WO2017220044A2 - 联苯苄唑的药物组合物及其保肝作用 - Google Patents

联苯苄唑的药物组合物及其保肝作用 Download PDF

Info

Publication number
WO2017220044A2
WO2017220044A2 PCT/CN2017/097769 CN2017097769W WO2017220044A2 WO 2017220044 A2 WO2017220044 A2 WO 2017220044A2 CN 2017097769 W CN2017097769 W CN 2017097769W WO 2017220044 A2 WO2017220044 A2 WO 2017220044A2
Authority
WO
WIPO (PCT)
Prior art keywords
compound
bifonazole
liver
ethanol
pharmaceutical composition
Prior art date
Application number
PCT/CN2017/097769
Other languages
English (en)
French (fr)
Other versions
WO2017220044A3 (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 WO2017220044A2 publication Critical patent/WO2017220044A2/zh
Publication of WO2017220044A3 publication Critical patent/WO2017220044A3/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
    • A61K31/33—Heterocyclic compounds
    • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/365—Lactones
    • 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
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
    • A61K31/4164—1,3-Diazoles
    • A61K31/4174—Arylalkylimidazoles, e.g. oxymetazolin, naphazoline, miconazole
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D493/00—Heterocyclic compounds containing oxygen atoms as the only ring hetero atoms in the condensed system
    • C07D493/02—Heterocyclic compounds containing oxygen atoms as the only ring hetero atoms in the condensed system in which the condensed system contains two hetero rings
    • C07D493/04—Ortho-condensed systems
    • 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)
    • A61K36/88—Liliopsida (monocotyledons)
    • A61K36/896—Liliaceae (Lily family), e.g. daylily, plantain lily, Hyacinth or narcissus
    • A61K36/8966—Fritillaria, e.g. checker lily or mission bells

