WO2017215679A2 - 苯丁酸氮芥的药物组合物及其抗抑郁的医药用途 - Google Patents

苯丁酸氮芥的药物组合物及其抗抑郁的医药用途 Download PDF

Info

Publication number
WO2017215679A2
WO2017215679A2 PCT/CN2017/097016 CN2017097016W WO2017215679A2 WO 2017215679 A2 WO2017215679 A2 WO 2017215679A2 CN 2017097016 W CN2017097016 W CN 2017097016W WO 2017215679 A2 WO2017215679 A2 WO 2017215679A2
Authority
WO
WIPO (PCT)
Prior art keywords
chlorambucil
compound
ethanol
pharmaceutical composition
extract
Prior art date
Application number
PCT/CN2017/097016
Other languages
English (en)
French (fr)
Other versions
WO2017215679A3 (zh
Inventor
崔坤峰
Original Assignee
赵吉永
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by 赵吉永 filed Critical 赵吉永
Publication of WO2017215679A2 publication Critical patent/WO2017215679A2/zh
Publication of WO2017215679A3 publication Critical patent/WO2017215679A3/zh

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
    • A61K31/19—Carboxylic acids, e.g. valproic acid
    • A61K31/195—Carboxylic acids, e.g. valproic acid having an amino group
    • A61K31/196—Carboxylic acids, e.g. valproic acid having an amino group the amino group being directly attached to a ring, e.g. anthranilic acid, mefenamic acid, diclofenac, chlorambucil
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/336—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having three-membered rings, e.g. oxirane, fumagillin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/24—Antidepressants
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D301/00—Preparation of oxiranes
    • C07D301/32—Separation; Purification
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D303/00—Compounds containing three-membered rings having one oxygen atom as the only ring hetero atom
    • C07D303/02—Compounds containing oxirane rings
    • C07D303/12—Compounds containing oxirane rings with hydrocarbon radicals, substituted by singly or doubly bound oxygen atoms
    • C07D303/32—Compounds containing oxirane rings with hydrocarbon radicals, substituted by singly or doubly bound oxygen atoms by aldehydo- or ketonic radicals

