WO2011066683A1 - 用以治疗肝炎的医药组合物 - Google Patents

用以治疗肝炎的医药组合物 Download PDF

Info

Publication number
WO2011066683A1
WO2011066683A1 PCT/CN2009/075284 CN2009075284W WO2011066683A1 WO 2011066683 A1 WO2011066683 A1 WO 2011066683A1 CN 2009075284 W CN2009075284 W CN 2009075284W WO 2011066683 A1 WO2011066683 A1 WO 2011066683A1
Authority
WO
WIPO (PCT)
Prior art keywords
pharmaceutical composition
concentration
sesamin
serum
pharmaceutical combination
Prior art date
Application number
PCT/CN2009/075284
Other languages
English (en)
French (fr)
Inventor
侯建维
曾焕中
廖哲逸
黄乔志
黄介纬
贾维凡
Original Assignee
乔志亚生技股份有限公司
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 乔志亚生技股份有限公司 filed Critical 乔志亚生技股份有限公司
Priority to PCT/CN2009/075284 priority Critical patent/WO2011066683A1/zh
Publication of WO2011066683A1 publication Critical patent/WO2011066683A1/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
    • A61K31/357—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having two or more oxygen atoms in the same ring, e.g. crown ethers, guanadrel
    • A61K31/36—Compounds containing methylenedioxyphenyl groups, e.g. sesamin
    • 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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00

