WO2006094456A1 - An extract of styrax linn, the preparation and uses thereof - Google Patents

An extract of styrax linn, the preparation and uses thereof Download PDF

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Publication number
WO2006094456A1
WO2006094456A1 PCT/CN2006/000341 CN2006000341W WO2006094456A1 WO 2006094456 A1 WO2006094456 A1 WO 2006094456A1 CN 2006000341 W CN2006000341 W CN 2006000341W WO 2006094456 A1 WO2006094456 A1 WO 2006094456A1
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Prior art keywords
benzoin
extract
plant
benzofuran
jasmine
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PCT/CN2006/000341
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French (fr)
Chinese (zh)
Inventor
Qilin Li
Guolin Zhang
Jufang Yan
Xin Chen
Yu Huang
Bogang Li
Zhongrong Liu
Yuanqiao Ji
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Chengdu Di'ao Jiuhong Pharmaceutical Factory
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Priority claimed from CNB2005100204796A external-priority patent/CN100420690C/en
Priority claimed from CN200510020478A external-priority patent/CN1830435B/en
Priority claimed from CNB2005100204777A external-priority patent/CN100509803C/en
Application filed by Chengdu Di'ao Jiuhong Pharmaceutical Factory filed Critical Chengdu Di'ao Jiuhong Pharmaceutical Factory
Publication of WO2006094456A1 publication Critical patent/WO2006094456A1/en

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P5/00—Drugs for disorders of the endocrine system
    • A61P5/24—Drugs for disorders of the endocrine system of the sex hormones
    • A61P5/30—Oestrogens
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D307/00—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom
    • C07D307/77—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom ortho- or peri-condensed with carbocyclic rings or ring systems
    • C07D307/78—Benzo [b] furans; Hydrogenated benzo [b] furans
    • C07D307/79—Benzo [b] furans; Hydrogenated benzo [b] furans with only hydrogen atoms, hydrocarbon or substituted hydrocarbon radicals, directly attached to carbon atoms of the hetero ring
    • C07D307/80—Radicals substituted by oxygen atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D407/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having oxygen atoms as the only ring hetero atoms, not provided for by group C07D405/00
    • C07D407/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having oxygen atoms as the only ring hetero atoms, not provided for by group C07D405/00 containing two hetero rings
    • C07D407/04—Heterocyclic compounds containing two or more hetero rings, at least one ring having oxygen atoms as the only ring hetero atoms, not provided for by group C07D405/00 containing two hetero rings directly linked by a ring-member-to-ring-member bond
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D407/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having oxygen atoms as the only ring hetero atoms, not provided for by group C07D405/00
    • C07D407/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having oxygen atoms as the only ring hetero atoms, not provided for by group C07D405/00 containing three or more hetero rings

Definitions

  • the present invention relates to a traditional Chinese medicine extract, a preparation method and use thereof, and more particularly to a succulent plant extract, a preparation method thereof and use thereof. Background technique
  • Estrogen (especially estrogen E 2 ) is one of the most important hormones in humans and other higher animals and has a wide range of physiological functions. 90% of women before menopause have sputum 2 , 50% from the ovary and adrenal gland, women after menopause, due to ovarian atrophy, ovarian function is significantly reduced, so estrogen secretion is greatly reduced, when the estrogen produced by fat cells becomes estrogen in the body The main source, if this source is insufficient, E 2 in the blood is only equivalent to the level of early follicular phase of women before menopause, which may cause bone density to decrease and lead to osteoporotic fracture.
  • the average life expectancy of a woman is 80 years old, then one third of a woman's life is after menopause, and about a quarter of people will get osteoporosis after age 65. As the population ages, the number of patients suffering from menopausal syndrome and osteoporosis will increase. In addition, some non-menopausal women may have premature ovarian failure or ovarian dysfunction, insufficient estrogen levels, and some diseases such as polyposis, infertility and other diseases are also associated with a relative reduction in estrogen levels. Therefore, drugs that seek to improve ovarian function and promote estrogen synthesis help prevent and treat these diseases.
  • Si racflcefle also known as wild jasmine, oleanaceae, about 11 genera, 180 species. China produces 9 genera, 50 species, 9 varieties, and is widely distributed.
  • Sty rax Linn also known as wild jasmine, has about 130 species. There are about 30 species and 7 varieties in China, which are mainly produced in the provinces and regions south of the Yangtze River basin. Most of the benzoin plants are available for viewing, and some are available for medicinal purposes. Its resin contains more scented acid, called “benzoin", which has the effect of clearing the spirit, promoting blood circulation and relieving pain. It is a valuable medicine in medicine; this resin can also be used to make high-grade aromatic oil.
  • the technical proposal of the present invention is to provide a benzoin plant extract and a preparation method thereof, and a pharmaceutical preparation processed from the extract for preventing or treating premature ovarian failure, climacteric syndrome, osteoporosis and ovarian dysfunction in adolescence, Use in multidrug ovarian syndrome, infertility, and pharmaceutical preparations for diseases caused by estrogen deficiency or relative deficiency.
  • the present invention provides a benzoin plant which contains a benzofuran total benzofuran in a weight percentage of 40 to 98%.
  • the benzofuran extract of the benzoin plant contains one or more benzofuran biomonomers having the following chemical structures: wherein 3, 6, 7, 8, 12 are novel compounds.
  • the extract also contains one or more benzofuran derivative monomers having the following chemical structure:
  • the benzofuran extract of the benzoin plant provided by the present invention is obtained by extracting plants of the genus [styrax]. More specifically, it is into the foot of Shishan Anxiang [Styrax perkinsiaej, Luchun benzoin 5. macranthus Pert], Chuxiong benzoin 5. limprichtii Lingelsh et BorzaJ, Vietnamese benzoin 5. tonkinensis (Pierre) Craib et Hartw.], Zhejiang benzoin 5 ⁇ Zhejiangensis SM Hwang et LL Yu], Fragrant benzoin 5.
  • the present invention also provides a method for preparing the above-mentioned benzofuran extract of benzoin, comprising the following steps:
  • extract with water, alcohol (methanol or ethanol), or aqueous alcohol, room temperature or heating (including reflux) for a certain period of time can be repeatedly extracted 1 ⁇ 3 times, and then filter the dregs , the extract liquid is concentrated under reduced pressure to obtain a total extract;
  • c, b step After the extracted aqueous layer is adsorbed by a macroporous resin column, it is eluted with a certain amount of water to remove the sugar component, and then the column is eluted with 30 to 95% ethanol, and the eluate is concentrated under reduced pressure. The extract is then vacuumed or freeze-dried into a dry powder of the benzofuran extract of benzofuran.
  • the extraction solvent is preferably a 50-90% ethanol solution.
  • the amount of the ethanol solution is 6-12 times the weight of the medicinal material, and 8 times is preferred.
  • Macroporous resin models are: D101, HPD100, HPD200, HPD300, LD140.
  • the latter step of the macroporous resin column is preferably 95% ethanol.
  • the dry powder prepared above is a light brown solid substance with a slight bitter taste.
  • the total benzofuran contained therein is 40% - 98% of the total weight.
  • the present invention also provides a method for preparing the above-mentioned benzofuran derivative monomer of the genus Benzoin, which is characterized by comprising the following steps:
  • extract with water, methanol or ethanol, or aqueous alcohol, room temperature or heating (including reflux) for a certain period of time can be repeatedly extracted 1 ⁇ 3 times, and then filter the dregs, the filtrate Concentration under reduced pressure gave a total extract.
  • the present invention also provides an extract of the genus Benzoin prepared by the above method.
  • the extract of the genus benzoin is Watzan benzoin "Sz ⁇ ra perkinsiaej, S. macranthus Perk., S. limprichtii Lingelsh et Borza", Vietnamese benzoin [S. tonkinensis (Pierre) Craib Et Hartw.], S. zhejiangensis SM Hwang et LL Yu, S. odoratissimus Champ., S. chinehsis H et SY Liang, S. benzoinoides Graib S. benzoin Dry and], S. argentifolius HL Li, ash 1 "S.
  • serrulatus Roxb sapphire scent [ An extract of any one or more of S. suberifolius Hook, et Am], S. wilsonii Rehd, S. ferrugineus, S. officinalis, S. obassia Sieb. et Zucc.
  • tile The extract of S. macranthus Perk. is an extract of the fruit of the sage of the scent of the scent of the scent of the scent of the scent of the scent of the scent of the scent of the scent of the scent of the scent of the scent of the sage Extract.
  • the present invention also provides the benzoin plant of the present invention or the total benzofuran extract thereof or the benzofuran derivative monomer of the benzoin plant for the prevention or treatment of premature ovarian failure, climacteric syndrome, osteoporosis and ovarian ovarian Uses in drugs with low function, multiple ovarian syndrome, infertility, and diseases caused by estrogen deficiency or relative deficiency.
  • the active ingredient of the medicament comprises the above-mentioned benzoin plant prodrug or its total benzofuran extract or benzoin benzofuran derivative monomer, or may contain a benzoin plant prodrug or its total benzofuran extract.
  • a compound of a monomer or a benzofuran derivative monomer that is, a western medicine or a traditional Chinese medicine active ingredient which is compatible with the synthesis of estrogen.
  • the preparation is an oral tablet, a capsule, an oral solution or an injection.
  • 1 tablet including ordinary tablets, film tablets, enteric tablets and the like.
  • dry powder of total benzofuran extract add appropriate amount of diluent such as starch, dextrin, nectar, microcrystalline fiber, etc., appropriate amount of binder such as water, ethanol, starch slurry, gelatin, cellulose, etc., moderate amount of disintegration
  • diluent such as starch, dextrin, nectar, microcrystalline fiber, etc.
  • binder such as water, ethanol, starch slurry, gelatin, cellulose, etc.
  • moderate amount of disintegration Such as dry starch, sodium methyl starch, sodium alginate, etc., and appropriate amount of lubricants such as magnesium stearate, talc, polyethylene glycol, etc., according to conventional wet granulation, drying, whole or dry granulation After pressing.
  • the film-coated tablets may be coated into a sealed bottle or an aluminum-plastic plate by conventional coating, such as cellulose, polyethylene glycol or the like
  • 2 capsules including ordinary capsules, intestinal sols and so on.
  • the above-mentioned total benzofuran extract dry powder is added with an appropriate amount of auxiliary materials such as starch, carbonic acid, mannitol, magnesium oxide, micro-silica gel, etc., and a suitable amount of lubricants such as talc, magnesium stearate, ethylene glycol ester, polysilicone, etc.
  • a suitable amount of adhesives such as mineral oil, edible oil, etc., mixed into a dry powder or granules, filled into a rubber sleeve, and packed in a closed aluminum plastic packaging.
  • a suitable amount of a sweetener such as D-xylose, xylitol, maltitol, stevioside, aspartame or the like may be added to each of the above dosage forms.
  • a medicinal additive such as a suspending agent (such as sorbitol syrup, cellulose derivative, glucose/sucrose syrup, gelatin, aluminum stearate) Or hydrogenated edible fat); emulsifiers (such as lecithin, acacia or sorbitan-oleate); non-aqueous carriers (such as almond oil, oily esters, ethanol or refined vegetable oils); and preservatives (for example, p-hydroxyl) Osteric acid decyl ester or propyl paraben or sorbic acid).
  • the liquid preparation may further contain a known buffering agent, a flavoring agent and an aroma component, a dye, and a sweetener, as needed.
  • injection using the above total benzofuran extract dry powder, adding pharmaceutical adjuvants, such as suspending agents, stabilizers and / or dispersing agents and / or reagents to adjust the osmotic pressure of the solution, can be formulated into injections, preferably muscles, veins Injection.
  • pharmaceutical adjuvants such as suspending agents, stabilizers and / or dispersing agents and / or reagents to adjust the osmotic pressure of the solution
  • the present invention also provides a pharmaceutical composition
  • a pharmaceutical composition comprising the benzoin plant or the benzoin plant extract according to any one of claims 1 to 14 or the compound according to any one of claims 2 to 12.
  • a preferred pharmaceutical composition wherein the active ingredient is a benzoin plant according to claims 1-14
  • the above-mentioned benzofuran extract of the benzoin plant can be combined with other traditional Chinese medicines and/or other chemical drugs to form a traditional Chinese medicine compound or a combination of Chinese and Western medicines, and the tablets are prepared according to a conventional method.
  • a variety of dosage forms such as oral liquid and injection.
  • the in vitro activity test showed that the benzofuran extract of benzoin plant has a good effect of promoting estrogen synthesis. It can be made into oral tablets, capsules and other dosage forms, suitable for premature ovarian failure, menopausal syndrome, Osteoporosis and ovarian dysfunction in adolescence, ovarian syndrome, infertility and treatment of diseases caused by estrogen deficiency or relative deficiency have the characteristics of remarkable curative effect, low price and convenient use.
  • the filtrate was combined twice and concentrated under reduced pressure to give 253 g of the total extract. Dissolve the total extract with 2L of hot water; take 2.5kg of D101 macroporous resin, and absorb the hot water dissolved extract with macroporous resin, elute with 20L of water, discard the eluate, then use 95%
  • the column was eluted with ethanol.
  • the eluate was concentrated under reduced pressure and then applied to a 1.5 kg polyamide column. The column was eluted with 95% ethanol.
  • the fruit of Washan benzoin is 2.2 kg (dry weight), and after pulverization, it is leached 3 times for 7 days at room temperature with 80-90% industrial alcohol 25 L.
  • the extract was concentrated in vacuo to obtain 550 g of total extract. It was suspended in 5 L of hot water, and extracted with petroleum ether, chloroform and n-butanol for 5 times, each time 5 L of solvent, and the extracts were separately concentrated.
  • Paste 16 g of petroleum ether, 200 g of chloroform, and 120 g of n-butanol. Sampling For in vitro activity screening, both chloroform and n-butanol fractions were shown to promote estrogen E2 production.
  • Infrared spectroscopy The main characteristic bands are located at 3456, 2946, 2919, 1600, 1504, 1482, 1459, 1236, 1042, 928 cm- 1 ;
  • Nuclear magnetic resonance carbon spectrum (150 MHz, DMSO-d6): ⁇ 32.2 (C-2"), 35.2 (Cl"), 60.6 (C-3") : 101.9 (-0-C3 ⁇ 4-0- ), 105.4 (C-2'), 111.0 (C-5'), 119.1 (C-6'), 124.8 (C-1'), 148.2 (C-4') , 148.4 (C-3'), 101.5 (C-3), 109.3 (C-6), 111.6 (C-4), 131.1 (C-9), 138.4 (C-5), 141.8 (C-8) 142.3 (C-7), 155.3 (C-2).
  • Color and shape pale yellow solid; melting point mp 76-79 ° C;
  • Infrared spectroscopy The main characteristic bands are located at 2959, 1729, 1599, 1504, 1477, 1449, 1236, 1186, 1042, 930 cm" 1 ;
  • the UV absorption of oleanol is similar, suggesting a benzofuran derivative.
  • Color and shape colorless needle crystals (acetone);
  • Infrared spectroscopy The main characteristic bands are located at 2923, 1673, 1620, 1474, 1234, 1129, 1038 cm" 1 ;
  • H-1 and C-4, C-6, and H- 4.
  • H-6 is related to Cl" with HMBC (as shown in the figure below), indicating that the 3-oxypropenyl group is located at C-5. Therefore, the structure of compound 7 is determined to be 5-(3-oxypropenyl)-7- Methoxy-2-(3,4-methylenedioxyphenyl)benzofuran.
  • Infrared spectroscopy The main characteristic bands are located at 3430, 2925, 2854, 1613, 1469, 1459, 1337, 1161, 1117, 1053, 963 cm" 1 ;
  • Compound 12 is characterized by:
  • Infrared spectroscopy The main characteristic i bands are located at 3427, 2924, 1620, 1503, 1474, 1363, 1256, 1232, 1074, 1038, 929, 810 cm" 1 ;
  • Magnesium stearate lg (total 1000 tablets) Benzoin total benzofuran extract, HPMC, lactose, mixed, wet granules with 75% ethanol as binder, dried through 22 mesh, 5CTC dried for 3 h, 22 mesh sieved whole, add 4 parts of stearic acid and mix well, each piece weighs O.lg, for patients with estrogen deficiency, oral, three times a week, one tablet each time, January is a course of treatment.
  • composition of the capsule The Luchun benzoin extract prepared in Example 3 0.5g
  • Magnesium stearate lg (total 1000 tablets) Benzoin total benzofuran extract, starch, magnesium stearate, mixed, rubber sleeve. 100mg per capsule, for estrogen deficiency patients, oral, three times a day, one tablet each time, one month for one course.
  • a benzoin extract 0.5 g, a soybean isoflavone extract 200 g, and a tanshinone extract 200 g were added to the corresponding excipients according to a conventional method to prepare tablets or capsules. 200mg per capsule, for estrogen deficiency patients, oral, three times a day, one tablet each time, one month for a course of treatment.
  • the mouse ovarian granulosa cells were collected, cultured in a 96-well culture plate, and E 2 was used to synthesize the desired substrate testosterone (0.5 mM), and the test drug at a concentration of 100 g/ml, and the drug was added without testosterone.
