-300目；洗脱剂，石油醚/乙酸乙酯=20:1）纯化，得白色固体粉末0.523g，产率78.2%； m.p.103.8~ 105.2 ℃; ¹HNMR (CDCl₃) δ(ppm): 0.686 (s, 3H), 0.788~0.799 (m,6H), 0.845 (d, J = 6.6Hz,3H), 1.010~1.135 (m, 7H), 1.151~1.162 (m, 1H), 1.170~1.253 (m, 3H), 1.429~1.441 (m, 2H), 1.450~1.469 (m, 4H), 1.510~1.519 (m, 2H), 1.528~1.543 (m, 2H), 1.555~1.603 (m, 2H), 1.893~1.910 (m, 1H), 1.967~2.094 (m, 3H), 2.495 (d, J = 7.8Hz,1H), 4.196~4.229 (m, 1H), 5.013~5.041 (m, 1H), 5.125~5.165 (m, 1H), 5.399~5.414 (m, 1H), 7.527~7.545 (m, 3H), 7.718~7.734 (m, 2H); ESI-MS m/z 544.38 ([M+H]⁺,80%). Anal.Calcd for C₃₅H₅₂O₃S: C, 76.04; H, 9.48. Found: C, 75.99; H, 9.50。Take 0.5g (1.21mmol) of stigmasterol in a 50mL three-necked flask protected by nitrogen, add 20mL of dry chloroform, add 0.6 mL (about 4.3mmol) of dry triethylamine, control the temperature at 60°C, and slowly add 1ml of benzenesulfonate dropwise The mixture of acid chloride (6.0 mmol) and 3 mL of chloroform was added dropwise for half an hour, and then continued to stir. The reaction progress was tracked by thin-layer chromatography, and the reaction was terminated after 7 hours. Filter, wash the filtrate 3 times with saturated sodium chloride solution, dry the organic phase with anhydrous sodium sulfate overnight, filter again, evaporate the filtrate to dryness, perform silica gel column chromatography (silica gel, 200-300 mesh; eluent, petroleum ether/acetic acid ethyl ester=20:1) to obtain 0.523g of white solid powder with a yield of 78.2%; mp103.8~ 105.2 ℃;¹ HNMR (CDCl₃ ) δ(ppm): 0.686 (s, 3H), 0.788~0.799 ( m,6H), 0.845 (d,J = 6.6Hz,3H), 1.010~1.135 (m, 7H), 1.151~1.162 (m, 1H), 1.170~1.253 (m, 3H), 1.429~1.441 (m, 2H), 1.450~1.469 (m, 4H), 1.510~1.519 (m, 2H), 1.528~1.543 (m, 2H), 1.555~1.603 (m, 2H), 1.893~1.910 (m, 1H), 1.967~ 2.094 (m, 3H), 2.495 (d,J = 7.8Hz,1H), 4.196~4.229 (m, 1H), 5.013~5.041 (m, 1H), 5.125~5.165 (m, 1H), 5.399~5.414 ( m, 1H), 7.527~7.545 (m, 3H), 7.718~7.734 (m, 2H); ESI-MS m/z 544.38 ([M+H]⁺ ,80%). Anal. Calcd for C₃₅ H₅₂ O₃ S: C, 76.04; H, 9.48. Found: C, 75.99; H, 9.50.

实施例2。3-乙酰氧基-5,22-二烯-24-乙基-胆甾（AB-2）的合成。Example 2. Synthesis of 3-Acetoxy-5,22-diene-24-ethyl-cholesteric (AB-2).