Definitions

  • the present invention relates to a pharmaceutical composition for treating hepatitis, and more particularly to a method for treating liver inflammation caused by lipopolysaccharide and lead. Background technique
  • Hepatitis is a common disease in Taiwan, and refers to liver inflammation caused by viral infection, heavy metal poisoning or other factors. Hepatitis may cause a liver disease that causes high mortality such as chronic liver disease, liver cirrhosis, and liver cancer. According to the top ten causes of death of the Taiwan Health Department, people who die from chronic liver disease, liver cirrhosis and liver cancer are among the best, and about 10,000 people die each year from related liver diseases. Among them, excessive fatigue, alcohol abuse, long-term medication, diabetes or overweight are high risk groups for liver disease.
  • the main object of the present invention is to provide the effect of sesamin on lowering the concentration of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and proinflammatory cytokines in blood.
  • ALT alanine aminotransferase
  • AST aspartate aminotransferase
  • proinflammatory cytokines in blood.
  • the present invention provides a pharmaceutical composition for treating hepatitis comprising: an effective amount of sesamin as an active ingredient and a pharmaceutically acceptable carrier.
  • the hepatitis refers to a phenomenon of liver inflammation caused by lipopolysaccharide and lead.
  • the proportion of the sesamin in the pharmaceutical composition is
  • the proportion of the carrier is 97 ⁇ 99.998wt%. More preferably, the proportion of the sesamin in the pharmaceutical composition is 2 to 3 wt%; and the proportion of the carrier is 97 to 98 wt%.
  • the effective amount of sesamin is at a dose of 5 to 50 mg/kg/day. In a preferred embodiment, the sesamin has a purity of 30 to 95%.
  • the pharmaceutical composition is further added with vitamins, minerals or amino acids.
  • the pharmaceutically acceptable carrier is corn starch, sesame oil, lactose, glucose, microcrystalline cellulose, magnesium stearate, polyvinylpyrrolidone, carboxymethylcellulose or a mixture thereof.
  • the pharmaceutical composition is prepared as an oral or parenteral dosage form.
  • the pharmaceutical composition is formulated as a powder, a pill, a lozenge, a capsule, a suspension, a solution or an injection.
  • the pharmaceutical composition further comprises a biocompatible additive, the additive being a preservative, an antioxidant, a stabilizer, an excipient, or a mixture thereof.
  • the invention further provides a method of reducing the concentration of alanine aminotransferase and aspartate aminotransferase in blood comprising contacting a sample with a pharmaceutical composition of the invention.
  • the invention further provides a method of reducing the concentration of pro-inflammatory cytokines in blood comprising contacting a sample with a pharmaceutical composition of the invention.
  • the pro-inflammatory cytokine is interleukin-1, interleukin-6 or tumor necrosis factor- ⁇ .
  • the sample is a mammal.
  • the mammal is a mouse, pig, cow or human.
  • the invention further provides a method for treating hepatitis, comprising providing the medical group of the subject
  • the present invention relates to a pharmaceutical combination 4 which is based on sesamin and relates to a method for treating hepatitis using the pharmaceutical composition.
  • Figure 1A shows the change in the concentration of aspartate aminotransferase in the serum of experimental rats.
  • Figure 1B shows the change in alanine aminotransferase concentration in the serum of experimental mice.
  • Figure 2A shows the change in interleukin-1 concentration in the serum of experimental mice.
  • Figure 2B shows the change in interleukin-6 concentration in the serum of experimental rats.
  • FIG. 3 shows the concentration of tumor necrosis factor- ⁇ in the serum of experimental rats 4 hours after administration.
  • Figure 4 shows the concentration of nitric oxide in the serum of experimental rats 4 hours after application.
  • Figure 5 shows the concentration of active oxide in the serum of experimental rats 4 hours after application.
  • liver disease There are many causes of hepatitis. Excessive fatigue, alcohol abuse, long-term medication, diabetes or overweight are high risk groups for liver disease. In addition, viral infections and heavy metal poisoning are also important factors in triggering hepatitis.
  • ALT and AST are enzymes mainly present in liver cells. When liver cells are damaged and broken, they are released into the blood, so the serum is in the form of alanine. The concentration of acid transaminase and aspartate aminotransferase is an important indicator of liver cell damage.
  • the inflammatory response is often accompanied by an increase in the concentration of proinflammatory cytokines (such as interleukin-1, interleukin-6 or tumor necrosis factor- ⁇ ), nitric oxide and active oxides in the serum, thus, in combination with the alanine aminotransferase,
  • the concentration data of aspartate aminotransferase, proinflammatory cytokines, nitric oxide and active oxides in serum can be used for the diagnosis of hepatitis and the degree of inflammation of the liver.
  • Lipopolysaccharides lipopolysaccharides
  • lipopolysaccharides are macromolecules linked by covalent bonds of lipids and sugars, mainly distributed on the cell surface of gram-negative cells, which are endotoxin of pathogenic bacteria. It is currently known that lipopolysaccharide triggers a fierce immune inflammatory response in the host, and it is also common to cause inflammation of the liver.
  • Lead is a metal that is widely used in human society. Lead poisoning can cause disorder in the circulatory system, urinary system, nervous system and endocrine system, but it does not directly cause liver damage. However, studies have shown that the synergistic effect of lead and lipopolysaccharide can cause severe inflammation in the liver, and a significant increase in the concentration of pro-inflammatory cytokines in the serum is observed.
  • the present invention induces inflammation of the liver of the experimental mouse by the synergistic action of lead and lipopolysaccharide, and tests for the inhibition of inflammatory cytokines, nitric oxide, active oxides, alanine aminotransferase and aspartic acid in the experimental mice.
  • administration is defined as providing a pharmaceutical composition of the present invention to a subject by oral or parenteral administration.
  • the pharmaceutical composition comprises, but is not limited to: a powder, a pill, a tablet, a capsule, a suspension or a solution; wherein, in a non-oral manner, the form of the pharmaceutical composition comprises , but not limited to: Injection, for subcutaneous, skin, muscle, joint, vein, artery, or intracranial injection.
  • the term "effective amount” is defined as the dose that effectively reduces the concentration of proinflammatory cytokines, nitric oxide, active oxides, alanine aminotransferase, and aspartate aminotransferase in the blood, ⁇ , to achieve hepatitis. .
