WO2023051025A1 - Application of pulsatilla saponin b4 in preparation of medicine for preventing or treating non-alcoholic fatty liver diseases - Google Patents
Application of pulsatilla saponin b4 in preparation of medicine for preventing or treating non-alcoholic fatty liver diseases Download PDFInfo
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- WO2023051025A1 WO2023051025A1 PCT/CN2022/110354 CN2022110354W WO2023051025A1 WO 2023051025 A1 WO2023051025 A1 WO 2023051025A1 CN 2022110354 W CN2022110354 W CN 2022110354W WO 2023051025 A1 WO2023051025 A1 WO 2023051025A1
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- Prior art keywords
- medicine
- liver
- pulsatilla saponin
- fatty liver
- alcoholic fatty
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- 208000008338 non-alcoholic fatty liver disease Diseases 0.000 title claims abstract description 86
- 239000003814 drug Substances 0.000 title claims abstract description 49
- OUHBKBTZUPLIIA-OTEDBJMHSA-N [(2s,3r,4s,5s,6r)-6-[[(2r,3r,4r,5s,6r)-3,4-dihydroxy-6-(hydroxymethyl)-5-[(2s,3r,4r,5r,6s)-3,4,5-trihydroxy-6-methyloxan-2-yl]oxyoxan-2-yl]oxymethyl]-3,4,5-trihydroxyoxan-2-yl] (1r,3as,5ar,5br,7ar,8r,9s,11ar,11br,13ar,13br)-9-[(2s,3r,4s,5s)-4,5-dihydroxy- Chemical compound O[C@@H]1[C@H](O)[C@@H](O)[C@H](C)O[C@H]1O[C@@H]1[C@@H](CO)O[C@@H](OC[C@@H]2[C@H]([C@H](O)[C@@H](O)[C@H](OC(=O)[C@@]34[C@H]([C@@H](CC3)C(C)=C)[C@@H]3[C@@]([C@]5(C)CC[C@H]6[C@](C)(CO)[C@@H](O[C@H]7[C@@H]([C@@H](O)[C@@H](O)CO7)O[C@H]7[C@@H]([C@H](O)[C@@H](O)[C@H](C)O7)O)CC[C@]6(C)[C@H]5CC3)(C)CC4)O2)O)[C@H](O)[C@H]1O OUHBKBTZUPLIIA-OTEDBJMHSA-N 0.000 title claims abstract description 44
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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
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7028—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages
- A61K31/7034—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin
- A61K31/704—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin attached to a condensed carbocyclic ring system, e.g. sennosides, thiocolchicosides, escin, daunorubicin
-
- 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
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/06—Antihyperlipidemics
Definitions
- the invention belongs to the pharmaceutical technology, and in particular relates to the application of Pulsatilla saponin B4 in the preparation of medicaments for treating nonalcoholic fatty liver.
- Nonalcoholic fatty liver disease is a metabolic stress liver injury closely related to insulin resistance and genetic susceptibility. Its pathological changes are similar to those of alcoholic liver disease (ALD), but the patients have no history of excessive drinking. According to the progress of the disease course, it is divided into nonalcoholic simple fatty liver (NAFL), nonalcoholic steatohepatitis (NASH), cirrhosis and hepatocellular carcinoma (HCC).
- NAFL nonalcoholic simple fatty liver
- NASH nonalcoholic steatohepatitis
- HCC hepatocellular carcinoma
- MetS metabolic syndrome
- Metabolic syndrome includes increased waist circumference (such as obesity), hyperglycemia, hyperlipidemia, and hypertension.
- waist circumference such as obesity
- MetS increases the risk of NAFLD
- NAFLD may also aggravate several clinical features and comorbidities of MetS, so effective treatment of NASH is also beneficial to improve MetS symptoms.
- MetS is also an important driver of adverse cardiovascular disease (CVD) and overall mortality in NAFLD patients. Diabetes has the clearest biological link to the progression of NAFLD, with up to 75% of patients with type 2 diabetes having NAFLD.
- CVD adverse cardiovascular disease
- insulin resistance is also considered to be an integral part of the pathogenesis of NAFLD, and it worsens with disease progression. 50% of hypertensive patients are accompanied by NAFLD. Hypertension is closely related to the progression of fibrosis, and there has been a proven correlation between NAFLD and arteriosclerosis, myocardial remodeling, heart failure, and kidney disease.
- NAFLD non-alcoholic fatty liver disease
- NASH is considered a progressive form of NAFLD characterized by hepatic steatosis, inflammation, hepatocellular injury, and varying degrees of fibrosis.
- the occurrence of NASH is the result of multiple factors, including extrahepatic factors such as dietary factors (carbohydrates, metal ions, saturated fat/trans fat and cholesterol, etc.), metabolic dysfunction (insulin resistance and adipose tissue inflammation, adipokines, etc.) ), hepatic-gut axis (intestinal inflammation and intestinal barrier dysfunction, bile acids), etc.; intrahepatic factors such as hepatocyte stress (oxidative stress and endoplasmic reticulum stress, lipotoxicity, mitochondrial dysfunction), hepatic cell death ( Apoptosis, necroptosis, pyroptosis), nuclear receptors (PPAR ⁇ , FXR, CAR, etc.), liver cytokines, etc.; in addition to innate immunity, host genetics and epigenetics.
- Metformin which is widely used clinically, mainly plays a role in the treatment of non-alcoholic fatty liver disease (especially NASH) by activating AMPK, but this drug is an inhibitor of mitochondrial complex I, there is a risk of lactic acidosis, and there is no liver tissue Learning to improve.
- the current clinically recommended drugs for the treatment of NASH include insulin sensitizers such as pioglitazone, etc., but the above-mentioned drugs have limited clinical efficacy, and they have increased risks of prostate cancer and pancreatic cancer, weight gain, fluid retention, and fractures and cardiovascular diseases in female patients. Increased incidence of events, etc.
- New anti-NASH drugs under clinical development include farnesoid X receptor (FXR) agonists (such as obeticholic acid), acetyl-CoA carboxylase (ACC) inhibitors (such as GS-0976) and peroxisome Proliferator-activated receptors (PPARs) agonists (such as pemafibrate), etc.
- FXR farnesoid X receptor
- ACC acetyl-CoA carboxylase
- PPARs peroxisome Proliferator-activated receptors
- the invention discloses a safe and effective new drug for treating non-alcoholic fatty liver disease (especially NASH). Specifically, the invention adopts the following technical scheme: Application of Pulsatilla saponin B4 in the preparation of a drug for preventing non-alcoholic fatty liver .
- Pulsatilla saponin B4 in the preparation of medicine for treating non-alcoholic fatty liver.
- Pulsatilla saponin B4 in the preparation of medicine for treating nonalcoholic steatohepatitis.
- Pulsatilla saponin B4 in the preparation of medicaments for preventing non-alcoholic steatohepatitis.