取豆甾醇0.5 g（1.21 mmol）于氮气保护的50 mL三口瓶中，加入干燥的吡啶18 mL，控制温度在60℃，缓慢滴加0.6ml 醋酸酐（6.0 mmol）与3 mL 氯仿的混合液，半小时滴加完毕，再继续搅拌，薄层色谱跟踪反应进程，10小时后终止反应。冷却至室温，用1.0 N HCl调节pH至2。过滤，滤液用乙酸乙酯萃取3次，有机相用无水硫酸钠干燥过夜，再次过滤，蒸干滤液，硅胶柱层析（硅胶，200-300目；洗脱剂，石油醚/乙酸乙酯=20:1）纯化，得白色固体粉末0.473g，产率86.1%；m.p.140.9~142.4℃;¹HNMR (CDCl₃) δ(ppm): 0.696 (s, 3H), 0.790~0.800 (m, 6H), 0.846 (d, J = 6.0Hz, 3H), 0.911~0.974 (m, 1H), 1.016~1.127 (m, 7H), 1.129~1.137 (m, 1H), 1.142~1.191 (m, 4H), 1.225~1.248 (m, 1H), 1.469~1.510 (m, 9H), 1.520~1.529 (m, 1H), 1.853~1.873 (m, 2H), 1.978~2.035 (m, 6H), 2.300~2.347 (m, 2H), 4.603~4.612 (m, 1H), 4.992~5.032 (m, 1H), 5.130~5.169 (m, 1H), 5.374 (t, J = 2.4Hz, 1H); ESI-MS m/z 456 ([M+H]⁺, 20%). Anal.Calcd for C₃₁H₅₀O₂: C, 81.88; H, 11.08. Found: C, 81.90; H, 11.07。Take 0.5 g (1.21 mmol) of stigmasterol in a 50 mL three-necked flask protected by nitrogen, add 18 mL of dry pyridine, control the temperature at 60 °C, and slowly add dropwise a mixture of 0.6 ml of acetic anhydride (6.0 mmol) and 3 mL of chloroform After half an hour, the dropwise addition was completed, and the stirring was continued, and the reaction progress was tracked by thin-layer chromatography, and the reaction was terminated after 10 hours. Cool to room temperature and adjust pH to 2 with 1.0 N HCl. Filtration, the filtrate was extracted 3 times with ethyl acetate, the organic phase was dried overnight with anhydrous sodium sulfate, filtered again, the filtrate was evaporated to dryness, silica gel column chromatography (silica gel, 200-300 mesh; eluent, petroleum ether/ethyl acetate =20:1) to obtain white solid powder 0.473g, yield 86.1%; mp140.9~142.4℃;¹ HNMR (CDCl₃ ) δ(ppm): 0.696 (s, 3H), 0.790~0.800 (m, 6H), 0.846 (d,J = 6.0Hz, 3H), 0.911~0.974 (m, 1H), 1.016~1.127 (m, 7H), 1.129~1.137 (m, 1H), 1.142~1.191 (m, 4H) , 1.225~1.248 (m, 1H), 1.469~1.510 (m, 9H), 1.520~1.529 (m, 1H), 1.853~1.873 (m, 2H), 1.978~2.035 (m, 6H), 2.300~2.347 ( m, 2H), 4.603~4.612 (m, 1H), 4.992~5.032 (m, 1H), 5.130~5.169 (m, 1H), 5.374 (t,J = 2.4Hz, 1H); ESI-MS m/z 456 ([M+H]⁺ , 20%). Anal. Calcd for C₃₁ H₅₀ O₂ : C, 81.88; H, 11.08. Found: C, 81.90; H, 11.07.

实施例3。3-氯乙酰氧基-5,22-二烯-24-乙基-胆甾（AB-3）的合成。Example 3. Synthesis of 3-chloroacetoxy-5,22-diene-24-ethyl-cholesteric (AB-3).