Definitions

  • the invention belongs to the field of biomedicine and relates to a new use of bifonazole, in particular to a pharmaceutical composition of bifonazole and a medical use thereof for protecting liver.
  • Bifonazole is an imidazole antifungal agent with broad-spectrum antifungal activity and is effective against dermatophytes, yeasts, molds and other fungi such as F. sphaeroides and Corynebacterium parvum.
  • dermatophytes yeasts, molds and other fungi
  • F. sphaeroides F. sphaeroides
  • Corynebacterium parvum F. sphaeroides and Corynebacterium parvum.
  • In vitro tests have shown that the effect of this drug on skin fungi (such as hairpin) is mainly sterilization, and the effect on yeast is mainly inhibition.
  • the drug works well through the infected skin and acts quickly and lasts longer than 48 hours.
  • Indications acute and chronic dermatophytosis, especially the genus Trichophyton, microsporum, flocculent sputum; yeast, such as Candida albicans and other kinds of yeast; mold, such as Aspergillus Dermatological fungal disease; pityriasis caused by P. stipitis; red sputum caused by coryneform bacteria; superficial skin infection caused by the above fungi.
  • a pharmaceutical composition of bifonazole comprising bifonazole, compound (I) as described above, and a pharmaceutically acceptable carrier.
  • the preparation method of the compound (I) as described above comprising the following steps: (a) drying the bulb powder of Fritillaria Crushed, extracted with 85 ⁇ 95% ethanol under reflux, combined with extracts, concentrated to an alcohol-free taste, extracted with petroleum ether, ethyl acetate and water-saturated n-butanol to obtain petroleum ether extract and ethyl acetate extract respectively. And n-butanol extract; (b) the n-butanol extract in step (a) is removed with macroporous resin, first eluted with 35% ethanol and 8 column volumes, and then eluted with 90% ethanol for 12 column volumes.
  • step (b) The 90% eluate was collected and concentrated under reduced pressure to obtain a 90% ethanol eluting concentrate; (c) the 90% ethanol eluting concentrate in step (b) was separated by normal phase silica gel, and the volume ratio was 120:1, 60 in sequence.
  • step (c) component 3 is further separated by normal phase silica gel in a volume ratio of 40: Gradient elution of 1, 30:1 and 10:1 by dichloromethane-methanol to obtain 3 components;
  • component 2 in step (d) separated by octadecylsilane-bonded reversed phase silica gel, The aqueous solution of 85% by volume was eluted isocratically, and 14 to 18 column volumes of the eluate were collected, and the eluate was concentrated under reduced pressure to give Compound (I).
  • step (a) is carried out by hot reflux extraction with 90% ethanol, and the extracts are combined.
  • the macroporous resin is a D101 type macroporous adsorption resin.
  • the pharmaceutical composition of bifonazole provided by the present invention comprises bifonazole and a novel natural product isolated from the dried bulb of Fritillaria thunbergii.
  • bifonazole and the natural product act alone, It has a hepatoprotective effect; when the two are combined, the liver-protecting effect is further improved, and a drug for liver protection can be developed.
  • the present invention has outstanding substantial features and significant advances over the prior art.
  • step (b) 90% ethanol eluting concentrate in step (b) is separated by normal phase silica gel, sequentially using a volume ratio of 120:1 (11 column volumes), 60 :1 (9 column volumes), 30:1 (9 column volumes) and 15:1 (8 column volumes) of dichloromethane-methanol gradient elution to give 4 components;
  • Step (c) The middle component 3 is further separated by normal phase silica gel, and the volume ratio is 40:1 in sequence.
  • the 1710 cm -1 absorption band in the infrared spectrum and the 224 nm absorption band in the UV spectrum indicate that the compound contains an ⁇ , ⁇ -unsaturated carbonyl structure.
  • the H-4/H-10/H-11/H-12/H-13/H 2 -14 related signal is present in the other 1 H- 1 H COSY spectrum in combination with H-1 and C-4a shown in the HMBC spectrum.
  • ICR mice were randomly divided into six groups at (22 ⁇ 1) °C, humidity 40% ⁇ 60%, 12h day and night, free feeding and water feeding conditions for one week.