  • the drug control group was continuously cultured for 72 hours at 37 ° C and 5% C0 2 , and the culture solution was collected, centrifuged at 1000 rpm for 3 min, and the supernatant was taken.
  • the estradiol was detected by an enzyme-free kit (E 2 ). )content.
  • the differentiated mouse 3T3-L1 adipocytes were inoculated into 24-well culture medium at 2.4 ⁇ 105/well, 1 mL/well. After the cells were grown into a monolayer, E 2 was used to synthesize the desired substrate testosterone (0.5 mM). Concentration is ltX ⁇ g/ml of the drug to be tested, and a drug control group containing no drug and testosterone alone, and continuing to culture at 37 ° C, 5% C0 2 for 72 hours, collecting the culture solution, 1000 rpm After centrifugation for 3 min, the supernatant was taken, and the content of estradiol (E 2 ) therein was measured using an enzyme-free kit. The content of the synthetic E 2 in the control group in which only 0.5 mM of testosterone was added was regarded as 100%, and the synthesis ratio of the other sample groups E 2 was calculated based on this, and the results are shown in Table 2.
  • Compound 12 251.79 It is apparent from Table 3 that Compound 1-12 promotes estrogen production in rat ROS 17/2.8 osteosarcoma cells.
  • mice 50 adult female mice were divided into 5 groups, and 4 groups were surgically resected bilateral ovaries (castration). The remaining group underwent sham surgery (the same procedure was performed but the mouse ovaries were not removed). On the 4th day after surgery, vaginal smear was performed on each castration group until no vaginal epithelial keratinocytes were observed, which proved that oophorectomy was complete and met the "menopause" performance.
  • the model group was given normal saline by intragastric administration; the experimental group was intragastrically administered with compounds 10 and 11 (50 mg / kg / d); the positive control group was injected with estradiol benzoate (0.03113 ⁇ 4/13 ⁇ 4/(1), The sham operation group was used as a normal control group, and normal saline was administered by gavage. All the mice were subjected to blood sampling for 30 days after administration, and estrogen E 2 levels were determined by separating serum. The results are shown in the following table.
  • the present invention not only provides a novel plant extract of benzofuran benzofuran, and a novel benzofuran derivative monomer of the benzoin plant, but also provides a new drug for treating diseases associated with estrogen deficiency.
  • Composition due to the nature of the natural botanical itself, the present invention is for the treatment of premature ovarian failure, climacteric syndrome, osteoporosis and ovarian dysfunction in adolescence, polycystic ovary syndrome, infertility and estrogen deficiency or relative deficiency
  • the disease provides a safe, inexpensive, effective, and easy-to-use route of administration.

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Abstract

The invention provides an extract of Styrax Linn, which comprises from 40 to 98 % by weight of total benzofurans of Styrax Linn. It also provides the preparation and uses of such extract. The extract according to the invention has the characteristic of potent therapeutic efficiency, lower price, convenient use and the like.

Description

安息香属植物提取物及其制备方法和用途 技术领域  Benzoin extract, preparation method and use thereof
本发明涉及一种中药提取物及其制备方法和用途，具体地说， 涉及一种安 息香属植物提取物及其制备方法和用途。 背景技术  The present invention relates to a traditional Chinese medicine extract, a preparation method and use thereof, and more particularly to a succulent plant extract, a preparation method thereof and use thereof. Background technique
雌激素 (特别是雌激素 E2 )是人及其它高等动物体内最重要的激素之一， 具有广泛的生理功能。 更年期前妇女 90%的 Ε2、 50% 分别来自卵巢和肾上 腺， 更年期后妇女， 由于卵巢萎缩， 卵巢功能显著降低， 因而雌激素分泌大 大减少， 这时脂肪细胞产生的雌激素成为体内雌激素的主要来源， 如果这个 来源不足， 血液中 、 E2仅相当于更年期前妇女早卵泡期的水平， 易引起骨 质密度降低， 导致骨质疏松型骨折。 殳妇女的平均寿命为 80岁， 那么妇女 一生有 1/3的时间处于更年期后， 且约有 1/4的人在 65岁后会得骨质疏松症。 随着人口老龄化的日趋严重， 患更年期综合症和骨质疏松症的患者将越来越 多。 另外， 一些非更年期女性可能出现卵巢功能早衰或卵巢功能低下， 雌激 素水平不足， 一些疾病如多嚢卵巢综合症、 不孕症等疾病也与雌激素水平相 对减少有关。 因此， 寻求改善卵巢功能、 促进雌激素合成的药物有助于预防 和治疗这些疾病。 Estrogen (especially estrogen E 2 ) is one of the most important hormones in humans and other higher animals and has a wide range of physiological functions. 90% of women before menopause have sputum 2 , 50% from the ovary and adrenal gland, women after menopause, due to ovarian atrophy, ovarian function is significantly reduced, so estrogen secretion is greatly reduced, when the estrogen produced by fat cells becomes estrogen in the body The main source, if this source is insufficient, E 2 in the blood is only equivalent to the level of early follicular phase of women before menopause, which may cause bone density to decrease and lead to osteoporotic fracture. The average life expectancy of a woman is 80 years old, then one third of a woman's life is after menopause, and about a quarter of people will get osteoporosis after age 65. As the population ages, the number of patients suffering from menopausal syndrome and osteoporosis will increase. In addition, some non-menopausal women may have premature ovarian failure or ovarian dysfunction, insufficient estrogen levels, and some diseases such as polyposis, infertility and other diseases are also associated with a relative reduction in estrogen levels. Therefore, drugs that seek to improve ovarian function and promote estrogen synthesis help prevent and treat these diseases.
安息香科（Si racflcefle ) 又名野茉莉科、 齐墩果科， 约 11属， 180种。 我国产 9属， 50种， 9变种， 分布很广。 安息香属 ( Sty rax Linn ) 又名野茉莉 属， 约 130种， 我国约有 30种， 7变种， 主产于长江流域以南各省区。 安息 香科植物大部份可供观赏用， 有的种类供材用或药用。 其树脂含较多的香脂 酸， 称 "安息香"， 有开窍清神行气、 活血、 止痛的作用， 是医药上的贵重药 材； 这种树脂还可用于制造高级芳香油。 果实的药用价值方面， 从越南安息 香（即白花树）的果实中提取出的油就是著名的 "白花油"， 供药用， 具有杀 菌、 止痛的作用。 现代研究表明， 安息香属植物果实主要含油、 糖类、 木质 素以及新木质素（苯并呋喃类）。 有研究报道了其苯并呋喃类成分具有一定的 抗菌活性， 尚无安息香属植物总苯并呋喃提取物及其治疗卵巢功能早衰、 更 年期综合征、 骨质疏松以及青春期卵巢功能低下、 多嚢卵巢综合症、 不孕症 以及因雌激素缺乏或相对不足引起的疾病的有关文献记载或研究报道。 发明内容 Si racflcefle (also known as wild jasmine, oleanaceae, about 11 genera, 180 species). China produces 9 genera, 50 species, 9 varieties, and is widely distributed. Sty rax Linn, also known as wild jasmine, has about 130 species. There are about 30 species and 7 varieties in China, which are mainly produced in the provinces and regions south of the Yangtze River basin. Most of the benzoin plants are available for viewing, and some are available for medicinal purposes. Its resin contains more scented acid, called "benzoin", which has the effect of clearing the spirit, promoting blood circulation and relieving pain. It is a valuable medicine in medicine; this resin can also be used to make high-grade aromatic oil. In terms of the medicinal value of the fruit, the oil extracted from the fruit of Vietnamese benzoin (ie, white flower tree) is the famous "white flower oil", which is used for medicinal purposes and has the functions of sterilization and pain relief. Modern studies have shown that the fruits of the genus Benzoin are mainly composed of oil, sugar, lignin and new lignin (benzofurans). Studies have reported that its benzofuran component has a certain antibacterial activity, there is no benzofuran extract of benzoin plant and its treatment of premature ovarian failure, climacteric syndrome, osteoporosis and ovarian dysfunction in adolescence, ovarian ovary Syndrome, infertility And related literature records or research reports of diseases caused by estrogen deficiency or relative deficiency. Summary of the invention
本发明的技术方案是提供一种安息香属植物提取物及其制备方法，以及由 该提取物加工成的药物制剂在预防或治疗卵巢功能早衰、 更年期综合征、 骨 质疏松以及青春期卵巢功能低下、 多嚢卵巢综合症、 不孕症以及因雌激素缺 乏或相对不足引起的疾病的药物制剂中的用途。  The technical proposal of the present invention is to provide a benzoin plant extract and a preparation method thereof, and a pharmaceutical preparation processed from the extract for preventing or treating premature ovarian failure, climacteric syndrome, osteoporosis and ovarian dysfunction in adolescence, Use in multidrug ovarian syndrome, infertility, and pharmaceutical preparations for diseases caused by estrogen deficiency or relative deficiency.
本发明提供安息香属植物是取物，含有安息香属植物总苯并呋喃，其重量 百分比为 40 ~ 98 %。  The present invention provides a benzoin plant which contains a benzofuran total benzofuran in a weight percentage of 40 to 98%.
所述的安息香属植物总苯并呋喃提取物中，含有一种或几种具有以下化学 结构的苯并呋喃 生物单体： 其中 3、 6、 7、 8、 12为新化合物。
Figure imgf000003_0001
The benzofuran extract of the benzoin plant contains one or more benzofuran biomonomers having the following chemical structures: wherein 3, 6, 7, 8, 12 are novel compounds.
Figure imgf000003_0001
5-(3-羟基)丙基 -7-甲氧基 -2-(3, 4-二氧亚甲基苯基）苯并呋喃 （即齐墩果醇） 5-(3-hydroxy)propyl-7-methoxy-2-(3,4-dioxymethylenephenyl)benzofuran (ie oleanol)
Figure imgf000003_0002
Figure imgf000003_0002
5-(3-羟基)丙基 -2-(3， 4-二氧亚甲基苯基）苯并呋喃 （即去曱氧基齐墩果醇）  5-(3-hydroxy)propyl-2-(3,4-dioxymethylenephenyl)benzofuran (ie demethoxyl oleanol)
Figure imgf000003_0003
Figure imgf000003_0003
10 齐墩果醇 -龙胆双糖苷  10 oleanol - gentian diglucoside
该提取物还含有一种或几种具有以下化学结构的苯并呋喃衍生物单体:
Figure imgf000004_0001
The extract also contains one or more benzofuran derivative monomers having the following chemical structure:
Figure imgf000004_0001
3 3
_(3_羟基)丙基 _7_羟基— 2- (3, 4-二氧亚曱基苯基）苯并呋喃（即去甲基齐墩果 醇） _( 3 _hydroxy)propyl- 7 7 -hydroxy-2-(3,4-dioxoindolylphenyl)benzofuran (ie demethyl oleanol)
Figure imgf000004_0002
Figure imgf000004_0003
Figure imgf000004_0002
Figure imgf000004_0003
Figure imgf000004_0004
Figure imgf000004_0004
7  7
5- (3-氧丙烯基） -7-曱氧基 -2- (3, 4-亚曱基二氧苯基） 苯并呋喃
Figure imgf000005_0001
5-(3-Oxopropenyl)-7-methoxy-2-(3,4-indolyldioxyphenyl)benzofuran
Figure imgf000005_0001
8  8
(trans)-5-(3-羟基丙基) -7-甲氧基 -2- ( 2, 3-二氢 -3-羟曱基 -7-甲氧基 -2- ( 3-甲氧 基 _4-羟基苯基 )苯并呋喃基）苯并呋喃  (trans)-5-(3-hydroxypropyl)-7-methoxy-2-(2,3-dihydro-3-hydroxyindol-7-methoxy-2-(3-methoxy) _4-hydroxyphenyl)benzofuranyl)benzofuran
Figure imgf000005_0002
Figure imgf000005_0002
9 齐墩果醇-葡萄糖苷  9 oleanol-glucoside
Figure imgf000005_0003
Figure imgf000005_0003
11 齐墩果醇 -龙胆三糖苷  11 oleanol - gentian triose
Figure imgf000005_0004
去曱氧基齐墩果醇 -龙胆双糖苷 本发明提供的安息香属植物总苯并呋喃提取物， 是由安息香属 [styrax]植 物提取获得的。 更具体的， 是入尺瓦山安 香 [Styrax perkinsiaej , 禄春安息香5. macranthus Pert] ,楚雄安息香 5. limprichtii Lingelsh et BorzaJ,越南安息香5. tonkinensis (Pierre) Craib etHartw.],浙江安息香5· zhejiangensis S.M. Hwang et L. L. Yu], 芬芳安息香 5. odoratissimus Champ.] , 中华安息香 S. chinehsis Hu et S. Y. Liang], 、真南安息香 51. benzoinoides Graib], 苏门答腊安息香 iS. benzoin Dryand] 花安^香 [S. argentifolius H. L. Li],灰叶野茉莉 5. calvescens Perk] , 嘉赐叶野茉莉 S. casearifolius Craib], 黄果安息香 chrysocarpus H. L. Li], 赛山? f条5. confusus Hemsl] ,垂珠^ j[S. dasyanthus Perk] ,白 H[S. faberi Perk] , 大花野茉莉 [S. grandifloms GriffJ, hemsleyanus Diels] , 西藏野茉莉 [S. hookeri C. B. Clarke] , 野茉莉5. japonicus Sieb. et Zucc], 大籽野茉莉5. macrospermus C. Y. Wu], 桐叶野茉莉 «S. mallotifoUus C. Y. Wu], 粉花野茉莉 /5. roseus Dunn] , 铍叶野茉莉 iS. rugosus KurzJ , 齿叶野茉莉 serrulatus Roxb], 栓叶安息香 suberifolius Hook, et Am], 小叶安息香 wilsonii Rehd], S. ferrugineus , S. officinalis, S. obassia Sieb. et Zucc等中任意一种或几种中提取 获得。 通过 TLC (薄层层析）、 HPLC等方法对上述各种植物进行成分预试验 显示， 它们的化学成分种类和含量基本一致。
Figure imgf000005_0004
Deoxylized oleanol-gentiobiose The benzofuran extract of the benzoin plant provided by the present invention is obtained by extracting plants of the genus [styrax]. More specifically, it is into the foot of Shishan Anxiang [Styrax perkinsiaej, Luchun benzoin 5. macranthus Pert], Chuxiong benzoin 5. limprichtii Lingelsh et BorzaJ, Vietnamese benzoin 5. tonkinensis (Pierre) Craib et Hartw.], Zhejiang benzoin 5· Zhejiangensis SM Hwang et LL Yu], Fragrant benzoin 5. odoratissimus Champ.] , Chinese benzoin S. chinehsis Hu et SY Liang], 真南安息香 5 1 . benzoinoides Graib], Sumatra benzoin iS. benzoin Dryand] [S. argentifolius HL Li], jasmine wild jasmine 5. calvescens Perk], jasmine jasmine S. casearifolius Craib], yellow fruit benzoin chrysocarpus HL Li], 赛山?f条5. confusus Hemsl] , 垂珠^ j[S. dasyanthus Perk] , white H[S. faberi Perk] , jasmine jasmine [S. grandifloms GriffJ, hemsleyanus Diels] , Tibet wild jasmine [S. Hookeri CB Clarke] , jasmine 5. japonicus Sieb. et Zucc], jasmine 5. macrospermus CY Wu], jasmine jasmine «S. mallotifoUus CY Wu], pink jasmine/5. roseus Dunn] , 铍野野茉莉 iS. rugosus KurzJ, jasmine serrulatus Roxb], sylvestris suberifolius Hook, et Am], lobular benzoin wilsonii Rehd], S. ferrugineus, S. officinalis, S. obassia Sieb. et Zucc One or several of the extractions are obtained. Pre-testing the ingredients of the above various plants by TLC (thin layer chromatography), HPLC or the like revealed that the chemical components and contents thereof were substantially the same.
本发明还提供上述安息香属植物总苯并呋喃提取物的制备方法， 包括以 下步骤：  The present invention also provides a method for preparing the above-mentioned benzofuran extract of benzoin, comprising the following steps:
a、 将安息香属植物果实经适当粉碎后， 用水、 醇（甲醇或乙醇）， 或含 水醇类， 室温或加热 （包括回流）提取一定时间， 可重复提取 1 ~ 3次， 然后 滤去药渣， 提取液液经减压浓缩得到总浸膏；  a. After properly pulverizing the fruits of the genus benzoin, extract with water, alcohol (methanol or ethanol), or aqueous alcohol, room temperature or heating (including reflux) for a certain period of time, can be repeatedly extracted 1 ~ 3 times, and then filter the dregs , the extract liquid is concentrated under reduced pressure to obtain a total extract;
b、 将上述提取浸膏用适量热水溶散后， 用一定量的工业用石油醚萃取数 次， 以除掉其中的油性成分；  b. Dissolving the above extracted extract with an appropriate amount of hot water, and extracting it with a certain amount of industrial petroleum ether several times to remove the oily component therein;
c、 b 步骤萃取后的水层用大孔树脂柱吸附后， 用一定量的水洗脱以除去 其中的糖成分， 然后用 30 ~ 95 %乙醇洗脱柱子， 洗脱液经减压浓缩后得浸膏， 再经真空或冷冻干燥成安息香属植物总苯并呋喃提取物干粉。  c, b step After the extracted aqueous layer is adsorbed by a macroporous resin column, it is eluted with a certain amount of water to remove the sugar component, and then the column is eluted with 30 to 95% ethanol, and the eluate is concentrated under reduced pressure. The extract is then vacuumed or freeze-dried into a dry powder of the benzofuran extract of benzofuran.