Definitions

  • the invention belongs to the field of biomedicine and relates to a new use of chlorambucil, and particularly relates to a pharmaceutical composition of chlorambucil and its antidepressant medical use.
  • Chlorambucil is a nitrogen mustard derivative, similar to cyclophosphamide, and has inhibitory effects on various tumors. It is clinically used for chronic lymphocytic leukemia, lymphosarcoma, He Jinjie's disease, ovarian cancer, breast cancer, and villus epithelioma. , multiple myeloma and so on.
  • Chlorambucil has a good effect on Chet syndrome (genital ulcer, aphthous and uveitis syndrome), lupus erythematosus, Wegener's granulomatosis; for the treatment of rheumatoid arthritis complicated with vasculature Inflammation and autoimmune hemolytic anemia with cold lectin have a good effect; patients with nephrotic syndrome relying on corticosteroids can achieve complete remission, and prednisolone and frequent nephrotic syndrome can be significantly reduced Its recurrence rate; scleroderma can quickly prevent its development, so that skin ulcers heal, lung function improved.
  • a pharmaceutical composition of chlorambucil comprising chlorambucil, a compound (I) as described above, and a pharmaceutically acceptable carrier.
  • the preparation method of the compound (I) as described above comprises the following steps: (a) pulverizing the dried roots of Radix Pseudostellariae, extracting with 75-85% ethanol under reflux, combining the extracts, concentrating to an alcohol-free taste, and sequentially using Extraction of petroleum ether, ethyl acetate and water-saturated n-butanol to obtain petroleum ether extract, ethyl acetate extract and n-butanol extract, respectively; (b) step (a) The n-butanol extract was removed by macroporous resin, first eluted with 10% ethanol, 8 column volumes, and then eluted with 70% ethanol for 10 column volumes, 70% eluate was collected, and concentrated under reduced pressure to obtain 70% ethanol.
  • step (c) 70% ethanol elution concentrate in step (b) separated by normal phase silica gel, sequentially using dichloromethane in a volume ratio of 60:1, 30:1, 15:1 and 5:1 - methanol gradient elution to obtain 4 components;
  • component (c) in step (c) is further separated by normal phase silica gel, sequentially using dichloromethane in a volume ratio of 8:1, 4:1 and 2:1 - The methanol gradient elutes to obtain 3 components;
  • step (e) component 2 is separated by octadecylsilane-bonded reversed phase silica gel and eluted isocratically with 75% by volume aqueous methanol solution. 10-15 column volumes of eluate were collected, and the eluate was concentrated under reduced pressure to give compound (I).
  • step (a) the extract is extracted by hot reflux with 80% ethanol, and the extracts are combined.
  • the macroporous resin is a D101 type macroporous adsorption resin.
  • the pharmaceutical composition of the chlorambucil provided by the present invention comprises chlorambucil and a novel natural product isolated from the ginseng, and the chlorambucil and the natural product have a single action. Certain antidepressant effects; when the two are combined, the antidepressant effect is further improved and can be developed into an antidepressant drug.
  • the present invention has outstanding substantial features and significant advances over the prior art.
  • Reagent sources ethanol, petroleum ether, ethyl acetate, n-butanol, dichloromethane for analytical grade, purchased from Shanghai Lingfeng Chemical Reagent Co., Ltd., methanol, analytically pure, purchased from Jiangsu Hanbang Chemical Reagent Co., Ltd.
  • step (b) the n-butanol extract is decontaminated with D101 macroporous resin, first eluted with 10 column volumes of 10% ethanol, and then eluted with 70% ethanol for 10 column volumes to collect 70% eluate. The concentrate is concentrated by pressure to obtain a 70% ethanol elution; (c) the 70% ethanol elution concentrate in step (b) is separated by normal phase silica gel in a volume ratio of 60:1 (10 column volumes), 30:1.
  • the infrared spectrum indicated that the compound contained a carbonyl group (1740 cm -1 and 1672 cm -1 ) and an olefin (1624 cm -1 ) group; and it had ultraviolet absorption at 244 nm, indicating that it contained an ⁇ , ⁇ -unsaturated carbonyl unit.
  • 13 C-NMR, DEPT and HSQC spectra show 20 carbon signals, including three methyl groups, three methylene groups (two olefin carbons), seven methine groups (two olefin carbons and one oxygenated carbon) And seven quaternary carbons (two carbonyl carbons, one oxycarbon carbon and four olefin carbons), and the above functional structure combined with the number of unsaturation indicates that the compound has a tetracyclic structure.
  • H-12/H-13/H-14/H-8/H-7, H-8/H-9 and H 2 -3/H-2/H 3 -19 are present in the 1 H- 1 H COSY spectrum
  • H 2 -3 and C-1, C-4 and C-10, H-7 and C-5, C-6, C-8 and C-14, H-8 are shown in the HMBC spectrum.
  • the C-7-related signal can be constructed by the related information in the above NMR spectrum, and the compound can be determined to be a rhamnofolane-type diterpenoid. Correlation between H-7 and C-5, C-6, C-8 and C-14, H 3 -20 and C-5, C-6 and C-7 in HMBC spectra and C-6 and C-7 Chemical shifts indicate that C-6 and C-7 form an epoxy structure.
  • mice Male and female, weighing 18-22 g, were purchased from the Experimental Animal Center of Guangzhou University of Traditional Chinese Medicine.
  • the chlorambucil was purchased from the China National Institute for the Control of Pharmaceutical and Biological Products, and the compound (I) was prepared. The preparation method is shown in Example 1. Fluoxetine Hydrochloride Capsule (Prozac) (Lilly Suzhou Pharmaceutical Co., Ltd.
  • the hanging tail experimental device was made by itself, and the wooden stick was separated into two zones by cardboard, and the rat tail was fixed with tape; the swimming experimental device was a rectangular glass cylinder (19.4cm ⁇ 20.4cm); the diamond mechanical stopwatch was 3, and the Shanghai base instrument was limited.
  • mice Fifty mice were randomly divided into 5 groups of 10 animals each. They were blank control group (normal saline), positive control group (fluoxetine hydrochloride 4 mg/mL), chlorambucil group (80 mg/kg), compound (I) group (80 mg/kg), phenylbutyrate nitrogen.
  • mice were placed in a rectangular container with a water depth of 10 cm and a water temperature of (27 ⁇ 3) °C for a total of 6 minutes. After 2 minutes of adaptation, the mice were accumulating within 4 minutes (floating and not moving, only showing the nostrils to keep breathing, limbs) Occasionally, to keep your body from sinking.)
  • the results are shown in Table 1.
  • the immobile state of the mouse in the tail-hanging model reflects its desperate behavior.
  • the antidepressant activity of the drug-administered group began to manifest from the third day, compared with the blank control group, the chlorambucil and the compound (I) composition group.
  • the time of mouse tail suspension was significantly reduced (P ⁇ 0.01), and the time of suspension of mice in the chlorambucil group and compound (I) group decreased (P ⁇ 0.05).

