WO2012006864A1 - 一种三萜皂苷类化合物，制备方法及用途 - Google Patents

一种三萜皂苷类化合物，制备方法及用途 Download PDF

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WO2012006864A1
WO2012006864A1 PCT/CN2011/000970 CN2011000970W WO2012006864A1 WO 2012006864 A1 WO2012006864 A1 WO 2012006864A1 CN 2011000970 W CN2011000970 W CN 2011000970W WO 2012006864 A1 WO2012006864 A1 WO 2012006864A1
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李滨生
刘沭军
姜生
宋建平
邱峰
于丽丽
魏星
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哈尔滨三乐生物工程有限公司
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7028—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages
    • A61K31/7034—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin
    • A61K31/704—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin attached to a condensed carbocyclic ring system, e.g. sennosides, thiocolchicosides, escin, daunorubicin
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    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P13/00—Drugs for disorders of the urinary system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P13/00—Drugs for disorders of the urinary system
    • A61P13/10—Drugs for disorders of the urinary system of the bladder
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P13/00—Drugs for disorders of the urinary system
    • A61P13/12—Drugs for disorders of the urinary system of the kidneys
    • A—HUMAN NECESSITIES
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    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P21/00—Drugs for disorders of the muscular or neuromuscular system
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    • A—HUMAN NECESSITIES
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    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/14—Drugs for disorders of the nervous system for treating abnormal movements, e.g. chorea, dyskinesia
    • A61P25/16—Anti-Parkinson drugs
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/04—Antibacterial agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07J—STEROIDS
    • C07J63/00—Steroids in which the cyclopenta(a)hydrophenanthrene skeleton has been modified by expansion of only one ring by one or two atoms
    • C07J63/008—Expansion of ring D by one atom, e.g. D homo steroids

Definitions

  • the present invention relates to the field of medical technology, and in particular to a class of pentacyclic triterpenoids and a preparation method thereof.
  • BACKGROUND OF THE INVENTION The fruit of the genus Xanthoceras sorbifolia Bunge is a fruit of the perennial herb of the genus Uncle.
  • Xanthoceras is a perennial plant native to northern China and is now cultivated. Xanthoceras is mainly used for the treatment of rheumatoid arthritis, rheumatoid arthritis, and skin rheumatism. It was included in the Chinese Pharmacopoeia in 1977.
  • Chinese patents (CN1092992A) and (CN1092991A) disclose the powder of the genus Pleurotus ostreatus, and the Chinese patent (CN1052636C).
  • the ethanol extract of the powder of the genus Corydalis can effectively treat enuresis and may improve brain function and increase intelligence. .
  • R1 and R2 represent the same or different monosaccharides, disaccharides or polysaccharides;
  • R3 and R4 represent the same or different hydrogen, C1-C6 sulfhydryl, monosaccharide, disaccharide or polysaccharide;
  • R5 represents hydrogen, C1-C6 Anthracenyl, C1-C7 3 ⁇ 4 hydrocarbon, methylene, carbonyl, sulfonyl or acyl;
  • R6 and R7 represent the same or different hydrogen, halogen, hydroxy, thiol, C1-C6 fluorenyl or acyl group.
  • the monosaccharide, the disaccharide or the polysaccharide is a glycoside of 1 to 5 units, and the glycoside is a sugar of glucose, galactose, rhamnose, xylose, arabinose or glucuronic acid.
  • R1 and R2 represent the same or different 1 to 3 units of ⁇ -D-glucose.
  • R3 and R4 represent hydrogen.
  • R5 represents a carbonyl group.
  • R1 and R2 represent the same or different monosaccharides, disaccharides or polysaccharides;
  • R3 and R4 represent the same or different hydrogen, C1-C6 sulfhydryl, monosaccharide, disaccharide or polysaccharide;
  • R5 represents hydrogen, C1-C6 Mercapto, C1-C7 olefin, methylene, carbonyl, sulfonyl or acyl;
  • R6 and R7 represent the same or different hydrogen, halogen, light, thiol, C1-C6 fluorenyl or acyl group.
  • the monosaccharide, disaccharide or polysaccharide is a glycoside of 1 to 5 units, and the saccharide is a sugar of glucose, galactose, rhamnose, xylose, arabinose or glucuronic acid.
  • R1 and R2 represent the same or different 1 to 3 units of ⁇ -D-glucose.
  • R3 and R4 represent hydrogen.
  • R5 represents a carbonyl group.