取豆甾醇0.5 g（1.21 mmol）于氮气保护的50 mL三口瓶中，加入干燥的氯仿20 mL，加入干燥的三乙胺0.6 mL（约4.3mmol），室温下缓慢滴加0.6 ml 氯乙酰氯（6.0 mmol）与3 mL 氯仿的混合液，半小时滴加完毕，再继续搅拌，薄层色谱跟踪反应进程，2小时后终止反应。过滤，蒸干滤液，硅胶柱层析（硅胶，200-300目；洗脱剂，石油醚/乙酸乙酯=25:1）纯化，得白色固体粉末0.544g，产率92.0%； m.p.175.1~176.9℃;¹HNMR (CDCl₃) δ(ppm): 0.696 (s, 3H), 0.789~0.816 (m, 6H), 0.846 (d, J = 6.6Hz, 3H), 0.912~0.977 (m, 1H), 1.016~1.094 (m, 7H), 1.127~1.136 (m, 1H), 1.144~1.296 (m, 6H), 1.375~1.455 (m, 2H), 1.494~1.571 (m, 5H), 1.611~1.624 (m, 1H), 1.693~1.721 (m, 1H), 1.821~1.891 (m, 2H), 1.932~2.002 (m, 3H), 2.353~2.365 (m, 2H), 4.040 (s, 2H), 4.684~4.723 (m, 1H), 4.964~5.033 (m, 1H), 5.129~5.169 (m, 1H), 5.389 (t, J = 1.8 Hz, 1H); ESI-MS m/z 490 ([M+H]⁺, 10%). Anal.Calcd for C₃₁H₄₉O₂Cl: C, 76.11; H, 10.10. Found: C, 76.10; H, 10.09。Take 0.5 g (1.21 mmol) of stigmasterol in a 50 mL three-necked flask protected by nitrogen, add 20 mL of dry chloroform, add 0.6 mL (about 4.3 mmol) of dry triethylamine, and slowly add 0.6 ml of chloroacetyl chloride dropwise at room temperature (6.0 mmol) and 3 mL of chloroform, the mixture was added dropwise for half an hour, and the stirring was continued, and the reaction progress was tracked by thin-layer chromatography, and the reaction was terminated after 2 hours. Filtrate, evaporate the filtrate to dryness, and purify by silica gel column chromatography (silica gel, 200-300 mesh; eluent, petroleum ether/ethyl acetate=25:1) to obtain 0.544 g of white solid powder with a yield of 92.0%; mp175.1 ~176.9℃;¹ HNMR (CDCl₃ ) δ(ppm): 0.696 (s, 3H), 0.789~0.816 (m, 6H), 0.846 (d,J = 6.6Hz, 3H), 0.912~0.977 (m, 1H ), 1.016~1.094 (m, 7H), 1.127~1.136 (m, 1H), 1.144~1.296 (m, 6H), 1.375~1.455 (m, 2H), 1.494~1.571 (m, 5H), 1.611~1.624 (m, 1H), 1.693~1.721 (m, 1H), 1.821~1.891 (m, 2H), 1.932~2.002 (m, 3H), 2.353~2.365 (m, 2H), 4.040 (s, 2H), 4.684 ~4.723 (m, 1H), 4.964~5.033 (m, 1H), 5.129~5.169 (m, 1H), 5.389 (t,J = 1.8 Hz, 1H); ESI-MS m/z 490 ([M+H ]⁺ , 10%). Anal. Calcd for C₃₁ H₄₉ O₂ Cl: C, 76.11; H, 10.10. Found: C, 76.10; H, 10.09.

实施例4。3-苯甲酰基-5,22-二烯-24-乙基-胆甾（AB-4）的合成。Example 4. Synthesis of 3-benzoyl-5,22-diene-24-ethyl-cholesteric (AB-4).

