WO2021073397A1 - 制备含去壳台湾红藜萃取物而可治疗非酒精性脂肪肝的医药组合物及其应用 - Google Patents

制备含去壳台湾红藜萃取物而可治疗非酒精性脂肪肝的医药组合物及其应用 Download PDF

Info

Publication number
WO2021073397A1
WO2021073397A1 PCT/CN2020/117553 CN2020117553W WO2021073397A1 WO 2021073397 A1 WO2021073397 A1 WO 2021073397A1 CN 2020117553 W CN2020117553 W CN 2020117553W WO 2021073397 A1 WO2021073397 A1 WO 2021073397A1
Authority
WO
WIPO (PCT)
Prior art keywords
extract
pharmaceutical composition
fatty liver
application according
fat
Prior art date
Application number
PCT/CN2020/117553
Other languages
English (en)
French (fr)
Inventor
许竣杰
蔡黛华
张书豪
Original Assignee
财团法人医药工业技术发展中心
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 财团法人医药工业技术发展中心 filed Critical 财团法人医药工业技术发展中心
Publication of WO2021073397A1 publication Critical patent/WO2021073397A1/zh

Links

Images

Classifications

    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L7/00—Cereal-derived products; Malt products; Preparation or treatment thereof
    • A23L7/10—Cereal-derived products
    • 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)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • 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 present invention relates to the application of shelled red quinoa extract, in particular to the application of shelled Taiwan red quinoa water extract, including the use of shelled Taiwan red quinoa water extract to treat or prevent, reduce oil droplets in fat cells in the liver, Reduce the volume of fat cells or reduce the accumulation of fat cells, especially the use of shelled Taiwan red quinoa water extract to inhibit the formation of non-alcoholic fatty liver.
  • fatty liver can be divided into many reasons, such as: alcoholism, obesity, abnormal fat metabolism, drugs, viruses, rapid weight loss or malnutrition. In the medical profession, it is often used to be divided into two categories, namely alcoholic fatty liver caused by alcohol abuse, and nonalcoholic fatty liver (NAFLD) (Nonalcoholic Fatty Liver Disease).
  • NAFLD nonalcoholic fatty liver
  • non-alcoholic fatty liver disease is currently the most common chronic liver disease in the world. Its incidence is about 15-30% in Western countries, while Eastern countries are increasing year by year. 15-45% has also attracted the attention of Asian governments.
  • the current treatment of fatty liver is to control weight, control blood sugar, control blood fat and moderate exercise, and reduce calorie intake according to the menu designed by nutritionist, and through weight control, it can ultimately improve liver function and fatty liver. Degree.
  • drugs tested at this stage such as insulin sensitizer pioglitazone, biguanide insulin sensitizer metformin, pentoxifylline, etc., larger trials are still needed to prove that they can effectively improve the effect of fatty liver.
  • one of the objectives of the present invention is to provide a use of a pharmaceutical composition containing an extract of Chenopodium formosanum, which is used for at least one of the following:
  • the oil droplets of fat cells reduce the volume of fat cells or reduce the accumulation of fat cells.
  • the hulled red quinoa extract is a water extract of hulled red quinoa
  • the composition is a pharmaceutical composition or a food composition.
  • Another object of the present invention is to provide a use of a pharmaceutical composition containing Chenopodium formosanum extract, which is used to prepare a composition for the prevention/treatment of non-alcoholic fatty liver.
  • the shelled red quinoa water extract is used to reduce the accumulation of fat in liver cells, effectively protect the liver and reduce the influence caused by fat accumulation.
  • the present invention provides an application for preparing a pharmaceutical composition containing shelled Taiwan red quinoa extract and capable of treating non-alcoholic fatty liver, wherein the pharmaceutical composition comprises Taiwan red quinoa extract.
  • Taiwan red quinoa extract is an extract obtained by extraction with water as a solvent.
  • Taiwan red quinoa extract is a shelled Taiwan red quinoa extract.
  • the treatment of non-alcoholic fatty liver is to improve the accumulation of fat and oil droplets in the liver.