- Pulsatilla saponin B4 in preparation of medicine for treating abnormal elevation of fatty acid ester and cholesterol in blood caused by high-fat diet.
- non-alcoholic fatty liver refers to non-alcoholic fatty liver caused by high-fat diet
- non-alcoholic steatohepatitis refers to non-alcoholic steatohepatitis caused by high-fat diet
- the medicine for preventing or treating non-alcoholic fatty liver is injection medicine or oral medicine.
- the dosage of injection medicine is 1-20 mg/kg, preferably 2.5-20 mg/kg; the dosage of oral medicine is 10-100 mg/kg, preferably 12-50 mg/kg. All are calculated with Pulsatilla saponin B4.
- the invention discloses a new drug for treating non-alcoholic fatty liver.
- the active ingredient is Pulsatilla saponin B4.
- Pulsatilla saponin B4 can effectively reduce lipid accumulation in rat liver caused by high-fat diet, and the curative effect is comparable to positive control It can avoid the side effects of fenofibrate leading to liver toxicity, so it can be used for the prevention and treatment of non-alcoholic fatty liver disease (NAFLD).
- Figure 2 is the change curve of the body weight of rats in different administration groups.
- Figure 3 is the effect of different administration groups on the food intake of rats.
- Figure 4 shows the effect on the liver of NAFLD rats after 4 weeks of administration.
- Figure 5 shows HE-stained and oil red-stained sections of the liver of NAFLD rats after 4 weeks of administration.
- Figure 6 shows the effect of administration for 4 weeks on serum biochemical indicators in NAFLD rats.
- This example demonstrates the effect of Pulsatilla saponin B4 in treating non-alcoholic fatty liver by the protective effect of Pulsatilla saponin B4 on the NAFLD rat model induced by high-fat diet (HFD).
- HFD high-fat diet
- Instruments animal weight scale; automatic blood biochemical analyzer; frozen microtome; inverted microscope.
- Pulsatilla saponin B4 Pulsatilla saponin B4 (purity: 99.4%) was isolated and purified from Pulsatillae medicinalis according to existing methods (He, et. al., Phytomedicine 56 (2019) 136–146); fenofibrate (fenofibrat) was purchased from Shanghai Titan Technology Co., Ltd.; high-fat feed was purchased from Wuxi Fanbo Biotechnology Co., Ltd. (D12492, 60kcal%); common feed was purchased from Suzhou Shuangshi Experimental Animal Feed Technology Co., Ltd.
- Example Rat experiment animal grouping and modeling: After a one-week adaptation period, all rats were randomly divided into two groups according to body weight for modeling: normal group and high-fat diet model group. The normal group was given normal feed and normal drinking water, and the high-fat feed model group was given high-fat feed and normal drinking water, and the model was established for 13 weeks.
- the normal group was randomly divided into two groups according to body weight, normal control group (N), normal + Pulsatilla saponin B4 5mg/kg intraperitoneal injection group (N+B4 5mg/kg), and the hyperlipidemia rats were randomly divided into two groups according to body weight.
- hyperlipidemia model group M
- positive drug fenofibrate 20 mg/kg gavage group Pulsatilla saponin B4 5mg/kg high-dose intraperitoneal injection group (B4H i.p)
- Pulsatilla saponin B4 2.5mg/kg kg low-dose intraperitoneal injection group B4L i.p
- Pulsatilla saponin B4 25mg/kg high-dose intragastric administration group B4H i.g
- Pulsatilla saponin B4 12.5mg/kg low-dose intragastric administration group B4L i.g.
- liver tissue was fixed with 4% paraformaldehyde and used for HE-stained sections and frozen sections. The remaining liver tissues were stored in a -80°C refrigerator for subsequent detection of other indicators.
- TG triglyceride
- CHO total cholesterol
- LDL low-density lipoprotein cholesterol
- HDL high-density lipoprotein cholesterol
- Liver tissue section Frozen sections were stained with Oil Red. HE stained sections were made from the pretreated tissues.
- the liver volume of the rats in group N was normal, reddish-brown in color, smooth in surface, soft and elastic, with sharp edges, and the cut surface of the liver was smooth and dense.
- the liver volume of the liver rats in the M group was significantly enlarged, and it was earthy yellow in color, tough in texture, blunt in edge, tense in the capsule on the surface of the liver, scattered yellow fat spots, and loose and greasy on the cut surface.
- the liver of Pulsatilla saponin B4 treatment group was close to that of normal group.
- HE staining showed that the morphology of liver cells and the structure of hepatic lobules in group N were normal, the cytoplasm was abundant, and the hepatic cords were arranged neatly and radially.
- the lesions in each administration group were alleviated to varying degrees, the damage area was significantly reduced, intracellular lipid droplets and inflammatory cell infiltration were reduced, and the lobular structure was restored to varying degrees.
- the fat content in the liver can be visually observed by Oil Red O staining.
- the fat in the M group increased significantly, and a large number of orange-red lipid droplets appeared.
- the orange-red lipid droplets in the liver decreased, and the curative effect was equivalent.
- Pulsatilla saponin B4 can effectively reduce the lipid accumulation in rat liver caused by high-fat diet, and its efficacy is comparable to that of the positive control drug fenofibrate. It is worth noting that clinical studies and animal experiments have found that although fenofibrate reduces liver lipid accumulation, liver function should be tested regularly when using this drug to be alert to its liver toxicity, while Pulsatilla saponin B4 does not have this effect in animal experiments. class side effects. This suggests that Pulsatilla saponin B4 may avoid the hepatotoxic side effects of fenofibrate in clinical application, so it can be used for the prevention and treatment of nonalcoholic fatty liver disease (NAFLD).
- NAFLD nonalcoholic fatty liver disease
- Acute liver injury refers to acute liver dysfunction caused by various factors on the basis of patients without chronic liver disease.
- the main causes of acute liver injury are viral infection, hepatotoxic drugs, food additives, excessive intake of ethanol, ingestion of poisonous food, and radiation damage.
- the mechanism of liver injury can be divided into chemical and immune.
- the chemical mechanism mainly produces damage through cytochrome P450 and the intermediate metabolites produced by the binding reaction, such as changing the integrity of the plasma membrane, mitochondrial dysfunction, changes in intracellular ion concentration, activity of degrading enzymes, and the role of free radicals; the immune mechanism is Injuries are generated through cytokines, nitric oxide, complement, immune allergy, and pathological apoptosis.
- the main drugs for the treatment of acute liver injury are diammonium glycyrrhizinate, reduced glutathione, S-adenosylmethionine, silibinin, and ursodeoxycholic acid.
- Non-alcoholic fatty liver disease is a chronic liver disease.
- a high-calorie diet rich in saturated fatty acids and fructose and a sedentary lifestyle are also important factors that induce NAFLD.