Landscapes

  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Epidemiology (AREA)
  • Psychiatry (AREA)
  • Pain & Pain Management (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biomedical Technology (AREA)
  • Neurology (AREA)
  • Neurosurgery (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Medicines Containing Plant Substances (AREA)
  • Steroid Compounds (AREA)

Abstract

本发明公开了苯丁酸氮芥的药物组合物及其抗抑郁的医药用途，本发明提供的苯丁酸氮芥的药物组合物中含有苯丁酸氮芥和一种从太子参的干燥块根中分离得到的结构新颖的天然产物化合物(Ⅰ)，苯丁酸氮芥和该天然产物单独作用时，具有一定的抗抑郁作用；二者联合作用时，抗抑郁作用进一步提高，可以开发成抗抑郁的药物。本发明与现有技术相比具有突出的实质性特点和显著的进步。

Description

苯丁酸氮芥的药物组合物及其抗抑郁的医药用途 技术领域
本发明属于生物医药领域，涉及苯丁酸氮芥的新用途，具体涉及一种苯丁酸氮芥的药物组合物及其抗抑郁的医药用途。
背景技术
苯丁酸氮芥为氮芥衍生物，作用与环磷酰胺相似，对多种肿瘤有抑制作用，临床用于慢性淋巴细胞白血病、淋巴肉瘤、何金杰氏病、卵巢癌、乳腺癌、绒毛上皮瘤、多发性骨髓瘤等。苯丁酸氮芥对切特综合征(生殖器溃疡、口疮及眼色素层炎综合征)、红斑狼疮、韦格内肉芽肿病有较好疗效；用于治疗类风湿性关节炎并发的脉管炎和伴有寒冷凝集素的自身免疫性溶血性贫血有良好效果；用于依赖皮质激素的肾病综合征患者可得到完全的缓解，与氢化泼尼松并用于频发的肾病综合征可显著降低其复发率；对硬皮病可迅速阻止其发展，使皮肤溃疡痊愈，肺功能改善。
迄今为止，尚未见苯丁酸氮芥及其药物组合物与抗抑郁的相关性报道。
发明内容
本发明的目的在于提供一种苯丁酸氮芥的药物组合物，该药物组合物中含有苯丁酸氮芥和一种从草本中分离得到的结构新颖的天然产物，苯丁酸氮芥和该天然产物可以协同抗抑郁。
本发明的上述目的是通过下面的技术方案得以实现的：
一种具有下述结构式的化合物(Ⅰ)，
Figure PCTCN2017097016-appb-000001
一种苯丁酸氮芥的药物组合物，包括苯丁酸氮芥、如上所述的化合物(Ⅰ)和药学上可以接受的载体。
如上所述的化合物(Ⅰ)的制备方法，包含以下操作步骤：(a)将太子参的干燥块根粉碎，用75～85％乙醇热回流提取，合并提取液，浓缩至无醇味，依次用石油醚、乙酸乙酯和水饱和的正丁醇萃取，分别得到石油醚萃取物、乙酸乙酯萃取物和正丁醇萃取物；(b)步骤(a) 中正丁醇萃取物用大孔树脂除杂，先用10％乙醇洗脱8个柱体积，再用70％乙醇洗脱10个柱体积，收集70％洗脱液，减压浓缩得70％乙醇洗脱浓缩物；(c)步骤(b)中70％乙醇洗脱浓缩物用正相硅胶分离，依次用体积比为60:1、30:1、15:1和5:1的二氯甲烷-甲醇梯度洗脱得到4个组分；(d)步骤(c)中组分4用正相硅胶进一步分离，依次用体积比为8:1、4:1和2:1的二氯甲烷-甲醇梯度洗脱得到3个组分；(e)步骤(d)中组分2用十八烷基硅烷键合的反相硅胶分离，用体积百分浓度为75％的甲醇水溶液等度洗脱，收集10～15个柱体积洗脱液，洗脱液减压浓缩得到化合物(Ⅰ)。
进一步地，步骤(a)中用80％乙醇热回流提取，合并提取液。
