WO2017220051A2 - 盐酸苄丝肼的药物组合物及其降血糖的医药用途 - Google Patents

盐酸苄丝肼的药物组合物及其降血糖的医药用途 Download PDF

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WO2017220051A2
WO2017220051A2 PCT/CN2017/097789 CN2017097789W WO2017220051A2 WO 2017220051 A2 WO2017220051 A2 WO 2017220051A2 CN 2017097789 W CN2017097789 W CN 2017097789W WO 2017220051 A2 WO2017220051 A2 WO 2017220051A2
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compound
ethanol
blood sugar
benserazide hydrochloride
pharmaceutical composition
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WO2017220051A3 (zh
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崔坤峰
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赵吉永
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C49/00—Ketones; Ketenes; Dimeric ketenes; Ketonic chelates
    • C07C49/527—Unsaturated compounds containing keto groups bound to rings other than six-membered aromatic rings
    • C07C49/557—Unsaturated compounds containing keto groups bound to rings other than six-membered aromatic rings having unsaturation outside the rings
    • 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
    • A61K31/12—Ketones
    • A61K31/122—Ketones having the oxygen directly attached to a ring, e.g. quinones, vitamin K1, anthralin
    • 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
    • A61K31/16—Amides, e.g. hydroxamic acids
    • A61K31/165—Amides, e.g. hydroxamic acids having aromatic rings, e.g. colchicine, atenolol, progabide
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
    • A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C45/00—Preparation of compounds having >C = O groups bound only to carbon or hydrogen atoms; Preparation of chelates of such compounds
    • C07C45/78—Separation; Purification; Stabilisation; Use of additives

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  • the invention belongs to the field of biomedicine and relates to a new use of benserazide hydrochloride, in particular to a pharmaceutical composition of benserazide hydrochloride and a medical use thereof for lowering blood sugar.
  • Benzoin hydrochloride is a peripheral decarboxylase inhibitor and is combined with levodopa to prepare a compound preparation, dopas.
  • Levodopa is an effective drug for the treatment of Parkinson's disease. Its pharmacological action is achieved by the conversion of decarboxylase into dopamine in the brain. Dopamine in the brain can improve Parkinson's disease. However, most of levodopa is converted to dopamine by aromatic amino acid decarboxylase (AADC) in the periphery, and only a small amount can enter the brain.
  • AADC aromatic amino acid decarboxylase
  • benzylhydrazine hydrochloride can be used in combination with levodopa to inhibit levodopa. Peripheral conversion to dopamine increases the amount of levodopa entering the brain and effectively improves the symptoms of Parkinson's disease.
  • Diabetes is a common endocrine and metabolic syndrome.
  • the main feature is the disorder of glucose metabolism.
  • the clinical manifestations are polydipsia, polyuria, and weight loss.
  • the incidence of diabetes has increased year by year, and it has become the third serious threat to human health after malignant tumors and cardiovascular and cerebrovascular diseases.
  • a pharmaceutical composition of benserazide hydrochloride comprising benserazide hydrochloride, compound (I) as described above, and a pharmaceutically acceptable carrier.
  • the preparation method of the compound (I) as described above comprising the steps of: (a) pulverizing the dried tubers of the gastrodia elata, Extracted by hot reflux with 80-90% ethanol, the extracts were combined, concentrated to an alcohol-free taste, and extracted with petroleum ether, ethyl acetate and water-saturated n-butanol to obtain petroleum ether extract, ethyl acetate extract and positive Butanol extract; (b) The n-butanol extract in step (a) is removed with macroporous resin, first eluted with 30% ethanol in 6 column volumes, and then eluted with 85% ethanol in 85% column to collect 85 % eluent, concentrated under reduced pressure to obtain a concentrate of 85% ethanol; (c) 85% ethanol elution concentrate in step (b) separated by normal phase silica gel, sequentially using a volume ratio of 100:1, 50:1 , 25:1 and 12:1 dichloromethane
  • step (a) is extracted by hot reflux with 85% ethanol, and the extracts are combined.