- the drugs for the treatment of NAFLD mainly include pioglitazone, metformin, fenofibrate, orlistat, etc., which mainly play a role in the treatment of NAFLD by improving insulin resistance, lowering blood lipids, and reducing weight, but none of them have the effect of improving liver histology, or even Can induce rare but serious liver damage.
- the symptoms and treatment mechanism of non-alcoholic fatty liver are not the same as those of acute liver injury, and cannot be regarded as the same as acute liver injury.
- the invention discloses for the first time the application of Pulsatilla saponin B4 in the preparation of medicaments for preventing and treating non-alcoholic fatty liver.
- the protective effect of Pulsatilla saponin B4 on NAFLD rat model induced by high-fat diet (HFD) illustrates the effect of Pulsatilla saponin B4 in the treatment of non-alcoholic fatty liver disease.
- the results showed that Pulsatilla saponin B4 can effectively reduce the lipid accumulation in the liver of rats induced by high-fat diet, and its efficacy is comparable to that of the positive control drug fenofibrate.
- clinical studies and animal experiments have found that although fenofibrate reduces liver lipid accumulation, there is a problem of liver toxicity when using this drug, while Pulsatilla saponin B4 does not have such side effects in animal experiments.
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Abstract
An application of Pulsatilla saponin B4 in the preparation of medicine for treating non-alcoholic fatty liver diseases. A medicine for treating non-alcoholic fatty liver diseases, which has an active ingredient of Pulsatilla saponin B4. Experiments confirm that Pulsatilla saponin B4 may effectively reduce the lipid accumulation in rat liver caused by a high-fat diet, and the curative effect thereof is equivalent to that of a positive control medicine. The present invention may avoid the side effects of fenofibrate leading to hepatotoxicity, and therefore may be used for the prevention and treatment of non-alcoholic fatty liver diseases (NAFLDs).
Description
本发明属于药物技术，具体涉及白头翁皂苷B4在制备治疗非酒精性脂肪肝的药物中的应用。The invention belongs to the pharmaceutical technology, and in particular relates to the application of Pulsatilla saponin B4 in the preparation of medicaments for treating nonalcoholic fatty liver.
非酒精性脂肪性肝病（NAFLD）是一种与胰岛素抵抗和遗传易感密切相关的代谢应激性肝损伤，其病理学改变与酒精性肝病（ALD）相似，但患者无过量饮酒史，随着病程的进展依次分为非酒精性单纯性脂肪肝(NAFL)、非酒精性脂肪性肝炎（NASH）、肝硬化和肝细胞癌（HCC）。Nonalcoholic fatty liver disease (NAFLD) is a metabolic stress liver injury closely related to insulin resistance and genetic susceptibility. Its pathological changes are similar to those of alcoholic liver disease (ALD), but the patients have no history of excessive drinking. According to the progress of the disease course, it is divided into nonalcoholic simple fatty liver (NAFL), nonalcoholic steatohepatitis (NASH), cirrhosis and hepatocellular carcinoma (HCC).
目前研究发现代谢综合症（MetS）是NAFLD、NASH发生的最强风险因素，代谢综合症包括腰围增加（如肥胖）、高血糖、高血脂、高血压等。MetS与NAFLD之间的关联可能是双向的，特别是糖尿病和高血压，MetS会增加NAFLD的风险，而NAFLD也可能会加重MetS的几种临床特征和合并症，因此有效治疗NASH也有利于改善MetS症状。此外MetS也是NAFLD患者出现不良心血管疾病（CVD）和总死亡率的重要驱动因素。糖尿病与NAFLD的进展具有最清晰的生物学联系，高达75%的2型糖尿病患者伴有NAFLD。目前胰岛素抵抗也被认为是NAFLD发病机制的组成部分，且随着疾病进展而恶化。50%的高血压患者伴有NAFLD，高血压与纤维化进展密切相关，且NAFLD与动脉硬化、心肌重塑、心力衰竭、肾脏疾病之间已证明存在相关性。Current studies have found that metabolic syndrome (MetS) is the strongest risk factor for NAFLD and NASH. Metabolic syndrome includes increased waist circumference (such as obesity), hyperglycemia, hyperlipidemia, and hypertension. The association between MetS and NAFLD may be bidirectional, especially diabetes and hypertension, MetS increases the risk of NAFLD, and NAFLD may also aggravate several clinical features and comorbidities of MetS, so effective treatment of NASH is also beneficial to improve MetS symptoms. In addition, MetS is also an important driver of adverse cardiovascular disease (CVD) and overall mortality in NAFLD patients. Diabetes has the clearest biological link to the progression of NAFLD, with up to 75% of patients with type 2 diabetes having NAFLD. At present, insulin resistance is also considered to be an integral part of the pathogenesis of NAFLD, and it worsens with disease progression. 50% of hypertensive patients are accompanied by NAFLD. Hypertension is closely related to the progression of fibrosis, and there has been a proven correlation between NAFLD and arteriosclerosis, myocardial remodeling, heart failure, and kidney disease.
据统计全球约有13亿肝病患者，而中国就有4亿之多，占全球肝病患者的30%以上。调查显示我国乙肝患者9600万、丙肝患者1000万、脂肪肝患者2亿、酒精肝患者6000万，其他肝病患者3700万。近年来，以非酒精性脂肪性肝病（NAFLD）为代表的非感染性肝病的发病率显著上升，其患病率高达15%～40%，已取代病毒性肝病，成为全球第一大肝病，可见NAFLD的临床防治工作任重道远，而临床基础研究更是迫在眉睫。According to statistics, there are about 1.3 billion liver disease patients in the world, and there are as many as 400 million in China, accounting for more than 30% of the global liver disease patients. According to the survey, there are 96 million hepatitis B patients, 10 million hepatitis C patients, 200 million fatty liver patients, 60 million alcoholic liver patients and 37 million other liver disease patients in my country. In recent years, the incidence of non-infectious liver disease represented by non-alcoholic fatty liver disease (NAFLD) has increased significantly, and its prevalence rate is as high as 15% to 40%. It has replaced viral liver disease and has become the world's largest liver disease. It can be seen that the clinical prevention and treatment of NAFLD has a long way to go, and clinical basic research is even more urgent.
NAFLD发病机制目前仍处于争论阶段。在传统的“二次打击”学说中，“第一次打击”是肝脏脂肪堆积；“第二次打击”是肝脏脂肪堆积引发的氧化应激和炎性细胞因子的过度释放，导致肝脏炎症和肝纤维化，进而导致肝硬化和肝癌。但根据最新的研究发现，这种观点被认为已经过时。很多分子途径在NASH的发病过程中都起着很重要的作用，甚至不能确定NASH是否总是继发于NAFLD。此外，致病因素在所有患者中并不相同，导致疾病的发病机制和临床表现具有高度的异质性。The pathogenesis of NAFLD is still under debate. In the traditional "two-hit" theory, the "first hit" is fat accumulation in the liver; the "second hit" is the oxidative stress and excessive release of inflammatory cytokines caused by fat accumulation in the liver, leading to liver inflammation and Liver fibrosis, which in turn leads to cirrhosis and liver cancer. But that view is considered outdated, according to new research findings. Many molecular pathways play an important role in the pathogenesis of NASH, and it is not even certain whether NASH is always secondary to NAFLD. In addition, causative factors are not the same in all patients, resulting in a high degree of heterogeneity in the pathogenesis and clinical manifestations of the disease.