进一步地，所述大孔树脂为D101型大孔吸附树脂。
如上所述的化合物(Ⅰ)在制备抗抑郁的药物中的应用。
如上所述的苯丁酸氮芥的药物组合物在制备抗抑郁的药物中的应用。
本发明的优点：
本发明提供的苯丁酸氮芥的药物组合物中含有苯丁酸氮芥和一种从太子参中分离得到的结构新颖的天然产物，苯丁酸氮芥和该天然产物单独作用时，具有一定的抗抑郁作用；二者联合作用时，抗抑郁作用进一步提高，可以开发成抗抑郁的药物。本发明与现有技术相比具有突出的实质性特点和显著的进步。
具体实施方式
下面结合实施例进一步说明本发明的实质性内容，但并不以此限定本发明保护范围。尽管参照较佳实施例对本发明作了详细说明，本领域的普通技术人员应当理解，可以对本发明的技术方案进行修改或者等同替换，而不脱离本发明技术方案的实质和范围。
实施例1：化合物(Ⅰ)分离制备及结构确证
试剂来源：乙醇、石油醚、乙酸乙酯、正丁醇、二氯甲烷为分析纯，购自上海凌峰化学试剂有限公司，甲醇，分析纯，购自江苏汉邦化学试剂有限公司。
分离方法：(a)将太子参的干燥块根(2kg)粉碎，用80％乙醇热回流提取(15L×3次)，合并提取液，浓缩至无醇味(3L)，依次用石油醚(3L×3次)、乙酸乙酯(3L×3次)和水饱和的正丁醇(3L×3次)萃取，分别得到石油醚萃取物、乙酸乙酯萃取物和正丁醇萃取物；(b)步骤(a)中正丁醇萃取物用D101型大孔树脂除杂，先用10％乙醇洗脱8个柱体积，再用70％乙醇洗脱10个柱体积，收集70％洗脱液，减压浓缩得70％乙醇洗脱浓缩物；(c)步骤(b)中70％乙醇洗脱浓缩物用正相硅胶分离，依次用体积比为60:1(10个柱体积)、30:1(8个柱体积)、15:1(10个柱体积)和5:1(8个柱体积)的二氯甲烷-甲醇梯度洗脱得到4个组分；(d)步骤(c)中组分4用正相硅胶进一步分离，依次用体积比为8:1(10个 柱体积)、4:1(8个柱体积)和2:1(6个柱体积)的二氯甲烷-甲醇梯度洗脱得到3个组分；(e)步骤(d)中组分2用十八烷基硅烷键合的反相硅胶分离，用体积百分浓度为75％的甲醇水溶液等度洗脱，收集10～15个柱体积洗脱液，洗脱液减压浓缩得到化合物(Ⅰ)(365mg，HPLC归一化纯度大于98％)。
结构确证：白色粉末，HR-ESI-MS显示[M+Na]+为m/z 333.1460，结合核磁特征可得分子式为C20H22O3，不饱和度为10。核磁共振氢谱数据δH(ppm，CDCl3，500MHz)：H-2(2.48，dd，J＝13.9，6.6Hz)，H-3a(3.05，dd，J＝18.3，6.7Hz)，H-3b(1.91，dd，J＝18.3，2.1Hz)，H-7(2.73，m)，H-8(1.93，dd，J＝9.1，4.2Hz)，H-9(2.47，d，J＝4.2Hz)，H-12(6.14，d，J＝10.2Hz)，H-13(6.71，dd，J＝10.2，6.3Hz)，H-14(2.64，dd，J＝9.1，6.3Hz)，H-16a(4.85，s)，H-16b(4.87，s)，H-17(1.66，s)，H-18a(4.83，s)H-18b(4.37，s)，H-19(1.25，d，J＝7.8Hz)，H-20(2.33，s)；核磁共振碳谱数据δC(ppm，CDCl3，125MHz)：209.8(C，1-C)，40.5(CH，2-C)，34.2(CH2，3-C)，167.6(C，4-C)，207.6(C，5-C)，66.7(C，6-C)，69.4(CH，7-C)，46.3(CH，8-C)，36.8(CH，9-C)，140.3(C，10-C)，146.3(C，11-C)，128.3(CH，12-C)，136.5(CH，13-C)，49.7(CH，14-C)，144.6(C，15-C)，115.2(CH2，16-C)，19.4(CH3，17-C)，111.2(CH2，18-C)，18.1(CH3，19-C)，18.7(CH3，20-C)。