  • the macroporous resin is a D101 type macroporous adsorption resin.
  • the pharmaceutical composition of benserazide hydrochloride provided by the present invention comprises benserazide hydrochloride and a novel natural product isolated from dried tubers of Gastrodia elata, and the natural product of B. benthamiana and the natural product have a lowering effect.
  • the present invention has outstanding substantial features and significant advances over the prior art.
  • the concentrate was concentrated by pressure to obtain 85% ethanol elution; (c) the 85% ethanol elution concentrate in step (b) was separated by normal phase silica gel in a volume ratio of 100:1 (12 column volumes), 50:1. (10 column volumes), 25:1 (8 column volumes) and 12:1 (8 column volumes) of dichloromethane-methanol gradient elution to give 4 components; (d) Group (c) The fraction 4 was further separated by normal phase silica gel, and the volume ratio was 20:1.
  • IR spectrum indicates that the compound containing a carbonyl group (1735cm -1 to 1675cm -1) and the olefin (1632cm -1) group; and which has a 245nm UV absorption, shown to contain ⁇ , ⁇ - unsaturated carbonyl unit.
  • 13 C-NMR, DEPT and HSQC spectra show 20 carbon signals, including three methyl groups, two methylene groups (two olefin carbons), seven methine groups (four olefin carbons), and eight For the quaternary carbon (two carbonyl carbons and six olefin carbons), the above functional structure combined with the number of unsaturation indicates that the compound has a tricyclic structure.
  • H-12/H-13/H-14/H-8/H-7 and H-8/H-9 related signals are present in the 1 H- 1 H COSY spectrum, and H-3 and C are shown in the HMBC spectrum.
  • the C-6 and C-7 related signals can be constructed by the related information in the above NMR spectrum, and the compound can be determined to be a rhamnofolane type diterpenoid.
  • SW-CJ-2FD ultra-clean workbench Suzhou Purification Equipment Co., Ltd.
  • QYC-2112 large double-layer full-temperature culture shaker Haitian Experimental Instrument Manufacturing Co., Ltd.
  • mice were randomly selected (20 ⁇ 2) g, fasted for 16 hours, and intraperitoneally injected with 1% STZ 150 mg ⁇ kg -1 .
  • a 20% glucose solution was injected after 4 hours.
  • the rats were fasted for 3 hours, and the blood glucose level was measured by a glucose kit.
  • the blood glucose level was ⁇ 11.1 mmol ⁇ L -1 and determined as a diabetic model rat.
  • mice Thirty diabetic rats were randomly divided into 4 groups, 10 in each group: model group, benserazide hydrochloride group (100 mg ⁇ kg -1 ), compound (I) group (100 mg ⁇ kg -1 ), benserazide hydrochloride
  • the compound (I) composition group [50 mg ⁇ kg -1 benserazide hydrochloride + 50 mg ⁇ kg -1 compound (I)]
  • Another 10 healthy mice were used as a normal control group.
  • Each group was intragastrically administered (model group and normal control group were intragastrically administered with equal volume of normal saline) for 30 days, blood glucose level was measured, and the basic signs of hair color, water consumption, dampness of the litter and body weight were monitored.
  • mice Another batch of mice was taken, modeled and administered as above. After 1 hour of the last administration, 2 g ⁇ kg -1 glucose solution was administered, and blood glucose levels were measured at 0, 30, 60, and 120 minutes after gavage, and the area under the blood glucose curve was calculated. AUC).
  • A, B, C, and D are 0, 30, 60, and 120 min blood glucose levels, respectively.
  • Blood glucose began to increase after gavage of glucose in each group of mice. Among them, the combination of benserazide hydrochloride and compound (I) had the lowest AUC value, showing the strongest glucose tolerance (P ⁇ 0.01), followed by benserazide hydrochloride and compound (I) (P ⁇ 0.05). .
  • the composition is more tolerant to sugar than a single group. The results are shown in Table 1.