NASH被认为是NAFLD的进行性形式，其特征在于肝脏脂肪变性、炎症、肝细胞损伤和不同程度的纤维化。NASH的发生是多因素综合作用的结果，包括肝外因素如饮食因素（碳水化合物、金属离子、饱和脂肪/反式脂肪和胆固醇等）、代谢功能紊乱（胰岛素抵抗和脂肪组织炎症、脂肪因子等）、肝肠轴（肠道炎症和肠道屏障功能障碍、胆汁酸）等；肝内因素如肝细胞压力（氧化应激和内质网应激、脂毒性、线粒体功能障碍）、肝细胞死亡（细胞凋亡、坏死性凋亡、细胞焦亡）、核受体（PPARα、FXR、CAR等）、肝细胞因子等；除此之外还有先天性免疫、宿主遗传学和表观遗传学。NASH is considered a progressive form of NAFLD characterized by hepatic steatosis, inflammation, hepatocellular injury, and varying degrees of fibrosis. The occurrence of NASH is the result of multiple factors, including extrahepatic factors such as dietary factors (carbohydrates, metal ions, saturated fat/trans fat and cholesterol, etc.), metabolic dysfunction (insulin resistance and adipose tissue inflammation, adipokines, etc.) ), hepatic-gut axis (intestinal inflammation and intestinal barrier dysfunction, bile acids), etc.; intrahepatic factors such as hepatocyte stress (oxidative stress and endoplasmic reticulum stress, lipotoxicity, mitochondrial dysfunction), hepatic cell death ( Apoptosis, necroptosis, pyroptosis), nuclear receptors (PPARα, FXR, CAR, etc.), liver cytokines, etc.; in addition to innate immunity, host genetics and epigenetics.
运动与改善不良的生活方式是治疗NAFLD的基础，由于非酒精性脂肪肝病的肝损伤的细胞和分子机制尚不十分清楚，对该病缺少有效的药物治疗方法，故积极探索安全有效的药物治疗是当前的研究热点。Exercise and improving unhealthy lifestyles are the basis for the treatment of NAFLD. Since the cellular and molecular mechanisms of liver damage in non-alcoholic fatty liver disease are not yet clear, and there is no effective drug treatment for the disease, safe and effective drug treatment is actively explored is a current research hotspot.
临床上广泛使用的二甲双胍主要是通过激活AMPK而发挥治疗非酒精性脂肪性肝病(特别是NASH）的作用，但这个药物是线粒体复合物I的抑制剂，存在乳酸中毒的风险，并且没有肝脏组织学改善作用。目前临床推荐使用的NASH治疗药物包括胰岛素增敏剂如吡格列酮等，但上述药物的临床疗效有限，且其存在前列腺癌和胰腺癌发生风险增加、体重增加、液体潴留以及女性患者的骨折和心血管事件发生率增加等。Metformin, which is widely used clinically, mainly plays a role in the treatment of non-alcoholic fatty liver disease (especially NASH) by activating AMPK, but this drug is an inhibitor of mitochondrial complex I, there is a risk of lactic acidosis, and there is no liver tissue Learning to improve. The current clinically recommended drugs for the treatment of NASH include insulin sensitizers such as pioglitazone, etc., but the above-mentioned drugs have limited clinical efficacy, and they have increased risks of prostate cancer and pancreatic cancer, weight gain, fluid retention, and fractures and cardiovascular diseases in female patients. Increased incidence of events, etc.
临床在研的抗NASH新药包括法尼酯X受体（FXR）激动剂（如奥贝胆酸）、乙酰辅酶A羧化酶（ACC）抑制剂（如GS-0976）和过氧化物酶体增殖体激活受体（PPARs）激动剂（如pemafibrate）等。然而上述抗NASH新药的疗效和安全性有待进一步验证。综上所述，临床上迫切需要安全、有效的新型治疗非酒精性脂肪性肝病（尤其是NASH）的药物。New anti-NASH drugs under clinical development include farnesoid X receptor (FXR) agonists (such as obeticholic acid), acetyl-CoA carboxylase (ACC) inhibitors (such as GS-0976) and peroxisome Proliferator-activated receptors (PPARs) agonists (such as pemafibrate), etc. However, the efficacy and safety of the above new anti-NASH drugs need to be further verified. To sum up, there is an urgent need for safe and effective new drugs for the treatment of nonalcoholic fatty liver disease (especially NASH).
本发明公开了安全、有效的新型治疗非酒精性脂肪性肝病（尤其是NASH）的药物，具体的，本发明采用如下技术方案：白头翁皂苷B4在制备预防非酒精性脂肪肝的药物中的应用。The invention discloses a safe and effective new drug for treating non-alcoholic fatty liver disease (especially NASH). Specifically, the invention adopts the following technical scheme: Application of Pulsatilla saponin B4 in the preparation of a drug for preventing non-alcoholic fatty liver .
白头翁皂苷B4在制备治疗非酒精性脂肪肝的药物中的应用。Application of Pulsatilla saponin B4 in the preparation of medicine for treating non-alcoholic fatty liver.
白头翁皂苷B4在制备治疗非酒精性脂肪性肝炎的药物中的应用。Application of Pulsatilla saponin B4 in the preparation of medicine for treating nonalcoholic steatohepatitis.
白头翁皂苷B4在制备预防非酒精性脂肪性肝炎的药物中的应用。Application of Pulsatilla saponin B4 in the preparation of medicaments for preventing non-alcoholic steatohepatitis.
白头翁皂苷B4在制备治疗高脂饮食所致的血液中脂肪酸酯和胆固醇异常升高的药物中的应用。Application of Pulsatilla saponin B4 in preparation of medicine for treating abnormal elevation of fatty acid ester and cholesterol in blood caused by high-fat diet.
本发明中，非酒精性脂肪肝为高脂饮食所致的非酒精性脂肪肝；非酒精性脂肪性肝炎为高脂饮食所致的非酒精性脂肪性肝炎。In the present invention, non-alcoholic fatty liver refers to non-alcoholic fatty liver caused by high-fat diet; non-alcoholic steatohepatitis refers to non-alcoholic steatohepatitis caused by high-fat diet.