  • the experimental rats (Sprague-Dawley rats) were provided by the National Laboratory Animal Center (Taipei, Taiwan, PRC) and hosted at the Animal Center of Chinese Medical University. Taichung, Taiwan, PRC randomly assigned the same number of experimental rats to three groups: negative control group, positive control group, and experimental group 0 .
  • the positive control group and the experimental group were injected intraperitoneally with lipopolysaccharide (50 g/kg) and lead (5 mg/kg) to synergistically induce liver inflammation; the experimental group was fed a dose of sesamin. It was 10 mg/kg/day; the negative control group was replaced with an equal weight of physiological saline.
  • the sesamin has a purity of 60%.
  • mice At 4, 6, 12 and 24 hours, 0.8 ml blood samples from each group of mice were collected. The collected blood sample was placed in a microtube for 20 minutes, and then centrifuged at 2,000 g for 15 minutes at 4 ° C to collect serum.
  • the concentration of alanine aminotransferase and aspartate aminotransferase in the serum was measured by a blood biochemical analyzer to estimate liver function.
  • the experiment was carried out using a conventional enzyme-linked immunosorbent assay kit (R&D Systems, Inc. Minnesota, USA), and the serum was measured at a wavelength of 450 nm using a microplate reader. Concentrations of interleukin-1 and interleukin-6. [Detection of tumor necrosis factor- ⁇ concentration]
  • 100 ⁇ serum samples were placed in a 96-well ELISA reader plate and assayed using a conventional enzyme-linked immunosorbent assay kit (R&D Systems, Inc. Minnesota, USA).
  • the concentration of tumor necrosis factor- ⁇ in the serum was measured at a wavelength of 450 nm using a microplate reader.
  • nitric oxide (NO) becomes a stable N0 2 , and then a microplate reader is used.
  • the concentration of N0 2 can be determined by detecting the absorbance at a wavelength of 540 nm to determine the concentration of nitric oxide.
  • Active oxide is a highly active molecule centered on an oxygen atom, such as superoxide anion radical (0 2 — ), hydrogen peroxide (H 2 0 2 ) hydrogen.
  • concentration of active oxide is determined by reference to the experimental method. 0.1 ml of serum is placed in a 96-well culture dish, and 1 ⁇ of 10 is added. mM oxidation-sensitive fluorescent calibration (3 ⁇ 4DCFDA, 2',7'-dichlorofluorescein diacetate) is allowed to react at 37 ° C for 60 minutes.
  • Example 2 The effect of sesamin on lowering the concentration of alanine aminotransferase and aspartate aminotransferase in blood
  • every 10 experimental rats were randomly assigned to the negative control group.
  • the positive control group and the experimental group Blood samples were taken after 0.5, 1, 1.5, 2, 4, and 6 hours of the experiment to detect the concentrations of alanine aminotransferase and aspartate aminotransferase in the serum.
  • Figure 1A the concentration of aspartate aminotransferase (AST) in serum is shown;
  • Figure 1B shows the concentration of alanine aminotransferase (ALT) in serum.
  • the concentration of alanine aminotransferase and aspartate aminotransferase in the serum of the positive control group and the experimental group was significantly increased about 1 hour after the administration, indicating that the liver cells were being subjected to damage.
  • This increased trend lasted for 6 hours, and it was observed that the concentrations of alanine aminotransferase and aspartate aminotransferase in the serum of the experimental group were significantly lower than those in the positive control group.
  • sesamin reduces the increase in alanine aminotransferase and aspartate aminotransferase concentrations in the serum of rats, which means that liver cell damage is alleviated.
  • Example 3 The effect of sesamin on reducing the concentration of interleukin-1 and interleukin-6 in serum
  • every 10 experimental rats were randomly assigned to the negative control group, the positive control group and the The experimental group. Blood samples were taken at 0.5, 1, 1.5, 2, 4, and 6 hours after the experiment to detect the concentrations of interleukin-1 and interleukin-6 in the serum. See Figure 2A for the concentration of interleukin-1 in serum; Figure 2B shows the concentration of interleukin-6 in serum.
  • the concentrations of interleukin-1 and interleukin-6 in the serum of the positive control group and the experimental group were significantly higher than those of the negative control group, indicating that a severe inflammatory reaction was occurring.
  • the concentration of interleukin-1 and interleukin-6 in the serum of the experimental group was significantly lower than that of the positive control group, indicating that sesamin can decrease the serum interleukin-1 and interleukin-6 concentrations.
  • the degree of elevation, gp relieves the extent of the inflammatory response.
  • Example 4 The effect of sesamin on reducing the concentration of tumor necrosis factor- ⁇ in serum
  • Example 1 According to the experimental design of Example 1, every 10 experimental mice were randomly assigned to the negative control group, the positive control group, and the experimental group. After 4 hours of the experiment, the concentration of tumor necrosis factor- ⁇ in the serum of each group of mice was examined.
  • Example 1 According to the experimental design of Example 1, every 10 experimental mice were randomly assigned to the negative control group, the positive control group, and the experimental group. After 4 hours of the experiment, the concentration of nitric oxide in the serum of each group of mice was examined.
  • nitric oxide is also a message molecule in the living body
  • the basal content of nitric oxide is present in the serum of the negative control group.
  • the concentration of nitric oxide in the serum increased significantly due to the induction of hepatitis.
  • the concentration of nitric oxide in the serum returned to the negative control group after 4 hours of experiment due to the action of sesamin. The same base content. From this data, it can be seen that after 4 hours of the experiment, sesamin has significantly relieved the increase in the concentration of nitric oxide, and it has been judged that the effect of reducing inflammation has been achieved.
  • Example 6 The utility of sesamin in reducing the concentration of active oxides in serum
  • every 6 experimental mice were randomly assigned to the negative control group, the positive control group, and the experimental group. After 4 hours of the experiment, the concentrations of active oxides in the serum of each group of mice were examined.
  • the concentration of active oxide is expressed as an oxidation-sensitive fluorescent label (2',7'-dichlorofluorescin).
  • Active oxides are a must-have metabolite in organisms undergoing aerobic respiration, so it can be seen from the figure that basal levels of active oxide are present in the serum of mice in the negative control group.
  • the concentration of the active oxide increased due to the induction of hepatitis, but the mice in the experimental group greatly reduced the concentration of the active oxide in the serum due to the action of sesamin. It was evident that sesamin significantly relieved the inflammation after 4 hours of the experiment.