  • the pharmaceutical composition can effectively restore the degree of fatty liver (Steatosis) degeneration of 35% to 60%.
  • the dehulled Taiwan Red Chenopodium extract is administered in an effective dose to the desired biological individual or biological cell, and the effective dose of the pharmaceutical composition is 800 to 1200 mg per administration to the desired biological individual.
  • the effective dose concentration of the pharmaceutical composition for each administration of the required biological cells is 10-100 ⁇ g/mL.
  • the organism is selected from mammals and companion animals.
  • the desired biological individual is an obese individual
  • the body mass index (BMI) of the obese individual is 24.0-27.9 or/and greater than or equal to 28.0.
  • administration of the pharmaceutical composition to the obese individual can inhibit the accumulation of body fat.
  • the body fat includes subcutaneous fat and visceral fat.
  • the dehulled red quinoa water extract of the present invention for preparing a composition for reducing the oil droplets of fat cells in the liver, reducing the volume of fat cells or reducing the accumulation of fat cells and/or preventing/treating non-alcoholic fatty liver, wherein the composition It is used to improve and/or condition the liver.
  • the composition can also be a food or a medical and health care product, where the food includes, but is not limited to, drinks, jelly, dairy products, meal replacement packages, and the like.
  • Fig. 1 is the extraction flow chart of the water extract of Taiwan red quinoa from the shell of the present invention.
  • FIG. 2A to 2B are high performance liquid chromatograms of the shelled Taiwan red quinoa extract (as shown in FIG. 2A) and the shelled Taiwan red quinoa extract (as shown in FIG. 2B) according to another embodiment of the present invention.
  • the detection wavelength of Fig. 2A and Fig. 2B is 203nm.
  • Figure 3 shows the experimental results of another embodiment of the present invention in which the water extract of Taiwan red quinoa inhibits the accumulation of fat in liver cells.
  • the foregoing experiment is based on the relative fluorescence intensity percentage obtained by Nile Red staining compared with DAPI staining (Relative fluorescence intensity(%)) for numerical analysis.
  • Figures 4A to 4D are haematoxylin-eosin (H&E) staining diagrams of liver tissue of a non-alcoholic fatty liver model animal fed with 0.1% shelled Taiwan red quinoa extract, etc. in another embodiment of the present invention .
  • H&E haematoxylin-eosin
  • Fig. 5 is a time course diagram of a test for evaluating 3T3-L1 intracellular lipid accumulation in another embodiment of the present invention.
  • Figure 6A ⁇ Figure 6Q are the present invention by administering various extraction methods such as 10 ⁇ g/mL, 30 ⁇ g/mL, 100 ⁇ g/mL and other extraction methods.
  • Hoechst staining diagram where Figure 6A is the control group (CTL), Figure 6B is the blank group (Vehicle), Figure 6C ⁇ Figure 6E are the PDC-2749 group, Figure 6F ⁇ Figure 6H are the PDC-2899 group, Figure 6I ⁇ Figure 6K
  • Figure 6L ⁇ Figure 6N are for the PDC-2948 group, Figure 6O Fig. 6Q is the silymarin group, Fig. 6C, Fig. 6F, Fig. 6I, Fig. 6L, Fig.
  • Fig. 6D administers the aforementioned groups at a dose concentration of 10 ⁇ g/mL
  • Fig. 6D administers the aforementioned groups at a dose concentration of 10 ⁇ g/mL
  • Fig. 6D administers the aforementioned groups at a dose concentration of 10 ⁇ g/mL
  • Fig. 6G administers the aforementioned groups
  • Fig. 6J administers the aforementioned groups
  • the dose concentration of the group was 30 ⁇ g/mL
  • Fig. 6E, Fig. 6H, Fig. 6K, Fig. 6N, and Fig. 6Q were administered to the aforementioned groups at a concentration of 100 ⁇ g/mL.
  • Fig. 7 is a numerical analysis diagram of the relative fluorescence intensity (%) of the Nile Red staining of lipid accumulation of 3T3-L1 cells of the decapsulated Taiwan red Chenopodium extract administered with different dosages of various extraction methods according to the present invention.