一种预防或者治疗非酒精性脂肪肝的药物，或者预防或者治疗非酒精性脂肪性肝炎的药物，其活性成分为白头翁皂苷B4，还包括常规药物辅料，比如生理盐水。A medicine for preventing or treating non-alcoholic fatty liver, or a medicine for preventing or treating non-alcoholic steatohepatitis, the active ingredient of which is Pulsatilla saponin B4, and conventional pharmaceutical excipients, such as normal saline.
本发明中，预防或者治疗非酒精性脂肪肝的药物为注射药物或者口服药物。优选的，注射药物的用量为1～20mg/kg，优选2.5～20mg/kg；口服药物的用量为10～100mg/kg，优选12～50mg/kg。都以白头翁皂苷B4计。In the present invention, the medicine for preventing or treating non-alcoholic fatty liver is injection medicine or oral medicine. Preferably, the dosage of injection medicine is 1-20 mg/kg, preferably 2.5-20 mg/kg; the dosage of oral medicine is 10-100 mg/kg, preferably 12-50 mg/kg. All are calculated with Pulsatilla saponin B4.
由于非酒精性脂肪肝病的肝损伤的细胞和分子机制尚不十分清楚，对该病缺少有效的药物治疗方法，故积极探索安全有效的药物治疗是当前的研究热点。现有药物的疗效和安全性有待进一步改善或者验证。本发明公开了新的治疗非酒精性脂肪肝的药物，活性成分为白头翁皂苷B4，实验证实，白头翁皂苷B4可以有效地降低高脂饮食导致的大鼠肝脏的脂质堆积，且疗效与阳性对照药相当，可避免非诺贝特导致肝毒性的副作用，因此可用于预防和治疗非酒精性脂肪性肝病（NAFLD）。Since the cellular and molecular mechanisms of liver damage in NAFLD are still unclear, and there is no effective drug treatment for the disease, active exploration of safe and effective drug treatment is a current research hotspot. The efficacy and safety of existing drugs need to be further improved or verified. The invention discloses a new drug for treating non-alcoholic fatty liver. The active ingredient is Pulsatilla saponin B4. Experiments have proved that Pulsatilla saponin B4 can effectively reduce lipid accumulation in rat liver caused by high-fat diet, and the curative effect is comparable to positive control It can avoid the side effects of fenofibrate leading to liver toxicity, so it can be used for the prevention and treatment of non-alcoholic fatty liver disease (NAFLD).
图
为NAFLD造模6周正常组和高脂饲料模型组的肝组织及HE染色。
picture Liver tissue and HE staining of the normal group and the high-fat diet model group for NAFLD modeling for 6 weeks.
图 2为不同给药组大鼠体重的变化曲线。Figure 2 is the change curve of the body weight of rats in different administration groups.
图 3为不同给药组对大鼠摄食量的影响。Figure 3 is the effect of different administration groups on the food intake of rats.
图 4为给药4周后对NAFLD大鼠肝脏的影响。Figure 4 shows the effect on the liver of NAFLD rats after 4 weeks of administration.
图 5为给药4周后NAFLD大鼠肝脏HE染色和油红染色切片。Figure 5 shows HE-stained and oil red-stained sections of the liver of NAFLD rats after 4 weeks of administration.
图 6为给药4周对NAFLD大鼠血清生化指标的影响。Figure 6 shows the effect of administration for 4 weeks on serum biochemical indicators in NAFLD rats.
本实施例通过白头翁皂苷B4对高脂饮食（HFD）诱导NAFLD大鼠模型的保护作用说明白头翁皂苷B4治疗非酒精性脂肪肝的效果。This example demonstrates the effect of Pulsatilla saponin B4 in treating non-alcoholic fatty liver by the protective effect of Pulsatilla saponin B4 on the NAFLD rat model induced by high-fat diet (HFD).
动物：雄性SD大鼠75只，SPF级，6周龄，初始体重200±20g；ICR小鼠，6周龄；都购于上海斯莱克实验动物有限责任公司。在苏州大学实验动物中心SPF级动物房饲养，所有动物保持12小时交替的昼夜节律，自由摄食饮水。Animals: 75 male SD rats, SPF grade, 6 weeks old, initial body weight 200±20g; ICR mice, 6 weeks old; all purchased from Shanghai Slack Experimental Animal Co., Ltd. All animals were raised in the SPF grade animal room of the Experimental Animal Center of Soochow University. All animals kept a 12-hour alternating circadian rhythm and had free access to food and water.
仪器：动物体重秤；全自动血生化分析仪；冷冻切片机；倒置显微镜。Instruments: animal weight scale; automatic blood biochemical analyzer; frozen microtome; inverted microscope.
试剂：白头翁皂苷B4（纯度为99.4%）是根据现有方法从白头翁药材中分离纯化得到（He, et. al., Phytomedicine 56 (2019)
136–146）；非诺贝特（fenofibrat）购自上海泰坦科技股份有限公司；高脂饲料购自无锡帆泊生物技术有限公司（D12492，60kcal%）；普通饲料购自苏州双狮实验动物饲料科技有限公司。Reagent: Pulsatilla saponin B4 (purity: 99.4%) was isolated and purified from Pulsatillae medicinalis according to existing methods (He, et. al., Phytomedicine 56 (2019)
136–146); fenofibrate (fenofibrat) was purchased from Shanghai Titan Technology Co., Ltd.; high-fat feed was purchased from Wuxi Fanbo Biotechnology Co., Ltd. (D12492, 60kcal%); common feed was purchased from Suzhou Shuangshi Experimental Animal Feed Technology Co., Ltd.
实施例 大鼠实验：动物分组及造模：一周适应期以后，所有大鼠和按体重随机分为2组进行造模：正常组和高脂饲料模型组。正常组给予普通饲料，正常饮水，高脂饲料模型组给予高脂饲料，正常饮水，造模13周。Example Rat experiment: animal grouping and modeling: After a one-week adaptation period, all rats were randomly divided into two groups according to body weight for modeling: normal group and high-fat diet model group. The normal group was given normal feed and normal drinking water, and the high-fat feed model group was given high-fat feed and normal drinking water, and the model was established for 13 weeks.
13周之后，按体重将正常组随机分为2组，正常对照组（N）、正常+白头翁皂苷B4 5mg/kg腹腔注射组（N+B4 5mg/kg），将高脂大鼠按体重随机分为6组，即高脂模型组（M）、阳性药非诺贝特20 mg/kg灌胃组、白头翁皂苷B4 5mg/kg高剂量腹腔注射组（B4H i.p）、白头翁皂苷B4 2.5mg/kg低剂量腹腔注射组（B4L i.p）、白头翁皂苷B4 25mg/kg高剂量灌胃组（B4H i.g）和白头翁皂苷B4 12.5mg/kg低剂量灌胃组（B4L i.g）。After 13 weeks, the normal group was randomly divided into two groups according to body weight, normal control group (N), normal + Pulsatilla saponin B4 5mg/kg intraperitoneal injection group (N+B4 5mg/kg), and the hyperlipidemia rats were randomly divided into two groups according to body weight. Divided into 6 groups, namely hyperlipidemia model group (M), positive drug fenofibrate 20 mg/kg gavage group, Pulsatilla saponin B4 5mg/kg high-dose intraperitoneal injection group (B4H i.p), Pulsatilla saponin B4 2.5mg/kg kg low-dose intraperitoneal injection group (B4L i.p), Pulsatilla saponin B4 25mg/kg high-dose intragastric administration group (B4H i.g) and Pulsatilla saponin B4 12.5mg/kg low-dose intragastric administration group (B4L i.g).