  • Compounds of formula I and II can be extracted from the shell, stem, stem, leaf, seed, seed, root or bark of Xanthoceras, or can be isolated from other plants, or synthesized. The method is obtained.
  • the compounds of the present invention are generally present in a mixture of stereoisomeric forms, and in these stereoisomeric forms, usually in one form, which can be obtained according to conventional techniques known to those of ordinary skill in the art. isomer.
  • Representative structures of compounds I and II can be seen in the table below. Representative structure of compound I
  • the invention further provides the compound of the formula I, II as an active ingredient, which has anti-inflammatory, anti-tumor, antibacterial and anti-viral activity, and can be used for treating frequent urination, urgency, enuresis, urinary incontinence and overactive bladder. It can also prevent brain aging, enhance memory, improve brain function, treat mental retardation, dementia, senile dementia, autism, brain trauma, Parkinson's disease and other diseases caused by impaired brain function or disorders, and can also be used to treat arthritis. Rheumatism, circulatory dysfunction, arteriosclerosis, Raynaud's disease, angina pectoris, cardiac dysfunction, headache, dizziness, renal dysfunction, impotence and premature ejaculation. These compounds have a variety of pharmaceutical and medical uses.
  • the effect of the compound of the present invention on isolated bladder smooth muscle of rats antagonizes the effect of Ach (acetylcholine) contraction on isolated bladder smooth muscle of rats, has a relaxing effect on bladder detrusor, and is related to antagonizing choline-like action.
  • the effect of Compound I on contraction of bladder sphincter was demonstrated by the effects of female infertility and post-natal recovery of rat bladder sphincter and the bioelectricity of extracellular electrodes of rabbit bladder external sphincter.
  • the effects of the extract of Xanthoceras sorbifolia L The effects of the extract of Xanthoceras sorbifolia L.
  • compositions of the present invention comprise a therapeutically effective amount of a compound of formula I, II and compositions and monomers thereof as an active ingredient, and one or more pharmaceutically acceptable carriers.
  • the compounds and pharmaceutical compositions of this invention are useful in the preparation of pharmaceutical compositions for the treatment of urinary incontinence and overactive bladder.
  • the pharmaceutically acceptable carrier described above refers to a conventional pharmaceutical carrier in the pharmaceutical field, for example, a diluent, an excipient such as water, a filler such as starch, sucrose, etc.; a binder such as a cellulose derivative, alginic acid Salts, gelatin and polyvinylpyrrolidone; wetting agents such as glycerin; disintegrants such as agar, calcium carbonate and sodium hydrogencarbonate; absorption enhancers such as quaternary compounds; surfactants such as hexadecanol; adsorption carriers such as kaolin And soap clay; lubricants such as talc, calcium and magnesium stearate, and polyethylene glycol.
  • a diluent an excipient such as water, a filler such as starch, sucrose, etc.
  • a binder such as a cellulose derivative, alginic acid Salts, gelatin and polyvinylpyrrolidone
  • wetting agents such as
  • the compounds of the invention may be administered to a patient in need of such treatment by oral, nasal inhalation, rectal or parenteral administration in the form of a composition.
  • oral administration it can be formulated into conventional solid preparations such as tablets, powders, granules, capsules, etc., into liquid preparations such as water or oil suspensions or other liquid preparations such as syrups; for parenteral administration In this case, it can be made into a solution for injection, water or an oily suspension, and the like.
  • Preferred forms are tablets, coated tablets, capsules, suppositories, nasal sprays and injections.
  • the various dosage forms of the pharmaceutical compositions of the present invention can be prepared according to conventional methods of manufacture in the pharmaceutical arts.
  • the active ingredient is mixed with one or more carriers which are then brought into the preparations required.
  • the pharmaceutical composition of the present invention preferably contains from 0.1% to 99.5% by weight of the active ingredient, and most preferably from 0.5% to 95% by weight of the active ingredient.
  • Example 1 8 kg of the fruit seed powder was extracted twice with 70 L of 70% ethanol, respectively, for 2 hours, filtered while hot, and the filtrate was concentrated under reduced pressure to a non-alcoholic taste to obtain 20 L of concentrated liquid. The supernatant was eluted with a D101 macroporous resin column with different concentrations of ethanol water to obtain water, 30%, 60%, 70% and 90% ethanol. The 60% ethanol elution fraction was mixed with 140 mesh silica gel to
  • ODS column was used in different proportions of methanol aqueous solution (10%, 20%, 25%). , 30%, 35%, 40%, 45%, 100%) Gradient elution, according to liquid phase detection, 17 ⁇ 25 fractions were prepared by preparative high performance liquid phase with 40% methanol aqueous solution as mobile phase. , 3 compounds yw517-l ⁇ yw517-3 were obtained.