  • FIG. 8 is a numerical analysis diagram of the relative fluorescence intensity (%) of Hoechst staining of lipid accumulation of 3T3-L1 cells in 3T3-L1 cells obtained by applying various extraction methods with different dosages of different extraction methods of the present invention.
  • Figure 9 is the relative fluorescence intensity percentage obtained by Nile Red staining of 3T3-L1 cell lipid accumulation of the decapsulated Taiwan red Chenopodium extract with different doses of various extraction methods compared with Hoechst staining (Relative fluorescence intensity( %)) Numerical analysis graph.
  • the extract can be implemented by foods, health foods, health foods, functional foods, nutritional supplements, special nutritional foods, dietary supplements, or medicines.
  • the extract can also be combined with other foods or beverages, and the organism can be taken orally in a form suitable for consumption.
  • the term "individual” as used herein refers to a mammal, which may be a human or a non-human animal. In another embodiment of the present invention, the individual is selected from a companion animal, such as dogs, cats, hamsters, and the like.
  • treatment should not be interpreted as treating an individual until complete recovery, but should include maintaining an individual's disease progression or symptoms to a substantially static level and increasing the recovery rate of an individual , Improve the severity of a specific condition, or improve the quality of life of a patient.
  • prevention refers to inhibiting or preventing the onset of a specific disease, or maintaining the good health of a sensitive individual or establishing the individual's tolerance to the disease.
  • composition provided according to the present invention can be consumed at different frequencies such as once a day, multiple times a day, or once a few days, depending on the age, weight, and health status of the individual being administered.
  • the content of the extract in the composition provided by the present invention can also be adjusted according to the needs of specific ethnic groups, for example, adjusted to the amount that should be taken daily.
  • the biological individual is an obese individual, and the body mass index (BMI) of the aforementioned obese individual is 24.0-27.9 or/and greater than or equal to 28.0.
  • BMI body mass index
  • the individual's Body Mass Index (BMI) is 24.0-27.9, it is defined as overweight; if the individual's body mass index is greater than or equal to 28.0, it is defined as obesity.
  • the present invention is not limited to this, and the pharmaceutical composition administered to the obese individual can inhibit the accumulation of body fat.
  • the body fat includes subcutaneous fat and visceral fat.
  • the scoring criteria for assessing the extent of liver steatosis are shown in Table 1.
  • the extraction process of the water extract of the dehulled Taiwan red quinoa in the present invention the dehulled Taiwan red quinoa is added with 10 times equal weight of reverse osmosis water (Reverse Osmosis Water), heated in a heating bag to boil, and extracted for 0.5-2 hours. Use a sieve to filter and collect the first extraction filtrate. Add 10 times the weight of reverse osmosis water (Reverse Osmosis Water) to the shelled Taiwan red quinoa residue, heat it with a heating bag and boil for 0.5 to 2 hours, filter with a sieve, and collect the second extraction filtrate.
  • reverse Osmosis Water reverse Osmosis Water
  • the extraction process of the water extract of Taiwan red quinoa (ie, the shelled Taiwan red quinoa) also follows the extraction process of the aforementioned unshelled Taiwan red quinoa water extract.
  • Example Three Model test of dehulled Taiwan red Chenopodium extract in non-alcoholic fatty liver cells
  • a hepatocyte cell line (HepG2 cell) was used to evaluate whether the dehulled Taiwan red Chenopodium extract of the present invention has the effect of reducing lipid accumulation in liver cells.
  • the administration of palmitic acid (PA) and oleic acid (OA) to the hepatocyte cell line (HepG2 cell) induces lipid production in the hepatocyte cell line, and the dosage of the dehulled Taiwan red pineapple extract It is 10-100 ⁇ g/mL, and then Nile Red staining is used to analyze the lipid accumulation of hepatocytes.
  • DAPI staining is also used to evaluate the survival of HepG2 cell. analysis.
  • the extract of Red Chenopodium from its shell has the effect of reducing lipid accumulation in liver cells at medium dose (30 ⁇ g/mL) and high dose (100 ⁇ g/mL).