  • the unit of blood glucose at 0 min, 30 min, 60 min and 120 min was mmol ⁇ L -1
  • the unit of AUC was mmol ⁇ h ⁇ L -1 .
  • the ⁇ -glucosidase activity of diabetic mice after administration is shown in Table 2.
  • the ⁇ -glucosidase activity of the administration group was between 0.3 and 0.7 U ⁇ mg -1 .
  • the intestinal mucosal glucosidase activity of the group of the compound of the compound (I) and the compound (I) was significantly down-regulated (P ⁇ 0.01), and the small intestinal mucosal glucose of the mice in the group consisting of the benserazide hydrochloride group and the compound (I) group.
  • Glycosidase activity was also significantly down-regulated (P ⁇ 0.05).
  • the antioxidant activity of diabetic mice is shown in Table 3. Compared with the model group, the SOD activity of the group of the benzylhydrazine hydrochloride and the compound (I) was significantly increased (P ⁇ 0.01), and the MDA content was significantly decreased (P ⁇ 0.01), and this effect was stronger than that of the benzyl chloride. Or a group of compounds (I).
  • benserazide hydrochloride and compound (I) can significantly increase the glucose tolerance, inhibit ⁇ -glucosidase activity, and improve the anti-oxidative damage ability of diabetic mice.
  • benserazide hydrochloride When combined with compound (I), benserazide hydrochloride has better pharmacological activity, and the two have synergistic effects and can be developed into hypoglycemic drugs.

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  • Animal Behavior & Ethology (AREA)
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Abstract

本发明公开了盐酸苄丝肼的药物组合物及其降血糖的医药用途，本发明提供的盐酸苄丝肼的药物组合物中含有盐酸苄丝肼和一种从天麻的干燥块茎中分离得到的结构新颖的天然产物化合物(Ⅰ)，盐酸苄丝肼和该天然产物化合物(Ⅰ)单独作用时，具有降血糖作用；二者联合作用时，降血糖效果进一步提高，可以开发成降血糖的药物，与现有技术相比具有突出的实质性特点和显著的进步。

Description

盐酸苄丝肼的药物组合物及其降血糖的医药用途 技术领域
本发明属于生物医药领域，涉及盐酸苄丝肼的新用途，具体涉及盐酸苄丝肼的药物组合物及其降血糖的医药用途。
背景技术