Landscapes

  • Health & Medical Sciences (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Veterinary Medicine (AREA)
  • Chemical & Material Sciences (AREA)
  • Public Health (AREA)
  • General Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Animal Behavior & Ethology (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Organic Chemistry (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • General Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Epidemiology (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

本发明是关于一种用以治疗肝炎的医药组合物，其包含有效量的芝麻素做为活性成分以及医药上可接受的载体，可降低血清中丙氨酸转氨酶、天门冬氨酸转氨酶和促发炎细胞素的浓度。

Description

用以治疗肝炎的医药组合物 技术领域
本发明是关于一种用以治疗肝炎的医药组合物， 尤其是指一种用以治 疗由脂多糖与铅所协同引发的肝脏发炎现象。 背景技术
肝炎是中国台湾常见的疾病， 泛指于病毒感染、 重金属中毒或其它因 素所引发的肝脏发炎现象。 肝炎可能进一歩引发慢性肝病、 肝硬化、 肝癌 等死亡率高的肝脏疾病。 根据中国台湾卫生署的十大死因统计， 因慢性肝 病、 肝硬化和肝癌死亡者名列前茅， 每年约略有一万人死于相关的肝脏疾 病。 其中， 疲劳过度、 有酗酒习惯、 长期服药、 糖尿病或体重过重者， 为 肝脏疾病的高危险群。
近年来， 芝麻中所含的木质酚 Gignans)如芝麻素 (sesamin；)、 芝麻醚素 (sesamolin,或称芝麻酚林、芝麻林素)、芝麻酚 (sesamol)和芝麻醇 (sesaminol) 已引起学术与产业上广泛的兴趣， 尤其是它们于抗氧化、 降血脂、 降血压 和降胆固醇的功效。 其中， 芝麻素亦是一个极具潜力的抗发炎药剂， 但目 前尚未有任何研究文献指出其于肝炎的效用。 发明内容
本发明的主要目的为提供芝麻素于降低血液中丙氨酸转氨酶 (alanine aminotransferase , ALT)、天门冬氨酉變转氨酶 (aspartate aminotransferase, AST) 和促发炎细胞素 (proinflammatory cytokines)浓度的效用， 并进一歩提供一种 以芝麻素为活性成分的治疗肝炎的医药组合物。
为达上述目的， 本发明提供了一种用以治疗肝炎的医药组合物， 其包 含： 有效量的芝麻素做为活性成分以及医药上可接受的载体。 于一较佳实施例中， 所述肝炎是指由脂多糖与铅所协同引发的肝脏发 炎现象。
于一较佳实施例中， 所述芝麻素于所述医药组合物中所占比例为
0.002~3wt%; 所述载体所占比例为 97~99.998wt%。 更佳地， 所述芝麻素于 所述医药组合物中所占比例为 2~3wt%； 所述载体所占比例为 97~98wt%。
于一较佳实施例中， 所述有效量的芝麻素的剂量为 5~50 mg/kg/day。 于一较佳实施例中， 所述芝麻素的纯度为 30~95%。
于一较佳实施例中， 所述医药组合物进一歩添加维生素、 矿物质或氨 基酸。
于一较佳实施例中， 所述医药上可接受的载体为玉米淀粉、 麻油、 乳 糖、 葡萄糖、 微结晶纤维素、 硬脂酸镁、 聚乙烯吡咯酮、 羧甲基纤维素或 其混合物。
于一较佳实施例中， 所述医药组合物是制备为口服或非口服剂型。 于一较佳实施例中， 所述医药组合物是配制为粉剂、 丸剂、 锭剂、 胶 囊、 悬浮液、 溶液或注射剂。
于一较佳实施例中， 所述医药组合物进一歩包含生物可相容的添加剂， 该添加剂为防腐剂、 抗氧化剂、 安定剂、 赋形剂或其混合。
本发明另提供一种降低血液中丙氨酸转氨酶和天门冬氨酸转氨酶浓度 的方法， 其包含使样本与本发明的医药组合物接触。
本发明又提供一种降低血液中促发炎细胞素浓度的方法， 其包含使样 本与本发明的医药组合物接触。
于一较佳实施例中， 所述促发炎细胞素为白介素 -1、 白介素 -6 或肿瘤 坏死因子 -α。
于一较佳实施例中， 所述样本为哺乳动物。 其中， 该哺乳动物为老鼠、 猪、 牛或人。
本发明更提供一种治疗肝炎的方法， 其包含提供受测对象所述医药组 综上所述， 本发明是关于一种医药组合 4 ， 其是以芝麻素为主要成分， 并关于使用该医药组合物于治疗肝炎的方法
酶、 天门冬氨酸转氨酶和促发炎细胞素浓度。 附图说明
图 1A呈现实验鼠血清中天门冬氨酸转氨酶浓度的变化。
图 1B呈现实验鼠血清中丙氨酸转氨酶浓度的变化。
图 2A呈现实验鼠血清中白介素 -1浓度的变化。
图 2B呈现实验鼠血清中白介素 -6浓度的变化。
图 3呈现施药 4小时后， 实验鼠血清中肿瘤坏死因子 -α的浓度。
图 4呈现施药 4小时后， 实验鼠血清中一氧化氮的浓度。