红外波谱表明该化合物含有羰基(1740cm-1与1672cm-1)和烯烃(1624cm-1)基团；且其在244nm有紫外吸收，表明含有α,β-不饱和羰基单元。13C-NMR、DEPT和HSQC谱中显示有20个碳信号，包括三个甲基，三个亚甲基(两个烯烃碳)，七个次甲基(两个烯烃碳和一个连氧碳)，以及七个季碳(两个羰基碳、一个连氧碳和四个烯烃碳)，以上功能结构再结合不饱和数表明该化合物为四环结构。1H-NMR谱结合HSQC谱显示三个甲基质子信号δH 1.66(3H，s)、1.25(3H，d，J＝7.8Hz)、2.33(3H，s)，一组亚甲基质子信号δH 3.05(1H，dd，J＝18.3，6.7Hz)与1.91(1H，dd，J＝18.3，2.1Hz)，两组端烯烃质子信号δH 4.85(1H，s)与4.87(1H，s)、4.83(1H，s)与4.37(1H，s)，一对烯烃质子信号δH 6.14(1H，d，J＝10.2Hz)与6.71(1H，dd，J＝10.2，6.3Hz)，一个含氧次甲基质子信号δH2.73(1H，m)，四个次甲基质子信号δH 2.48(1H，dd，J＝13.9，6.6Hz)、1.93(1H，dd，J＝9.1，4.2Hz)、2.47(1H，d，J＝4.2Hz)、2.64(1H，dd，J＝9.1，6.3Hz)。1H-1H COSY谱中存在H-12/H-13/H-14/H-8/H-7、H-8/H-9以及H2-3/H-2/H3-19相关信号，同时HMBC谱中显示有H2-3与C-1、C-4和C-10，H-7与C-5、C-6、C-8和C-14，H-8与C-6、C-7、C-9和C-14，H-13与C-8、C-12、C-14和C-15，H2-16与C-14、C-15和C-17，H2-18与C-9、C-11和C-12，H3-19与C-1、C-2和C-3，H3-20与C-5、C-6和C-7相关信号，通过上述NMR谱中的相关信息可以构建该化合物的连接方式，并且可以确定该化合物为 rhamnofolane型二萜类化合物。HMBC谱中H-7与C-5、C-6、C-8和C-14，H3-20与C-5、C-6和C-7的相关性以及C-6与C-7化学位移表明C-6和C-7形成环氧结构。此外，通过H-12与H-13化学位移及偶合常数可知C-12和C-13之间为双键。ROESY谱中H-7和H-8与Me-20相关信号可知环氧结构为α构型。综合氢谱、碳谱、HMBC谱和ROESY谱，以及文献关于相关类型核磁数据，可基本确定该化合物如下所示，立体构型进一步通过ECD试验确定，理论值与实验值基本一致。化学结构式和碳原子编号如下：
Figure PCTCN2017097016-appb-000002
实施例2：抗抑郁作用
1、材料与方法
1.1动物
实验动物SPF级昆明小鼠，雌雄各半，体重18～22g，购自广州中医药大学实验动物中心。
1.2药物与试剂
苯丁酸氮芥购自中国药品生物制品检定所，化合物(Ⅰ)自制，制备方法见实施例1。盐酸氟西汀胶囊(百忧解)(礼来苏州制药有限公司产品，规格：20mg/片)；重酒石酸去甲肾上腺素(NE)；肾上腺素(E)；盐酸多巴胺(DA)；5-羟色胺盐酸盐(5-HT)；以上5种对照品均购自中国药品生物制品检定所(供含量测定用)；乙腈(色谱纯，澳洲移民局科技有限公司)；辛烷磺酸钠(B8，分析纯，天津市大茂化学试剂厂)；高氯酸(分析纯，广州化学试剂厂)；磷酸二氢钠(分析纯，广州化学试剂厂)。
1.3主要仪器