取豆甾醇0.5 g（1.21 mmol）于氮气保护的50 mL三口瓶中，加入干燥的氯仿20 mL，加入无水碳酸钾 0.4 g（2.7 mmol）。控制温度在80℃，缓慢滴加0.7 ml 苯甲酰氯（6.0 mmol）与3 mL 氯仿的混合液，半小时滴加完毕，再继续搅拌，薄层色谱跟踪反应进程，2小时后终止反应。过滤，滤液用饱和氯化钠溶液洗3次，有机相用无水硫酸钠干燥过夜，再次过滤，蒸干滤液，硅胶柱层析（硅胶，200-300目；洗脱剂，石油醚/乙酸乙酯=25:1）纯化，得白色固体粉末0.415g，产率66.4%；m.p.156.4~157.7℃;¹HNMR (CDCl₃) δ(ppm): 0.712 (s, 3H), 0.797~0.811 (m, 6H), 0.852 (d, J = 6.0Hz, 3H), 0.954~1.117 (m, 9H), 1.145~1.348 (m, 5H), 1.395~1.677 (m, 8H), 1.690~1.726 (m, 2H), 1.909~1.931 (m, 1H), 1.989~2.099 (m, 4H), 2.461~2.474 (m, 2H), 4.821~4.899 (m, 1H), 5.031~5.045 (m, 1H), 5.144~5.169 (m, 1H), 5.420 (t, J = 2.4 Hz,1H), 7.429 (t, J = 7.8 Hz, 2H), 7.540 (t, J = 7.8 Hz, 1H), 8.043 (d, J = 7.8 Hz, 2H); ESI-MS m/z 518 ([M+H]⁺, 10%). Anal.Calcd for C₃₆H₅₂O₂: C, 83.67; H, 10.14. Found: C, 83.65; H, 10.13。Take 0.5 g (1.21 mmol) of stigmasterol in a 50 mL three-necked flask protected by nitrogen, add 20 mL of dry chloroform, and add 0.4 g (2.7 mmol) of anhydrous potassium carbonate. Control the temperature at 80°C, slowly add a mixture of 0.7 ml of benzoyl chloride (6.0 mmol) and 3 mL of chloroform dropwise, the dropwise addition is completed in half an hour, then continue to stir, follow the reaction progress by thin-layer chromatography, and stop the reaction after 2 hours. Filter, wash the filtrate 3 times with saturated sodium chloride solution, dry the organic phase with anhydrous sodium sulfate overnight, filter again, evaporate the filtrate to dryness, perform silica gel column chromatography (silica gel, 200-300 mesh; eluent, petroleum ether/acetic acid ethyl ester=25:1) to obtain 0.415g of white solid powder, yield 66.4%; mp156.4~157.7℃;¹ HNMR (CDCl₃ ) δ(ppm): 0.712 (s, 3H), 0.797~0.811 ( m, 6H), 0.852 (d,J = 6.0Hz, 3H), 0.954~1.117 (m, 9H), 1.145~1.348 (m, 5H), 1.395~1.677 (m, 8H), 1.690~1.726 (m, 2H), 1.909~1.931 (m, 1H), 1.989~2.099 (m, 4H), 2.461~2.474 (m, 2H), 4.821~4.899 (m, 1H), 5.031~5.045 (m, 1H), 5.144~ 5.169 (m, 1H), 5.420 (t,J = 2.4 Hz, 1H), 7.429 (t,J = 7.8 Hz, 2H), 7.540 (t,J = 7.8 Hz, 1H), 8.043 (d,J = 7.8 Hz, 2H); ESI-MS m/z 518 ([M+H]⁺ , 10%). Anal. Calcd for C₃₆ H₅₂ O₂ : C, 83.67; H, 10.14. Found: C, 83.65; H , 10.13.

实施例5。3-羟基-5,6,22,23-四溴-24-乙基-胆甾（AB-5）的合成。Example 5. Synthesis of 3-hydroxy-5,6,22,23-tetrabromo-24-ethyl-cholesteric (AB-5).