图 5呈现施药 4小时后， 实验鼠血清中活性氧化物的浓度。 具体实施方式
引发肝炎的原因有很多， 疲劳过度、 有酗酒习惯、 长期服药、 糖尿病 或体重过重者， 为肝病的高危险群。 除此之外， 病毒感染和重金属中毒也 是引发肝炎的重要因素。
丙氨酸转氨酶（alanine aminotransferase， ALT)和天门冬氨酸转氨酶 (aspartate aminotransferase, AST)是主要存在于肝脏细胞的酶， 当肝脏细胞 受损破碎时， 会释放至血液中， 因此血清中丙氨酸转氨酶和天门冬氨酸转 氨酶的浓度为肝脏细胞受损的重要指标。 而发炎反应往往伴随着血清中促 发炎细胞素（如白介素 -1、 白介素 -6或肿瘤坏死因子 -α)、 一氧化氮和活性 氧化物浓度的提高， 因此， 综合所述丙氨酸转氨酶、 天门冬氨酸转氨酶、 促发炎细胞素、 一氧化氮和活性氧化物于血清中的浓度数据， 可用于肝炎 的诊断以及肝脏发炎程度的判断。 脂多糖 (lipopolysaccharides;)是由脂质和糖类经共价键链接的大分子，主 要分布于革兰氏阴菌 (gram-negative;)的细胞表面， 为病原菌的内毒素。 目前 已知， 脂多糖会引发宿主体内激烈的免疫发炎反应， 也常见造成肝脏发炎 的现象。
铅为广泛运用于人类社会中的金属。 铅中毒会造成循环系统、 泌尿系 统、 神经系统和内分泌系统失序， 但并不会直接造成肝脏的损伤。 然而， 研究指出铅与脂多糖的协同作用会引发肝脏严重的发炎现象， 并观察到血 清中促发炎细胞素浓度的显著提升。
据此， 本发明以铅与脂多糖的协同作用引发实验鼠肝脏发炎， 并测试 芝麻素于降低实验鼠体内促发炎细胞素、 一氧化氮、 活性氧化物、 丙氨酸 转氨酶和天门冬氨酸转氨酶浓度的效用， 以定义其治疗肝炎的效果。
名词 "服用" 的定义为以口服或非口服的方式提供本发明的医药组合 物给一受测对象。 其中， 以口服的方式， 所述医药组合物的形式包含， 但 不限于： 粉剂、 丸剂、 锭剂、 胶囊、 悬浮液或溶液； 其中， 以非口服的方 式， 所述医药组合物的形式包含， 但不限于： 注射剂， 用以于皮下、 皮肤、 肌肉、 关节、 静脉、 动脉、 或颅内注射。
名词 "有效量" 的定义为， 该剂量能充分降低血液中促发炎细胞素、 一氧化氮、 活性氧化物、 丙氨酸转氨酶和天门冬氨酸转氨酶的浓度， δ卩， 达到治疗肝炎的效果。
以下实施例是用于进一歩了解本发明的优点， 并非用于限制本发明的 请求保护范围。 实施例一： 实验设计
[实验鼠]
实验鼠 (Sprague-Dawley rats)是由中国台北国家实验研究院实验动物中 心 (National Laboratory Animal Center, Taipei, Taiwan, PRC)提供， 并寄存于 台中中国医药学院动物中心 (Animal Center of Chinese Medical University, Taichung, Taiwan, PRC 随机将同数量的实验鼠分配为三个组别： 负对照 组 (negative control group)、 正对照组 (positive control group)禾口实验组 (experimental group) 0 请参下表一， 其中， 正对照组和实验组分别经腹腔内 注射的方式注射脂多糖 (50 g/kg)和铅 (5 mg/kg)以协同引发肝脏发炎现象； 实验组并进一歩喂食芝麻素， 其剂量为 10 mg/kg/day; 负对照组则喂以等 重量的生理食盐水替代。
Figure imgf000007_0001
+表示有喂食； - 表示没有喂食。 其中， 所述芝麻素的纯度为 60%。
[收集血清样本]
于注射脂多糖和醋酸铅、 喂食芝麻素或生理食盐水后 0.5、 1、 1.5、 2、
4、 6、 12和 24小时， 分别采集各组别的老鼠 0.8 ml 的血液样本。 将采集 的血液样本置于离心小管 (microtube)中静置 20分钟后，再以 2,000 g的速度 于 4°C离心 15分钟以收集血清。
[肝脏功能侦测]
以血液生化分析仪 (blood biochemical analyzer)测量所述血清中丙氨酸 转氨酶和天门冬氨酸转氨酶的浓度以推算肝脏功能。
[白介素 -l(IL-l)和白介素 -6(IL-6)的浓度侦测]
使用习知的酶联免疫吸附分析试剂盒 (enzyme-linked immunosorbent assay kit, R&D Systems, Inc. Minnesota, USA)进行实验， 再以微量盘分析仪 (microplate reader)于波长 450 nm测定所述血清中白介素 -1和白介素 -6的浓 度。 [肿瘤坏死因子 -α的浓度侦测]
将 100 μΐ血清样品置于 96 孔盘 (96-well ELISA reader plate)中，使用习 知的酶联免疫吸附分析试剂盒 (enzyme-linked immunosorbent assay kit, R&D Systems, Inc. Minnesota, USA)进行实验， 再以微量盘分析仪 (microplate reader)于波长 450 nm测定所述血清中肿瘤坏死因子 -α的浓度。
[一氧化氮的浓度侦测]