  • Yw517-3 is a colorless transparent crystal.
  • the 10% sulfuric acid ethanol solution was purple and the Liebermann-Burchard reaction was positive.
  • the structure of the compound yw517-3 is 3-O-yff-D-glucopyranosyl-(l-
  • Yw517-2 colorless transparent crystal.
  • the 10% sulfuric acid ethanol solution was purple, and the Liebermann-Burchard reaction was positive.
  • ESI-MS gives the excimer ion peak m/z 1321 [M+Na] + , and the structure of compound yw517-2 is 3 -0-5-D-glucopyranosyl-( 1 ⁇ 6)-ff-D-glucopyranosyl, 28 -O-yff-D-glucopyranosyl [( 1 -6)- 9-D-glucopyranosyl-(l-2)]- yi-D-glucopyranosyl 16-oxobarringtogenol C.
  • the 1H-NMR, 13 C-NMR data are shown in Table 3.
  • H-1 "" (S 4.97) and C-28 5 73.4) are remotely related, H-1 "'" (S 5.10) and C-6
  • '" ( ⁇ 70.5) is determined to be 3-0-?-D-glucopyranosyl[(l-6)-yff-D-glucopyranosyl-(l -*2)] - y9-D-glucopyranosyl, 28-O-y9-D-glucopyranosyl-(l ⁇ 6)-ff-D-glucopyranosyl- 16-oxobarringtogenol C.
  • the ⁇ -NMR, 13 C-NMR data are shown in Table 4.
  • Example 2 dried fruit dried fruit shell 15K g , crushed, with 70% ethanol (each time 8 times the amount was refluxed three times for 2 hours each time, and the extract was evaporated under reduced pressure to give a solvent.
  • Example 3 The cytotoxic activity of different compounds in human melanoma (A375-S2) and human cervical cancer cells (HeLa) was compared by the MTT method. The IC 5Q values of the above two cell lines are shown in Table 6. Place

Abstract

本发明公开一类五环三萜类化合物及其制备方法，该化合物具有抗炎、抗肿瘤、抗菌、抗病毒活性，能够治疗尿失禁和膀胱过度活动症。

Description

一种三萜皂苷类化合物， 制备方法及用途 技术领域 本发明属于医药技术领域， 具体涉及一类五环三萜类化合物及其制备方法。 背景技术 文冠果别名文官果、岩木瓜，为无患子科文冠果属多年生草本植物 Xanthoceras sorbifolia Bunge的果实。 文冠果为多年生植物， 原产于我国北方,现多栽培。 文 冠果主要用于祛风湿性关节炎、风湿内热、皮肤风湿等症的治疗， 1977 年收载 于中国药典。 文冠果种子在民间用于治遗尿症， 茎叶在蒙药中治风湿， 已有很 长的历史。 但是， 对文冠果药用方面的深入幵发研究才是近年来的事。 中国专利（CN1092992A)和 （CN1092991A)公开文冠果种仁的粉， 和中国 专利 （CN1052636C) 公开文冠果种仁的粉的乙醇提取物可以有效的治疗遗尿， 并可能改善脑功能和增智。 沈阳药科大学学报 （2001， 18 (1): 53-56) 报导了文 冠果的木材的正丁醇提取物和它的抗炎性作用。 美国专利申请公开书（NO20030096030)公开了文冠果果壳提取物和所含的 四种皂苷和两个甾醇化合物， 以及它们改善功能和抗衰老的功能， 和用于治疗 夜尿， 尿频， 尿失禁， 弱智， 痴呆和增强人体抗御糖尿病的方法。 发明内容： 本发明公开如 I式的五环三萜类化合物， 本发明提供从文冠果中提取得到的 I式化合物的制备方法。 本发明进一步提供 I式化合物为活性成分的， 具有抗炎、 抗肿瘤、 抗菌、 抗 病毒活性， 能够用于治疗尿失禁和膀胱过度活动症。 具有 I 式的化合物