  • the hepatic cell line (HepG2 cell) was also given palmitic acid and oleic acid to induce lipid production on the hepatic cell line.
  • the present invention uses a high-fat diet to induce non-alcoholic fatty liver injury in mice to evaluate whether the dehulled Taiwan red quinoa extract of the present invention has the effect of improving the fat accumulation of individual fatty liver.
  • All model animals were given 0.1% Hulled Taiwan red quinoa extract, 0.1% Taiwan red quinoa extract, or deionized water (such as the blank group) 1 week before the high-fat diet, and then given high-fat diet for 8 weeks. Fatty liver.
  • blood was collected and the animal weight was weighed. The final test was completed when the liver injury was expected to be successfully induced.
  • the liver was collected to measure its weight.
  • the largest right lobe liver was cut into a liver tissue block of about 0.5 cm square and fixed in 10%.
  • the test group of model animals includes a control group (no high-fat feed, as shown in Figure 4A), a blank group (as shown in Figure 4B), and 0.1% shelled Taiwan red
  • the quinoa extract group (as shown in Figure 4C) and the 0.1% unshelled Taiwan red quinoa extract group (as shown in Figure 4D) have corresponding steatosis scores of 0.00 ⁇ 0.00, 2.14 ⁇ 0.69, 1.13 ⁇ 0.83, and 1.38 ⁇ 0.74, where the dehulled Taiwan red quinoa extract of the present invention has the effect of reducing the amount of liver fat in the interpretation of liver tissue slices and the corresponding steatosis score.
  • the effective dose of the dehulled Taiwan red quinoa extract of the present invention is 1000 mg/kg body weight per administration to the desired biological individual, such as a non-alcoholic fatty liver individual.
  • the aforementioned husked red quinoa extract can effectively restore fatty liver. (Steatosis)
  • the degree of degeneration is 47.2%.
  • Example 5 Evaluation test of dehulled Taiwan red quinoa extract in reducing somatic fat
  • somatic cells are used to evaluate the effect of a variety of unshelled Taiwan red pineapple extracts on inhibiting intracellular adipogenesis.
  • the evaluation test method is based on insulin, ground Dexamethasone (dexamethasone) and rosiglitazone (rosiglitazone) induce somatic cell line 3T3-L1 cell line differentiation and lipid production, and then administered 10 ⁇ g/mL, 30 ⁇ g/mL, 100 ⁇ g/mL and other different doses of PDC-2747 (to Shell Taiwan red quinoa water extract), PDC-2749 (50E extract of Taiwan red quinoa shelled), PDC-2898 (50E-EA layer-not partitioned hexane layer extract of Taiwan red quinoa from shell), PDC-2899 (50E-Butanol layer-not partitioned hexane layer extract from Taiwan red quinoa), PDC-2900 (50E-H 2 O layer-not partition
  • the aforementioned 50E refers to 50% ethanol
  • the aforementioned EA refers to ethyl acetate.
  • CTL is the control group
  • Vehicle is the blank group
  • the aforementioned blank group is based on the somatic 3T3-L1 cell line. Insulin, dexamethasone and rosiglitazone are also used to induce differentiation and lipid generate.
  • the experimental steps of the aforementioned evaluation test method are as follows: (1) Plant the 3T3-L1 mouse embryonic fibroblast cell line in a 96-well microculture plate with a black hole and a transparent bottom. The cells are evenly dispersed in the multi-well plate, and cultured in DMEM with 10% NBCS (new born calf serum) in an incubator temperature of 37°C and 5% CO 2.