将 100 μΐ血清样品置于 96 孔盘 (96-well ELISA reader plate)中，再加入 100 μΐ的格里斯试剂 (Griess reagent, 是将溶于 5%磷酸盐溶液的 1%磺氨溶 液 (sulfanilamide solution)与 0.1% NED 溶液 (N- 1 -napthylenediamine dihydrochloride solution )以 1 : 1 的体积混合）， 其中， 一氧化氮 (NO)会变成 稳定的 N02， 然后再利用微量盘分析仪 (microplate reader)于波长 540 nm 侦 测吸光值以测定 N02的浓度， 即可判断一氧化氮的浓度。
[活性氧化物的浓度侦测： I
活性氧化物 (； ROS， reactive oxygen species)是以氧原子为中心的高活性 分子， 如超氧阴离子自由基 (superoxide anion radical, 02— )、 过氧化氢 (hydrogenperoxide , H202) 氢氧自由基 (hydroxyl radical , OH") 及其所衍生 的氧化活性分子。活性氧化物的浓度测定是参考习知的实验方法，取 0.1 ml 血清置于 96 孔培养盘中， 加入 1 μΐ 的 10 mM 氧化敏感荧光标定物 (¾DCFDA， 2',7'-dichlorofluorescein diacetate) 使其于 37 °C下反应 60 分钟。 当 ¾DCFDA与 ¾02 作用会氧化成 DCF(2',7'-dichlorofluorescein)， 经荧光 侦测仪激发后可在波长 525 nm侦测到荧光， 即可判断活性氧化物的浓度。 实施例二： 芝麻素于降低血液中丙氨酸转氨酶和天门冬氨酸转氨酶浓 度的效用
依据实施例一的实验设计，随机将每 10只实验鼠分配为所述负对照组、 所述正对照组和所述实验组。 于实验 0.5、 1、 1.5、 2、 4、 6 小时之后采集 血液样本以侦测血清中丙氨酸转氨酶和天门冬氨酸转氨酶的浓度。 请参见 图 1A, 显示血清中天门冬氨酸转氨酶 (AST)的浓度； 图 1B则显示血清中丙 氨酸转氨酶 (ALT)的浓度。
由图 1A和图 1B可知， 于施药后 1个小时左右， 正对照组和实验组的 老鼠血清中丙氨酸转氨酶和天门冬氨酸转氨酶的浓度都有明显的提高， 显 示肝脏细胞正受到损伤。 这一增加的趋势持续到 6个小时的时候， 可以看 到实验组的老鼠血清中丙氨酸转氨酶和天门冬氨酸转氨酶的浓度显著地低 于正对照组。 这一结果显示芝麻素减轻了老鼠血清中丙氨酸转氨酶和天门 冬氨酸转氨酶浓度提升的程度， 意味着减轻了肝脏细胞受损的情况。 实施例三： 芝麻素于降低血清中白介素 -1和白介素 -6浓度的效用 依据实施例一的实验设计，随机将每 10只实验鼠分配为所述负对照组、 所述正对照组和所述实验组。 于实验 0.5、 1、 1.5、 2、 4、 6 小时之后采集 血液样本以侦测血清中白介素 -1和白介素 -6的浓度。请参见图 2A, 显示血 清中白介素 -1的浓度； 图 2B则显示血清中白介素 -6的浓度。
由图 2A和图 2B可知， 正对照组和实验组的老鼠血清中白介素 -1和白 介素 -6 的浓度相较于负对照组都有显著的上升， 代表剧烈的发炎反应正在 发生。 但于第 6个小时的数据可以看出， 实验组的老鼠血清中白介素 -1和 白介素 -6 的浓度显著地低于正对照组， 显示芝麻素可降低血清中白介素 -1 和白介素 -6浓度上升的程度， gp， 舒缓发炎反应的程度。 实施例四： 芝麻素于降低血清中肿瘤坏死因子 -α浓度的效用
依据实施例一的实验设计，随机将每 10只实验鼠分配为所述负对照组、 所述正对照组和所述实验组。 于实验 4 小时之后， 侦测各组别老鼠血清中 肿瘤坏死因子 -α的浓度。
比较诱发肝脏发炎的组别， 由图 3可知， 施予芝麻素的组别 （实验组） 的老鼠的肿瘤坏死因子 -α浓度显著低于未施予芝麻素的组别（正对照组） 。 显示芝麻素舒缓了老鼠体内肿瘤坏死因子 -α浓度的提升， 意味着减缓了因 脂多糖和铅所协同引发的肝脏发炎现象。 实施例五： 芝麻素于降低血清中一氧化氮浓度的效用
依据实施例一的实验设计，随机将每 10只实验鼠分配为所述负对照组、 所述正对照组和所述实验组。 于实验 4 小时之后， 侦测各组别老鼠血清中 一氧化氮的浓度。
请参见图 4， 由于一氧化氮同时亦为生物体内的讯息分子， 因此在负对 照组的老鼠的血清中， 即有基础含量的一氧化氮存在。 正对照组由于肝炎 的诱发， 血清中一氧化氮的浓度显著升高， 反观实验组由于芝麻素的作用， 血清中一氧化氮的浓度于实验 4个小时之后， 已回复至与负对照组差不多 相同的基础含量。 由此数据可知， 于实验 4个小时之后， 芝麻素已显著舒 缓了一氧化氮浓度的提升， 可判断已达到减轻发炎的效果。 实施例六： 芝麻素于降低血清中活性氧化物浓度的效用
依据实施例一的实验设计， 随机将每 6只实验鼠分配为所述负对照组、 所述正对照组和所述实验组。 于实验 4 小时之后， 侦测各组别老鼠血清中 活性氧化物的浓度。
请参见图 5， 以氧化敏感荧光标定物 (2',7'-dichlorofluorescin)表现活性氧 化物的浓度。 活性氧化物为行有氧呼吸的生物体内必定存在的代谢产物， 因此由图中可见， 负对照组的老鼠的血清中存在基础含量的活性氧化物。 在正对照组的老鼠的血清中， 活性氧化物的浓度因为肝炎的诱发而提高， 但实验组的老鼠因为芝麻素的作用， 大幅降低其血清中活性氧化物的浓度。 显见芝麻素于实验 4个小时之后， 显著的舒缓了发炎的现象。