Figure imgf000003_0001
I 其中 Rl及 R2表示相同或不同的单糖、 双糖或多糖； R3及 R4表示相同或 不同的氢、 C1-C6的垸基、单糖、双糖或多糖； R5表示氢、 C1-C6的垸基、 C1-C7 ¾烃、 亚甲基、 羰基、 磺酰基或酰基； R6及 R7表示相同或不同的氢、 卤素、 羟基、 硫醇基、 C1-C6的垸基或酰基。 上述 I 式的化合物中单糖、 双糖或多糖为 1至 5个单元的糖元， 糖元为葡萄 糖、 半乳糖、 鼠李糖、 木糖、 阿拉伯糖或葡萄糖醛酸的糖。 上述 I 式的化合物中 R1及 R2表示相同或不同的 1至 3个单元的 Ρ -D-葡萄 糖。 上述 I 式的化合物中 R3及 R4表示氢。 R5表示羰基。 具有 Π式的化合物
Figure imgf000004_0001
其中 Rl及 R2表示相同或不同的单糖、 双糖或多糖； R3及 R4表示相同或 不同的氢、 C1-C6的垸基、单糖、双糖或多糖； R5表示氢、 C1-C6的垸基、 C1-C7 烯烃、 亚甲基、 羰基、 磺酰基或酰基； R6及 R7表示相同或不同的氢、 卤素、 轻基、 硫醇基、 C1-C6的垸基或酰基。 上述 II式的化合物中单糖、双糖或多糖为 1至 5个单元的糖元，糖元为葡萄 糖、 半乳糖、 鼠李糖、 木糖、 阿拉伯糖或葡萄糖醛酸的糖。 上述 II 式的化合物中 R1及 R2表示相同或不同的 1至 3个单元的 β -D-葡萄 糖。 上述 II 式的化合物中 R3及 R4表示氢。 R5表示羰基。
I 式、 II式的化合物可以从文冠果的果壳、 果柄、 枝干、 叶、 种仁、 种壳、 根或树皮中提取得到， 也可以从其他植物中分离得到， 或用合成方法得到。 本发明的化合物通常是以立体异构形式的混合物存在的， 在这些立体异构体 形式中， 通常是以一种形式为主， 可依据本领域普通技术人员所掌握的常规技 术得到所需的异构体。 在下表中可见化合物 I、 II的代表性结构 合物 I的代表性结构
Figure imgf000005_0001
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Figure imgf000006_0001
本发明进一步提供 I、 II式化合物为活性成分的， 具有抗炎、 抗肿瘤、 抗菌、 抗病毒活性， 能够用于治疗尿频、 尿急、 遗尿、 尿失禁和膀胱过度活动症。 还 可防治脑衰老、 增强记忆、 改善脑功能， 治疗弱智、 痴呆、 老年痴呆， 孤独症、 脑创伤、 帕金森病和其他由于脑功能不健全或障碍引起的疾病， 还可用于治疗 关节炎、 风湿症、 循环不良、 动脉硬化、 雷诺氏症、 心绞痛、 心脏功能紊乱、 头痛、 头晕、 肾功能紊乱、 阳痿和早泄。 这些化合物具有各种药学和医疗学用 途。 经药理活性研究，本发明的化合物对大鼠离体膀胱平滑肌的影响，拮抗 Ach (乙酰胆碱） 收缩大鼠离体膀胱平滑肌的作用， 对膀胱逼尿肌有松弛作用， 和 拮抗胆碱样作用有关。 通过对雌性未育及育后恢复大鼠离体膀胱括约肌的影响 和对家兔膀胱外括约肌细胞外电极生物电的影响， 证明 I式化合物对膀胱括约 肌有收缩作用。 通过文冠果提取物对水负荷小鼠排尿次数的影响和大鼠阴部神 经损伤后漏尿点压的影响，证明本发明化合物能改善尿频、遗尿、尿失禁等 OAB 的症状。 本发明的药物组合物含有治疗有效量的 I、 II式化合物和组合物及其单体为 活性成分， 以及含有一种或多种药学上可接受的载体。 本发明的化合物和药物 组合物可用于制备治疗尿失禁和膀胱过度活动症的药物组合物。 上文所述药学 上可接受的载体是指药学领域常规的药物载体， 例如： 稀释剂、赋形剂如水等， 填充剂如淀粉、蔗糖等、； 粘合剂如纤维素衍生物、 藻酸盐、 明胶和聚乙烯毗咯 垸酮； 湿润剂如甘油； 崩解剂如琼脂、 碳酸钙和碳酸氢钠； 吸收促进剂如季按 化合物； 表面活性剂如十六垸醇； 吸附载体如高岭土和皂粘土； 润滑剂如滑石 粉、 硬脂酸钙和镁、 和聚乙二醇等。 