给药。连续给药4周（每天注射或者灌胃一次），给药期间正常对照组和正常+白头翁皂苷B4 5mg/kg腹腔注射组给予普通饲料和正常饮水，其余6组给予高脂饲料和正常饮水。实验期间，每周记录各组大鼠体重和摄食量，仔细观察并记录大鼠毛发、粪便及活动情况。medication. Continuous administration for 4 weeks (injection or gavage once a day), normal control group and normal + Pulsatilla saponin B4 5mg/kg intraperitoneal injection group were given normal feed and normal drinking water during the administration period, and the remaining 6 groups were given high-fat feed and normal drinking water. During the experiment, the body weight and food intake of the rats in each group were recorded every week, and the hair, feces and activities of the rats were carefully observed and recorded.
取材。提前12小时禁食不禁水，次日上午腹主动脉取血，处死取肝脏。肝脏右小叶组织用4%多聚甲醛固定，用于HE染色切片和冷冻切片。其余肝组织保存于-80℃冰箱内，以备后续其他指标检测。draw materials. They were fasted 12 hours in advance without food and water, and blood was collected from the abdominal aorta in the morning of the next day, and the liver was sacrificed. The right lobule tissue of the liver was fixed with 4% paraformaldehyde and used for HE-stained sections and frozen sections. The remaining liver tissues were stored in a -80°C refrigerator for subsequent detection of other indicators.
生化指标的测定。将全血室温静置2 h，3000 rpm离心15 min，收集血清。使用全自动生化分析仪测定血清中甘油三脂（TG）、总胆固醇（CHO）、低密度脂蛋白胆固醇（LDL）和高密度脂蛋白胆固醇（HDL）的水平。Determination of biochemical indicators. The whole blood was allowed to stand at room temperature for 2 h, centrifuged at 3000 rpm for 15 min, and the serum was collected. The levels of triglyceride (TG), total cholesterol (CHO), low-density lipoprotein cholesterol (LDL) and high-density lipoprotein cholesterol (HDL) in serum were measured using an automatic biochemical analyzer.
肝脏组织切片。冷冻切片进行油红染色。将预处理好的组织制作HE染色切片。Liver tissue section. Frozen sections were stained with Oil Red. HE stained sections were made from the pretreated tissues.
实验结果：如图1所示，NAFLD造模6周正常组和高脂饲料模型组的肝组织及HE染色。高脂饲料模型组HE染色切片可观察到肝细胞出现脂肪变性，胞浆中出现许多大小不等的空泡（脂肪滴)，边界清晰，将细胞核挤向一侧。肝细胞体积增大、肿胀，细胞核明显，细胞与细胞之间界限不清。Experimental results: As shown in Figure 1, the liver tissue and HE staining of the normal group and the high-fat diet model group after NAFLD modeling for 6 weeks. In the HE stained section of the high-fat diet model group, fatty degeneration of liver cells could be observed, and many vacuoles (fat droplets) of different sizes appeared in the cytoplasm, with clear boundaries, and the nuclei were squeezed to one side. Hepatocytes were enlarged and swollen, with obvious nuclei and unclear boundaries between cells.
如图2所示，不同给药组对大鼠体重的影响。经过13周的高脂饲料饲养，高脂饲料模型组大鼠的体重明显高于正常组大鼠，呈现出肥胖特征。腹腔注射和灌胃2种给药途径的白头翁皂苷B4和非诺贝特均减缓高脂饲料模型组大鼠的体重增长。As shown in Figure 2, the effects of different administration groups on the body weight of rats. After 13 weeks of high-fat diet feeding, the weight of the rats in the high-fat diet model group was significantly higher than that of the normal group, showing obesity characteristics. Both intraperitoneal injection and intragastric administration of Pulsatilla saponin B4 and fenofibrate slowed down the weight gain of rats in the high-fat diet model group.
如图3所示，不同给药组对大鼠摄食量的影响。与高脂饲料模型组相比，白头翁皂苷B4 5mg/kg高剂量腹腔注射组（B4H i.p）摄食量显著降低，具有显著性差异(P < 0.05)。As shown in Figure 3, the effects of different administration groups on the food intake of rats. Compared with the high-fat diet model group, the food intake of Pulsatilla saponin B4 5mg/kg high-dose intraperitoneal injection group (B4H i.p) was significantly reduced (P < 0.05).
如图4所示，N组大鼠肝脏体积正常，呈红褐色，表面光滑，质软富有弹性，边缘锐利，肝脏切面光滑致密。M组肝脏大鼠肝脏体积明显增大，呈土黄色，质韧，边缘圆钝，肝脏表面包膜紧张，可见散在黄色脂肪斑，切面疏松有油腻感。白头翁皂苷B4治疗组肝脏与正常组接近。As shown in Figure 4, the liver volume of the rats in group N was normal, reddish-brown in color, smooth in surface, soft and elastic, with sharp edges, and the cut surface of the liver was smooth and dense. The liver volume of the liver rats in the M group was significantly enlarged, and it was earthy yellow in color, tough in texture, blunt in edge, tense in the capsule on the surface of the liver, scattered yellow fat spots, and loose and greasy on the cut surface. The liver of Pulsatilla saponin B4 treatment group was close to that of normal group.