其中，提取溶剂以 50-90 %乙醇溶液较好。乙醇溶液用量为药材重量的 6-12 倍， 8倍为佳。大孔树脂型号为： D101、 HPD100 、 HPD200、 HPD300, LD140。 大孔树脂柱的后一步洗脱液以 95 %乙醇为佳。 以上制备的干粉， 为浅棕色固 体物质， 味微苦。 其中所含总苯并呋喃为总重量的 40 % -98 %。  Among them, the extraction solvent is preferably a 50-90% ethanol solution. The amount of the ethanol solution is 6-12 times the weight of the medicinal material, and 8 times is preferred. Macroporous resin models are: D101, HPD100, HPD200, HPD300, LD140. The latter step of the macroporous resin column is preferably 95% ethanol. The dry powder prepared above is a light brown solid substance with a slight bitter taste. The total benzofuran contained therein is 40% - 98% of the total weight.
本发明还提供上述的安息香属植物苯并呋喃衍生物单体的制备方法， 其 特征在于包括以下步骤:  The present invention also provides a method for preparing the above-mentioned benzofuran derivative monomer of the genus Benzoin, which is characterized by comprising the following steps:
a、 将安息香属植物果实经适当粉碎后，用水、甲醇或乙醇，或含水醇类， 室温或加热（包括回流 )提取一定时间， 可重复提取 1 ~ 3次， 然后滤去药渣， 滤液经减压浓缩得到总浸膏。  a. After properly pulverizing the fruits of the genus benzoin, extract with water, methanol or ethanol, or aqueous alcohol, room temperature or heating (including reflux) for a certain period of time, can be repeatedly extracted 1 ~ 3 times, and then filter the dregs, the filtrate Concentration under reduced pressure gave a total extract.
b、 将上述提取浸膏用适量热水溶散后，依次用一定量的石油醚、氯仿和 正丁醇各萃取数次， 浓缩后得各部分萃取物， 石油醚部分弃去。 c、 氯仿部分上硅胶层析柱， 先用石油醚 /丙酮体系进行梯度洗脱（依次 以石油醚:丙酮 = 10:1， 5:1, 0:1洗脱较好）， 得到化合物 1、 2、 4、 5、 6、 7, 再用氯仿 /甲醇体系洗脱（以 CHCl3:MeOH = 50:1洗脱较好）， 得到化合物 3 和 8。 b. Dissolve the above extract with an appropriate amount of hot water, and then extract with a certain amount of petroleum ether, chloroform and n-butanol several times, and concentrate to obtain each part of the extract, and the petroleum ether is partially discarded. c. The chloroform is partially chromatographed on a silica gel column, first eluted with a petroleum ether/acetone system (preferably eluted with petroleum ether: acetone = 10:1, 5:1, 0:1) to obtain compound 1. 2, 4, 5, 6, and 7 were eluted with a chloroform/methanol system (preferably eluted with CHCl 3 : MeOH = 50:1) to give compounds 3 and 8.
正丁醇部分先在大孔树脂上用水洗脱掉糖， 然后上硅胶层析柱， 用氯仿 / 甲醇体系进行梯度洗脱 (依次用 CHCl3:MeOH = 20:1, 10:1, 5:1， 0:1洗脱较好)， 得到化合物 9、 10、 11和 12。 The n-butanol fraction was first eluted with water on a macroporous resin, and then subjected to a gradient elution with a chloroform/methanol system using a silica gel column (using CHCl 3 : MeOH = 20:1, 10:1, 5: 1, 0:1 elutes well, and gives compounds 9, 10, 11 and 12.
本发明还提供一种由上述方法制备的安息香属植物提取物。 所述的安息 香属植物提取物为瓦山安息香" Sz^ra perkinsiaej, 禄春安息香 [S. macranthus Perk.], 楚雄安息香 [S. limprichtii Lingelsh et Borza] , 越南安息香 [S. tonkinensis (Pierre) Craib et Hartw.]， 浙江安息香 [S. zhejiangensis S.M. Hwang et L. L. Yu] , 芬芳安息香 [S. odoratissimus Champ.] , 中华安息香 [S. chinehsis H et S. Y. Liang] , 滇南安息香 [S. benzoinoides Graib] , 苏门答腊安息香 [S. benzoin Dry and] , ^艮花安息香 [S. argentifolius H. L. Li] ,灰1"野茉莉 [S. calvescens Perk] , 嘉赐叶野茉莉 [S. casearifolius Craib] , 黄果安息香 [S. chrysocarpus H. L. Li] , 赛山梅 [S. confusus Hemsl] ,垂^ ¾[S. dasyanthus Perk] ,白花龙 [S. faberi Perk] , 大花野茉莉 [S.
Figure imgf000007_0001
Griff], 老鴿铃 [S. /z.e s/eyamw Diels]， 西藏野茉莉 [S. hookeri C. B. Clarke] , 野茉莉 [S. japonicus Sieb. et Zucc] , 大籽野茉莉 [S. macrospermus C. Y. Wu] ,桐叶野茉莉 [S. mallotifolius C. Y. Wu] ,粉花野茉莉 [S. roseus Dunn] , 铍叶野茉莉 [S. rugosus Kurz] , 齿叶野茉莉 [S. serrulatus Roxb] , 栓叶安息香 [S. suberifolius Hook, et Am] , 小叶安息香 [S. wilsonii Rehd] , S. ferrugineus, S. officinalis , S. obassia Sieb. et Zucc等中任意一种或几种的提取 物。 The present invention also provides an extract of the genus Benzoin prepared by the above method. The extract of the genus benzoin is Watzan benzoin "Sz^ra perkinsiaej, S. macranthus Perk., S. limprichtii Lingelsh et Borza", Vietnamese benzoin [S. tonkinensis (Pierre) Craib Et Hartw.], S. zhejiangensis SM Hwang et LL Yu, S. odoratissimus Champ., S. chinehsis H et SY Liang, S. benzoinoides Graib S. benzoin Dry and], S. argentifolius HL Li, ash 1 "S. calvescens Perk", S. casearifolius Craib, yellow fruit benzoin [ S. chrysocarpus HL Li] , S. confusus Hemsl, ^^3⁄4[S. dasyanthus Perk], S. faberi Perk, Dahuaye jasmine [S.
Figure imgf000007_0001
Griff], old pigeon bell [S. /ze s/eyamw Diels], S. hookeri CB Clarke, S. japonicus Sieb. et Zucc, S. macrospermus CY Wu ], S. mallotifolius CY Wu, S. roseus Dunn, S. rugosus Kurz, S. serrulatus Roxb, sapphire scent [ An extract of any one or more of S. suberifolius Hook, et Am], S. wilsonii Rehd, S. ferrugineus, S. officinalis, S. obassia Sieb. et Zucc.
优选的是瓦
Figure imgf000007_0002
、 禄春安息香 [S . macranthus Perk.] 的提取物， 进一步的， 为瓦山安息香^ ra perkinsiaej 、 禄春安息香 [S. macranthus Perk.] 果实的提取物， 最优选的是瓦山安息香果实的提取物。 Preferred is tile
Figure imgf000007_0002
The extract of S. macranthus Perk., further, is an extract of the fruit of the sage of the scent of the scent of the scent of the scent of the scent of the scent of the scent of the scent of the scent of the scent of the scent of the sage Extract.
本发明还提供本发明所述的安息香属植物或其总苯并呋喃提取物或安息 香属植物苯并呋喃衍生物单体在制备预防或治疗卵巢功能早衰、 更年期综合 征、 骨质疏松以及青春期卵巢功能低下、 多嚢卵巢综合症、 不孕症以及因雌 激素缺乏或相对不足引起的疾病的药物中的用途。 所述药物的活性成分中含有上述安息香属植物原生药或其总苯并呋喃提 取物或安息香属植物苯并呋喃衍生物单体， 也可以是含有安息香属植物原生 药或其总苯并呋喃提取物或苯并呋喃衍生物单体的复方， 即配伍有其它促进 雌激素合成的西药或中药活性成分。 所述制剂是口服片剂、 胶嚢、 口服液或 注射液。 The present invention also provides the benzoin plant of the present invention or the total benzofuran extract thereof or the benzofuran derivative monomer of the benzoin plant for the prevention or treatment of premature ovarian failure, climacteric syndrome, osteoporosis and ovarian ovarian Uses in drugs with low function, multiple ovarian syndrome, infertility, and diseases caused by estrogen deficiency or relative deficiency. The active ingredient of the medicament comprises the above-mentioned benzoin plant prodrug or its total benzofuran extract or benzoin benzofuran derivative monomer, or may contain a benzoin plant prodrug or its total benzofuran extract. A compound of a monomer or a benzofuran derivative monomer, that is, a western medicine or a traditional Chinese medicine active ingredient which is compatible with the synthesis of estrogen. The preparation is an oral tablet, a capsule, an oral solution or an injection.
①片剂： 包括普通片、 薄膜片、 肠溶片等。 用上述总苯并呋喃提取物干 粉， 加入适量稀释剂如淀粉、 糊精、 甘露醉、 微晶纤维等， 适量的粘合剂如 水、 乙醇、 淀粉浆、 明胶、 纤维素等， 适量的崩解剂如干淀粉、 援甲基淀粉 钠、 海藻酸钠等， 以及适量的润滑剂如硬脂酸镁、 滑石粉、 聚乙二醇等， 按 常规湿法制粒， 干燥后整粒或干法制粒后压片。 如包薄膜衣片， 可用成膜材 料如纤维素类、 聚乙二醇类等， 按常规包衣， 分装入密闭瓶或铝塑板中。  1 tablet: including ordinary tablets, film tablets, enteric tablets and the like. Use the above dry powder of total benzofuran extract, add appropriate amount of diluent such as starch, dextrin, nectar, microcrystalline fiber, etc., appropriate amount of binder such as water, ethanol, starch slurry, gelatin, cellulose, etc., moderate amount of disintegration Such as dry starch, sodium methyl starch, sodium alginate, etc., and appropriate amount of lubricants such as magnesium stearate, talc, polyethylene glycol, etc., according to conventional wet granulation, drying, whole or dry granulation After pressing. For example, the film-coated tablets may be coated into a sealed bottle or an aluminum-plastic plate by conventional coating, such as cellulose, polyethylene glycol or the like.
②胶嚢剂： 包括普通胶嚢剂、 肠溶胶嚢剂等。 将上述总苯并呋喃提取物 干粉加入适量辅料如淀粉、 碳酸 、 甘露醇、 氧化镁、 微粉硅胶等， 适量润 滑剂如滑石粉、 硬脂酸镁、 乙二醇酯， 聚硅酮类等， 以及适量的黏合剂如矿 物油、 食用油等， 混合成干粉或制成颗粒， 填充入胶套， 分装如密闭铝塑包 装中。 以上各剂型可加入适量甜味剂， 如 D-木糖、 木糖醇、 麦芽糖醇、 甜叶 菊素、 天冬甜母等。  2 capsules: including ordinary capsules, intestinal sols and so on. The above-mentioned total benzofuran extract dry powder is added with an appropriate amount of auxiliary materials such as starch, carbonic acid, mannitol, magnesium oxide, micro-silica gel, etc., and a suitable amount of lubricants such as talc, magnesium stearate, ethylene glycol ester, polysilicone, etc. And a suitable amount of adhesives such as mineral oil, edible oil, etc., mixed into a dry powder or granules, filled into a rubber sleeve, and packed in a closed aluminum plastic packaging. A suitable amount of a sweetener such as D-xylose, xylitol, maltitol, stevioside, aspartame or the like may be added to each of the above dosage forms.
③口服液： 口服液制备过程中， 用上述总苯并呋喃提取物干粉加入药用 添加剂， 如悬浮剂 (如山梨醇糖浆、 纤维素衍生物、 葡萄糖 /蔗糖糖浆、 明胶、 硬脂酸铝胶或氢化食用脂肪)； 乳化剂（例如卵磷脂、 阿拉伯胶或失水山梨醇- 油酸酯)； 非水载体 (例如杏仁油、油状酯、 乙醇或精炼植物油）； 以及防腐剂（例 如对羟基笨甲酸曱酯或对羟基苯甲酸丙酯或山梨酸)。 根据需要， 液体制剂还 可以含有已知的緩冲剂、 矫味剂和芳香成分、 染料和甜味剂。  3 Oral solution: In the preparation of oral liquid, the above dry powder of total benzofuran extract is added with a medicinal additive such as a suspending agent (such as sorbitol syrup, cellulose derivative, glucose/sucrose syrup, gelatin, aluminum stearate) Or hydrogenated edible fat); emulsifiers (such as lecithin, acacia or sorbitan-oleate); non-aqueous carriers (such as almond oil, oily esters, ethanol or refined vegetable oils); and preservatives (for example, p-hydroxyl) Osteric acid decyl ester or propyl paraben or sorbic acid). The liquid preparation may further contain a known buffering agent, a flavoring agent and an aroma component, a dye, and a sweetener, as needed.
④注射液： 用上述总苯并呋喃提取物干粉， 加入药用辅剂， 如悬浮剂、 稳定剂和 /或分散剂和 /或调节溶液渗透压的试剂， 可配制成注射剂， 优选 肌肉、 静脉注射液。  4 injection: using the above total benzofuran extract dry powder, adding pharmaceutical adjuvants, such as suspending agents, stabilizers and / or dispersing agents and / or reagents to adjust the osmotic pressure of the solution, can be formulated into injections, preferably muscles, veins Injection.
本发明还提供一种药物组合物， 其活性成分含有安息香属植物原生药或 权利要求 1-14所述的安息香属植物提取物或权利要求 2-12任一项所述的化合 物。  The present invention also provides a pharmaceutical composition comprising the benzoin plant or the benzoin plant extract according to any one of claims 1 to 14 or the compound according to any one of claims 2 to 12.
优选的药物组合物， 其活性成分为权利要求 1-14所述的安息香属植物提 取物， 或其与调节雌激素合成的西药或其他中药活性成分配伍成的复方， 加 上药学上可接受的载体或辅料， 制备而成的制剂。 A preferred pharmaceutical composition, wherein the active ingredient is a benzoin plant according to claims 1-14 A preparation prepared by combining a substance, or a combination thereof with a western medicine or other traditional Chinese medicine for regulating the synthesis of estrogen, together with a pharmaceutically acceptable carrier or adjuvant.
按照本发明， 可以将上述安息香属植物总苯并呋喃提取物与其它中药和 /或其它化学药物共同组成中药复方或中西药复方制剂， 并按照常规方法制 成口月 I片剂、 胶嚢、 口月 液、 注射液等多种剂型。  According to the present invention, the above-mentioned benzofuran extract of the benzoin plant can be combined with other traditional Chinese medicines and/or other chemical drugs to form a traditional Chinese medicine compound or a combination of Chinese and Western medicines, and the tablets are prepared according to a conventional method. A variety of dosage forms such as oral liquid and injection.
经体外活性测试发现安息香属植物总苯并呋喃提取物具有良好的促进雌 激素合成的作用， 可将其制成口服片剂、 胶嚢等多种剂型， 适用于卵巢功能 早衰、 更年期综合征、 骨质疏松以及青春期卵巢功能低下、 多嚢卵巢综合症、 不孕症以及因雌激素缺乏或相对不足引起的疾病的治疗， 具有疗效显著、 价 格低廉、 使用方便等特点。  The in vitro activity test showed that the benzofuran extract of benzoin plant has a good effect of promoting estrogen synthesis. It can be made into oral tablets, capsules and other dosage forms, suitable for premature ovarian failure, menopausal syndrome, Osteoporosis and ovarian dysfunction in adolescence, ovarian syndrome, infertility and treatment of diseases caused by estrogen deficiency or relative deficiency have the characteristics of remarkable curative effect, low price and convenient use.
显然， 根据本发明的上述内容， 按照本领域的普通技术知识和惯用手段， 在不脱离本发明上述基本技术思想前提下， 还可以做出其它多种形式的修改、 替换或变更。  It is apparent that various other modifications, substitutions and changes can be made in the form of the above-described embodiments of the present invention.