  • DMEM fresh born calf serum
  • each well is about 8-9
  • insulin 1.5ug/mL
  • DMEM/F12 (1:1) with 10% fetal bovine serum albumin is configured in DMEM/F12 (1:1) with 10% fetal bovine serum albumin to become a Maintenance kit to induce cells Lipid accumulation, and add separately the above-mentioned 10 ⁇ g/mL, 30 ⁇ g/mL, 100 ⁇ g/mL and other different dosages of various extraction methods of dehulled Taiwan red quinoa extract.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Natural Medicines & Medicinal Plants (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Medicinal Chemistry (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • General Health & Medical Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Food Science & Technology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Organic Chemistry (AREA)
  • Obesity (AREA)
  • Hematology (AREA)
  • Diabetes (AREA)
  • Polymers & Plastics (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nutrition Science (AREA)
  • Alternative & Traditional Medicine (AREA)
  • Biotechnology (AREA)
  • Botany (AREA)
  • Medical Informatics (AREA)
  • Microbiology (AREA)
  • Mycology (AREA)
  • Epidemiology (AREA)
  • Medicines Containing Plant Substances (AREA)

Abstract

公开了一种制备含去壳台湾红藜（Chenopodium formosanum）萃取物而可治疗非酒精性脂肪肝的医药组合物及其应用，所述组合物包含台湾红藜萃取物，系用于以下至少一种用途：减少肝脏中脂肪细胞的油滴、缩小脂肪细胞体积或降低脂肪细胞累积，从而达到治疗或预防非酒精性脂肪肝的效果。

Description

制备含去壳台湾红藜萃取物而可治疗非酒精性脂肪肝的医药组合物及其应用 技术领域
本发明是关于去壳红藜萃取物的应用，尤其是关于去壳台湾红藜水萃取物的应用，包括使用去壳台湾红藜水萃取物治疗或预防、减少肝脏中脂肪细胞的油滴、缩小脂肪细胞体积或降低脂肪细胞累积的疾病，尤其是使用去壳台湾红藜水萃取物抑制非酒精性脂肪肝的形成。
背景技术
脂肪肝的形成能区分为许多种原因，例如：酗酒、肥胖、脂肪代谢异常、药物、病毒、急速减肥或营养不良等。而于医界中，常习惯分成两大类，分别为酗酒所造成的酒精性脂肪肝，另一类则为非酒精性脂肪肝(Nonalcoholic Fatty Liver Disease,NAFLD)。
[根据细则9.2更正 16.10.2020] 然而，非酒精性脂肪肝是目前全球最常见的慢性肝脏疾病，其发生率在西方国家约为15～30％，而东方国家等则逐年上升当中，约为15～45％，亦已引起亚洲各国政府的关注。