悬尾实验装置自制，木棒上用纸板隔离成2个区，用胶布固定鼠尾；游泳实验装置为矩形玻璃缸(19.4cm×20.4cm)；钻石牌机械秒表3个，上海垒固仪器有限公司；库仑阵列电化学高效液相色谱仪(美国ESA公司：Model5600A-16通道检测器，Model 582 solvent Delivery System，Model542自动进样器，Coularray Win工作站)；Dionex-Tcc-100型柱温箱(美国戴安公司)；Thermo D-37520型高速冷冻离心机(德国Heraeus公司)；Phs-25型pH计(上 海精密科学仪器有限公司)；GenPure超纯水系统(德国TKA公司)；KQ-500型超声波清洗器(昆山市超声仪器有限公司)；3K超滤管(PALL公司)；BP211D电子天平(德国Sartorious)；佳美SK-1快速混匀器(江苏金坛市佳美仪器厂)。
1.4小鼠悬尾实验(TST)
取小鼠50只，随机分成5组，每组10只。分别为空白对照组(生理盐水)、阳性对照组(盐酸氟西汀4mg/mL)及苯丁酸氮芥组(80mg/kg)、化合物(Ⅰ)组(80mg/kg)、苯丁酸氮芥与化合物(Ⅰ)组合物组【(40mg/kg)苯丁酸氮芥+(40mg/kg)化合物(Ⅰ)】，自由饮水进食灌胃1次/d，0.2mL/只，连续灌胃15d，第1、3、5、7天给药1h后，将小鼠尾部距末端约2cm处用胶布黏住固定，使其头部向下悬挂呈倒悬状，离箱底约5cm。中间以挡板隔离小鼠视线。总观察6min，前2min为适应时间，观察后4min内小鼠的不动(小鼠在空中停止挣扎，或仅有细小的肢体运动)时间。
1.5小鼠强迫游泳实验(FST)
实验前24h进行游泳训练15min，禁食不禁水，实验前12h禁食。第15天给药1h后进行游泳实验。将小鼠单只放入水深10cm，水温(27±3)℃的矩形容器内，总观察6min，适应2min后开始计算后4min内累计不动(漂浮不动状态，仅露出鼻孔保持呼吸，四肢偶尔划动以保持身体不至于沉下去)时间。
2、实验结果
2.1对小鼠悬尾不动时间的影响
结果如表1所示。小鼠在悬尾模型中出现的不动状态反映了其绝望行为，给药组抗抑郁活性从第3天开始体现，与空白对照组比，苯丁酸氮芥与化合物(Ⅰ)组合物组显著减少小鼠悬尾不动时间(P<0.01)，苯丁酸氮芥组、化合物(Ⅰ)组小鼠悬尾不动时间降低(P＜0.05)。
表1对悬尾小鼠不动时间的影响(s，x±s，n＝8)
组别 第3天 第7天
空白对照组 71.00±30.07 99.25±18.42
阳性对照组 29.62±13.96 40.37±16.16
苯丁酸氮芥组 46.37±21.05 57.37±22.30
化合物(Ⅰ)组 43.87±25.65 52.00±26.02
苯丁酸氮芥与化合物(Ⅰ)组合物组 32.75±21.10 43.62±28.37
2.2对小鼠强迫游泳不动时间的影响
结果如表2所示。与空白对照组比较，苯丁酸氮芥与化合物(Ⅰ)组合物组小鼠游泳的 不动时间均显著缩短(P＜0.01)，苯丁酸氮芥组、化合物(Ⅰ)组小鼠游泳不动时间降低(P<0.05)。
表2对小鼠强迫游泳不动时间的影响(x±s，n＝10)
组别 不动时间/s
空白对照组 123.90±44.33
阳性对照组 65.70±22.95
苯丁酸氮芥组 82.33±34.55
化合物(Ⅰ)组 89.56±28.70
苯丁酸氮芥与化合物(Ⅰ)组合物组 58.43±26.20
以上结果表明，苯丁酸氮芥和化合物(Ⅰ)均具有明显的抗抑郁作用，并且两者合用抗抑郁效果显著增强，可以开发成抗抑郁的药物。
上述实施例的作用在于说明本发明的实质性内容，但并不以此限定本发明的保护范围。本领域的普通技术人员应当理解，可以对本发明的技术方案进行修改或者等同替换，而不脱离本发明技术方案的实质和保护范围。