以下通过实施例形式的具体实施方式， 对本发明的上述内容再作进一步 的详细说明。 但不应将此理解为本发明上述主题的范围仅限于以下的实例。 凡基于本发明上述内容所实现的技术均属于本发明的范围。 发明的具体实施方式  The above content of the present invention will be further described in detail below by way of specific embodiments in the form of embodiments. However, the scope of the above-mentioned subject matter of the present invention should not be construed as being limited to the following examples. Any technique implemented based on the above description of the present invention is within the scope of the present invention. DETAILED DESCRIPTION OF THE INVENTION
【实施例 1】瓦山安息香提取物 1的制备  [Example 1] Preparation of Washan Benzoin Extract 1
瓦山安息香干燥果实 1000g， 粉碎后加 80 %乙醇溶液 8L, 加热回流提取 1.5小时， 滤渣再加 80 %乙醇溶液 6L, 加热回流提取 1小时， 过滤。 合并两次 滤液， 经减压浓缩得到总浸膏 250g。 将总浸膏用 2L热水溶散后， 用 3L工业 用石油醚萃取 5次。取 HPD100大孔树脂 2.5kg装柱， 将热水溶散的浸膏用大 孔树脂吸附后， 用 20L水洗脱， 洗脱液弃去， 然后用 95 %乙醇洗脱柱子， 洗 脱液经减压浓缩后得浸膏， 再经真空或冷冻干燥得干粉 180g, 采用 HPLC分 析并测定其总苯并呋喃含量约为总重量的 40 %。  Washan benzoin dried fruit 1000g, crushed and added 80% ethanol solution 8L, heated reflux extraction for 1.5 hours, filter residue plus 80% ethanol solution 6L, heated reflux extraction for 1 hour, filtered. The filtrate was combined twice and concentrated under reduced pressure to give a total extract 250 g. The total extract was dissolved in 2 L of hot water and extracted 5 times with 3 L of industrial petroleum ether. Take 2.5kg packed column of HPD100 macroporous resin, adsorb the hot water dissolved extract with macroporous resin, elute with 20L water, discard the eluent, then elute the column with 95% ethanol, and reduce the eluent. The extract was concentrated by pressure, and then dried or vacuum-dried to obtain a dry powder of 180 g, which was analyzed by HPLC and determined to have a total benzofuran content of about 40% by weight.
【实施例 2】瓦山安息香提取物 2的制备  [Example 2] Preparation of Washan Benzoin Extract 2
瓦山安息香干燥果实 1000g, 粉碎后加 80 %乙醇溶液 8L, 加热回流提取 1.5小时， 滤渣再加 80 %乙醇溶液 6L, 加热回流提取 1小时， 过滤。 合并两次 滤液， 经減压浓缩得到总浸膏 250g。 将总浸膏用 2L热水溶散后， 用 3L工业 用石油醚萃取 5次。取 HPD100大孔树脂 2.5kg装柱， 将热水溶散的浸膏用大 孔树脂吸附后， 用 20L水洗脱， 洗脱液弃去， 然后用 95 %乙醇洗脱柱子， 洗 脱液经减压浓缩后上 1.5kg聚酰胺柱， 用 95 %乙醇洗脱柱子， 洗脱液经减压 浓缩后得浸膏，再经真空或冷冻干燥得干粉 98g,采用 HPLC分析并测定其总 苯并呋喃含量约为总重量的 90 %。 Washan benzoin dried fruit 1000g, pulverized and added 8L of 80% ethanol solution, heated and refluxed for 1.5 hours, and the filter residue was further added with 80% ethanol solution 6L, heated and refluxed for 1 hour, and filtered. The filtrate was combined twice and concentrated under reduced pressure to give a total extract 250 g. After the total extract is dissolved in 2L of hot water, use 3L industrial Extracted 5 times with petroleum ether. Take 2.5kg packed column of HPD100 macroporous resin, adsorb the hot water dissolved extract with macroporous resin, elute with 20L water, discard the eluent, then elute the column with 95% ethanol, and reduce the eluent. After pressure concentration, 1.5 kg of polyamide column was applied, and the column was eluted with 95% ethanol. The eluate was concentrated under reduced pressure to obtain an extract, and then vacuum or freeze-dried to obtain 98 g of dry powder. The total benzofuran was analyzed by HPLC. The content is about 90% of the total weight.
【实施例 3】禄春安息香提取物的制备  [Example 3] Preparation of Luchun benzoin extract
禄春安息香干燥果实 1000g, 粉碎后加 80 %乙醇溶液 8L, 加热回流提取 1.5小时， 滤渣再加 80 %乙醇溶液 6L， 加热回流提取 1小时， 过滤。 合并两次 滤液， 经减压浓缩得到总浸膏 248g。 将总浸膏用 2L热水溶散； 取 LD140大 孔树脂 2.5kg装柱， 将热水溶散的浸膏用大孔树脂吸附后， 用 20L水洗脱, 洗 脱液弃去， 然后用 95 %乙醇洗脱柱子， 洗脱液经减压浓缩后得浸膏， 再经真 空或冷冻干燥得干粉 110g, 采用 HPLC分析并测定其总苯并呋喃含量约为总 重量的 80 %。  Luchun benzoin dried fruit 1000g, crushed and added 80% ethanol solution 8L, heated reflux extraction for 1.5 hours, filter residue plus 80% ethanol solution 6L, heated reflux extraction for 1 hour, filtered. The filtrate was combined twice and concentrated under reduced pressure to give 248 g of total extract. Dissolve the total extract with 2L of hot water; take LD140 macroporous resin 2.5kg column, adsorb the hot water dissolved extract with macroporous resin, elute with 20L water, discard the eluate, then use 95% The column was eluted with ethanol, and the eluate was concentrated under reduced pressure to obtain an extract, which was then subjected to vacuum or lyophilization to obtain a dry powder of 110 g, which was analyzed by HPLC and determined to have a total benzofuran content of about 80% by weight.
【实施例 4】楚雄安息香提取物的制备  [Example 4] Preparation of Chuxiong Benzoin Extract
楚雄安息香干燥果实 1000g, 粉碎后加 80 %乙醇溶液 8L, 加热回流提取 1.5小时， 滤渣再加 80 %乙醇溶液 6L， 加热回流提取 1小时， 过滤。 合并两次 滤液， 经减压浓缩得到总浸膏 253g。 将总浸膏用 2L热水溶散； 取 D101大孔 树脂 2.5kg装柱，将热水溶散的浸膏用大孔树脂吸附后， 用 20L水洗脱， 洗脱 液弃去， 然后用 95 %乙醇洗脱柱子， 洗脱液经减压浓缩后上 1.5kg聚酰胺柱， 用 95 %乙醇洗脱柱子， 洗脱液经减压浓缩后得浸膏， 再经真空或冷冻干燥得 干粉 98g, 采用 HPLC分析并测定其总苯并呋喃含量约为总重量的 60 %。 【实施例 5】滇南安息香提取物的制备  Chuxiong benzoin dried fruit 1000g, crushed and added 80% ethanol solution 8L, heated reflux extraction for 1.5 hours, filter residue plus 80% ethanol solution 6L, heated reflux extraction for 1 hour, filtered. The filtrate was combined twice and concentrated under reduced pressure to give 253 g of total extract. Dissolve the total extract with 2L of hot water; take 2.5kg of D101 macroporous resin, and absorb the hot water dissolved extract with macroporous resin, elute with 20L of water, discard the eluate, then use 95% The column was eluted with ethanol. The eluate was concentrated under reduced pressure and then applied to a 1.5 kg polyamide column. The column was eluted with 95% ethanol. The eluate was concentrated under reduced pressure to obtain an extract, and then dried or vacuum dried to obtain 98 g of dry powder. It was analyzed by HPLC and determined to have a total benzofuran content of about 60% by weight. [Example 5] Preparation of Benzoin benzoin extract
滇南安息香干燥果实 1000g, 粉碎后加 80 %乙醇溶液 8L, 加热回流提取 1.5小时， 滤渣再加 80 %乙醇溶液 6L, 加热回流提取 1小时， 过滤。 合并两次 滤液， 经减压浓缩得到总浸膏 253g。 将总浸膏用 2L热水溶散； 取 D101大孔 树脂 2.5kg装柱， 将热水溶散的浸膏用大孔树脂吸附后， 用 20L水洗脱， 洗脱 液弃去， 然后用 95 %乙醇洗脱柱子， 洗脱液经减压浓缩后上 1.5kg聚酰胺柱， 用 95 %乙醇洗脱柱子， 洗脱液经减压浓缩后得浸膏， 再经真空或冷冻干燥得 干粉 98g, 采用 HPLC分析并测定其总苯并呋喃含量约为总重量的 70 %。 【实施例 6】 瓦山安息香植物苯并呋喃衍生物单体的制备 Minnan benzoin dried fruit 1000g, pulverized and added 8L of 80% ethanol solution, heated and refluxed for 1.5 hours, and the filter residue was further added with 80% ethanol solution 6L, heated and refluxed for 1 hour, and filtered. The filtrate was combined twice and concentrated under reduced pressure to give 253 g of the total extract. Dissolve the total extract with 2L of hot water; take 2.5kg of D101 macroporous resin, and absorb the hot water dissolved extract with macroporous resin, elute with 20L of water, discard the eluate, then use 95% The column was eluted with ethanol. The eluate was concentrated under reduced pressure and then applied to a 1.5 kg polyamide column. The column was eluted with 95% ethanol. The eluate was concentrated under reduced pressure to obtain an extract, and then dried or vacuum dried to obtain 98 g of dry powder. It was analyzed by HPLC and determined to have a total benzofuran content of about 70% by weight. [Example 6] Preparation of benzofuran derivative monomer of Washan benzoin plant
瓦山安息香的果实 2.2 kg (干重)， 粉碎后用 80-90%工业酒精 25 L在室温 下浸提 3次，每次 7天。浸提液经真空浓缩后得总浸膏 550 g，将其悬浮于 5 L 热水中， 依次用石油醚、 氯仿和正丁醇各萃取 5次， 每次 5 L溶剂， 分别浓缩 萃取液得浸膏： 石油醚部分 16 g， 氯仿部分 200 g， 正丁醇部分 120 g。 取样 进行体外活性筛选， 氯仿和正丁醇部分均显示具有促进雌激素 E2生成活性。 将氯仿部分上硅胶层析柱， 依次用石油醚：丙酮 = 10:1, 5:1， 0:1洗脱， 将其分 为三段 Fr.l- 3。 Fr.l(30 g)上硅胶柱， 用石油醚:丙酮 = 10:1洗脱， 得到化合物 6 ( 60 mg ) 4 ( 30 mg )和 5 ( 20 mg )。 Fr.2 (150 g)上硅胶柱， 用石油醚:丙酮 = 5:1洗脱， 得到化合物 1(95 g)、 2 ( 5 g )和 7 ( 60 mg )。 Fr. 3 (15 g)上硅胶 柱， 用 CHCB:MeOH = 50:1洗脱， 得到化合物 3 (45 mg)和 8 ( 15mg )。 正丁 醇部分先用 D101 大孔树脂脱糖后得 55 g 浸膏， 上硅胶层析柱， 依次用 CHC13:MeOH = 20:1, 10:1, 5:1， 0:1洗脱， 得到化合物 9 ( 4,2 g )、 10 ( 33 g )、 11 ( 2.8 g )和 12 ( 50 mg )。 化合物的结构鉴定 化合物 3 5-(3-羟基)丙基 -7-羟基 -2-(3 , 4-二氧亚甲基苯基） 苯并呋喃， 即去曱基齐墩果醇：  The fruit of Washan benzoin is 2.2 kg (dry weight), and after pulverization, it is leached 3 times for 7 days at room temperature with 80-90% industrial alcohol 25 L. The extract was concentrated in vacuo to obtain 550 g of total extract. It was suspended in 5 L of hot water, and extracted with petroleum ether, chloroform and n-butanol for 5 times, each time 5 L of solvent, and the extracts were separately concentrated. Paste: 16 g of petroleum ether, 200 g of chloroform, and 120 g of n-butanol. Sampling For in vitro activity screening, both chloroform and n-butanol fractions were shown to promote estrogen E2 production. The chloroform fraction was applied to a silica gel column, and eluted successively with petroleum ether: acetone = 10:1, 5:1, 0:1, and then divided into three portions of Fr.l-3. Fr.l (30 g) was applied to a silica gel column eluting with petroleum ether: acetone = 10:1 to give compound 6 (60 mg) 4 (30 mg) and 5 (20 mg). Fr. 2 (150 g) was applied to a silica gel column eluting with petroleum ether: acetone = 5:1 to give compound 1 (95 g), 2 (5 g) and 7 (60 mg). Fr. 3 (15 g) on a silica gel column eluting with CHC:MeOH = 50:1 to afford compound 3 (45 mg) and 8 (15 mg). The n-butanol fraction was first degreased with D101 macroporous resin to obtain 55 g of extract, which was eluted with CHC13:MeOH = 20:1, 10:1, 5:1, 0:1. Compound 9 (4,2 g), 10 (33 g), 11 (2.8 g) and 12 (50 mg). Structural identification of the compound Compound 3 5-(3-hydroxy)propyl-7-hydroxy-2-(3,4-dioxymethylenephenyl)benzofuran, ie dethiol oleanol:
颜色和形状： 白色无定形粉末； 熔点 mp 196-199 °C;  Color and shape: white amorphous powder; melting point mp 196-199 °C;
红外光谱： 主要的特征 带位于 3456， 2946, 2919, 1600, 1504, 1482, 1459， 1236, 1042， 928 cm-1; Infrared spectroscopy: The main characteristic bands are located at 3456, 2946, 2919, 1600, 1504, 1482, 1459, 1236, 1042, 928 cm- 1 ;
紫外光 (MeOH): λπι&χ (log ε) = 204 (4.69)， 302 (4.47), 317 (4.54) nm; 质谱（ HRESIMS ): m/z = 335.0886 [M + Na]+ (calcd.: 335.0890); Ultraviolet light (MeOH): λπι & χ (log ε) = 204 (4.69), 302 (4.47), 317 (4.54) nm; mass spectrum (HRESIMS): m/z = 335.0886 [M + Na] + (calcd.: 335.0890) ;
核磁共振氢傳 (600 MHz, DMSO- d6): δ 6.83 (1H， s, H-3)， 7.45 (1H， d， J = 1.6 Hz, H-4), 6.57 (1H， d, J = 1.6 Hz, H-6), 9.88 (1H， s， H-7-OH), 7.17 (1H, d, J = 1.6 Hz, H-2'), 7.05 (1H, d， J = 8.0 Hz, H-5'), 7.42 (1H， dd, J = 8.0, 1.6 Hz, H-6'), 2.60 (2H， t， J = 7.4 Hz, Η- ), 1.72 (2H， m， H-2"), 3.42 (2H, m， H-3")， 4.46 (1H, t， J = 5.0 Hz, H-3"-OH), 6.10 (2H, s， H-(-0-CH2-0-)); Nuclear magnetic resonance (600 MHz, DMSO-d6): δ 6.83 (1H, s, H-3), 7.45 (1H, d, J = 1.6 Hz, H-4), 6.57 (1H, d, J = 1.6 Hz, H-6), 9.88 (1H, s, H-7-OH), 7.17 (1H, d, J = 1.6 Hz, H-2'), 7.05 (1H, d, J = 8.0 Hz, H- 5'), 7.42 (1H, dd, J = 8.0, 1.6 Hz, H-6'), 2.60 (2H, t, J = 7.4 Hz, Η- ), 1.72 (2H, m, H-2"), 3.42 (2H, m, H-3"), 4.46 (1H, t, J = 5.0 Hz, H-3"-OH), 6.10 (2H, s, H-(-0-CH 2 -0-)) ;
核磁共振碳谱 (150MHz,DMSO-d6): δ 32.2 (C-2"), 35.2 (C-l"), 60.6 (C-3"): 101.9 (-0-C¾-0- ), 105.4 (C-2'), 111.0 (C- 5')， 119.1 (C-6'), 124.8 (C- 1')， 148.2 (C-4'), 148.4 (C- 3')， 101.5 (C-3), 109.3 (C-6), 111.6 (C-4), 131.1 (C-9), 138.4 (C-5)， 141.8 (C-8); 142.3 (C-7)， 155.3 (C-2)。 Nuclear magnetic resonance carbon spectrum (150 MHz, DMSO-d6): δ 32.2 (C-2"), 35.2 (Cl"), 60.6 (C-3") : 101.9 (-0-C3⁄4-0- ), 105.4 (C-2'), 111.0 (C-5'), 119.1 (C-6'), 124.8 (C-1'), 148.2 (C-4') , 148.4 (C-3'), 101.5 (C-3), 109.3 (C-6), 111.6 (C-4), 131.1 (C-9), 138.4 (C-5), 141.8 (C-8) 142.3 (C-7), 155.3 (C-2).