  • 12 blank control groups no treatment during the experiment, free feeding; 12 in the model group: 0.9% saline; 12 in the bifonazole group: concentration 100 mg/mL; compound (I) Group 12: concentration of 100 mg / mL; bifonazole and compound (I) composition group 12: concentration of 50 mg / mL bifonazole + 50 mg / mL of compound (I); positive control group of 12: concentration It is a 25 mg/mL Haiwang gold sputum solution.
  • a 20 mL/kg ⁇ BW test sample was administered orally by oral gavage daily.
  • the blank control group did not receive any intragastric administration, and was fed with free water.
  • the model control group was given normal saline by body weight for 30 days. Weigh once a week and adjust the dosage according to body weight.
  • the model control group and each sample group were intragastrically administered with 5 mL/kg ⁇ BW of white wine, and the blank control group was given distilled water.
  • liver tissue was cut 0.1 g of liver tissue into a small beaker (in ice water), prepare 9 times (0.9 mL) of 0.9% physiological saline in liver tissue weight, transfer 2/3 of the total amount of normal saline in a beaker, and cut as soon as possible.
  • Mouse body weight The body weight of the mice was weighed once a week until the end of the experiment to examine the effect of the test samples on the body weight of each group of mice.
  • Determination of liver tissue index The contents of GSH, MDA and TG in the liver tissues of each group, and the determination of ADH activity were carried out according to the procedure of each kit manual.
  • the total content of liver glutathione (GSH) is an important indicator for evaluating the function of hangover and liver protection. It can be seen from Table 1 that compared with the blank group, the GSH content in the liver of the model group was significantly decreased after oral administration of liquor (P ⁇ 0.01), indicating that the model was successfully modeled.
  • the GSH content in the liver homogenate of the positive control group and the bifonazole and compound (I) composition group was significantly higher than that of the model group (P ⁇ 0.01), the bifonazole group and the compound (I) group.
  • the GSH content in mouse liver homogenate was also significantly increased (P ⁇ 0.05).
  • the composition can maintain the liver GSH content, resist the liver GSH depletion caused by ethanol damage, and improve the body's ability to scavenge free radicals, thereby protecting the liver of mice. Since GSH can prevent lipid peroxidation in liver tissue to a certain extent, and malondialdehyde (MDA) is one of the products of lipid peroxidation, the decrease of GSH concentration will indirectly lead to an increase in liver MDA content. As can be seen from Table 1, compared with the blank group, the mice in the model group were administered with stomach. The MDA content in the liver was significantly increased (P ⁇ 0.01), indicating that the model was successfully modeled.
  • MDA malondialdehyde
  • the MDA content in the liver homogenate of the positive control group and the bifonazole and compound (I) composition group decreased significantly compared with the model group P ( ⁇ 0.01), and the bifonazole group and the compound (I) group also showed The MDA content decreased significantly and the level of significance was significant (P ⁇ 0.05).
  • TG triglyceride
  • Table 2 compared with the blank group, the TG content in the liver tissue of the model group was extremely significantly increased (P ⁇ 0.01), indicating that the model was successfully modeled.
  • the TG content in the liver homogenate of the bifonazole and the compound (I) composition group and the positive control group was extremely significantly decreased (P ⁇ 0.01).
  • each drug group showed a decrease in liver TG content in drunken mice, and the composition group reached extremely significant level (P ⁇ 0.01), indicating that the composition can reduce liver tissue TG content and reduce liver fat in drunken mice. Stacked to protect the liver.
  • bifonazole and compound (I) can increase the content of GSH in mouse liver tissue, decrease the content of MDA, inhibit the accumulation of TG, and have a certain liver-protecting effect; the combination of bifonazole and compound (I) When the treatment is better, it can be developed into a liver-protecting drug.