近年于非酒精性脂肪肝疾病的研究显示，此族群中约八九成的病人都是单纯的肝脏脂肪堆积状态，对于肝脏的影响不大；但其中有约一成的非酒精性脂肪肝患者会合并发炎甚至坏死的情况，意即脂肪肝炎。这些患者的肝脏会由肝纤维化慢慢进展成肝硬化，约有0.5％发生肝癌的机率。
目前脂肪肝的治疗方式以控制体重、控制血糖、控制血脂肪及适度的运动并依照营养师所设计的菜单降低所摄入热量，而藉由体重的控制，最终可达到改善肝功能和脂肪肝的程度。然而，现阶段测试的药物中，如胰岛素致敏剂pioglitazone、双胍类药物的胰岛素致敏剂metformin、pentoxifylline等仍需更大型的试验来证实可有效改善脂肪肝的效果。
有鉴于此，如何开发可一方面长期给予患者服用，另一方面又不致于导致副作用，而患者接收度高的饮食补充方式来预防或缓解非酒精性脂肪肝症状，进而解决现有技术的缺失，实为相关技术领域者目前所迫切需要解决问题。
发明内容
鉴于所述技术问题，本发明目的之一，在于提供一种含去壳台湾红藜(Chenopodium formosanum)萃取物的医药组合物的用途，所述组合物是用于以下至少一者：减少肝脏中脂肪细胞的油滴、缩小脂肪细胞体积或降低脂肪细胞累积。较佳地，所述去壳红藜萃取物是去壳红藜水萃取物，且所述组合物是一医药组合物或食品组合物。
本发明的另一目的是提供一种含去壳台湾红藜(Chenopodium formosanum)萃取物的医药组合物的用途，其是用于制备预防/治疗非酒精性脂肪肝的组合物。所述去壳红藜水萃取物用于减少肝脏细胞中脂肪的累积，有效保护肝脏降低因脂肪累积而造成的影响。
本发明提供一种制备含去壳台湾红藜萃取物而可治疗非酒精性脂肪肝的医药组合物的应用，其中所述医药组合物包含台湾红藜萃取物。
其中，所述台湾红藜萃取物是以水为溶剂进行萃取获得的萃取物。
特别是，所述台湾红藜萃取物为去壳台湾红藜萃取物。
其中，所述治疗非酒精性脂肪肝为改善肝脏中脂肪累积、油滴累积。
特别是，所述医药组合物能有效恢复脂肪肝(Steatosis)变性程度为35％～60％。
尤其是，所述去壳台湾红藜萃取物是以有效剂量施予所需生物个体或生物细胞，所述医药组合物每次施予所述所需生物个体的有效剂量为800～1200毫克/公斤体重，所述医药组合物每次施予所述所需生物细胞的有效剂量浓度为10～100μg/mL。
特别是，所述生物选自哺乳动物、伴侣动物。
尤其是，所述所需生物个体是为肥胖个体，所述肥胖个体的身体质量指数(Body Mass Index，BMI)为24.0～27.9或/及大于或等于28.0。
特别是，所述医药组合物施予所述肥胖个体能抑制其体脂肪累积。
尤其是，所述体脂肪包括皮下脂肪与内脏脂肪。
本发明去壳红藜水萃取物用于制备减少肝脏中脂肪细胞的油滴、缩小脂肪细胞体积或降低脂肪细胞累积及/或预防/治疗非酒精性脂肪肝组合物的用途，其中该组合物系用于改善及/或调理肝脏。此外，该组合物亦可为一食品或一医疗保健品，其中该食品包括但不限于饮品、果冻、乳制品、代餐包等。
附图说明
图1是本发明去壳台湾红藜水萃取物的萃取流程图。
图2A～图2B是本发明又一实施例的去壳台湾红藜萃取物(如图2A所示)与带壳台湾红藜萃取物(如图2B所示)高效能液相层析图。图2A与图2B检测波长为203nm。
图3是本发明再一实施例去壳台湾红藜水萃取物抑制肝脏细胞内脂肪累积的实验结果，前述实验是以Nile Red(尼罗红)染色相较DAPI染色所获得的相对荧光强度百分比(Relative fluorescence intensity(％))进行数值分析。
图4A～图4D是本发明另一实施例非酒精性脂肪肝模型动物经喂食如0.1％去壳台湾红藜萃取物等的肝脏组织苏木紫-伊红(haematoxylin-eosin,H&E)染色图。
图5是本发明再一实施例评评估3T3-L1细胞内脂质堆积试验时程图。
图6A～图6Q是本发明藉由施予10μg/mL、30μg/mL、100μg/mL不同剂量等多种萃取方法的去壳台湾红藜萃取物的3T3-L1细胞脂质堆积的Nile Red/Hoechst染色图，其中图6A为控制组(CTL)，图6B为空白组(Vehicle)，图6C～图6E为PDC-2749组，图6F～图6H为PDC-2899组，图6I～图6K为PDC-2900组，图6L～图6N为PDC-2948组，图6O