从化合物 3的高分辨质谱准分子离子峰 m/z = 335.0886 [M+Na]+推算出其 分子式为 C18H1605; 紫外光 i "在 Ainax = 204， 302 and 317 nm处出现最大吸收 峰， 与齐墩果醇（egonol )的紫外吸收光语相似， 提示为苯并呋喃衍生物。 化 合物 3的 1H-NMR信号也与齐墩果醇的相似， 只是比齐墩杲醇多一个酚羟基 信号 δ 9.88 (IH, s, Η-7-ΟΗ), 而少一个曱基信号。 化合物 3 的曱基化反应 ( KOH-(CH3)2S04 )得到了齐墩果醇， 表明其酚羟基在 C-7位。 因此化合物 3 为 5-(3-羟基)丙基 -7-羟基 -2-(3，4-二氧亚曱基苯基） 苯并呋喃， 即去甲基齐墩果 醇。 化合物 6 齐墩果醇丁酸酯： From the high-resolution mass spectrometry ion peak of compound 3 m/z = 335.0886 [M+Na]+, the molecular formula is C 18 H 16 0 5 ; the ultraviolet light i "maximum at Ainax = 204, 302 and 317 nm The absorption peak, similar to the UV absorption spectroscopy of edolol, suggests a benzofuran derivative. The 1H-NMR signal of compound 3 is similar to that of oleanol, but one more than oleanol. The phenolic hydroxyl signal δ 9.88 (IH, s, Η-7-ΟΗ), and one less sulfhydryl signal. The thiolation reaction of compound 3 ( KOH-(CH 3 ) 2 S0 4 ) gave oleanol, indicating Its phenolic hydroxyl group is at the C-7 position. Therefore, compound 3 is 5-(3-hydroxy)propyl-7-hydroxy-2-(3,4-dioxoindolylphenyl)benzofuran, ie demethylation Oleanol. Compound 6 Oleanol butyrate:
颜色和形状： 浅黄色固体； 熔点 mp 76-79 °C;  Color and shape: pale yellow solid; melting point mp 76-79 ° C;
红外光谱： 主要的特征谱带位于 2959, 1729, 1599, 1504， 1477， 1449, 1236, 1186, 1042, 930 cm"1; Infrared spectroscopy: The main characteristic bands are located at 2959, 1729, 1599, 1504, 1477, 1449, 1236, 1186, 1042, 930 cm"1;
紫外光谱 (MeOH): ληΐ Χ (log ε) = 220 (3.97), 300 (3.86), 315 (4.01)nm; 质谱（ HRESIMS ): mlz = 419.1459 [M + Na]+ (calcd.: 419.1465);  Ultraviolet spectrum (MeOH): ληΐ Χ (log ε) = 220 (3.97), 300 (3.86), 315 (4.01) nm; mass spectrum (HRESIMS): mlz = 419.1459 [M + Na]+ (calcd: 419.1465);
核磁共振氢谱 (600 MHz, CDC13): δ 6.74 (IH, s, H-3), 7.28 (IH, d, J = 1.2 Hz, H-4), 6.55 (IH, d, J = 1.2 Hz, H-6), 6.90 (IH, d, J = 1.6 Hz, H-2'), 6.82 (IH, d, J = 8.2 Hz, H-5')， 7.35 (IH, dd, J = 8.2, 1.6 Hz, H-6'), 2.70 (2H， t, J = 7.4 Hz, H-l")， 1.95 (2H, m, H-2"), 4.07 (2H, t， J = 6.5 Hz, H-3"), 5.96 (2H, s, H-(-0-CH2-0-)), 3.98 (3H， s， H-7-OMe), 2.26 (2H, t, J = 7.8 Hz, H- 2"')， 1.62 (2H, m, H-3'"), 0.92 (3H， t, J = 8.5 Hz, H-4'"); Nuclear magnetic resonance spectroscopy (600 MHz, CDC13): δ 6.74 (IH, s, H-3), 7.28 (IH, d, J = 1.2 Hz, H-4), 6.55 (IH, d, J = 1.2 Hz, H-6), 6.90 (IH, d, J = 1.6 Hz, H-2'), 6.82 (IH, d, J = 8.2 Hz, H-5'), 7.35 (IH, dd, J = 8.2, 1.6 Hz, H-6'), 2.70 (2H, t, J = 7.4 Hz, Hl"), 1.95 (2H, m, H-2"), 4.07 (2H, t, J = 6.5 Hz, H-3" ), 5.96 (2H, s, H-(-0-CH 2 -0-)), 3.98 (3H, s, H-7-OMe), 2.26 (2H, t, J = 7.8 Hz, H- 2"'), 1.62 (2H, m, H-3'"), 0.92 (3H, t, J = 8.5 Hz, H-4'");
核磁共振碳语(150 MHz, CDC13): δ 13.9 (C-4'"), 18.7 (C-3'"), 36.5 (C-2'"), 31.0 (C-2"), 32.7 (C- ), 63.8 (C-3"), 56.4 (OMe-7), 101.5 (-0-CH2-0-)， 105.8 (C-2'), 108.8 (C-5')， 119.4 (C-6'), 124.9 (C-l')， 148.2 (C-4'), 148.3 (C-3'), 100.6 (C-3), 107.6 (C-6), 112.6 (C-4), 131.3 (C-9), 137.2 (C-5), 142.7 (C-8); 145.0 (C-7), 156.3 (C-2), 174.0 (C-l"')。 Nuclear Magnetic Resonance Carbon (150 MHz, CDC1 3 ): δ 13.9 (C-4'"), 18.7 (C-3'"), 36.5 (C-2'"), 31.0 (C-2"), 32.7 ( C- ), 63.8 (C-3"), 56.4 (OMe-7), 101.5 (-0-CH 2 -0-), 105.8 (C-2'), 108.8 (C-5'), 119.4 (C -6'), 124.9 (C-l'), 148.2 (C-4'), 148.3 (C-3'), 100.6 (C-3), 107.6 (C-6), 112.6 (C-4), 131.3 (C-9), 137.2 (C-5), 142.7 (C-8); 145.0 (C-7), 156.3 (C-2), 174.0 (Cl"').
化合物 6的紫外光谱在 max = 220, 300， 315 nm处出现最大吸收峰，与齐 墩果醇的紫外吸收光讲相似，提示为苯并呋喃衍生物。化合物 6的大部分 1H、 13C-NMR信号与齐墩果醇的相似， 只是比齐墩果醇多出 -NMR信号 δ 0.92 (3Η, J = 8.2 Hz, H-4'")， 2.26 (2H, t, J = 7.8 Hz, H-2'"), 1.62 (2H, m， H-3'")和相 应的 13C-NMR信号 δ 13.9 (C-4'"), 36.5 (C-2'"), 18.7 (C-3'"), 174.0 (C-l'") (据 HMQC ), 表明化合物 6 比齐墩果醇多出一个丁酸酯基。 化合物 6碱水解 ( 10 % KOH乙醇溶液）得到齐墩果醇， 因此， 该化合物的结构为 5- ( 3-丁酰 氧丙基） _7—曱氧基— 2_ ( 3 , 4-二氧亚曱基苯基） 苯并呋喃， 即齐墩果醇丁酸 酯。 高分辨质谱准分子离子峰 mlz = 419.1459 [M+Na]+推算出其分子式为 C23H2406， 进一步确证了该结构的正确性。 化合物 7 5- ( 3-氧丙烯基） -7-甲氧基 -2- ( 3 , 4-亚甲基二氧苯基） 苯并 呋喃 （7 ) 的特征在于： The UV spectrum of Compound 6 shows a maximum absorption peak at max = 220, 300, 315 nm. The UV absorption of oleanol is similar, suggesting a benzofuran derivative. Most of the 1H, 13 C-NMR signals of Compound 6 were similar to those of oleanol, except that the NMR signal was δ 0.92 (3Η, J = 8.2 Hz, H-4'"), 2.26 (in comparison with oleanol). 2H, t, J = 7.8 Hz, H-2'"), 1.62 (2H, m, H-3'") and the corresponding 13 C-NMR signal δ 13.9 (C-4'"), 36.5 (C- 2'"), 18.7 (C-3'"), 174.0 (C-l'") (according to HMQC), indicating that compound 6 has one more butyrate group than oleanol. Compound 6 alkaline hydrolysis (10%) KOH ethanol solution) to give oleanolic alcohol, therefore, the structure of the compound is 5- (3-butyryl-oxopropyl) _ 7-- Yue oxy - 2 _ (3,4-dioxo alkylene Yue phenyl) Benzofuran, oleanol butyrate. High-resolution mass spectrometry excimer ion peak mlz = 419.1459 [M+Na] + The formula is calculated as C 2 3H 24 0 6 , which further confirms the correctness of the structure. Compound 7 5-(3-Oxopropenyl)-7-methoxy-2-(3,4-methylenedioxyphenyl)benzofuran (7) is characterized by:
颜色和形状： 无色针状晶体（丙酮）；  Color and shape: colorless needle crystals (acetone);
熔点： 187-189 °C;  Melting point: 187-189 °C;
红外光谱： 主要的特征谱带位于 2923, 1673, 1620, 1474, 1234, 1129, 1038 cm"1; Infrared spectroscopy: The main characteristic bands are located at 2923, 1673, 1620, 1474, 1234, 1129, 1038 cm"1;
紫外光谱 (MeOH): max (log ε) = 216 (4.08), 299 (4.30), 319 (4.40) nm; 质谱（ HRESIMS ): mlz = 345.0730 [M + Na]+ (calcd.: 345.0733);  Ultraviolet spectrum (MeOH): max (log ε) = 216 (4.08), 299 (4.30), 319 (4.40) nm; mass spectrum (HRESIMS): mlz = 345.0730 [M + Na]+ (calcd.: 345.0733);
核磁共振氢谱 (600 MHz, CDC13): δ 6.87 (1H, s, H-3), 7.38 (1H, d, J = 1.6 Hz, H-4)， 6.99 (1H, d, J = 1.6 Hz, H-6)， 7.33 (1H， d， J = 1.4 Hz, H-2'), 6.90 (1H， d， J = 8.2 Hz, H- 5')， 7.42 (1H， dd, J = 8.2, 1.4 Hz, H-6'), 7.54 (1H， d, J = 15.8 Hz, H-l"), 6.72 (1H， dd, J = 15.8， 7.6 Hz, H-2"), 9.71 (1H， d, J = 7.6 Hz, H-3"), 6.03 (2H, s, H-(- 0-CH2-0-))， 4.08 (3H, s， H-7-OMe); Nuclear magnetic resonance spectroscopy (600 MHz, CDC13): δ 6.87 (1H, s, H-3), 7.38 (1H, d, J = 1.6 Hz, H-4), 6.99 (1H, d, J = 1.6 Hz, H-6), 7.33 (1H, d, J = 1.4 Hz, H-2'), 6.90 (1H, d, J = 8.2 Hz, H-5'), 7.42 (1H, dd, J = 8.2, 1.4 Hz, H-6'), 7.54 (1H, d, J = 15.8 Hz, Hl"), 6.72 (1H, dd, J = 15.8, 7.6 Hz, H-2"), 9.71 (1H, d, J = 7.6 Hz, H-3"), 6.03 (2H, s, H-(- 0-CH 2 -0-)), 4.08 (3H, s, H-7-OMe);
核磁共振碳谱 (150 MHz, CDC13) : δ 56.2 (OMe-7), 100.4 (C-3), 101.4 (-0-CH2-0- )， 105.6 (C-6), 105.7 (C-2')， 108.7 (C-5')， 115.2 (C-4), 119.6 (C-6')， 124.0 (C-l'), 127.6 (C-2"), 130.2 (C-5), 131.5 (C-9), 145.7 (C-7)， 145.6 (C-8), 148.2 (C-4'), 148.5 (C-3'), 153.7 (C-l"), 157.3 (C-2), 193.6 (C-3")。 Nuclear Magnetic Resonance Carbon Spectra (150 MHz, CDC13): δ 56.2 (OMe-7), 100.4 (C-3), 101.4 (-0-CH 2 -0- ), 105.6 (C-6), 105.7 (C-2 '), 108.7 (C-5'), 115.2 (C-4), 119.6 (C-6'), 124.0 (C-l'), 127.6 (C-2"), 130.2 (C-5), 131.5 (C-9), 145.7 (C-7), 145.6 (C-8), 148.2 (C-4'), 148.5 (C-3'), 153.7 (Cl"), 157.3 (C-2), 193.6 (C-3").
从化合物 Ί的高分辨质谱准分子离子峰 m/z = 345.0730 [M + Na]+推算出 其分子式为 C19H1405; -NMR信号 δ 6.72 (1Η, dd, J = 7.6, 15.8 Hz, H-2"), 7.54 (1H, d， J = 15.8 Hz, H-l"), 9.71 (1H, d, J = 7.6 Hz, H-3"), 以及相应的 13C-NMR 信号 δ 127.6 (C-2")， 153.7 (C-l"), 193.6 (C-3") (据 HMQC )提示分子中有一 个 3-氧丙烯基存在;化合物 7的大部分 ¾、 13C-NMR信号与齐墩果醇的相似， 不同之处在于化合物 7含 3-氧丙錄基， 而齐墩果醇含正丙醇基。 H- 1"与 C-4、 C-6, 以及 H-4、 H-6与 C-l"有 HMBC相关（如下图所示）， 说明 3-氧丙烯基 位于 C-5。 因此， 化合物 7的结构被确定为 5- ( 3-氧丙烯基） -7-甲氧基 -2- ( 3， 4-亚甲基二氧苯基） 苯并呋喃。 From the high-resolution mass spectrometry ion peak m/z of the compound Ί m/z = 345.0730 [M + Na] + , the molecular formula is C 19 H 14 0 5 ; -NMR signal δ 6.72 (1Η, dd, J = 7.6, 15.8 Hz , H-2"), 7.54 (1H, d, J = 15.8 Hz, Hl"), 9.71 (1H, d, J = 7.6 Hz, H-3"), and the corresponding 13 C-NMR The signals δ 127.6 (C-2"), 153.7 (Cl"), 193.6 (C-3") (according to HMQC) suggest that a 3-oxypropenyl group exists in the molecule; most of the compound 7 is 3⁄4, 13 C-NMR The signal is similar to oleanol, except that compound 7 contains a 3-oxopropyl group and oleanol contains a n-propanol group. H-1" and C-4, C-6, and H- 4. H-6 is related to Cl" with HMBC (as shown in the figure below), indicating that the 3-oxypropenyl group is located at C-5. Therefore, the structure of compound 7 is determined to be 5-(3-oxypropenyl)-7- Methoxy-2-(3,4-methylenedioxyphenyl)benzofuran.
Figure imgf000014_0001
Figure imgf000014_0001
8  8
Major HMBC Correlations ( ~ of 7 and 8 化合物 8 (ira«s)-5-(3-羟基丙基) -7-曱氧基 -2- ( 2, 3-二氢 -3-羟甲基 -7-甲 氧基 -2- ( 3-曱氧基 -4-羟基苯基）苯并呋喃基）苯并呋喃：  Major HMBC Correlations ( ~ of 7 and 8 Compound 8 (ira«s)-5-(3-hydroxypropyl)-7-decyloxy-2-( 2, 3-dihydro-3-hydroxymethyl-7 -Methoxy-2-(3-decyloxy-4-hydroxyphenyl)benzofuranyl)benzofuran:
颜色和形状： 白色无定形粉末； 熔点 mp 214-216 °C ;  Color and shape: white amorphous powder; melting point mp 214-216 °C;
红外光谱： 主要的特征谱带位于 3430， 2925, 2854， 1613, 1469， 1459， 1337, 1161， 1117, 1053, 963 cm"1; Infrared spectroscopy: The main characteristic bands are located at 3430, 2925, 2854, 1613, 1469, 1459, 1337, 1161, 1117, 1053, 963 cm"1;
紫外光 i| MeOH): ληι χ (log ε) = 222 (3.84), 316 (3.81) nm;  Ultraviolet light i| MeOH): ληι χ (log ε) = 222 (3.84), 316 (3.81) nm;
质谱（ HRESIMS ): m/z = 529.1849 [M + Na]+ (calcd.: 529.1833); Mass spectrum (HRESIMS): m/z = 529.1849 [M + Na] + (calcd.: 529.1833);
核磁共振氢 #(600 MHz, DMS0-d6): δ = 7.18 (IH, s， H-3), 6.97 (IH, d， J = 1.4 Hz, H-4), 6.74 (1H， d, J = 1.4 Hz, H-6), 5.51 (IH, d, J = 6.4 Hz, H-2')， 3.56 (1H， dt， J = 8.6, 6.4 Hz, H-3'), 7.43 (IH, d, J = 1.2 Hz, H-4'), 7.36 (IH, d, J = 1.2 Hz, H-6')， 3.73 (2H， d， J = 8.6 Hz, H-10'), 5.10 (IH, s, H-IO'-OH), 6.94 (IH, d, J = 1.6 Hz, H-2"), 9.06 (1H， s， H-l"-OH), 6.76 (IH, d, J = 8.2 Hz, H-5"), 6.79 (IH, dd J = 8.2, 1.6 Hz, H-6"), 3.95 (3H, s, H-7-OMe), 3.88 (3H, s， H-7'-OMe), 3.75 (3H, s: H-3"-OMe), 2.67 (2H, t, J = 7.6 Hz, Η- ')， 1.77 (2H, m, H-2'"), 3.44 (2H, t, J = 6.2 Hz, H-3'"), 4.49 (IH, s， H-3"'-OH); Nuclear Magnetic Resonance Hydrogen # (600 MHz, DMS0-d6): δ = 7.18 (IH, s, H-3), 6.97 (IH, d, J = 1.4 Hz, H-4), 6.74 (1H, d, J = 1.4 Hz, H-6), 5.51 (IH, d, J = 6.4 Hz, H-2'), 3.56 (1H, dt, J = 8.6, 6.4 Hz, H-3'), 7.43 (IH, d, J = 1.2 Hz, H-4'), 7.36 (IH, d, J = 1.2 Hz, H-6'), 3.73 (2H, d, J = 8.6 Hz, H-10'), 5.10 (IH, s , H-IO'-OH), 6.94 (IH, d, J = 1.6 Hz, H-2"), 9.06 (1H, s, Hl"-OH), 6.76 (IH, d, J = 8.2 Hz, H -5"), 6.79 (IH, dd J = 8.2, 1.6 Hz, H-6"), 3.95 (3H, s, H-7-OMe), 3.88 (3H, s, H-7'-OMe), 3.75 (3H, s : H-3"-OMe), 2.67 (2H, t, J = 7.6 Hz, Η- '), 1.77 (2H, m, H-2'"), 3.44 (2H, t, J = 6.2 Hz, H-3'"), 4.49 (IH, s, H-3"'-OH);
核磁共振碳谱 (150MHz，DMSO-d6): δ 156.4 (C-2)， 100.8 (C-3), 112.4 (C-4)， 138.6 (C-5), 107.8 (C-6), 144.8 (C-7), 141.9 (C-8)， 123.7 (C-9), 88 (C-2')， 53.3 (C-3')， 114.2 (C-4')， 131.1 (C-5'), 109.4 (C-6'), 144.5 (C-7 148.9 (C-8')， 130.7 (C- 9'), 63.2 (C-10')， 132.4 (C-l"), 110.9 (C-2"), 148.1 (C-3"), 147.0 (C-4"), 115.8 (C-5"), 119.2 (C-6"), 56.1 (OCH3-7), 56.4 (OC¾-7')， 56.2 (OCH3-3"), 32.5 (C-l'"), 35.2 (C-2'"), 60.6 (C-3"')。 Nuclear Magnetic Resonance Spectroscopy (150 MHz, DMSO-d6): δ 156.4 (C-2), 100.8 (C-3), 112.4 (C-4), 138.6 (C-5), 107.8 (C-6), 144.8 ( C-7), 141.9 (C-8), 123.7 (C-9), 88 (C-2'), 53.3 (C-3'), 114.2 (C-4'), 131.1 (C-5') , 109.4 (C-6'), 144.5 (C-7 148.9 (C-8'), 130.7 (C-9'), 63.2 (C-10'), 132.4 (Cl"), 110.9 (C-2"), 148.1 (C-3"), 147.0 (C-4"), 115.8 (C-5 "), 119.2 (C-6"), 56.1 (OCH 3 -7), 56.4 (OC3⁄4-7'), 56.2 (OCH 3 -3"), 32.5 (C-l'"), 35.2 (C-2 '"), 60.6 (C-3"').