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material 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)
  • Gastroenterology & Hepatology (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

本发明公开了联苯苄唑的药物组合物及其保肝作用，本发明提供的联苯苄唑的药物组合物中含有联苯苄唑和一种从浙贝母的干燥鳞茎中分离得到的结构新颖的天然产物化合物(Ⅰ)，联苯苄唑、化合物(Ⅰ)可以提高小鼠肝组织中GSH含量，降低MDA含量，抑制TG的积累，具有一定的保肝作用；联苯苄唑和化合物(Ⅰ)联合使用时，治疗效果更好，可开发成保肝药物，与现有技术相比具有突出的实质性特点和显著的进步。

Description

联苯苄唑的药物组合物及其保肝作用 技术领域
本发明属于生物医药领域，涉及联苯苄唑的新用途，具体涉及联苯苄唑的药物组合物及其保肝的医药用途。
背景技术
联苯苄唑为咪唑类抗真菌剂，具有广谱抗真菌作用，对皮肤真菌、酵母菌、霉菌及其它真菌，如秕糠状鳞斑霉菌，微小棒状杆菌有效。体外试验表明本药对皮肤真菌(如发癣菌)的作用主要是杀菌，而对酵母菌的作用主要是抑菌。本药能很好地透过被感染的皮肤，作用迅速并持续时间长，维持时间超过48hr。
适应症：急性及慢性皮肤真菌病，特别是皮肤毛癣菌属，小孢子菌属，絮状表皮癣菌；酵母菌，如白色念珠菌及其它种类的酵母菌；霉菌，如曲霉菌所致的皮肤真菌病；秕糠状鳞斑霉所引起的花斑糠疹；棒状杆菌所致的红癣；上述真菌所致皮肤浅表感染。
目前尚未见联苯苄唑与保肝的相关性报道。
发明内容
本发明的目的在于提供一种联苯苄唑的药物组合物，该药物组合物中含有联苯苄唑和一种从草本中分离得到的结构新颖的天然产物，联苯苄唑和该天然产物可以协同保肝。
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 PCTCN2017097769-appb-000001
一种联苯苄唑的药物组合物，包括联苯苄唑、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)将浙贝母的干燥鳞茎粉 碎，用85～95％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35％乙醇洗脱8个柱体积，再用90％乙醇洗脱12个柱体积，收集90％洗脱液，减压浓缩得90％乙醇洗脱浓缩物；(c)步骤(b)中90％乙醇洗脱浓缩物用正相硅胶分离，依次用体积比为120:1、60:1、30:1和15:1的二氯甲烷-甲醇梯度洗脱得到4个组分；(d)步骤(c)中组分3用正相硅胶进一步分离，依次用体积比为40:1、30:1和10:1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85％的甲醇水溶液等度洗脱，收集14～18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90％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)在制备保肝的药物中的应用。
如上所述的联苯苄唑的药物组合物在制备保肝的药物中的应用。
本发明的优点：
本发明提供的联苯苄唑的药物组合物中含有联苯苄唑和一种从浙贝母的干燥鳞茎中分离得到的结构新颖的天然产物，联苯苄唑和该天然产物单独作用时，具有保肝作用；二者联合作用时，保肝效果进一步提高，可以开发成保肝的药物。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)将浙贝母的干燥鳞茎(2kg)粉碎，用90％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35％乙醇洗脱8个柱体积，再用90％乙醇洗脱12个柱体积，收集90％洗脱液，减压浓缩得90％乙醇洗脱浓缩物；(c)步骤(b)中90％乙醇洗脱浓缩物用正相硅胶分离，依次用体积比为120:1(11个柱体积)、60:1(9个柱体积)、30:1(9个柱体积)和15:1(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3用正相硅胶进一步分离，依次用体积比为40:1 (6个柱体积)、30:1(8个柱体积)和10:1(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85％的甲醇水溶液等度洗脱，收集14～18个柱体积洗脱液，洗脱液减压浓缩得到化合物(Ⅰ)(553mg，HPLC归一化纯度大于98％)。