盐酸苄丝肼是外周脱羧酶抑制药，与左旋多巴联合应用制成复方制剂多巴丝肼。左旋多巴是治疗帕金森病的有效药物，其药理作用是通过在脑中经脱羧酶作用转化为多巴胺实现的，脑中的多巴胺可改善帕金森病症。但是大多数左旋多巴在外周经芳香族氨基酸脱羧酶(AADC)作用转化为多巴胺，只有少量能进入脑内，盐酸苄丝肼作为AADC抑制剂与左旋多巴联合使用，可抑制左旋多巴在外周转化为多巴胺，从而使进入脑内的左旋多巴的量增多，有效地改善帕金森病症状。
糖尿病是一种常见的内分泌代谢综合症，主要特征为糖代谢紊乱，临床表现为多饮多食、多尿，体重减轻。目前糖尿病发病率逐年升高，已成为继恶性肿瘤、心脑血管病后第3位严重威胁人类健康的疾病。
目前尚未见盐酸苄丝肼与降血糖的相关性报道。
发明内容
本发明的目的在于提供一种盐酸苄丝肼的药物组合物，该药物组合物中含有盐酸苄丝肼和一种从草本中分离得到的结构新颖的天然产物，盐酸苄丝肼和该天然产物可以协同降血糖。
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 PCTCN2017097789-appb-000001
一种盐酸苄丝肼的药物组合物，包括盐酸苄丝肼、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)将天麻的干燥块茎粉碎， 用80～90％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)中正丁醇取物用大孔树脂除杂，先用30％乙醇洗脱6个柱体积，再用85％乙醇洗脱12个柱体积，收集85％洗脱液，减压浓缩得85％乙醇洗脱浓缩物；(c)步骤(b)中85％乙醇洗脱浓缩物用正相硅胶分离，依次用体积比为100:1、50:1、25:1和12:1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20:1、12:1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88％的甲醇水溶液等度洗脱，收集13～16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)用85％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)在制备降血糖的药物中的应用。
如上所述的盐酸苄丝肼的药物组合物在制备降血糖的药物中的应用。
本发明的优点：
本发明提供的盐酸苄丝肼的药物组合物中含有盐酸苄丝肼和一种从天麻的干燥块茎中分离得到的结构新颖的天然产物，盐酸苄丝肼和该天然产物单独作用时，具有降血糖作用；二者联合作用时，降血糖效果进一步提高，可以开发成降血糖的药物。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
分离方法：(a)将天麻的干燥块茎(2kg)粉碎，用85％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)中乙酸乙酯萃取物用D101型大孔树脂除杂，先用30％乙醇洗脱6个柱体积，再用85％乙醇洗脱12个柱体积，收集85％洗脱液，减压浓缩得85％乙醇洗脱浓缩物；(c)步骤(b)中85％乙醇洗脱浓缩物用正相硅胶分离，依次用体积比为100:1(12个柱体积)、50:1(10个柱体积)、25:1(8个柱体积)和12:1(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20:1 (6个柱体积)、12:1(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88％的甲醇水溶液等度洗脱，收集13～16个柱体积洗脱液，洗脱液减压浓缩得到化合物(Ⅰ)(514mg，HPLC归一化纯度大于98％)。
结构确证：白色针晶，HR-ESI-MS显示[M+H]+为m/z 293.1492，结合核磁特征可得分子式为C20H20O2，不饱和度为11。核磁共振氢谱数据δH(ppm，CDCl3，500MHz)：H-3(6.68，s)，H-7(5.53，dd，J＝7.7，1.8Hz)，H-8(2.04，dt，J＝11.5，7.2Hz)，H-9(3.26，d，J＝11.5Hz)，H-12(6.15，d，J＝10.5Hz)，H-13(6.62，dd，J＝10.5，6.7Hz)，H-14(2.42，dd，J＝6.7，7.2Hz)，H-16a(4.73，s)，H-16b(4.74，s)，H-17(1.53，s)，H-18a(5.03，s)H-18b(4.82，s)，H-19(1.88，s)，H-20(1.63，s)；核磁共振碳谱数据δC(ppm，CDCl3，125MHz)：198.2(C，1-C)，144.2(C，2-C)，146.4(CH，3-C)，160.3(C，4-C)，207.2(C，5-C)，138.1(C，6-C)，133.9(CH，7-C)，47.4(CH，8-C)，42.8(CH，9-C)，151.5(C，10-C)，148.9(C，11-C)，129.7(CH，12-C)，135.8(CH，13-C)，51.4(CH，14-C)，146.4(C，15-C)，114.3(CH2，16-C)，18.5(CH3，17-C)，109.7(CH2，18-C)，10.5(CH3，19-C)，19.3(CH3，20-C)。