取豆甾醇0.5 g（1.21 mmol）于氮气保护的50 mL三口瓶中，加入干燥的氯仿20 mL，室温下，缓慢滴加2 ml 苯甲酰氯（12.1 mmol）与3 mL 氯仿的混合液，半小时滴加完毕，再继续搅拌，薄层色谱跟踪反应进程，3.5小时后终止反应。过滤，滤液用饱和碳酸氢钠溶液洗3次，有机相用无水硫酸钠干燥过夜，再次过滤，蒸干滤液，硅胶柱层析（硅胶，200-300目；洗脱剂，石油醚/乙酸乙酯/甲醇=5：1: 0.05）纯化，得红棕色固体粉末0.377g，产率42.6%；m.p.101.9~103.8℃;¹HNMR (CDCl₃) δ(ppm): 0.713 (s, 1H), 0.732 (s, 2H),0.896~1.191 (m, 7H), 1.199~1.397 (m, 8H), 1.401~1.810 (m, 18H), 1.839~1.924 (m, 2H), 1.940~2.012 (m, 2H), 2.197~2.225 (m, 1H), 2.289~2.345 (m, 1H), 2.486~2.511 (m, 1H), 2.813~2.841 (m, 1H), 4.190~4.198 (m, 1H), 4.394~4.416 (m, 1H), 4.480~4.534 (m, 1H), 4.842~4.871 (m, 1H); ESI-MS m/z 733 ([M+H]⁺, 60%). Anal.Calcd for C₂₉H₄₈BrO: C, 47.56; H, 6.61. Found: C, 47.60; H,6.60。Take 0.5 g (1.21 mmol) of stigmasterol in a 50 mL three-neck flask protected by nitrogen, add 20 mL of dry chloroform, and slowly add a mixture of 2 ml of benzoyl chloride (12.1 mmol) and 3 mL of chloroform dropwise at room temperature, half After the dropwise addition was completed within 1 hour, the stirring was continued, and the reaction progress was tracked by thin-layer chromatography, and the reaction was terminated after 3.5 hours. Filter, wash the filtrate 3 times with saturated sodium bicarbonate solution, dry the organic phase with anhydrous sodium sulfate overnight, filter again, evaporate the filtrate to dryness, perform silica gel column chromatography (silica gel, 200-300 mesh; eluent, petroleum ether/acetic acid Ethyl ester/methanol=5:1:0.05) to obtain 0.377g of reddish-brown solid powder with a yield of 42.6%; mp101.9~103.8℃;¹ HNMR (CDCl₃ ) δ(ppm): 0.713 (s, 1H) , 0.732 (s, 2H),0.896~1.191 (m, 7H), 1.199~1.397 (m, 8H), 1.401~1.810 (m, 18H), 1.839~1.924 (m, 2H), 1.940~2.012 (m, 2H), 2.197~2.225 (m, 1H), 2.289~2.345 (m, 1H), 2.486~2.511 (m, 1H), 2.813~2.841 (m, 1H), 4.190~4.198 (m, 1H), 4.394~ 4.416 (m, 1H), 4.480~4.534 (m, 1H), 4.842~4.871 (m, 1H); ESI-MS m/z 733 ([M+H]⁺ , 60%). Anal. Calcd for C₂₉ H₄₈ BrO: C, 47.56; H, 6.61. Found: C, 47.60; H, 6.60.

实施例6。3-乙氧基-5,22-二烯-24-乙基-胆甾（AB-6）的合成。Example 6. Synthesis of 3-ethoxy-5,22-diene-24-ethyl-cholesteric (AB-6).