如图5所示，为进一步观察高脂饲料对肝脏的影响，对肝脏进行了病理切片观察。HE染色显示N组肝细胞形态、肝小叶结构正常，胞质丰富，肝索排列整齐呈放射状。M组存在明显的肝细胞脂肪变性，肝小叶界限不清，肝索结抅紊乱，肝组织可见弥漫性、混合性的空泡及气球样改变，细胞肿胀变圆，胞浆疏松，胞内含有大、小脂滴并有融合，部分胞核被挤到胞膜，汇管区和小叶内可见大量中性粒细胞浸润呈灶状积聚，脂变细胞以腺泡区明显。与M组相比，各给药组病变有不同程度减轻，损伤面积明显减小，胞内脂滴及炎性细胞浸润减少，小叶结构不同程度恢复。通过油红O染色能直观地观察到肝脏内的脂肪含量。M组较N组脂肪明显增加，出现大量橘红色脂滴。给予白头翁皂苷B4和非诺贝特治疗后，肝脏内橘红色脂滴减少，疗效相当。As shown in Figure 5, in order to further observe the effect of high-fat diet on the liver, the pathological section of the liver was observed. HE staining showed that the morphology of liver cells and the structure of hepatic lobules in group N were normal, the cytoplasm was abundant, and the hepatic cords were arranged neatly and radially. In group M, there were obvious fatty degeneration of hepatic cells, unclear boundary of hepatic lobules, disordered hepatic cord knots, diffuse and mixed vacuoles and balloon-like changes in liver tissue, swelling and rounding of cells, loose cytoplasm, and intracellular Large and small lipid droplets were fused, some nuclei were extruded to the cell membrane, a large number of neutrophils infiltrated and accumulated in the portal area and lobules, and the steatotic cells were evident in the acinar area. Compared with the M group, the lesions in each administration group were alleviated to varying degrees, the damage area was significantly reduced, intracellular lipid droplets and inflammatory cell infiltration were reduced, and the lobular structure was restored to varying degrees. The fat content in the liver can be visually observed by Oil Red O staining. Compared with the N group, the fat in the M group increased significantly, and a large number of orange-red lipid droplets appeared. After treatment with Pulsatilla saponin B4 and fenofibrate, the orange-red lipid droplets in the liver decreased, and the curative effect was equivalent.
如图6所示，与正常组比较，高脂饲料模型组血清CHO、HDL和LDL水平显著升高；经过腹腔注射和灌胃2种给药途径的白头翁皂苷B4和非诺贝特治疗后，血清CHO、HDL和LDL水平均显著下降。以上结果提示白头翁皂苷B4可通过降低血清CHO、HDL和LDL水平而对肝脏脂质积累发挥一定有益作用。As shown in Figure 6, compared with the normal group, the serum CHO, HDL and LDL levels in the high-fat diet model group were significantly increased; Serum CHO, HDL and LDL levels were significantly decreased. The above results suggest that Pulsatilla saponin B4 can play a beneficial role in hepatic lipid accumulation by reducing serum CHO, HDL and LDL levels.
以上结果表明，白头翁皂苷B4可以有效地降低高脂饮食导致的大鼠肝脏的脂质堆积，且疗效与阳性对照药非诺贝特相当。值得注意的是，临床研究和动物实验发现非诺贝特虽然降低肝脏脂质堆积，但使用此药，应定期检测肝功能，警惕其肝毒性，而白头翁皂苷B4在动物实验中则不具有此类副作用。这提示白头翁皂苷B4在临床应用中有可能避免非诺贝特导致肝毒性的副作用，因此可用于预防和治疗非酒精性脂肪性肝病（NAFLD）。The above results show that Pulsatilla saponin B4 can effectively reduce the lipid accumulation in rat liver caused by high-fat diet, and its efficacy is comparable to that of the positive control drug fenofibrate. It is worth noting that clinical studies and animal experiments have found that although fenofibrate reduces liver lipid accumulation, liver function should be tested regularly when using this drug to be alert to its liver toxicity, while Pulsatilla saponin B4 does not have this effect in animal experiments. class side effects. This suggests that Pulsatilla saponin B4 may avoid the hepatotoxic side effects of fenofibrate in clinical application, so it can be used for the prevention and treatment of nonalcoholic fatty liver disease (NAFLD).
现有技术公开了白头翁皂苷B4在制备治疗LPS诱导的小鼠急性肝损伤中的应用。急性肝损伤与非酒精性脂肪肝发病机制不同，急性肝损伤是指患者在无慢性肝病基础上，由各种因素导致的急性肝脏功能异常。临床上造成急性肝损伤的原因主要有病毒感染、肝毒性药物、食物添加剂、乙醇摄入过量、误服有毒食物、放射线损伤等。肝损伤的发生机制可分为化学性和免疫性。化学机制主要通过细胞色素P450及结合反应产生的中间代谢产物产生损伤，如改变质膜的完整性、线粒体功能失调、细胞内离子浓度变化、降解酶的活性和自由基的作用等；免疫机制则通过细胞因子、一氧化氮、补体及免疫变态反应、病理性细胞凋亡等产生损伤。目前治疗急性肝损伤的药物主要有甘草酸二铵、还原型谷胱甘肽、S-腺苷甲硫氨酸、水飞蓟宾、熊去氧胆酸，此类药物作为NAFLD患者的辅助治疗，尤其对NASH的治疗效果仍需进一步的临床试验证实。而非酒精性脂肪肝是一种慢性肝病，富含饱和脂肪酸和果糖的高热量膳食结构和久坐少动的生活方式也是诱发NAFLD的重要因素。目前治疗NAFLD的药物主要有吡格列酮、二甲双胍、非诺贝特、奥利司他等，主要通过改善胰岛素抵抗，降血脂，减轻体重来发挥治疗NAFLD的效果，但是均没有肝脏组织学改善作用，甚至会诱发罕见但严重的肝损害。综上非酒精性脂肪肝的病症和治疗机理与急性肝损伤并不相同，不能与急性肝损伤等同视之。The prior art discloses the application of Pulsatilla saponin B4 in the preparation and treatment of LPS-induced acute liver injury in mice. The pathogenesis of acute liver injury is different from that of non-alcoholic fatty liver disease. Acute liver injury refers to acute liver dysfunction caused by various factors on the basis of patients without chronic liver disease. Clinically, the main causes of acute liver injury are viral infection, hepatotoxic drugs, food additives, excessive intake of ethanol, ingestion of poisonous food, and radiation damage. The mechanism of liver injury can be divided into chemical and immune. The chemical mechanism mainly produces damage through cytochrome P450 and the intermediate metabolites produced by the binding reaction, such as changing the integrity of the plasma membrane, mitochondrial dysfunction, changes in intracellular ion concentration, activity of degrading enzymes, and the role of free radicals; the immune mechanism is Injuries are generated through cytokines, nitric oxide, complement, immune allergy, and pathological apoptosis. At present, the main drugs for the treatment of acute liver injury are diammonium glycyrrhizinate, reduced glutathione, S-adenosylmethionine, silibinin, and ursodeoxycholic acid. These drugs are used as adjuvant therapy for patients with NAFLD , especially the therapeutic effect on NASH still needs to be confirmed by further clinical trials. Non-alcoholic fatty liver disease is a chronic liver disease. A high-calorie diet rich in saturated fatty acids and fructose and a sedentary lifestyle are also important factors that induce NAFLD. At present, the drugs for the treatment of NAFLD mainly include pioglitazone, metformin, fenofibrate, orlistat, etc., which mainly play a role in the treatment of NAFLD by improving insulin resistance, lowering blood lipids, and reducing weight, but none of them have the effect of improving liver histology, or even Can induce rare but serious liver damage. To sum up, the symptoms and treatment mechanism of non-alcoholic fatty liver are not the same as those of acute liver injury, and cannot be regarded as the same as acute liver injury.