Figure PCTCN2020117553-appb-000001
图6Q为silymarin组，图6C、图6F、图6I、图6L、图6O施予前述各组剂量浓度为10μg/mL，图6D、图6G、图6J、图6M、图6P施予前述各组剂量浓度为30μg/mL，图6E、图6H、图6K、图6N、图6Q施予前述各组剂量浓度为100μg/mL。
图7是本发明施予不同剂量的多种萃取方法之去壳台湾红藜萃取物的3T3-L1细胞脂质堆积的Nile Red染色相对荧光强度百分比(Relative fluorescence intensity(％))数值分析图。
图8是本发明施予不同剂量的多种萃取方法去壳台湾红藜萃取物之3T3-L1细胞脂质堆积的Hoechst染色相对荧光强度百分比(Relative fluorescence intensity(％))数值分析图。
图9是本发明施予不同剂量的多种萃取方法之去壳台湾红藜萃取物的3T3-L1细胞脂质堆积的Nile Red染色相较Hoechst染色所获得的相对荧光强度百分比(Relative fluorescence intensity(％))数值分析图。
具体实施方式
以下藉由特定的具体实施形态说明本发明之技术内容，熟悉此技艺的人士可由本说明书所揭示的内容而了解本发明的优点与功效。然在不背离本发明精神下，本发明尚可以多种不同形式的态样来实践或加以应用。
于本发明的一具体实施例中，萃取物可藉由食品、保健食品、健康食品、机能性食品、营养补充品、特殊营养食品、膳食补充物、或药品等实施例予以实施。
于本发明再一实施例中，亦可将萃取物与其他食品或饮料组合，以适于食用的样态供生物体以口服方式服用。
再者，本文中所使用的术语「个体」是指哺乳动物，该哺乳动物可以为人类或非人动物。于本发明另一实施例中，所述个体选自一伴侣动物，例如包括狗、猫、仓鼠等。
另外，本文中所使用的术语「治疗」，不应被解释为治疗一个体直至完全恢复，而应包括将一个体的疾病进展或症状维持在一实质上静态的程度、增加一个体的恢复速率、改善一具体病况的严重性、或提高一患者的生命质量。除此之外，本文中所使用的所谓「预防」是指抑制或防止一具体病 况的发作、或维持敏感个体的良好健康状态或建立该个体对疾病的耐受性。
不仅如此，根据本发明所提供的组合物是可以一日一次、一日多次、或数日一次等不同频率食用，端视投予生物个体的年龄、体重、及健康状况而异。亦可针对特定族群的需要，调整据本发明所提供的组合物中的萃取物的含量，例如，调整至每日应服用的量。
进一步言，所述生物个体是为肥胖个体，前述肥胖个体的身体质量指数(Body Mass Index，BMI)为24.0～27.9或/及大于等于28.0。换句话说，所述个体的身体质量指数(Body Mass Index，BMI)为24.0～27.9者，则定义为过重；所述个体的身体质量指数为大于或等于28.0者，则定义为肥胖。
当然，本发明不限于此，所述医药组合物施予所述肥胖个体能抑制其体脂肪累积。所述体脂肪包括皮下脂肪与内脏脂肪。
于本发明一实施例中，评估肝脏脂肪变性范围评分标准如表一所示。
表一
Figure PCTCN2020117553-appb-000002
兹以下列实施例进一步例示说明本发明，其中该等实施例于此仅提供作为说明之用。
实施例
实施例一、去壳台湾红藜萃取物制备方法
如图1，本发明去壳台湾红藜水萃取物的萃取流程，去壳台湾红藜加入10倍等重量的反渗透水(Reverse Osmosis Water)，以加热包加热沸腾，萃取0.5～2小时，使用筛网过滤，收集第一次萃取滤液。去壳台湾红藜残渣继续加入10倍药材等重量的逆渗透水(Reverse Osmosis Water)，以加热包加热沸腾萃取0.5～2小时，使用筛网过滤，收集第二次萃取滤液。前述二次萃取滤液收集混合均匀后，进行下述指纹图谱(HPLC)分析，同时利用大型减压浓缩机浓缩与冷冻干燥机冻干，进而获得本发明去壳台湾红藜水萃取物的干粉率相关数据。于本发明另一实施例中，台湾红藜(即带壳台湾红藜)水萃取物的萃取流程也依循前述去壳台湾红藜水萃取物的萃取流程进行。