从化合物 8的高分辨质谱准分子离子峰 /z = 529.1849 [M + Na]+推算出其 分子式为 C29H30O8; ^-NMR信号 δ 3.95 (3H， s， H3-7-OMe), 3.88 (3H， s, H3-7'- OMe)，及 3.75 (3H， s， H3-3"-OMe)提示分子中含 3个曱氧基； 两个羟基 信号分別在 δ 4.49 (1H, s, Η- 3"'-ΟΗ)和 5.10 (1H, s, H-10'-OH); -NMR信号 δ 3.44 (2Η, t, J = 6.2 Hz, Η-3'" )、 3.73 (2H, d， J = 8.6 Hz, H-10'), 以及它们相应的 13C-NMR信号 δ 60.6 (C-3'", CH2)， 63.2 (C-10'， CH2) (据 HMQC )提示分子中有 两个羟甲基； 1H- NMR信号 δ 6.76 (1H, d, J = 8.2 Hz, H-5"), 6.94 (1H, d, J = 1.6 Hz, H-2")及 6.79 (1H， dd， J = 8.2, 1.6 Hz, H-6")表明有一个 1，3,4-三取代苯环 存在； 其它 5个芳环质子信号分别在 δ 7.18 (1H， s， H- 3)， 6.97 (1H, d, J = 1.4 Hz, H-4), 6.74 (1H, d, J = 1.4 Hz, H-6), 7.43 (1H， d, J = 1.2 Hz, H-4'), 7.36 (1H, d, J = 1.2 Hz, H-6'); 根据 HMQC和 HMBC (见上图）， 化合物 8的结构被确定为 (trans)-5-(3-羟基丙基) -7-曱氧基 -2- ( 2，3-二氢 -3-羟甲基 -7-曱氧基 -2- ( 3-曱氧基 -4-羟基苯基）苯并呋喃基）苯并呋喃， 它是 dihydrodehydrodiconiferyl alcohol 与苯并呋喃通过 C-2'和 C-3'的偶联产物， H-2'与 H-3'之间的偶合常数 ( J = 6.4 Hz ), 以及 H-2'与 H-10'之间有 NOESY相关信号决定了 C-2'与 C-3'之间的反 式相对构型。 From the high-resolution mass spectrometry ion peak of compound 8 /z = 529.1849 [M + Na] + , the molecular formula is C 29 H 30 O 8 ; ^-NMR signal δ 3.95 (3H, s, H3-7-OMe) , 3.88 (3H, s, H3-7'- OMe), and 3.75 (3H, s, H3-3"-OMe) suggest that the molecule contains three oxime groups; the two hydroxyl signals are at δ 4.49 (1H, s, Η- 3"'-ΟΗ) and 5.10 (1H, s, H-10'-OH); -NMR signal δ 3.44 (2Η, t, J = 6.2 Hz, Η-3'"), 3.73 (2H , d, J = 8.6 Hz, H-10'), and their corresponding 13 C-NMR signals δ 60.6 (C-3'", CH 2 ), 63.2 (C-10', CH 2 ) (according to HMQC) There are two hydroxymethyl groups in the molecule; 1H-NMR signal δ 6.76 (1H, d, J = 8.2 Hz, H-5"), 6.94 (1H, d, J = 1.6 Hz, H-2") and 6.79 (1H, dd, J = 8.2, 1.6 Hz, H-6") indicates the presence of a 1,3,4-trisubstituted benzene ring; the other five aromatic ring proton signals are at δ 7.18 (1H, s, H-, respectively) 3), 6.97 (1H, d, J = 1.4 Hz, H-4), 6.74 (1H, d, J = 1.4 Hz, H-6), 7.43 (1H, d, J = 1.2 Hz, H-4' ), 7.36 (1H, d, J = 1.2 Hz, H-6'); According to HMQC and HMBC (see above), the structure of compound 8 was determined to be (trans)-5-(3-hydroxypropyl) -7-decyloxy-2-( 2,3-dihydro-3-hydroxymethyl-7-decyloxy-2-(3-decyloxy-4-hydroxyphenyl)benzofuranyl) Benzofuran, which is the coupling product of dihydrodehydrodiconiferyl alcohol with benzofuran via C-2' and C-3', the coupling constant between H-2' and H-3' (J = 6.4 Hz), and H The NOESY related signal between -2' and H-10' determines the trans relative configuration between C-2' and C-3'.
化合物 12的特征在于：  Compound 12 is characterized by:
颜色和形状： 白色无定形粉末； 熔点 mp 110-113 °C;  Color and shape: white amorphous powder; melting point mp 110-113 °C;
红外光谱： 主要的特征 i 带位于 3427， 2924, 1620, 1503， 1474， 1363, 1256， 1232, 1074， 1038, 929, 810 cm"1; Infrared spectroscopy: The main characteristic i bands are located at 3427, 2924, 1620, 1503, 1474, 1363, 1256, 1232, 1074, 1038, 929, 810 cm"1;
紫外光语 (MeOH): max (log ε) = 216 (4.52), 302 (4.47), 317 (4.10), 331 (3.72) nm;  UV language (MeOH): max (log ε) = 216 (4.52), 302 (4.47), 317 (4.10), 331 (3.72) nm;
质谱（ HRESIMS ): m/z = 620.0546 [M + Na]+ (calcd.: 620.0539); Mass spectrum (HRESIMS): m/z = 620.0546 [M + Na] + (calcd.: 620.0539);
核磁共振氢谱 (600 MHz, DMSO-d6): δ 7.41 (1H, d, J = 8.4 Hz, H-7), 7.38 (1H， dd， J = 7.9, 1.8 Hz, H-6'), 7.39 (1H, d, J = 1.6 Hz, H-4), 7.17 (1H， s， H-2'), 7.01 (1H, d, J = 8.0 Hz, H-5'), 6.99 (1H, s， H-3), 6.77 (1H, s， H-6), 6.07 [2H, s， H-(- 0-CH2-0-)]， 3.45 (2H, t, J = 6.4 Hz, H-3"), 2.71 (2H, t， J = 7.6 Hz, Η- ), 1.86 (2H， m， H-2"), 5.04 (1H， d， J = 5.0 Hz, glc-H-1'), 4.90 (1H， d, J = 4.8 Hz, glc-H-1"); Nuclear magnetic resonance spectroscopy (600 MHz, DMSO-d6): δ 7.41 (1H, d, J = 8.4 Hz, H-7), 7.38 (1H, dd, J = 7.9, 1.8 Hz, H-6'), 7.39 (1H, d, J = 1.6 Hz, H-4), 7.17 (1H, s, H-2'), 7.01 (1H, d, J = 8.0 Hz, H-5'), 6.99 (1H, s, H-3), 6.77 (1H, s, H-6), 6.07 [2H, s, H-(- 0-CH 2 -0-)], 3.45 (2H, t, J = 6.4 Hz, H-3 "), 2.71 (2H, t, J = 7.6 Hz, Η- ), 1.86 (2H, m, H-2"), 5.04 (1H, d, J = 5.0 Hz, glc-H-1'), 4.90 (1H, d, J = 4.8 Hz, glc-H-1");
核磁共振碳语 (150MHz, DMSO-d6): δ 155.6 (C-2)， 101.5 (C-3), 112.6 (C-4)， Nuclear magnetic resonance carbon (150MHz, DMSO-d6): δ 155.6 (C-2), 101.5 (C-3), 112.6 (C-4),
138.3 (C-5), 108.3 (C-6), 120.2 (C-7), 142.1 (C-8), 130.9 (C-9), 124.5 (C- 1')，138.3 (C-5), 108.3 (C-6), 120.2 (C-7), 142.1 (C-8), 130.9 (C-9), 124.5 (C-1'),
105.4 (C-2')， 148.4 (C-3'), 148.2 (C- 4')， 109.3 (C-5'), 119.2 (C-6'), 101.9 [C-(-0-CH2-0-)]， 32.3 (C-l"); 31.9 (C-2"), 68.3 (C-3"), 103.3 (glc- 1')， 73.9 (glc-2'), 77.3 (glc-3'), 70.5 (glc-4'), 76.2 (glc-5'), 68.8 (glc-6')3 103.8 (glc-1"), 74.0 (glc-2"), 77.4 (glc-3"), 70.5 (glc-4"), 77.3 (glc-5"), 61.6 (glc-6"); 105.4 (C-2'), 148.4 (C-3'), 148.2 (C-4'), 109.3 (C-5'), 119.2 (C-6'), 101.9 [C-(-0-CH2- 0-)], 32.3 (Cl"); 31.9 (C-2"), 68.3 (C-3"), 103.3 (glc-1'), 73.9 (glc-2'), 77.3 (glc-3') , 70.5 (glc-4'), 76.2 (glc-5'), 68.8 (glc-6') 3 103.8 (glc-1"), 74.0 (glc-2"), 77.4 (glc-3"), 70.5 (glc-4"), 77.3 (glc-5"), 61.6 (glc-6");
化合物 12的相对旋光值为 [ot] D： -27.6。（c 0.1， MeOH); ESI-MS中的分子 离子峰 m/z 621 [M+H]+和 619 [M-H]-提示其分子量为 620， 碎片峰 m/z 459 [M+H-162]+, m/z 297 [M+H- 162- 162]+提示分子中存在 2个六碳糖单元。 UV 谱中出现与齐墩果醇的相似的特征吸收（ λπΐ3χ = 218, 302, 318 nm ),提示其为 齐墩果醇类苷。 该化合物经薄层酸水解后， 只检测出葡萄糖和去甲氧基齐墩 果醇。 NMR谱中 103.3, 103.8和 δΗ 5.04 (1H, d, J = 5.0 Hz), 4.90 (1H， d, J = 4.8 Hz)显示化合物 12存在 2个 β构型的葡萄糖基。化合物 12的 13C NMR数据 与化合物 2 (去曱氧基齐墩果醇）的比较， 多出二个葡萄糖基的 12个碳信号， 其余部分化学位移基本一致。直接与去曱氧基齐墩果醇相连的葡萄糖基中 C-6 向低场位移 δ 7.2 ( 61.6→68.8 )， C-5向高场位移 δ 1.0 ( 77.2—76.2 ), 表明两 个糖基的联结方式为 1→6，是龙胆双糖苷。 因此将化合物 12鉴定为去曱氧基 齐 i敦果醇龙胆双糖苷。 The relative optical rotation of Compound 12 was [ot] D: -27.6. (c 0.1, MeOH); molecular ion peak m/z 621 [M+H] + and 619 [MH]- in ESI-MS suggesting a molecular weight of 620, fragment peak m/z 459 [M+H-162] + , m/z 297 [M+H- 162- 162]+ indicates that there are two hexasaccharide units in the molecule. Similar characteristic absorption (λπΐ3χ = 218, 302, 318 nm) with oleanol appeared in the UV spectrum, suggesting that it is an oleanol glycoside. After the thin layer acid hydrolysis of the compound, only glucose and demethoxy oleanol were detected. In the NMR spectrum, 103.3, 103.8 and δ Η 5.04 (1H, d, J = 5.0 Hz), 4.90 (1H, d, J = 4.8 Hz) showed that the compound 12 has two β-configuration glucosyl groups. The 13 C NMR data of compound 12 was compared with that of compound 2 (deoxylized oleanol) by 12 carbon signals of two glucose groups, and the rest of the chemical shifts were basically the same. In the glucose group directly linked to demethoxyl oleanol, the C-6 shifted to the low field δ 7.2 (61.6→68.8), and the C-5 shifted to the high field δ 1.0 (77.2-76.2), indicating two glycosyl groups. The connection method is 1→6, which is gentian diglucoside. Compound 12 was therefore identified as demethoxyl-i-iolol gentiobiose.
在现有技术及文献中未见有上述的化合物 3、 化合物 6、 化合物 7、 化合物 8、 化合物 12, 因此均为新化合物。  The above-mentioned Compound 3, Compound 6, Compound 7, Compound 8, and Compound 12 are not found in the prior art and literature, and are therefore all novel compounds.
【实施例 6】 片剂的制备  [Example 6] Preparation of tablets
片剂组成： 实施例 2制得的瓦山安息香提取物 0.5g  Tablet composition: Washan benzoin extract prepared in Example 2 0.5g
HPMCLV100 30g  HPMCLV100 30g
乳糖 70g  Lactose 70g
硬脂酸镁 lg (共制成 1000片） 安息香属植物总苯并呋喃提取物、 HPMC、 乳糖， 混匀， 以 75 %乙醇为 粘合剂制湿颗粒， 过 22 目筛， 5CTC干燥 3h, 22 目筛整粒， 加入硬脂酸 4美混 匀压片， 每片重 O.lg, 用于雌激素缺乏症患者， 口服， 每曰三次， 每次 1片， 一月为一疗程。 Magnesium stearate lg (total 1000 tablets) Benzoin total benzofuran extract, HPMC, lactose, mixed, wet granules with 75% ethanol as binder, dried through 22 mesh, 5CTC dried for 3 h, 22 mesh sieved whole, add 4 parts of stearic acid and mix well, each piece weighs O.lg, for patients with estrogen deficiency, oral, three times a week, one tablet each time, January is a course of treatment.
【实施例 7】 胶嚢剂的制备  [Example 7] Preparation of an anthraquinone
胶嚢組成： 实施例 3制得的禄春安息香提取物 0.5g  Composition of the capsule: The Luchun benzoin extract prepared in Example 3 0.5g
淀粉 100g  Starch 100g
硬脂酸镁 lg (共制成 1000粒） 安息香属植物总苯并呋喃提取物、 淀粉、 硬脂酸镁， 混匀， 装胶套。 每 粒装 100mg， 用于雌激素缺乏症患者， 口服， 每日三次， 每次 1片， 一月为一 疔程。  Magnesium stearate lg (total 1000 tablets) Benzoin total benzofuran extract, starch, magnesium stearate, mixed, rubber sleeve. 100mg per capsule, for estrogen deficiency patients, oral, three times a day, one tablet each time, one month for one course.
【实施例 8】 注射液的制备  [Example 8] Preparation of injection
注射液组成： 实施例 4制得的楚雄安息提取物 0.5g  Injection composition: Chuxiong rest extract prepared in Example 4 0.5g
吐温 80 10ml  Tween 80 10ml
氯化钠 8g (共制成 1000ml) 安息香属植物总苯并呋喃提取物， 加 10 % Na2CO3调 pH至 7.0-7.5, 冷藏 滤过， 加吐温 -80， NaCl, 加注射用水至 1000ml, G3垂熔漏斗 (玻璃)滤过， 分 装， 灌封， 100°C流通蒸气灭菌 30min即得。 Sodium chloride 8g (total 1000ml) benzoin total benzofuran extract, add 10% Na 2 CO 3 to adjust the pH to 7.0-7.5, refrigerate filtration, add Tween-80, NaCl, add water for injection 1000ml, G 3 fused funnel (glass) filtered, sub-packaged, potted, 100 ° C steam sterilization for 30 min.