本发明首次公开了白头翁皂苷B4在制备预防、治疗非酒精性脂肪肝的药物中的应用。通过白头翁皂苷B4对高脂饮食（HFD）诱导NAFLD大鼠模型的保护作用说明白头翁皂苷B4治疗非酒精性脂肪肝的效果。结果表明，白头翁皂苷B4可以有效地降低高脂饮食导致的大鼠肝脏的脂质堆积，且疗效与阳性对照药非诺贝特相当。尤其是，临床研究和动物实验发现非诺贝特虽然降低肝脏脂质堆积，但使用此药，存在肝毒性的问题，而白头翁皂苷B4在动物实验中则不具有此类副作用。The invention discloses for the first time the application of Pulsatilla saponin B4 in the preparation of medicaments for preventing and treating non-alcoholic fatty liver. The protective effect of Pulsatilla saponin B4 on NAFLD rat model induced by high-fat diet (HFD) illustrates the effect of Pulsatilla saponin B4 in the treatment of non-alcoholic fatty liver disease. The results showed that Pulsatilla saponin B4 can effectively reduce the lipid accumulation in the liver of rats induced by high-fat diet, and its efficacy is comparable to that of the positive control drug fenofibrate. In particular, clinical studies and animal experiments have found that although fenofibrate reduces liver lipid accumulation, there is a problem of liver toxicity when using this drug, while Pulsatilla saponin B4 does not have such side effects in animal experiments.
Claims (10)
- 白头翁皂苷B4在制备预防非酒精性脂肪肝的药物中的应用。Application of Pulsatilla saponin B4 in the preparation of medicaments for preventing non-alcoholic fatty liver.
- 白头翁皂苷B4在制备治疗非酒精性脂肪肝的药物中的应用。Application of Pulsatilla saponin B4 in the preparation of medicine for treating non-alcoholic fatty liver.
- 白头翁皂苷B4在制备治疗非酒精性脂肪性肝炎的药物中的应用。Application of Pulsatilla saponin B4 in the preparation of medicine for treating nonalcoholic steatohepatitis.
- 白头翁皂苷B4在制备预防非酒精性脂肪性肝炎的药物中的应用。Application of Pulsatilla saponin B4 in the preparation of medicaments for preventing non-alcoholic steatohepatitis.
- 白头翁皂苷B4在制备治疗高脂饮食所致的血液中脂肪酸酯和胆固醇异常升高的药物中的应用。Application of Pulsatilla saponin B4 in preparation of medicine for treating abnormal elevation of fatty acid ester and cholesterol in blood caused by high-fat diet.
- 根据权利要求1至5任意一项所述的应用，其特征在于，药物为注射药物或者口服药物。The use according to any one of claims 1 to 5, characterized in that the medicine is injection medicine or oral medicine.
- 根据权利要求1至5任意一项所述的应用，其特征在于，药物的活性成分为白头翁皂苷B4。The use according to any one of claims 1 to 5, characterized in that the active ingredient of the medicine is Pulsatilla saponin B4.
- 根据权利要求1或者2所述的应用，其特征在于，非酒精性脂肪肝为高脂饮食所致的非酒精性脂肪肝。The use according to claim 1 or 2, characterized in that the non-alcoholic fatty liver is non-alcoholic fatty liver caused by high-fat diet.
- 根据权利要求3或者4所述的应用，其特征在于，非酒精性脂肪性肝炎为高脂饮食所致的非酒精性脂肪性肝炎。The use according to claim 3 or 4, characterized in that the non-alcoholic steatohepatitis is non-alcoholic steatohepatitis caused by high-fat diet.
- 一种预防或者治疗非酒精性脂肪肝的药物，或者预防或者治疗非酒精性脂肪性肝炎的药物，其活性成分为白头翁皂苷B4。A medicine for preventing or treating nonalcoholic fatty liver, or a medicine for preventing or treating nonalcoholic steatohepatitis, the active ingredient of which is Pulsatilla saponin B4.
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| CN113730422A (en) * | 2021-09-30 | 2021-12-03 | 苏州大学 | Application of pulsatilla chinensis saponin B4 in preparation of medicine for preventing or treating non-alcoholic fatty liver disease |
| CN114469972B (en) * | 2022-04-06 | 2022-07-08 | 中国中医科学院中药研究所 | Application of pulsatilla saponin B4 and/or pulsatilla saponin B5 in preparation of drug for preventing or treating drug-induced liver injury |
| CN114931582B (en) * | 2022-05-11 | 2024-04-09 | 广西馨海药业科技有限公司 | Application of pulsatilla chinensis saponin composition in preparation of medicines for treating atherosclerosis |
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| CN106389408A (en) * | 2016-09-18 | 2017-02-15 | 中国药科大学 | Application of polyoxymethylflavone, polyoxymethylflavone composition and polyoxymethylflavone preparation for prevention or treatment of hyperlipidemia |
| CN110755441A (en) * | 2019-11-29 | 2020-02-07 | 苏州大学 | Application of pulsatilla saponin B4 in preparation of medicine for treating diabetes and complications thereof |
| CN113730422A (en) * | 2021-09-30 | 2021-12-03 | 苏州大学 | Application of pulsatilla chinensis saponin B4 in preparation of medicine for preventing or treating non-alcoholic fatty liver disease |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN106389408A (en) * | 2016-09-18 | 2017-02-15 | 中国药科大学 | Application of polyoxymethylflavone, polyoxymethylflavone composition and polyoxymethylflavone preparation for prevention or treatment of hyperlipidemia |
| CN110755441A (en) * | 2019-11-29 | 2020-02-07 | 苏州大学 | Application of pulsatilla saponin B4 in preparation of medicine for treating diabetes and complications thereof |
| CN113730422A (en) * | 2021-09-30 | 2021-12-03 | 苏州大学 | Application of pulsatilla chinensis saponin B4 in preparation of medicine for preventing or treating non-alcoholic fatty liver disease |
Non-Patent Citations (2)
| Title |
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| PEI, LIANG ET AL.: "Hepatoprotective effect of anemoside B4 against sepsis-induced acute liver injury through modulating the mTOR/p70S6K-mediated autophagy.", CHEMICO-BIOLOGICAL INTERACTIONS., vol. 345, 27 May 2021 (2021-05-27), XP086700974, ISSN: 0009-2797, DOI: 10.1016/j.cbi.2021.109534 * |
| SU DAN; LIAO ZHOU; FENG BINWEI; WANG TINGTING; SHAN BAIXI; ZENG QIANG; SONG JIAGUI; SONG YONGGUI: "Pulsatilla chinensis saponins cause liver injury through interfering ceramide/sphingomyelin balance that promotes lipid metabolism dysregulation and apoptosis", PHYTOMEDICINE, ELSEVIER, AMSTERDAM, NL, vol. 76, 15 June 2020 (2020-06-15), AMSTERDAM, NL , XP086232583, ISSN: 0944-7113, DOI: 10.1016/j.phymed.2020.153265 * |
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