Claims

权利要求书 一种用以治疗肝炎的医药组合物， 其包含：
有效量的芝麻素做为活性成分； 以及
医药上可接受的载体。
2. 如权利要求 1所述的医药组合物， 其中所述芝麻素于所述医药组合 中所占比例为 0.002~3wt%； 所述载体所占比例为 97~99.998wt%。
3. 如权利要求 1所述的医药组合 /, 其中所述肝炎是指由脂多糖与铅所 协同引发的肝脏发炎现象。
4. 如权利要求 1所述的医药组合 /, 其中所述有效量的芝麻素的剂量为 5-50 mg/kg/day。
如权利要求 1所述的医药组合 1，其中所述芝麻素的纯度为 30~95%。
6. 如权利要求 1所述的医药组合 /, 其进一歩添加维生素、 矿物质或氨
7 如权利要求 1所述的医药组合 /, 其中所述医药上可接受的载体为玉 米淀粉、 麻油、 乳糖、 葡萄糖、 微结晶纤维素、 硬脂酸镁、 聚乙烯吡 咯酮、 羧甲基纤维素或其混合
8. 如权利要求 1所述的医药组合 I， 其是制备为口服或非口服剂型。
9. 如权利要求 1所述的医药组合 /, 其是配制为粉剂、 丸剂、 锭剂、 胶 囊、 悬浮液、 溶液或注射剂。
10. 如权利要求 1所述的医药组合 其进一歩包含生物可相容的添加剂， 该添加剂为防腐剂、 抗氧化剂、 安定剂、 赋形剂或其混合。
11.一种降低血液中丙氨酸转氨酶和天门冬氨酸转氨酶浓度的方法， 其包 含使样本与权利要求 1至 10中任一项所述的医药组合物接触。
12. 如权利要求 11所述的方法， 其中所述样本为哺乳动物。
13. 如权利要求 12所述的方法， 其中所述哺乳动物包含老鼠、 猪、 牛或 人。
14.一种降低血液中促发炎细胞素浓度的方法， 其包含使样本与权利要求 1至 10中任一项所述的医药组合物接触。
15. 如权利要求 14所述的方法，其中所述促发炎细胞素为白介素 -1、 白介 素 -6或肿瘤坏死因子 -α。
16. 如权利要求 14所述的方法， 其中所述样本为哺乳动物。
17. 如权利要求 16所述的方法， 其中所述哺乳动物为老鼠、 猪、 牛或人。
18.一种治疗肝炎的方法， 其包含使受测对象服用权利要求 1至 10中任 一项所述的医药组合物。
PCT/CN2009/075284 2009-12-03 2009-12-03 用以治疗肝炎的医药组合物 WO2011066683A1 (zh)

Priority Applications (1)

Application Number Priority Date Filing Date Title
PCT/CN2009/075284 WO2011066683A1 (zh) 2009-12-03 2009-12-03 用以治疗肝炎的医药组合物