红外波谱表明该化合物含有羰基(1735cm-1与1675cm-1)和烯烃(1632cm-1)基团；且其在245nm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，两个亚甲基(两个烯烃碳)，七个次甲基(四个烯烃碳)，以及八个季碳(两个羰基碳和六个烯烃碳)，以上功能结构再结合不饱和数表明该化合物为三环结构。1H-NMR谱结合HSQC谱显示三个甲基质子信号δH 1.53(3H，s)、1.88(3H，s)、1.63(3H，s)，两组端烯烃质子信号δH 4.73(1H，s)与4.74(1H，s)、5.03(1H，s)与4.82(1H，s)，一对烯烃质子信号δH 6.15(1H，d，J＝10.5Hz)与6.62(1H，dd，J＝10.5，6.7Hz)，两个孤立烯烃质子信号δH 6.68(1H，s)、5.53(1H，dd，J＝7.7，1.8Hz)。1H-1H COSY谱中存在H-12/H-13/H-14/H-8/H-7以及H-8/H-9相关信号，同时HMBC谱中显示有H-3与C-1、C-2和C-4，H-7与C-5、C-6、C-8和C-14，H2-16与C-14、C-15和C-17，H3-17与C-14，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。通过C-4与C-10的碳化学位移δC 160.3与151.5可知C-4和C-10之间为双键。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。
该化合物化学式及碳原子编号如下：
Figure PCTCN2017097789-appb-000002
实施例2：药理作用
1、材料与方法
1.1材料与仪器
健康雄性ICR小鼠，沈阳医学院动物实验中心，体重18～22g；盐酸苄丝肼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。链脲佐菌素(STZ)美国Sigma化学公司，用前用柠檬酸缓冲液配成1％STZ溶液；其他试剂国产分析纯。SW-CJ-2FD超净工作台苏州净化设备有限公司；QYC-2112大型双层全温培养摇床上海天呈实验仪器制造有限公司；SHY-2A双功能水浴恒温振荡器江苏东鹏仪器制造有限公司；TGL20W台式高速冷冻离心机湖南湘仪实验室仪器开发有限公司；PB203-N电子天平瑞士，深圳市怡华新电子有限公司；FW177中草药粉碎机天津泰斯特有限责任公司；旋转蒸发仪日本，北京五洲东方科技发展有限公司；YRD-30.2A真空冷冻干燥机上海精宏实验设备有限公司；葡萄糖试剂盒，超氧化物歧化酶(SOD)和丙二醛(MDA)测定试剂盒四川迈克科技有限责任公司。
1.2STZ诱导糖尿病小鼠模型建立
随机抽取体重(20±2)g小鼠，禁食16h，腹腔注射1％STZ 150mg·kg-1。4h后注射20％葡萄糖溶液。正常饲养3d后，禁食3h，用葡萄糖试剂盒测定血糖值，血糖值≥11.1mmol·L-1者确定为糖尿病模型鼠。
1.3实验分组及给药
取糖尿病模型鼠40只，随机为4组，每组10只：模型组、盐酸苄丝肼组(100mg·kg-1)、化合物(Ⅰ)组(100mg·kg-1)、盐酸苄丝肼与化合物(Ⅰ)组合物组【50mg·kg-1盐酸苄丝肼+50mg·kg-1化合物(Ⅰ)】，另取健康鼠10只，作为正常对照组。各组分别灌胃给药(模型组和正常对照组灌胃等体积生理盐水)，连续给药30d，测血糖值，监测毛色，饮水量，垫料潮湿程度和体重的基本体征。
1.4葡萄糖耐量实验
另取一批小鼠，造模及给药同上，末次给药1h后，灌胃2g·kg-1葡萄糖溶液，灌胃后0、 30、60、120min测定血糖值，计算血糖曲线下面积(AUC)。
计算方法：mmol·h·L-1＝12A+B+C+12D；
其中，A、B、C、D分别为0、30、60、120min血糖值。
1.5α-葡萄糖苷酶活性的测定