另外还可以在组合物中加入其它辅剂如香 味剂、 甜味剂等。 本发明化合物可以组合物的形式通过口服、 鼻吸入、 直肠或 肠胃外给药的方式施用于需要这种治疗的患者。 用于口服时， 可将其制成常规 的固体制剂如片剂、 粉剂、 粒剂、 胶囊等， 制成液体制剂如水或油悬浮剂或其 它液体制剂如糖浆剂等； 用于肠胃外给药时， 可将其制成注射用的溶液、 水或 油性悬浮剂等。 优选的形式是片剂、 包衣片剂、 胶囊、 栓剂、 鼻喷雾剂和注射 剂。 本发明药物组合物的各种剂型可以按照药学领域的常规生产方法制备。 例如 使活性成分与一种或多种载体混合， 然后将其制成所需的剂型。 本发明的药物 组合物优选含有重量比为 0.1 %— 99.5 %的活性成分，最优选含有重量比为 0.5 %— 95 %的活性成分。 具体实施方式下面的实施例可以使本专业技术人员更全面地理解本发明，但 不以任何方式限制本发明。
实施例 1 文冠果种仁粉末 8公斤， 分 2次， 分别用 70%乙醇 30L提取 3次， 每次 2 小时， 趁热过滤， 滤液减压浓缩至无醇味， 得浓缩液 20L。 上清液采用 D101 大孔树脂柱以不同浓度的乙醇水梯度洗脱， 得到水， 30%、 60%、 70%和 90% 乙醇五个部分。 其中 60%乙醇洗脱部分以 140目硅胶拌样， 以
CHCl3:MeOH(4:l,2: l,l : l), CHCl3:MeOH:H20(6:3:l下层)为洗脱剂经硅胶柱色谱 分离得到 80个流分， 经薄层分析将成分一致部分合并。 其中 53~54部分再次 进行硅胶柱层析， 洗脱剂 CHCl3:MeOH:H2O(65:30: 10)下层， 等度洗脱， 得到 38 个流分。 其中 20~28合并以甲醇溶解进行 sephadexLH-20柱层析， 甲醇为洗脱 剂， 经薄层色谱检验将主含皂苷部分合并， 采用 ODS柱不同比例甲醇水溶液 ( 10%, 20%, 25%, 30%, 35%, 40%, 45%, 100%)梯度洗脱， 根据液相检测 对其中的 17~25流分分别采用制备高效液相以 40%甲醇水溶液为流动相进行了 分离制备， 得到 3个化合物 yw517-l~yw517-3。
Yw517-3无色透明晶体。 10%硫酸乙醇溶液显紫色， Liebermann-Burchard 反应 阳性。 化合物 yw517-3的结构为 3-O-yff-D-glucopyranosyl-(l—
6)-9-D-glucopyranosyl, 28-0-^-D-glucopyranosyl[(l—6)- y5-D-glucopyranosyl-(l -* 2)]- /?-D-glucopyranosyl 16-deoxybarringtogenol C。其 1H-NMR, 13C-NMR数据见 表 2。 (ui 'Hl)t76 'Ηΐ)38'ΐ '(o 'Hl)g8'l ZZ π
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Yw517-2 无色透明晶体。 10%硫酸乙醇溶液显紫色， Liebermann-Burchard 反应阳性。 ESI-MS给出准分子离子峰 m/z 1321 [M+Na]+, 化合物 yw517-2的结构为 3 -0-5-D-glucopyranosyl-( 1→ 6)-ff-D-glucopyranosyl, 28-O-yff-D-glucopyranosyl [( 1 —6)- 9-D-glucopyranosyl-(l—2)]- yi-D-glucopyranosyl 16-oxobarringtogenol C。其 1H-NMR, 13C-NMR数据见表 3。
Figure imgf000011_0001
表 3. 化合物 yw517-2 氢谱 （600MHz：)、 碳谱 (150MHz)数据
Yw-517-2
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Yw517-1 无色透明晶体。 10%硫酸乙醇溶液显紫色， Liebermann-Burchard 反应阳性。 结 合 1H NMR、 13C NMR推断其分子式为 C6oH9803o。