Claims (7)

  1. 一种具有下述结构式的化合物(Ⅰ)，
    Figure PCTCN2017097016-appb-100001
  2. 一种苯丁酸氮芥的药物组合物，其特征在于：包括苯丁酸氮芥、如权利要求1所述的化合物(Ⅰ)和药学上可以接受的载体。
  3. 权利要求1所述的化合物(Ⅰ)的制备方法，其特征在于，包含以下操作步骤：(a)将太子参的干燥块根粉碎，用75～85％乙醇热回流提取，合并提取液，浓缩至无醇味，依次用石油醚、乙酸乙酯和水饱和的正丁醇萃取，分别得到石油醚萃取物、乙酸乙酯萃取物和正丁醇萃取物；(b)步骤(a)中正丁醇萃取物用大孔树脂除杂，先用10％乙醇洗脱8个柱体积，再用70％乙醇洗脱10个柱体积，收集70％洗脱液，减压浓缩得70％乙醇洗脱浓缩物；(c)步骤(b)中70％乙醇洗脱浓缩物用正相硅胶分离，依次用体积比为60:1、30:1、15:1和5:1的二氯甲烷-甲醇梯度洗脱得到4个组分；(d)步骤(c)中组分4用正相硅胶进一步分离，依次用体积比为8:1、4:1和2:1的二氯甲烷-甲醇梯度洗脱得到3个组分；(e)步骤(d)中组分2用十八烷基硅烷键合的反相硅胶分离，用体积百分浓度为75％的甲醇水溶液等度洗脱，收集10～15个柱体积洗脱液，洗脱液减压浓缩得到化合物(Ⅰ)。
  4. 根据权利要求3所述的化合物(Ⅰ)的制备方法，其特征在于：步骤(a)中用80％乙醇热回流提取，合并提取液。
  5. 根据权利要求3所述的化合物(Ⅰ)的制备方法，其特征在于：所述大孔树脂为D101型大孔吸附树脂。
  6. 权利要求1所述的化合物(Ⅰ)在制备抗抑郁的药物中的应用。
  7. 权利要求2所述的苯丁酸氮芥的药物组合物在制备抗抑郁的药物中的应用。
PCT/CN2017/097016 2016-06-13 2017-08-11 苯丁酸氮芥的药物组合物及其抗抑郁的医药用途 WO2017215679A2 (zh)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN201610411967.8 2016-06-13
CN201610411967.8A CN105997981A (zh) 2016-06-13 2016-06-13 苯丁酸氮芥的药物组合物及其抗抑郁的医药用途

Publications (2)

Publication Number Publication Date
WO2017215679A2 true WO2017215679A2 (zh) 2017-12-21
WO2017215679A3 WO2017215679A3 (zh) 2018-02-15