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
PCT/CN2009/075284 WO2011066683A1 (zh) 2009-12-03 2009-12-03 用以治疗肝炎的医药组合物

Publications (1)

Publication Number Publication Date
WO2011066683A1 true WO2011066683A1 (zh) 2011-06-09

Family

ID=44114597

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2009/075284 WO2011066683A1 (zh) 2009-12-03 2009-12-03 用以治疗肝炎的医药组合物

Country Status (1)

Country Link
WO (1) WO2011066683A1 (zh)

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20010031275A1 (en) * 1998-02-09 2001-10-18 Forse R Armour Sesamol inhibitor of delta-5-desaturase activity and uses therefor
US20060228432A1 (en) * 2002-10-29 2006-10-12 Rao Janaswamy M alpha-glucosidase inhibitors from a natural source
CN1938317A (zh) * 2004-03-31 2007-03-28 三得利株式会社 脂连蛋白增强剂

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20010031275A1 (en) * 1998-02-09 2001-10-18 Forse R Armour Sesamol inhibitor of delta-5-desaturase activity and uses therefor
US20060228432A1 (en) * 2002-10-29 2006-10-12 Rao Janaswamy M alpha-glucosidase inhibitors from a natural source
CN1938317A (zh) * 2004-03-31 2007-03-28 三得利株式会社 脂连蛋白增强剂

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
CUI YUNSHAN ET AL.: "Sesamin Powder Protects Against Acute Hepatotoxicity Induced by Alcohol in Mice.", LISHIZHEN MEDICINE AND MATERIA MEDICA RESEARCH (CN)., June 2005 (2005-06-01), pages 116 - 117 *
WANG WUSAN ET AL.: "The Protective Effects of Sesamin against Liver Damage in Rodents", PHARMACOLOGY AND CLINICS OF CHINESE MATERIA MEDICA (CN), vol. 22, no. 3,4, April 2006 (2006-04-01), pages 27 - 32 *

Similar Documents

Publication Publication Date Title
US20210322407A1 (en) Use of Trimetazidine in Preparation of Drugs for Preventing and Treating Liver Diseases
Dong et al. Chlorogenic acid alleviates allergic inflammatory responses through regulating Th1/Th2 balance in ovalbumin-induced allergic rhinitis mice
WO2021216581A1 (en) Method of treating and preventing coronavirus disease 19 (covid-19) using a selenium administration
Billah et al. Pharmacology ofN-(3, 5-Dichloro-1-oxido-4-pyridinyl)-8-methoxy-2-(trifluoromethyl)-5-quinoline Carboxamide (SCH 351591), a Novel, Orally Active Phosphodiesterase 4 Inhibitor
Joo et al. Clinical Evaluation of Praziquantel (Embay 8440; Biltricide®) in the Treatment of Paragonimas westermani
CN111991378A (zh) 羟基-α-山椒素在制备调节血糖药物中的应用
WO2011066683A1 (zh) 用以治疗肝炎的医药组合物
WO2022033200A1 (zh) 索拉非尼在治疗1型糖尿病中的应用
CN109419787B (zh) 一种松香烷型二萜类化合物的用途
Zhu et al. Study on the clinical safe and effective methods of arsenic-containing compound-Qinghuang powder in the treatment of myelodysplastic syndrome
Lin et al. Metformin ameliorates neuronal necroptosis after intracerebral hemorrhage by activating AMPK
CN107617101B (zh) 含唑来膦酸和白细胞介素2的药物组合及其应用
CN113274393A (zh) Linsitinib化合物在制备抗EV71病毒药物中的应用
TW201119648A (en) Pharmaceutical composition for hepatitis treatment
CN113181152A (zh) Tiratricol化合物在制备抗EV71病毒药物中的应用
CN104605349A (zh) 一种含d-松醇并具有降血糖功能的保健品及功能学评价方法
CN111588021A (zh) 一株可缓解和治疗冠心病的乳双歧杆菌Probio-M8及其应用
WO2004032924A1 (fr) Emploi de berberine en tant que sensibilisant insulinique
CN114984094B (zh) 白头翁皂苷组合物在制备治疗car-t疗法引起crs的药物中的应用
CN115192592B (zh) 白头翁皂苷组合物在制备治疗糖尿病肾病的药物中的应用
EP0476391A2 (en) Anti-AIDS virus composition containing cepharanthine as active compound
CN114246880B (zh) 圣草次苷在制备lps诱导脓毒症急性肺损伤保护药物中的应用
CN113855670B (zh) A205804在制备治疗脓毒症药物中的应用
US11058718B2 (en) Method for treating non-alcoholic steatohepatitis (NASH) with the combination of polaprezinc and sodium selenite
CN111956680B (zh) 雷公藤多苷片在制备用于缓解和/或治疗噬血细胞综合征相关症状的药物上的应用

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: 09851783

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 09851783

Country of ref document: EP

Kind code of ref document: A1