结构确证：HR-ESI-MS显示[M+H]+为m/z 467.2378，结合核磁特征可得分子式为C28H34O6，不饱和度为12。核磁共振氢谱数据δH(ppm，CD3OD，500MHz)：H-1(6.81，s)，H-4(6.05，s)，H-5(5.38，s)，H-8(3.79，m)，H-9(1.36，m)，H-10(2.91，d，J＝12.4Hz)，H-11(2.08，m)，H-12(6.32，dd，J＝11.9，7.7Hz)，H-13(6.53，m)，H-14(2.37，m)，H-14(2.59，m)，H-16(1.02，d，J＝6.5Hz)，H-18(4.07，dd，J＝12.7Hz)，H-20(3.24，dqd，J＝13.2，7.4，6.5Hz)，H-21(1.34，m)，H-21(1.77，m)，H-22(1.20，m)，H-22(1.26，m)，H-23(1.24，m，2H)，H-24(1.24，m，2H)，H-25(1.25，m，2H)，H-26(0.88，t，J＝7.2Hz)，H-27(1.21，d，J＝7.2Hz)；核磁共振碳谱数据δC(ppm，CD3OD，125MHz)：143.3(CH，1-C)，156.6(C，3-C)，110.8(CH，4-C)，144.2(C，4a-C)，107.3(CH，5-C)，190.8(C，6-C)，82.4(C，7-C)，43.5(CH，8-C)，116.3(C，8a-C)，19.2(CH3，9-C)，62.1(CH，10-C)，21.1(CH，11-C)，133.6(CH，12-C)，123.2(CH，13-C)，43.3(CH2，14-C)，206.5(C，15-C)，19.5(CH3，16-C)，169.3(C，17-C)，51.1(CH，18-C)，206.3(C，19-C)，46.3(CH，20-C)，32.7(CH2，21-C)，27.6(CH2，22-C)，29.0(CH2，23-C)，31.3(CH2，24-C)，22.2(CH2，25-C)，14.2(CH3，26-C)，16.6(CH3，27-C)。红外波谱中的1710cm-1吸收带与UV谱中的224nm吸收带表明该化合物含有α,β-不饱和羰基结构。13C NMR、DEPT和HSQC谱中显示有28个碳信号，包括四个甲基，六个亚甲基，十个次甲基(五个烯烃碳)，以及八个季碳(一个连氧碳，四个羰基碳和三个烯烃碳)，以上功能结构再结合不饱和数表明该化合物为四环结构。核磁数据表明该化合物含有一个α,β-不饱和羰基结构[δH5.38(1H，s，H-5)；δC144.2(C-4a)，107.3(C-5)，190.8(C-6)]，一个五元内酯环[δH3.79(1H，m，H-8)，4.07(1H，dd，J＝12.7Hz，H-18)；δC82.4(C-7)，43.5(C-8)，169.3(C-17)，51.1(C-18)]，一个2-甲基辛酰基片段[δH3.24(1H，dqd，J＝13.2，7.4，6.5Hz，H-20)，1.34(1H，m，H-21)，1.77(1H，m，H-21)，1.20(1H，m，H-22)，1.26(1H，m，H-22)，1.24(2H，m，H-23)，1.24(2H，m，H-24)，1.25(2H，m，H-25)，0.88(3H，t，J＝7.2Hz，H-26)，1.21(3H，d，J＝7.2Hz，H-27)；δC206.3(C-19)，46.3(C-20)，32.7(C-21)，27.6(C-22)，29.0(C-23)，31.3(C-24)，22.2(C-25)，14.2(C-26)，16.6(C-27)]。1H-1H COSY谱中显示的H3-27/H-20/H2-21/H2-22/H2-23/H2-24/H2-25/H3-26相关信号与HMBC谱中显示的H-18与C-8 和C-19，H-20与C-19的相关性以及相关碳化学位移表明该化合物含有一个2-甲基辛酰基片段。另1H-1H COSY谱中存在H-4/H-10/H-11/H-12/H-13/H2-14相关信号结合HMBC谱中显示的H-1与C-4a、C-8a和C-8，H-4与C-3、C-4a和C-8a，H-5与C-4、C-4a和C-6，H-8与C-7、C-8a和C-18，H-10与C-3、C-11、C-15和C-16，H-12与C-11、C-13、C-14和C-16，H2-14与C-12、C-13和C-15，H-18与C-8和C-17相关信号，可以推测azaphilone骨架结构存，因此，根据上述结构片段构建化合物可以确认为Cohaerin衍生物。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 PCTCN2017097769-appb-000002
实施例2：药理作用
1、材料与方法
1.1材料与仪器
联苯苄唑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。ICR雌性小白鼠体重(25±5)g，上海斯莱克实验动物责任有限公司；考马斯亮蓝G-250、甘油三脂试剂盒、丙二醛测试盒、谷胱甘肽过氧化物试剂盒、乙醇脱氢酶测定试剂盒南京建成生物工程研究所；海王金樽，广东深圳海王药业有限公司；56°红星二锅头白酒，北京红星酿酒厂。Alpha-D2冻干机德国Christ公司；电子天平，上海天平仪器厂；UV752N紫外可见分光光度计，上海仪电分析仪器有限公司。
1.2动物分组
将72只ICR小鼠在(22±1)℃，湿度40％～60％，12h日夜交替，自由进食、进水的条件下适应性喂养一周后，随机分为六组。空白对照组12只：实验过程中不做任何处理，自由喂养；模型组12只：0.9％的生理盐水；联苯苄唑组12只：浓度为100mg/mL；化合物(Ⅰ) 组12只：浓度为100mg/mL；联苯苄唑与化合物(Ⅰ)组合物组12只：浓度为50mg/mL联苯苄唑+50mg/mL化合物(Ⅰ)；阳性对照组12只：浓度为25mg/mL的海王金樽溶液。
1.3给药
每日经口灌胃给予20mL/kg·BW受试样品。空白对照组不做任何灌胃，自由进食、进水；模型对照组按体重给予生理盐水，连续给药30d。每周称重一次，根据体重调整用药量。给予受试样品结束时，将模型对照组及各样品组一次灌胃给予白酒5mL/kg·BW，空白对照组给蒸馏水。
1.4肝脏样品制备