【实施例 9】 复方片剂的制备  [Example 9] Preparation of compound tablets
取安息香属植物提取物 0.5g,大豆异黄酮提取物 200g,丹参酮提取物 200g, 按常规方法， 加入相应辅料， 制成片剂或胶嚢。 每粒装 200mg, 用于雌激素缺 乏症患者， 口服， 每日三次， 每次 1片， 一月为一疗程。  A benzoin extract 0.5 g, a soybean isoflavone extract 200 g, and a tanshinone extract 200 g were added to the corresponding excipients according to a conventional method to prepare tablets or capsules. 200mg per capsule, for estrogen deficiency patients, oral, three times a day, one tablet each time, one month for a course of treatment.
以下通过安息香属植物提取物及部分有效成分的药效试验进一步说明本 发明的有益效果。  The beneficial effects of the present invention are further illustrated by the pharmacodynamic test of the extract of the genus Benzoin and a part of the active ingredient.
【实施例 10】 促进小鼠卵巢颗粒细胞合成雌激素：  [Example 10] Promoting the synthesis of estrogen in mouse ovarian granulosa cells:
收集小鼠卵巢颗粒细胞， 培养在 96孔培养板中， 加入 E2合成所需底物睾 酮 （0.5mM ), 和浓度为 lOO g/ml的待测药物， 同时设立不加药物、 仅含睾酮 的药物空白对照组， 继续在 37°C , 5% C02条件下培养 72小时， 收集培养液， 1000转 /分钟离心 3min, 取上清， 采用酶免试剂盒检测其中雌二醇（ E2 )含量。 将只加入 0.5mM睾酮的对照組合成 E2的含量视为 100%， 以此为基准计算其它 各样品组 E2的合成率， 结果见表 1。 表 1 促进小鼠卵巢颗粒细胞合成 E: The mouse ovarian granulosa cells were collected, cultured in a 96-well culture plate, and E 2 was used to synthesize the desired substrate testosterone (0.5 mM), and the test drug at a concentration of 100 g/ml, and the drug was added without testosterone. The drug control group was continuously cultured for 72 hours at 37 ° C and 5% C0 2 , and the culture solution was collected, centrifuged at 1000 rpm for 3 min, and the supernatant was taken. The estradiol was detected by an enzyme-free kit (E 2 ). )content. The content of the synthetic E 2 in the control group in which only 0.5 mM of testosterone was added was regarded as 100%, and the synthesis ratio of the other sample groups E 2 was calculated based on this, and the results are shown in Table 1. Table 1 Promotes mouse ovarian granulosa cell synthesis E:
样品 E2生成率 (％) Sample E 2 generation rate (%)
空白对照 100  Blank control 100
瓦山安息香提取物 1 174.75  Washan benzoin extract 1 174.75
瓦山安息香提取物 2 256.23  Washan benzoin extract 2 256.23
楚雄安息香提取物 144.21  Chuxiong benzoin extract 144.21
滇南安息香提取物 123.88  Minnan benzoin extract 123.88
禄春安息香提取物 179.21  Luchun benzoin extract 179.21
化合物 1 106.25  Compound 1 106.25
化合物 2 104.95  Compound 2 104.95
化合物 3 223.21  Compound 3 223.21
化合物 4 112.34  Compound 4 112.34
化合物 5 103.35  Compound 5 103.35
化合物 6 101.58  Compound 6 101.58
化合物 7 134.61  Compound 7 134.61
化合物 8 133.28  Compound 8 133.28
化合物 9 101.86  Compound 9 101.86
化合物 10 402.19  Compound 10 402.19
化合物 11 440.46  Compound 11 440.46
化合物 12 401.86  Compound 12 401.86
由表 1可以明显地看到瓦山安息香提取物 1、 2 (总苯并呋喃重量百分比分 别为 40%和 90% )、 楚雄安息香提取物（总苯并呋喃重量百分比为 60% )、 滇南 安息香提取物（总苯并呋喃重量百分比为 70% )、 禄春安息香提取物（总苯并 呋喃重量百分比为 80% )以及从瓦山安息香中分离的化合物 1-12对小鼠卵巢颗 粒细胞产生 E2有显箸的促进作用。 It can be clearly seen from Table 1 that the benzoin extract 1, 2 (the total benzofuran weight percentage is 40% and 90%, respectively), the Chuxiong benzoin extract (the total benzofuran weight percentage is 60%), Weinan Benzoin extract (70% by weight of total benzofuran), Luchun benzoin extract (80% by weight of total benzofuran), and compound 1-12 isolated from Wasabi benzoin produce mouse ovarian granulosa cells E 2 has a significant promoting effect.
【实施例 11】 促进小鼠脂肪细胞合成 E2: [Example 11] Promoting mouse fat cell synthesis E 2 :
将诱导分化的小鼠 3T3- L1脂肪细胞按 2.4x105/孔， lmL/孔接种到 24孔培养 液中， 待细胞生长为单层后， 加入 E2合成所需底物睾酮 （0.5mM )和浓度为 ltX^g/ml的待测药物， 同时设立不加药物、仅含睾酮的药物空白对照组， 继续 在 37°C , 5% C02条件下培养 72小^, 收集培养液， 1000转 /分钟离心 3min, 取 上清， 采用酶免试剂盒检测其中雌二醇（E2 )含量。 将只加入 0.5mM睾酮的对 照组合成 E2的含量视为 100%, 以此为基准计算其它各样品组 E2的合成率， 结 果见表 2。 The differentiated mouse 3T3-L1 adipocytes were inoculated into 24-well culture medium at 2.4×105/well, 1 mL/well. After the cells were grown into a monolayer, E 2 was used to synthesize the desired substrate testosterone (0.5 mM). Concentration is ltX^g/ml of the drug to be tested, and a drug control group containing no drug and testosterone alone, and continuing to culture at 37 ° C, 5% C0 2 for 72 hours, collecting the culture solution, 1000 rpm After centrifugation for 3 min, the supernatant was taken, and the content of estradiol (E 2 ) therein was measured using an enzyme-free kit. The content of the synthetic E 2 in the control group in which only 0.5 mM of testosterone was added was regarded as 100%, and the synthesis ratio of the other sample groups E 2 was calculated based on this, and the results are shown in Table 2.
表 2促进小鼠脂肪细胞合成 E:  Table 2 promotes mouse fat cell synthesis E:
Figure imgf000019_0001
由表 2可以明显地看到瓦山安息香提取物 1、 2以及楚雄安息香提取物、 滇南安息香提取物、 禄春安息香提取物和从瓦山安息香中分离的化合物 1-12 对 3T3-L1脂肪细胞产生 E2也有显著地促进作用。 【实施例 12】 化合物 1-12促进大鼠 ROS 17/2.8成骨样骨肉瘤细胞合成 E2 大鼠 ROS 17/2.8骨肉瘤细胞接种在 24孔培养板， 细胞生长为单层后，加入 E2合成所需底物睾酮（0.5mM )和浓度为 lOO g/ml的待测药物， 同时设立不加 药物、仅含睾酮的药物空白对照组， 继续在 37°C , 5% C02条件下培养 72小时， 收集培养液， 1000转 /分钟离心 3min, 取上清， 采用酶免试剂盒检测其中雌二 醇（E2 )含量。 将只加入 0.5mM睾酮的对照组合成 E2的含量视为 100%, 以此 为基准计算其它各样品组 E2的合成率， 结果见表 3。
Figure imgf000019_0001
From Table 2, it can be clearly seen that Washan benzoin extract 1, 2 and Chuxiong benzoin extract, Minnan benzoin extract, Luchun benzoin extract and compound 1-12 isolated from Washan benzoin to 3T3-L1 fat The production of E 2 by cells also has a significant promoting effect. [Example 12] Compound 1-12 promotes rat ROS 17/2.8 osteoblast-like osteosarcoma cell synthesis E 2 rat ROS 17/2.8 osteosarcoma cells are seeded in a 24-well culture plate, and after the cells are grown into a single layer, E is added. 2 Synthesis of the desired substrate testosterone (0.5 mM) and the concentration of 100 g / ml of the drug to be tested, while setting up a drug-free, testosterone-only drug blank control group, continue at 37 ° C, 5% C0 2 conditions After culturing for 72 hours, the culture solution was collected, centrifuged at 1000 rpm for 3 min, and the supernatant was taken, and the content of estradiol (E 2 ) therein was measured using an enzyme-free kit. The content of the synthetic E 2 in the control group in which only 0.5 mM of testosterone was added was regarded as 100%, and the synthesis ratio of the other sample groups E 2 was calculated based on this, and the results are shown in Table 3.
表 3.化合物 1-12促进大鼠 ROS 17/2.8骨肉瘤细胞合成 E2 Table 3. Compound 1-12 promotes rat ROS 17/2.8 osteosarcoma cell synthesis E 2
样品 E2生成率 (％) Sample E 2 generation rate (%)
空白对照 100  Blank control 100
化合物 1 118.19  Compound 1 118.19
化合物 2 112.43  Compound 2 112.43
化合物 3 180.06  Compound 3 180.06
化合物 4 138.73  Compound 4 138.73
化合物 5 131.61  Compound 5 131.61
化合物 6 114.84  Compound 6 114.84
化合物 7 140.66  Compound 7 140.66
化合物 8 139.15  Compound 8 139.15
化合物 9 120.30  Compound 9 120.30
化合物 10 280.39  Compound 10 280.39
化合物 11 314.67  Compound 11 314.67
化合物 12 251.79 由表 3可以明显看出化合物 1-12对大鼠 ROS 17/2.8骨肉瘤细胞产生雌激素 均有促进作用。  Compound 12 251.79 It is apparent from Table 3 that Compound 1-12 promotes estrogen production in rat ROS 17/2.8 osteosarcoma cells.
【实施例 13】齐墩杲醇龙胆二糖苷（化合物 10 )、 齐墩果醇龙胆三糖苷 [Example 13] Olean sterol gentioside (compound 10), oleanol gentiotriosine
(化合物 11 )提高卵巢去势小鼠血清雌激素水平， 促进小鼠体内雌激素 E2 的合成 (Compound 11) Improve serum estrogen levels in ovariectomized mice and promote the synthesis of estrogen E 2 in mice
成年雌性小鼠 50只，平均分为 5組，取 4组用手术切除双侧卵巢（去势）, 剩余一组进行假手术（进行同样步骤手术操作但不切除小鼠卵巢）。 术后第 4 天开始， 对去势各組逐只做阴道涂片检查， 直至观察到无阴道上皮角化细胞， 证明卵巢切除完全， 符合 "更年期" 表现。 然后， 模型组灌胃给予生理盐水； 实验组小鼠灌胃给予化合物 10和 11 ( 50mg /kg/d); 阳性对照组注射给予苯 曱酸雌二醇（0.0311¾/1¾/(1)，假手术组作为正常对照组，灌胃给予生理盐水。 所有小鼠在给药 30天后行眶静脉丛采血，分离血清测定其雌激素 E2水平， 结 果见下表。 50 adult female mice were divided into 5 groups, and 4 groups were surgically resected bilateral ovaries (castration). The remaining group underwent sham surgery (the same procedure was performed but the mouse ovaries were not removed). On the 4th day after surgery, vaginal smear was performed on each castration group until no vaginal epithelial keratinocytes were observed, which proved that oophorectomy was complete and met the "menopause" performance. Then, the model group was given normal saline by intragastric administration; the experimental group was intragastrically administered with compounds 10 and 11 (50 mg / kg / d); the positive control group was injected with estradiol benzoate (0.03113⁄4/13⁄4/(1), The sham operation group was used as a normal control group, and normal saline was administered by gavage. All the mice were subjected to blood sampling for 30 days after administration, and estrogen E 2 levels were determined by separating serum. The results are shown in the following table.
表 化合物 10、 11对卵巢去势小鼠血清 E2含量的影响 ±S,n = 10) Effects of Table Compounds 10 and 11 on Serum E 2 Content in Ovarian Ovariectomized Mice ±S, n = 10)
Figure imgf000021_0001
Figure imgf000021_0001
注： 与模型组比较 *P< 0.05  Note: Compared with the model group *P< 0.05
上表显示， 灌胃给予药化合物 10和 11一定时间后， 去势小鼠血清雌激素 E2水平明显升高， 说明化合物 10和 11能促进小鼠体内雌激素 E2的合成。 工业应用性 The above table shows that serum estrogen E2 levels in ovariectomized mice are significantly increased after a certain time of administration of drug compounds 10 and 11, indicating that compounds 10 and 11 can promote the synthesis of estrogen E 2 in mice. Industrial applicability
本发明不仅提供了一种新的安息香属总苯并呋喃植物提取物， 和新的安 息香属植物苯并呋喃衍生物单体， 还提供了一种新的治疗与雌激素缺乏相关 的疾病的药物组合物， 由于天然植物药本身的特性， 本发明为治疗卵巢功能 早衰、 更年期综合征、 骨质疏松以及青春期卵巢功能低下、 多囊卵巢综合症、 不孕症以及因雌激素缺乏或相对不足引起的疾病提供了一种安全价廉、 疗效 显著、 使用方便的用药途径。  The present invention not only provides a novel plant extract of benzofuran benzofuran, and a novel benzofuran derivative monomer of the benzoin plant, but also provides a new drug for treating diseases associated with estrogen deficiency. Composition, due to the nature of the natural botanical itself, the present invention is for the treatment of premature ovarian failure, climacteric syndrome, osteoporosis and ovarian dysfunction in adolescence, polycystic ovary syndrome, infertility and estrogen deficiency or relative deficiency The disease provides a safe, inexpensive, effective, and easy-to-use route of administration.

Claims

权利要求书 Claim
1、一种安息香属植物提取物，其特征是：含有安息香属植物总苯并呋喃， 其重量百分比为 40 ~ 98 %。 A benzoin plant extract characterized by containing a benzofuran total benzofuran in a weight percentage of 40 to 98%.
2、根据权利要求 1所述的安息香属植物提取物， 其特征是： 所述安息香 属植物总苯并呋喃包含通式为 I的齐 i袭果醇型苯并呋喃和 /或其苷
Figure imgf000022_0001
The benzoin plant extract according to claim 1, wherein: the benzoin plant total benzofuran comprises a quinone furan type benzofuran of the formula I and/or a glycoside thereof
Figure imgf000022_0001
其中： -OH, -OCH3; R2为 -H， -COCH3, -CO(CH2)2CH3, β-D- 葡萄糖基， 龙胆双糖基， 龙胆三糖基， R3-R7为 -H, -OH, -OCH3， 其他含氧 取代基团。 Wherein: -OH, -OCH 3 ; R 2 is -H, -COCH 3 , -CO(CH 2 ) 2 CH 3 , β-D-glucosyl, gentiobiose, gentiotriosyl, R 3 -R 7 is -H, -OH, -OCH 3 , other oxygen-containing substituent groups.
3、根据权利要求 2所述的安息香属植物提取物， 其特征在于： 所述通式 为 I的化合物， 是-0(：¾， R2是 -H， R4-R5为二氧亚曱基， R3、 R6、 R7均 为 -H， 即为 -5-(3-羟基)丙基 -7-曱氧基 -2-(3， 4-二氧亚曱基苯基） 苯并呋喃， 即齐墩果醇。 The benzoin plant extract according to claim 2, wherein: the compound of the formula I is -0 (: 3⁄4, R 2 is -H, and R 4 - R 5 is dioxy Sulfhydryl, R 3 , R 6 , and R 7 are all -H, which is -5-(3-hydroxy)propyl-7-decyloxy-2-(3,4-dioxoindenylphenyl) Benzofuran, oleanol.
4、 根据权利要求 2所述的安息香属植物提取物， 其特征在于： 是-11， R2是 -H， R4-R5为二氧亚曱基， R3、 R6、 R7均为 -H， 即为 -5-(3-羟基)丙基 -2-(3， 4-二氧亚曱基苯基）苯并呋喃， 即去曱氧基齐墩果醇。 The benzoin plant extract according to claim 2, which is characterized in that it is -11, R 2 is -H, R 4 -R 5 is a dioxyindenyl group, and R 3 , R 6 and R 7 are both It is -H, which is -5-(3-hydroxy)propyl-2-(3,4-dioxoindolylphenyl)benzofuran, that is, demethoxyl oleanol.
5、根据权利要求 2所述的安息香属植物提取物，其特征在于： 为-OH， R2为 -H， R4-R5为二氧亚甲基， R3、 R6、 R7均为 -H， 即为 5-(3-羟基)丙基 -7- 羟基 -2-(3， 4-二氧亚甲基苯基）苯并呋喃， 即去曱基齐墩果醇。 The benzoin plant extract according to claim 2, wherein: -OH, R 2 is -H, R 4 -R 5 is dioxymethylene, and R 3 , R 6 and R 7 are both It is -H, which is 5-(3-hydroxy)propyl-7-hydroxy-2-(3,4-dioxymethylenephenyl)benzofuran, that is, dethiol oleanol.