取氯代豆甾醇0.5 g（1.21 mmol）于氮气保护的50 mL三口瓶中，加入无水乙醇20 mL。控制温度在60℃，搅拌，薄层色谱跟踪反应进程，5小时后终止反应。过滤，蒸干滤液，硅胶柱层析（硅胶，200-300目；洗脱剂，石油醚/乙酸乙酯=20:1）纯化，得白色固体粉末0.174g，产率32.7%；m.p.132.5~133.9℃;¹HNMR (CDCl₃) δ(ppm): 0.696 (s, 3H), 0.790~0.824 (m, 6H), 0.845 (d, J = 6.6Hz, 3H), 0.897~0.945 (m, 1H), 0.986~1.094 (m, 9H), 1.187~1.213 (m, 6H), 1.292~1.342 (m, 1H), 1.345~1.601 (m, 8H), 1.645~1.723 (m, 1H), 1.847~1.979 (m, 2H), 1.998~2.104 (m, 3H), 2.178~2.224 (m, 1H), 2.347~2.392 (m, 1H), 3.152~3.199 (m, 1H), 3.516~3.534 (m, 2H), 5.023~5.038 (m, 1H), 5.133~5.172 (m, 2H), 5.342 (t, J = 3Hz, 1H); Cl, 8.22. ESI-MS m/z 442 ([M+H]⁺, 30%). Anal.Calcd for C₃₁H₅₂O: C, 84.48; H, 11.89. Found: C, 84.49; H, 11.87。Take 0.5 g (1.21 mmol) of chlorostigmasterol in a 50 mL three-necked flask protected by nitrogen, and add 20 mL of absolute ethanol. Control the temperature at 60° C., stir, follow the progress of the reaction by thin-layer chromatography, and terminate the reaction after 5 hours. Filtrate, evaporate the filtrate to dryness, and purify by silica gel column chromatography (silica gel, 200-300 mesh; eluent, petroleum ether/ethyl acetate=20:1) to obtain 0.174 g of white solid powder with a yield of 32.7%; mp132.5 ~133.9℃;¹ HNMR (CDCl₃ ) δ(ppm): 0.696 (s, 3H), 0.790~0.824 (m, 6H), 0.845 (d,J = 6.6Hz, 3H), 0.897~0.945 (m, 1H ), 0.986~1.094 (m, 9H), 1.187~1.213 (m, 6H), 1.292~1.342 (m, 1H), 1.345~1.601 (m, 8H), 1.645~1.723 (m, 1H), 1.847~1.979 (m, 2H), 1.998~2.104 (m, 3H), 2.178~2.224 (m, 1H), 2.347~2.392 (m, 1H), 3.152~3.199 (m, 1H), 3.516~3.534 (m, 2H) , 5.023~5.038 (m, 1H), 5.133~5.172 (m, 2H), 5.342 (t,J = 3Hz, 1H); Cl, 8.22. ESI-MS m/z 442 ([M+H]⁺ , 30 %). Anal. Calcd for C₃₁ H₅₂ O: C, 84.48; H, 11.89. Found: C, 84.49; H, 11.87.

实施例7。3-硝基-5,22-二烯-24-乙基-胆甾（AB-7）的合成。Example 7. Synthesis of 3-nitro-5,22-diene-24-ethyl-cholesteric (AB-7).