1H NMR谱中给出七个甲基信号： 0.89(3H, s), 1.08(3H, s), 1.11(3H, s), 1.21(3H, s), 1.23(3H, s), 1.25(3H, s)， 1.28(3H, s), 低场区给出 12位烯氢质子信 ： δ 5.51(lH, t，J= 4.0Hz), 5个糖端基质子信号： 4.86(1H, d, J= 8.2)， 4.97(1 H, d, J= 7.9)， 5.35(1H, d, J= 7.7 ), 5.10(1H, d, J= 2.9)， 5.11(1H, d, J= 2.8)。
13CNMR谱中给出 60个碳信号， 除去 30个糖的碳信号， 其余的 30个三萜母 核碳信号中， 在 370.0-90.0的连氧碳区给出 4个信号， 分子中存在四羟基取代， 3216.3表明母核上有一酮羰基。 在 HMBC谱中 (54.86(H- 1 ')与母核 3位碳信号 δ (89.4)存在远程相关， Η- Γ" 5.11 ) 和 C-6' (δ 70.7)存在远程相关， Η-Γ (S 5.35)和 C-2' (δ 83.3)存在远程相关。 H-1 "" (S 4.97)和 C-28 5 73.4)存在远程相关， H-1 "'" (S 5.10)和 C-6'" (δ 70.5)存在远程相 关 综上分析， 确定化合物 yw517-l的结构为 3-0- ?-D-glucopyranosyl[(l— 6)- yff-D-glucopyranosyl-(l -*2)]- y9-D-glucopyranosyl， 28-O-y9-D-glucopyranosyl-(l→ 6)-ff-D-glucopyranosyl- 16-oxobarringtogenol C。 其 ^-NMR, 13C-NMR数据见表 4。
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表 4. 化合物 yw517-l 氢谱 （600MHz)、 碳谱 (150MHz)数据 (£· Vn 'pp ' )sv u9 (o 'uq 'HOIS'S Ζ\
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实施例 3 采用 MTT法， 比较了实施例 2中不同化合物对人黑色素瘤 （A375-S2) 和 人宫颈癌细胞 （HeLa) 的细胞毒活性， 其对以上两种细胞株 IC5Q值见表 6所
表 6三萜类化合物体外肿瘤抑制活性测试结果 （IC5t) g/ml)
1 2 3 4 5 6 7 8 9 10 11 12 A375-S2 4.9 23.5 31.5 46.6 63.1 >100 47.4 13.8 35.9 37.8 42.0 42.0 HeLa 20.5 20.6 >100 >100 >100 30.0 34.4 52.6 >100 >100 >100 >100

Claims

权 利 要 求
1、 一种具有下式〖的化合物
Figure imgf000021_0001
其中 R1及 R2表示相同或不同的单糖、 双糖或多糖；
R3及 R4表示相同或不同的氢、 C1-C6的垸基、 单糖、 双糖或多糖；
R5表示氢、 C1-C6的烷基、 C1-C7烯烃、 亚甲基、 羰基、 磺酰基或酰基；
R6及 R7表示相同或不同的氢、 卤素、 羟基、 硫醇基、 C1-C6的烷基或酰基。
2、 一种具有下式 II的化合物
Figure imgf000021_0002
其中 R1及 R2表示相同或不同的单糖、 双糖或多糖；
R3及 R4表示相同或不同的氢、 C1-C6的烷基、 单糖、 双糖或多糖;
R5表示 0、 S 或 N; 、 如权利耍求 1或 2所述的化合物，其中单糖、双糖或多糖为 1至 5个单元的糖元，糖元为葡萄糖、 半乳糖、 鼠李糖、 木糖、 阿拉伯糖或葡萄糖醛酸的糖。 、 如权利耍求 3所述的化合物， 其中 R1及 R2表示相同或不同的 1至 3个单元的 β -D-葡萄糖。 、 如权利要求 3所述的化合物， 其中 R3及 R4表示氢。 、 如权利要求 1所述的化合物， 其中 R5表示羰基。 、 如权利要求 1或 2所述的化合物在制备治疗尿失禁药物中的应用。 、 如权利耍求 1或 2所述的化合物在制备治疗膀胱过度活动症药物中的应用。 、 一种药物组合物，所述组合物包含 0.1 %— 99.5 %的权利要求 1至 6任一所述化合物和药用载体。 、 一种制备权利耍求 1至 6任一所述化合物的方法， 所述方法包括将文冠果的果壳、 果柄、 枝 干、 叶、 种仁、 种壳、 根或树皮作为原料， 用溶剂提取并进行分离。
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