禁食处死小鼠，取小肠上段，0.01M PBS，pH7.4漂洗，反转小肠内腔，剥离小肠刷状缘膜，擦干，按W∶V＝1∶9加PBS后冰浴匀浆，4℃离心(4000r/min)取上清液，-34℃冷冻备用。小肠黏膜α-葡萄糖苷酶活性测定：用考马斯亮蓝测定提取液组织蛋白含量。稀释小肠粘膜提取液至最佳浓度，取0.2mL，分别加入pH6.8磷酸盐缓冲液0.7mL，37℃水浴10min后，加入0.5mol·L-1蔗糖溶液0.1mL，37℃水浴20min加入0.l mol·L-1Na2CO31mL终止反应。测定葡萄糖浓度，平行测定3管，取平均值，计算α-葡萄糖苷酶活力。
1.6抗氧化活性的测定
给药20d后，取血3mL，3500r/min离心5min，提取血清。采用黄嘌呤氧化酶法测定SOD活性，采用硫代巴比妥酸法测定MDA含量，按照SOD、MDA检测试剂盒说明书步骤操作。
1.7数据统计分析
用SPSS17.0软件处理数据，以x±s表示。
2、实验结果
2.1对小鼠糖耐受量的影响
各组小鼠灌胃葡萄糖后血糖开始升高。其中盐酸苄丝肼与化合物(Ⅰ)组合物组的AUC值最低，显示出最强的糖耐量作用(P<0.01)，其次是盐酸苄丝肼组、化合物(Ⅰ)组(P<0.05)。组合物糖耐受能力强于单一组。结果见表1，0min、30min、60min和120min血糖值单位为mmol·L-1，AUC单位为mmol·h·L-1。
表1小鼠糖耐受量
Figure PCTCN2017097789-appb-000003
2.2糖尿病小鼠α-葡萄糖苷酶活性的测定
给药后的糖尿病小鼠α-葡萄糖苷酶活性见表2。给药组α-葡萄糖苷酶活性在0.3～0.7U·mg-1 之间。与模型组相比，盐酸苄丝肼与化合物(Ⅰ)组合物组小鼠小肠黏膜葡萄糖苷酶活性显著下调(P<0.01)，盐酸苄丝肼组、化合物(Ⅰ)组小鼠小肠黏膜葡萄糖苷酶活性也有明显下调(P<0.05)。通过抑制该酶活性，降低了人体对摄入的淀粉、蔗糖等碳水化合物的吸收。
表2糖尿病小鼠α-葡萄糖苷酶活性的测定
组别 α-葡萄糖苷酶活性(U·mg-1)
正常对照组 0.8
模型组 1.4
盐酸苄丝肼组 0.6
化合物(Ⅰ)组 0.7
盐酸苄丝肼与化合物(Ⅰ)组合物组 0.3
2.3抗氧化活性的测定
糖尿病小鼠抗氧化活性见表3。与模型组相比，盐酸苄丝肼与化合物(Ⅰ)组合物组的SOD活性明显升高(P<0.01)，MDA含量有显著下降(P<0.01)，且这种作用强于盐酸苄丝肼组或化合物(Ⅰ)组。
表3对糖尿病小鼠SOD活性和MDA含量影响
组别 SOD(U·mL-1) MDA(nmol·mL-1)
正常对照组 98.76±7.23 5.33±1.24
模型组 86.54±6.26 8.42±1.82
盐酸苄丝肼组 93.25±6.21 6.21±1.66
化合物(Ⅰ)组 92.33±4.28 6.39±1.53
盐酸苄丝肼与化合物(Ⅰ)组合物组 99.87±6.38 5.04±1.42
试验表明，盐酸苄丝肼和化合物(Ⅰ)单独使用时，可以显著提高糖尿病模型小鼠的糖耐受量，抑制α-葡萄糖苷酶活性，同时提高糖尿病模型小鼠的抗氧化损伤能力。盐酸苄丝肼与化合物(Ⅰ)联合作用时，具有更好的药理活性，二者存在协同作用，可开发成降糖药物。
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 PCTCN2017097789-appb-100001
  2. 一种盐酸苄丝肼的药物组合物，其特征在于：包括盐酸苄丝肼、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)将天麻的干燥块茎粉碎，用80～90％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)中正丁醇取物用大孔树脂除杂，先用30％乙醇洗脱6个柱体积，再用85％乙醇洗脱12个柱体积，收集85％洗脱液，减压浓缩得85％乙醇洗脱浓缩物；(c)步骤(b)中85％乙醇洗脱浓缩物用正相硅胶分离，依次用体积比为100:1、50:1、25:1和12:1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20:1、12:1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88％的甲醇水溶液等度洗脱，收集13～16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)用85％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)在制备降血糖的药物中的应用。
  7. 权利要求2所述的盐酸苄丝肼的药物组合物在制备降血糖的药物中的应用。
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