末次给药后，禁食不禁水12h，将小鼠颈椎脱臼处死，解剖取出完整肝脏，预冷生理盐水清洗至洗液无血色后，吸干肝脏表面水分。剪取0.1g肝脏组织置于小烧杯中(于冰水)，准备9倍(0.9mL)于肝脏组织重量的0.9％的生理盐水，转移总量2/3的生理盐水于烧杯中，尽快剪碎组织块，倒入玻璃匀浆器，再将剩余的1/3生理盐水冲洗残留在烧杯中的碎组织块，一起倒入匀浆管进行匀浆，充分研磨后制成10％的组织匀浆，于离心机中以2000r/min离心15min，取上清液备用。
1.5检测指标
小鼠体重：每周称量一次小鼠体重，直到实验结束，以考察受试样品对各组小鼠体重的影响。肝组织指标测定：各组肝组织中GSH、MDA和TG含量，ADH活性测定根据各试剂盒说明书操作步骤进行。
1.3数据统计
数据均以(平均值±标准偏差)表示，组间差异采用SPSS16.0进行单因素分析(ANOVA)，以P＜0.05或P＜0.01确定差异是否有统计学意义。
2、实验结果
2.1对谷胱甘肽、丙二醛含量的影响
肝脏谷胱甘肽(GSH)的总含量是评价药物醒酒保肝功能的一个重要指标。由表1可知，与空白组比较，模型组小鼠经白酒灌胃后，肝脏GSH含量极显著降低(P<0.01)，说明该模型造模成功。阳性对照组和联苯苄唑与化合物(Ⅰ)组合物组小鼠肝匀浆中GSH含量与模型组相比极显著升高(P<0.01)，联苯苄唑组、化合物(Ⅰ)组小鼠肝匀浆中GSH含量也明显升高(P<0.05)。组合物能够维持肝脏GSH含量，对抗乙醇损伤引起的肝脏GSH耗竭，提高机体清除自由基的能力，从而对小鼠肝脏起到保护作用。由于GSH能在一定程度上阻止肝组织中脂质过氧化反应，而丙二醛(MDA)是脂质过氧化反应的产物之一，因此GSH浓度的降低会间接导致肝脏MDA含量的增加。由表1可知，与空白组比较，模型组小鼠灌胃后， 肝脏MDA含量极显著升高(P<0.01)，说明该模型造模成功。阳性对照组和联苯苄唑与化合物(Ⅰ)组合物组小鼠肝匀浆中MDA含量与模型组相比显著下降P(<0.01)，联苯苄唑组、化合物(Ⅰ)组也表现出MDA含量明显下降，显著性水平(P<0.05)。
表1受试样品对小鼠肝组织中GSH、MDA含量的影响
组别 GSH(mmol/L) MDA(nmol/mgprot)
空白组 27 5.2
模型组 17 11.7
阳性对照组 26 6.2
联苯苄唑组 24 8.7
化合物(Ⅰ)组 24 8.9
联苯苄唑与化合物(Ⅰ)组合物组 29 5.4
2.2对甘油三酯含量的影响
当机体摄入大量乙醇时，乙醇在乙醇脱氢酶的作用下大量氧化，使得三羧酸循环和脂肪代谢受阻，使得甘油三酯(TG)在肝脏中不断堆积，不但会诱发较为严重的心血管疾病，还会引发脂肪肝。由表2可知，与空白组比较，模型组小鼠肝脏组织TG含量极显著升高(P<0.01)，说明该模型造模成功。与模型组比较，联苯苄唑与化合物(Ⅰ)组合物组和阳性对照组小鼠肝匀浆中TG含量极显著降低(P<0.01)。本实验中，各药物组都表现出降低醉酒小鼠肝脏TG含量，并且组合物组都达到极显著水平(P<0.01)，说明此组合物可降低醉酒小鼠肝脏组织TG含量，减轻肝脏脂肪堆积，起到保护肝脏的作用。
表2受试样品对小鼠肝组织中TG含量的影响
组别 TG(mmol/L)
空白组 0.6±0.02
模型组 1.4±0.14
阳性对照组 0.7±0.01
联苯苄唑组 0.9±0.03
化合物(Ⅰ)组 0.9±0.02
联苯苄唑与化合物(Ⅰ)组合物组 0.7±0.02
上述试验表明，联苯苄唑、化合物(Ⅰ)可以提高小鼠肝组织中GSH含量，降低MDA含量，抑制TG的积累，具有一定的保肝作用；联苯苄唑和化合物(Ⅰ)联合使用时，治疗效果更好，可开发成保肝药物。
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 PCTCN2017097769-appb-100001
  2. 一种联苯苄唑的药物组合物，其特征在于：包括联苯苄唑、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)将浙贝母的干燥鳞茎粉碎，用85～95％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35％乙醇洗脱8个柱体积，再用90％乙醇洗脱12个柱体积，收集90％洗脱液，减压浓缩得90％乙醇洗脱浓缩物；(c)步骤(b)中90％乙醇洗脱浓缩物用正相硅胶分离，依次用体积比为120:1、60:1、30:1和15:1的二氯甲烷-甲醇梯度洗脱得到4个组分；(d)步骤(c)中组分3用正相硅胶进一步分离，依次用体积比为40:1、30:1和10:1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85％的甲醇水溶液等度洗脱，收集14～18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90％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)在制备保肝的药物中的应用。
  7. 权利要求2所述的联苯苄唑的药物组合物在制备保肝的药物中的应用。
PCT/CN2017/097769 2016-06-23 2017-08-17 联苯苄唑的药物组合物及其保肝作用 WO2017220044A2 (zh)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN201610463340.7 2016-06-23
CN201610463340.7A CN106083875A (zh) 2016-06-23 2016-06-23 联苯苄唑的药物组合物及其保肝作用