Family

ID=57091217

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2017/097016 WO2017215679A2 (zh) 2016-06-13 2017-08-11 苯丁酸氮芥的药物组合物及其抗抑郁的医药用途

Country Status (2)

Country Link
CN (1) CN105997981A (zh)
WO (1) WO2017215679A2 (zh)

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105997981A (zh) * 2016-06-13 2016-10-12 崔坤峰 苯丁酸氮芥的药物组合物及其抗抑郁的医药用途
CN106045835A (zh) * 2016-06-23 2016-10-26 崔坤峰 盐酸苄丝肼的药物组合物及其降血糖的医药用途

Family Cites Families (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP4854198B2 (ja) * 2002-11-12 2012-01-18 ジューイッシュ・ジェネラル・ホスピタル 慢性リンパ性白血病の処置のためのナイトロジェンマスタードアナログとイマチニブの組み合わせ剤
CN105997981A (zh) * 2016-06-13 2016-10-12 崔坤峰 苯丁酸氮芥的药物组合物及其抗抑郁的医药用途

Also Published As

Publication number Publication date
CN105997981A (zh) 2016-10-12
WO2017215679A3 (zh) 2018-02-15

Similar Documents

Publication Publication Date Title
WO2017220042A2 (zh) 氨力农的药物组合物及其在高血压治疗中的应用
CA3080001C (en) Piper laetispicum extract and preparation method therefor and use thereof
CN108689851B (zh) 一类惕各烷型二萜化合物及其制备方法和应用
WO2017215679A2 (zh) 苯丁酸氮芥的药物组合物及其抗抑郁的医药用途
Liu et al. Sesquiterpenes and diterpenes from Euphorbia thymifolia
CN105669611A (zh) 一种新的环金合欢烷型倍半萜类化合物及其制备方法和医药用途
CN108659089B (zh) 一种具有抗氧化作用的甾醇化合物及其在制备药物中的应用
CN105820053A (zh) 头孢氨苄的药物组合物及其在生物医药中的应用
CN114702467B (zh) 金凤花芳香化cassane二萜类化合物、提取方法及应用
CN105949266A (zh) 睡茄交酯类化合物及提取方法及应用
CN105669690A (zh) 一种硫酸氢氯吡格雷的药物组合物及其医药用途
CN105949272B (zh) 魏察酸浆苦素y及提取方法及用途
CN109180632B (zh) 一种从雷公藤中分离出的化合物的制备方法
CN111329866B (zh) 一种五环三萜类化合物在制备抗偏头痛药物中的应用
CN105753681A (zh) 一种胞磷胆碱钠的药物组合物及其医药用途
CN105801541A (zh) 一种阿莫西林的药物组合物及其医药用途
CN105646406A (zh) 一种新的环金合欢烷型倍半萜类化合物及其制备方法和医药用途
CN105884716A (zh) 一种格列本脲的药物组合物及其医药用途
CN106008641A (zh) 睡茄交酯类化合物及提取方法及用途
CN111909228A (zh) 一种生物碱类化合物及其制备方法和应用
CN105078938A (zh) 大环吉马烷型倍半萜类化合物在制备抗补体药物中的用途
CN110680826A (zh) 一种三萜皂苷类化合物及其盐的应用
CN110680819A (zh) 一种三萜皂苷类化合物的应用
CN105078965A (zh) 吉马烷内酯倍半萜类化合物在制备抗补体药物中的用途
CN110204477B (zh) 一种具有抗氧化作用的二萜生物碱及其在制备药物中的应用

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 17812790

Country of ref document: EP

Kind code of ref document: A2

NENP Non-entry into the national phase in:

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 17812790

Country of ref document: EP

Kind code of ref document: A2

32PN Ep: public notification in the ep bulletin as address of the adressee cannot be established

Free format text: NOTING OF LOSS OF RIGHTS PURSUANT TO RULE 112(1) EPC (EPO FORM 1205N DATED 17/07/2019)

122 Ep: pct application non-entry in european phase

Ref document number: 17812790

Country of ref document: EP

Kind code of ref document: A2