6、 居权利要求 2 所述的安息香属植物提取物， 其特征在于： 为 -OCH3， R2为 -COCH3， R4-R5为二氧亚曱基， R3、 R6、 R7均为 -H, 即为齐墩 果醇乙酸酯。 The benzoin plant extract according to claim 2, which is characterized in that: -OCH 3 , R 2 is -COCH 3 , R 4 -R 5 is dioxyarylene, R 3 , R 6 , R 7 is -H, which is oleanol acetate.
7、 才 据权利要求 2所述的安息香属植物提取物， 其特征在于： 为-11， R2为 -COCH3， R4-R5为二氧亚甲基， R3、 R6、 R7均为 -H, 即为去曱氧基齐墩 果醇乙酸酯。 7. The extract of benzoin plant according to claim 2, wherein: -11, R 2 is -COCH 3 , R 4 -R 5 is dioxymethylene, R 3 , R 6 , R 7 is -H, which is demethoxyl oleanol acetate.
8、 根据权利要求 2 所述的安息香属植物提取物， 其特征在于： 为 -OCH3, R2为 -CO(CH2)2C¾， R4-R5为二氧亚甲基， R3、 R6、 R7均为 -H, 即 为齐 J¾果醇丁酸酯。 8. The benzoin plant extract according to claim 2, wherein: -OCH3, R 2 is -CO(CH 2 ) 2 C3⁄4, R 4 -R 5 is a dioxymethylene group, and R 3 , R 6 and R 7 are both -H, that is, a J3⁄4 fruit alcohol butyrate.
9、 根据权利要求 2 所述的安息香属植物提取物， 其特征在于： 为 -OCH3， R2为 β- D-葡萄糖基， R4-R5为二氧亚甲基， R3、 R6、 R7均为 -H, 即 为齐墩果醇 -葡萄糖苷。 The benzoin plant extract according to claim 2, wherein: -OCH 3 , R 2 is β-D-glucosyl, R 4 -R 5 is dioxymethylene, R 3 , R 6. R 7 is -H, which is oleanol-glucoside.
10、 根据权利要求 2所述的安息香属植物提取物， 其特征在于： 为 OCH3， R2为龙胆双糖基， R4-R5为二氧亚曱基， R3、 R6、 R7均为 -H, 即为齐 墩果醇-龙胆双糖苷。 The benzoin plant extract according to claim 2, wherein: OCH 3 , R 2 is a gentiobiose group, R 4 - R 5 is a dioxinylene group, R 3 , R 6 , R 7 is -H, which is oleanol-gentiobiose.
11、 根据权利要求 2所述的安息香属植物提取物， 其特征在于： 为 OCH3， R2为龙胆三糖基， R4-R5为二氧亚曱基， R3、 R6、 R7均为 -H, 即为齐 墩果醇-龙胆三糖苷。 The benzoin plant extract according to claim 2, wherein: OCH 3 , R 2 is gentiotriosyl, R4-R 5 is dioxyindenyl, R 3 , R 6 , R 7 is -H, which is oleanol-gentiotriosin.
12、根据权利要求 2所述的安息香属植物提取物，其特征在于： 为-11， R2为龙胆双糖基， R4-R5为二氧亚甲基， R3、 R6、 R7均为 -H, 即为去曱氧基 齐墩果醇-龙胆汉糖苷。 The benzoin plant extract according to claim 2, wherein: -11, R 2 is gentiobiose, R 4 - R 5 is dioxymethylene, R 3 , R 6 , R 7 is -H, which is demethoxyl oleanol - gentian glucoside.
13、 根据权利要求 1所述的安息香属植物提取物， 其特征是： 所述安息 香属植物包括： 天然植物瓦山安息香 [SZyrax perkinsiae] , 禄春安息香 [S. macranthus Perk.],楚雄安息香 [S. Umprichtii Lingelsh et Borza] ,越南安息香 [S. tonkinensis (Pierre) Craib et Hartw.] , 浙江安息香 [51· zhejiangensis S.M. Hwang et L. L. Yu]，芬芳安息香 [S. odoratissimus Champ.] ,中华安息香 [S. chinehsis Hu et S. Y. Liang] ,滇南安息香 [S. benzoinoides Graib] ,苏门答腊安息香 [S. benzoin Dry and] , 银花安息香 [>S. argentifolius H. L. Li] , 灰叶野茉莉 [S. calvescens Perk] ,嘉赐叶野茉莉 [>S. casearifolius Craib]，黄果安息香 chrysocarpus H. L. Li]， 赛山梅 conf sus Hemsl]， 垂珠花 dasyanthus Perk]， 白花龙 [»S. faberi Perk] , 大花野茉莉 gra"i¾y?o ¾s Griff] , [S. hemsleyanus Diels] , 西藏 野茉莉 [>S. hookeri C. B. Clarke]， 野茉莉 [S. japonicus Sieb. et Zucc]， 大籽野茉 莉 [S. macrospermus C. Y. Wu]， 桐叶野茉莉 [S. mallotifolius C. Y. Wu]， 粉花野 茉莉 [«S. roseus Dunn] , 皱叶野茉莉 [>S. rugosus K z] , 齿叶野茉莉 [>S. serrulatus Roxb] , 栓叶安息香 [S. suberifolius Hook, et Arn] , 小叶安息香 [S. wilsonii Rehd] , S. ferrugineus, S. officinalis, <S. o&im " Sieb. et Zucc。 13. The benzoin plant extract according to claim 1, wherein: the benzoin plant comprises: a natural plant, SZyrax perkinsiae, S. macranthus Perk., Chuxiong benzoin [ S. Umprichtii Lingelsh et Borza], Vietnamese benzoin [S. tonkinensis (Pierre) Craib et Hartw.], Zhejiang benzoin [5 1 · zhejiangensis SM Hwang et LL Yu], fragrant benzoin [S. odoratissimus Champ.], Chinese benzoin [ S. chinehsis Hu et SY Liang] , S. benzoinoides Graib, S. benzoin Dry and, S. argentifolius HL Li, S. calvescens Perk ], 赐氏野野茉莉[>S. casearifolius Craib], 黄果息息香 chrysocarpus HL Li], 赛山梅conf sus Hemsl], 垂珠花 dasyanthus Perk], 白花龙[»S. faberi Perk] , 大花野Jasmine gra"i3⁄4y?o 3⁄4s Griff] , [S. hemsleyanus Diels] , S. hookeri CB Clarke, S. japonicus Sieb. et Zucc, S. macrospermus CY Wu], Tongye wild jasmine [S. m Allotifolius CY Wu], «S. roseus Dunn】, sage jasmine [>S. rugosus K z], jasmine jasmine [>S. serrulatus Roxb], sage leaf scent [S. suberifolius Hook, Et Arn], S. wilsonii Rehd, S. ferrugineus, S. officinalis, <S. o&im "Sieb. et Zucc.
14、根据权利要求 13所述的安息香属植物提取物， 其特征是： 所述安息 香属植物是瓦山安息香 er ^ e]或禄春安息香 [5". macranthus Perk.]。 The benzoin plant extract according to claim 13, wherein the benzoin plant is Washan benzoin er ^ e] or Luchun benzoin [5". macranthus Perk.
15、一种制备权利要求 1 ~ 14任一项所述的安息香属植物提取物的方法， 其特征在于： 包括以下步骤：  A method of preparing a benzoin plant extract according to any one of claims 1 to 14, which comprises the steps of:
a、 将安息香属植物果实或叶粉碎后， 用水或醇， 或 20-95%的含水醇为 提取溶剂提取， 提取液经减压浓缩得到总浸膏；  a. After pulverizing the fruit or leaf of the benzoin plant, it is extracted with water or alcohol, or 20-95% of an aqueous alcohol as an extraction solvent, and the extract is concentrated under reduced pressure to obtain a total extract;
b、将上述总浸膏用热水溶散后，用石油醚萃取，以除掉其中的油性成分； c、 b步骤萃取后的水层用大孔树脂柱吸附后， 先用水洗脱，再用 30-95% 乙醇洗脱柱子， 收集乙醇洗脱液， 浓缩、 干燥， 即得安息香属植物提取物。  b. After dissolving the above total extract with hot water, extract it with petroleum ether to remove the oily component; c, b. The aqueous layer after the extraction is adsorbed by the macroporous resin column, first eluted with water, and then used. The column was eluted with 30-95% ethanol, and the ethanol eluate was collected, concentrated, and dried to obtain a benzoin extract.
16、 根据权利要求 15 所述的安息香属植物总苯并呋喃提取物的制备方 法， 其特征在于： a步驟所述提取溶剂为 50-90 %乙醇溶液， 乙醇溶液用量为 药材重量的 6-12倍； c步驟所述洗脱剂为 95 %乙醇。  The method for preparing a benzofuran extract of benzoin according to claim 15, wherein: the extraction solvent is a 50-90% ethanol solution, and the amount of the ethanol solution is 6-12 of the weight of the medicinal material. The c-eluent is 95% ethanol.
17、 一种安息香属植物总苯并呋喃提取物， 其特征是： 它是安息香属植 物由权利要求 15或 16所述的方法制备而成。  A benzofuran extract of benzoin, characterized in that it is a plant of the genus Benzoin prepared by the method of claim 15 or 16.
18、根据权利要求 17的安息香属植物提取物， 其特征是： 所述安息香属 植物为瓦山安息香 [SCra perkinsiae]，禄春安息香 [S. macranthus Perk.]， 楚雄 安息香 [>S. limprichtii Lingelsh et Borza] , 越南安息香 tonkinensis (Pierre) Craib et Hartw.] , 浙江安息香 zhejiangensis S.M. Hwang et L. L. Yu]， 芬芳 安息香 odoratissimus Champ.]， 中华安息香 chinehsis Hu et S. Y. Liang]， 滇南安息香 benzoinoides G ib] ,苏门答腊安息香 benzoin Όιγ and] ,银花 安息香 [*S. arge«ii[/ /z'Ms H. L. Li]， 灰叶野茉莉 [S. c v&scera Perk] , 嘉赐叶野 茉莉 casearifolius Craib] , 黄果安息香 [S. chrysocarpus H. L. Li] , 赛山梅 confusus Hemsl]， 垂珠花 [51· dasyanthus Perk]， 白花龙 [51· faberi Perk]， 大花野 茉莉 grandiflorus Griff] , 老舌乌铃 hemsleyanus Diels] , 西藏野茉莉 hookeri C. B. Clarke] , 野茉莉 japonicus Sieb. et Zucc] , 大籽野茉莉 macrospermus C. Y. Wu] ,桐叶野茉莉 [>S. mallotifolius C. Y. Wu] ,粉花野茉莉 [S. roseus Dunn]， 铍叶野茉莉 [S. rugosus Kurz]， 齿叶野茉莉 [S. serrulatus Roxb]， 栓叶安息香 [S. suberifolius Hook, et Arn] , 小叶安息香 [S. wilsonii Rehd] , S. ferrugineus, S. officinalis, S. obassia Sieb. et Zucc等中任意一种或几种。 The benzoin plant extract according to claim 17, wherein: the benzoin plant is SCra perkinsiae, S. macranthus Perk., Chuxiong benzoin [>S. limprichtii Lingelsh Et Borza] , Vietnamese benzoin tonkinensis (Pierre) Craib et Hartw.], Zhejiang benzoin zhejiangensis SM Hwang et LL Yu], fragrant benzoin odoratissimus Champ., Chinese benzoin chinehsis Hu et SY Liang], Minnan benzoinides G ib] , Sumatra benzoin benzoin Όιγ and], silver flower benzoin [*S. arge«ii[/ /z'Ms HL Li], jasmine jasmine [S. c v&scera Perk], jasmine jasmine casearifolius Craib], yellow fruit Benzoin [S. chrysocarpus HL Li], Saishanmei confusus Hemsl], deciduous flower [5 1 · dasyanthus Perk], white flower dragon [5 1 · faberi Perk], jasmine grandiflorus Griff], old tongue hemsleyanus Diels ] , Tibet wild jasmine hookeri CB Clarke] , jasmine japonicus Sieb. et Zucc] , jasmine macrospermus CY Wu], paulownia jasmine [>S. mallotifolius CY Wu], pink jasmine [S. Roseus Dunn], S. rugosus Kurz, S. serrulatus Roxb, S. suberifolius Hook, et Arn, S. wilsonii Rehd, S. Any one or more of ferrugineus, S. officinalis, S. obassia Sieb. et Zucc.
19、根据权利要求 18所述的安息香属植物提取物， 其特征是： 所述安息 香属植物是 山安息香 [S^ rax 或禄春安息香 macranthus Perk.]。 The benzoin plant extract according to claim 18, wherein the benzoin plant is benzoin [S^rax or cyclamate macranthus Perk.].
20、 安息香属植物在制备预防或治疗卵巢功能早衰、 更年期综合征、 骨 质疏松以及青春期卵巢功能低下、 多嚢卵巢综合症以及不孕症以及因雌激素 缺乏引起的疾病的药物中的用途。  20. Use of benzoin plants for the preparation of a medicament for the prevention or treatment of premature ovarian failure, climacteric syndrome, osteoporosis and ovarian dysfunction in adolescence, polypseudo-ovarian syndrome and infertility and diseases caused by estrogen deficiency.
21、 权利要求 1-14任意一项或权利要求 17所述的安息香属植物总苯并 呋喃提取物在制备预防或治疗卵巢功能早衰、 更年期综合征、 骨质疏松以及 青春期卵巢功能低下、 多嚢卵巢综合症以及不孕症以及因雌激素缺乏引起的 疾病的药物中的用途。  The benzoic acid total benzofuran extract according to any one of claims 1 to 14 or claim 17, wherein the preparation or the prevention of premature ovarian failure, climacteric syndrome, osteoporosis and ovarian dysfunction in adolescence, and sputum Use in ovarian syndrome and infertility and drugs for diseases caused by estrogen deficiency.
22、权利要求 2- 12任一项所述的化合物在制备治疗卵巢功能早衰、 更年 期综合征、 骨质疏松以及青春期卵巢功能低下、 多嚢卵巢综合症、 不孕症以 及因雌激素缺乏或相对不足引起的疾病的药物中的用途。  22. A compound according to any one of claims 2 to 12 for the treatment of premature ovarian failure, climacteric syndrome, osteoporosis and ovarian dysfunction in adolescence, ovarian syndrome, infertility and estrogen deficiency or relative Use in drugs for diseases caused by insufficient.
23、 一种药物组合物， 其活性成分含有安息香属植物原生药或权利要求 1-14所述的安息香属植物提取物或权利要求 2-12任一项所述的化合物。  A pharmaceutical composition, wherein the active ingredient comprises a benzoin plant or a benzoin plant extract according to any one of claims 1 to 14 or a compound according to any one of claims 2 to 12.
24、根据权利要求 23所述的药物组合物， 其特征在于： 其活性成分为权 利要求 1-14所述的安息香属植物提取物，或其与调节雌激素合成的西药或其 他中药活性成分配伍成的复方， 加上药学上可接受的载体或辅料， 制备而成 的制剂。  The pharmaceutical composition according to claim 23, wherein the active ingredient is the extract of the genus Benzoin according to claims 1-14, or it is distributed with the activity of western medicine or other traditional Chinese medicine which regulates the synthesis of estrogen. A formulated preparation, together with a pharmaceutically acceptable carrier or adjuvant, is prepared.
25、根据权利要求 23、 24所述的药物组合物， 其特征在于： 所述的制剂 是： 口服片剂、 胶嚢、 口服液或注射液。  The pharmaceutical composition according to Claim 23, 24, wherein the preparation is: an oral tablet, a capsule, an oral solution or an injection.
PCT/CN2006/000341 2005-03-08 2006-03-07 An extract of styrax linn, the preparation and uses thereof WO2006094456A1 (en)

Applications Claiming Priority (6)

Application Number Priority Date Filing Date Title
CN200510020478.1 2005-03-08
CN200510020477.7 2005-03-08
CNB2005100204796A CN100420690C (en) 2005-03-08 2005-03-08 A kind of benzofuran compound and its preparation method and application
CN200510020478A CN1830435B (en) 2005-03-08 2005-03-08 Application of oleanol-type benzofuran and its glycoside in the preparation of anti-estrogen deficiency drugs
CN200510020479.6 2005-03-08
CNB2005100204777A CN100509803C (en) 2005-03-08 2005-03-08 Extractive of total benzofuran of benzoin plants, prepn. method and use thereof

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Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
ANIL H.: "Four benzofuran glycosides from Styrax officinalis", PHYTOCHEMISTRY, vol. 19, no. 12, 1980, pages 2784 - 2786 *
MENDONCA P.P. ET AL.: "Nor-lignans from the leaves of Styras ferrugineus (Styracaceae) with antibacterial and antifungal activity", PHYTOCHEMISTRY, vol. 55, no. 6, 2000, pages 597 - 601 *
TAKANASHI M. ET AL.: "New benzofurans related to egonol from immature seeds of Styrax obassia", PHYTOCHEMISTRY, vol. 27, no. 4, 1988, pages 1224 - 1226 *
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