Publications (2)

Publication Number Publication Date
WO2017220044A2 true WO2017220044A2 (zh) 2017-12-28
WO2017220044A3 WO2017220044A3 (zh) 2018-02-15

Family

ID=57252211

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2017/097769 WO2017220044A2 (zh) 2016-06-23 2017-08-17 联苯苄唑的药物组合物及其保肝作用

Country Status (2)

Country Link
CN (1) CN106083875A (zh)
WO (1) WO2017220044A2 (zh)

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106083875A (zh) * 2016-06-23 2016-11-09 崔坤峰 联苯苄唑的药物组合物及其保肝作用
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 (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105709053A (zh) * 2016-04-01 2016-06-29 胡文杰 一种具有保肝作用的中药复方及其制备方法
CN105622563A (zh) * 2016-04-08 2016-06-01 庄立 盐酸溴己新的药物组合物及其医药用途
CN105837654A (zh) * 2016-04-18 2016-08-10 镇江高海生物药业有限公司 一种盐酸强力霉素的药物组合物及其生物医药用途
CN105796560A (zh) * 2016-04-23 2016-07-27 徐挺 环丙沙星的药物组合物及其在生物医药中的应用
CN105801540A (zh) * 2016-04-23 2016-07-27 黄亦琼 一种盐酸卡替洛尔的药物组合物及其医药用途
CN105884715A (zh) * 2016-04-23 2016-08-24 陈斌 一种苄达赖氨酸的药物组合物及其医药用途
CN105693668A (zh) * 2016-04-23 2016-06-22 贺玉皓 一种氯法齐明的药物组合物及其医药用途
CN106083875A (zh) * 2016-06-23 2016-11-09 崔坤峰 联苯苄唑的药物组合物及其保肝作用

Also Published As

Publication number Publication date
WO2017220044A3 (zh) 2018-02-15
CN106083875A (zh) 2016-11-09

Similar Documents

Publication Publication Date Title
US7897791B2 (en) Purifications of pomegranate ellagitannins and their uses thereof
WO2017220041A2 (zh) 苯扎贝特的药物组合物及其在类风湿性关节炎中的应用
CN108707179A (zh) 一种金樱子皂苷元a的制备方法
CN105837654A (zh) 一种盐酸强力霉素的药物组合物及其生物医药用途
WO2017220044A2 (zh) 联苯苄唑的药物组合物及其保肝作用
WO2017220050A2 (zh) 阿奇霉素的药物组合物及其止咳的医药用途
CN105693668A (zh) 一种氯法齐明的药物组合物及其医药用途
CN105693806A (zh) 一种二甲磺酸阿米三嗪的药物组合物及其医药用途
CN113999272A (zh) 香蜂草苷的制备方法及其应用
Yu et al. Secondary metabolites of petri-dish cultured Antrodia camphorata and their hepatoprotective activities against alcohol-induced liver injury in mice
WO2017215680A2 (zh) 氯普噻吨的药物组合物及对脑缺血再灌注损伤的保护作用
CN105801540A (zh) 一种盐酸卡替洛尔的药物组合物及其医药用途
CN101205249B (zh) 从黑刺菝葜植物中制备拉肖皂苷元的方法
CN106496245A (zh) 赖诺普利的药物组合物及其在生物医药中的应用
CN107880084B (zh) 从凌霄属植物花中提取制备高纯度毛蕊花糖苷的方法
CN105884715A (zh) 一种苄达赖氨酸的药物组合物及其医药用途
CN105796560A (zh) 环丙沙星的药物组合物及其在生物医药中的应用
CN105622563A (zh) 盐酸溴己新的药物组合物及其医药用途
CN110204589B (zh) 青葙子有效成分、提取方法及其在制备神经保护药物方面的应用
WO2017215679A2 (zh) 苯丁酸氮芥的药物组合物及其抗抑郁的医药用途
CN106109459A (zh) 盐酸地匹福林的药物组合物及其在生物医药中的应用
CN105777683A (zh) 双环醇的药物组合物及其医药用途
CN105906684A (zh) 一种氟哌啶醇的药物组合物及其医药用途
CN111329866A (zh) 一种五环三萜类化合物在制备抗偏头痛药物中的应用
WO2017220051A2 (zh) 盐酸苄丝肼的药物组合物及其降血糖的医药用途

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: 17814781

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: 17814781

Country of ref document: EP

Kind code of ref document: A2