取氯代豆甾醇0.5 g（1.21 mmol）于氮气保护的50 mL三口瓶中，加入无水乙醇20 mL。室温下，缓慢滴加1.0 ml 硝酸银的乙醇溶液（6.0 mmol），半个小时滴加完毕，继续搅拌，薄层色谱跟踪反应进程，2小时后终止反应。过滤，蒸干滤液，硅胶柱层析（硅胶，200-300目；洗脱剂，石油醚/乙酸乙酯=3:1）纯化，得白色固体粉末0.266g，产率48.1%；m.p.90.1~92.0℃;¹HNMR (CDCl₃) δ(ppm): 0.702 (s, 3H), 0.792~0.807 (m, 6H), 0.848 (d, J = 6.0Hz, 3H), 0.901~1.103 (m, 9H), 1.145~1.211 (m, 3H), 1.265~1.293 (m, 1H), 1.402~1.599 (m, 10H), 1.623~1.745 (m, 2H), 2.004~2.099 (m, 4H), 2.435~2.448 (m, 2H), 4.746~4.801 (m, 1H), 5.028~5.043 (m, 1H), 5.135~5.174 (m, 1H), 5.440 (t, J = 3Hz, 1H); ESI-MS m/z 458 ([M+H]⁺, 10%). Anal.Calcd for C₂₉H₄₇NO₃: C, 76.10; H, 10.35; N 3.06. Found: C, 76.07; H, 10.36, N 3.07。Take 0.5 g (1.21 mmol) of chlorostigmasterol in a 50 mL three-necked flask protected by nitrogen, and add 20 mL of absolute ethanol. At room temperature, slowly add 1.0 ml of ethanol solution of silver nitrate (6.0 mmol) dropwise. After half an hour, the dropwise addition is completed, and the stirring is continued. The reaction progress is tracked by thin-layer chromatography, and the reaction is terminated after 2 hours. Filtrate, evaporate the filtrate to dryness, and purify by silica gel column chromatography (silica gel, 200-300 mesh; eluent, petroleum ether/ethyl acetate=3:1) to obtain 0.266 g of white solid powder, with a yield of 48.1%; mp90.1 ~92.0℃;¹ HNMR (CDCl₃ ) δ(ppm): 0.702 (s, 3H), 0.792~0.807 (m, 6H), 0.848 (d,J = 6.0Hz, 3H), 0.901~1.103 (m, 9H ), 1.145~1.211 (m, 3H), 1.265~1.293 (m, 1H), 1.402~1.599 (m, 10H), 1.623~1.745 (m, 2H), 2.004~2.099 (m, 4H), 2.435~2.448 (m, 2H), 4.746~4.801 (m, 1H), 5.028~5.043 (m, 1H), 5.135~5.174 (m, 1H), 5.440 (t,J = 3Hz, 1H); ESI-MS m/z 458 ([M+H]⁺ , 10%). Anal. Calcd for C₂₉ H₄₇ NO₃ : C, 76.10; H, 10.35; N 3.06. Found: C, 76.07; H, 10.36, N 3.07.

实施例8。豆甾醇及其衍生物对肿瘤细胞株生长的抑制作用。Example 8. Inhibitory effect of stigmasterol and its derivatives on the growth of tumor cell lines.

选择本发明的衍生物以及天然豆甾醇对肝癌上皮细胞（HEPG-2）、人胃癌细胞（MKN45）、人乳腺癌细胞（MDA-MB-231）、人卵巢癌细胞（SKOV3）的细胞毒作用。采用MTT法，测定其吸光度，分别计算抑制细胞生长达50%时化合物浓度，以IC₅₀表示。实验结果见下表。Selected derivatives of the present invention and the cytotoxic effect of natural stigmasterol on liver cancer epithelial cells (HEPG-2), human gastric cancer cells (MKN45), human breast cancer cells (MDA-MB-231), human ovarian cancer cells (SKOV3) . The absorbance was measured by MTT method, and the concentration of the compound when the cell growth was inhibited by 50% was calculated respectively, expressed as IC₅₀ . The experimental results are shown in the table below.

the

表1 不同样品对抑制NKM45细胞增殖的IC₅₀Table 1 IC₅₀ of different samples on inhibiting the proliferation of NKM45 cells

表2 不同样品对抑制SKOV3细胞增殖的IC₅₀Table 2 IC₅₀ of different samples on inhibiting the proliferation of SKOV3 cells

表3 不同样品对抑制HEPG-2细胞增殖的IC₅₀Table 3 IC₅₀ of different samples on inhibiting the proliferation of HEPG-2 cells

表4 不同样品对抑制MDA-MB-231细胞增殖的IC₅₀Table 4 IC₅₀ of different samples on inhibiting the proliferation of MDA-MB-231 cells

Claims

1. a Stigmasterol derivative is characterized in that having as shown in the formula structure shown in I or formula II:

The formula I

The group of the R representative in the formula I is: SO₂c₆h₅, COCH₃, COCH₂cl, COC₆h₅, CH₂cH₃, NO₂;

The formula II.

2. the application of Stigmasterol derivative claimed in claim 1 in the medicine of preparation treatment cancer.

3. a medicine that is used for the treatment of cancer, is characterized in that containing Stigmasterol derivative claimed in claim 1 and pharmaceutically acceptable auxiliary.

4. medicine according to claim 3, is characterized in that this medicine is injection, tablet, pill, capsule, suspension agent or emulsion.