实施例二、去壳台湾红藜萃取物的HPLC成分分析
如图2A～图2B，藉由高效液相层析仪高效能液相层析(high performance liquid chromatography,HPLC)化学指纹图谱分析去壳台湾红藜萃取物(如图2A所示)与台湾红藜萃取物(如图2B所示)，前述图2A～图2B的HPLC 203nm分析图谱其可得知去壳台湾红藜萃取物与台湾红藜萃取物二者的层析峰(chromatographic peak)的面积及峰高等具有差异性，如其中去壳台湾红藜萃取物于保留时间4分钟(min)与24分钟(min)左右的层析峰面积及高度与台湾红藜萃取物均不相同，因此可推知去壳台湾红藜萃取物及台湾红藜萃取物中的化学物质不同。
实施例三、去壳台湾红藜萃取物于非酒精性脂肪肝细胞模式试验
于另一实施例中，系藉肝细胞株(HepG2 cell)来评估本发明去壳台湾红藜萃取物是否具降低肝细胞内脂质堆积的功效。如图3，肝细胞株(HepG2 cell)给予棕榈酸(Palmitic acid；PA)及油酸(Oleic acid；OA)诱发对肝细胞株脂质生成，其中去壳台湾红藜萃取物的施予剂量为10～100μg/mL，进而利用尼罗红(Nile Red)染色进行肝细胞脂质堆积分析，同时也藉由DAPI染色进行评估去壳台湾红藜萃取物对肝细胞株(HepG2 cell)的存活分析。其中去壳红藜萃取物于中剂量(30μg/mL)、高剂量(100μg/mL)具降低肝细胞内脂质堆积的功效。其中空白组是以肝细胞株(HepG2 cell)同样给予棕榈酸及油酸诱发对肝细胞株脂质生成。
实施例四、去壳台湾红藜萃取物于非酒精性脂肪肝模型动物实验
于再一实施例中，本发明藉高脂饲料诱导小鼠产生非酒精性脂肪肝损伤的试验模式，来评估本发明去壳台湾红藜萃取物是否具改善个体脂肪肝的脂肪堆积功效。所有模型动物于喂食高脂饲料的前1周开始施予0.1％去壳台湾红藜萃取物、0.1％台湾红藜萃取物或去离子水(如空白组)，再给予8周高脂饲料诱导脂肪肝。试验期间定期进行采血、秤量动物体重，最后试验于预期可诱导肝损伤成功时结束，并采集肝脏测量其重量，将最大右叶肝切割出约0.5公分见方的肝组织块，固定于10％的中性福尔马林溶液，接着以石蜡包埋、切片、贴附在载玻片上，进而以苏木紫-伊红(haematoxylin-eosin,H&E)染色，来进行肝脏组织病理学观察与肝脏脂肪变性评分。综言之，于本发明的实施例中，模型动物测试组别包括控制组(无喂食高脂饲料，如图4A所示)、空白组(如图4B所示)、0.1％去壳台 湾红藜萃取物组(如图4C所示)、0.1％带壳台湾红藜萃取物组(如图4D所示)，其对应脂肪变性评分分别为0.00±0.00、2.14±0.69、1.13±0.83、1.38±0.74，其中本发明去壳台湾红藜萃取物于肝脏组织切片判读与对应脂肪变性评分具有效降低肝脏脂肪数量的功效。
换言之，本发明去壳台湾红藜萃取物每次施予所需生物个体如非酒精性脂肪肝个体的有效剂量为1000毫克/公斤体重，其中前述去壳台湾红藜萃取物可有效回复脂肪肝(Steatosis)变性程度为47.2％。
实施例五、去壳台湾红藜萃取物于降低体细胞脂肪评估测试
如图5～图9，本发明于另一实施例系利用体细胞来评估多种去壳台湾红藜萃取物抑制体细胞内脂肪生成的效果，该评估测试方法系以胰岛素(insulin)、地塞米松(dexamethasone)和罗格列酮(rosiglitazone)诱导体细胞3T3-L1细胞株分化与脂质生成，进而施予10μg/mL、30μg/mL、100μg/mL等不同剂量的PDC-2747(去壳台湾红藜水萃物)、PDC-2749(去壳台湾红藜的50E萃物)、PDC-2898(去壳台湾红藜的50E-EA layer-not partitioned hexane layer萃物)、PDC-2899(去壳台湾红藜的50E-Butanol layer-not partitioned hexane layer萃物)、PDC-2900(去壳台湾红藜的50E-H 2O layer-not partitioned hexane layer萃物)、PDC-2948(去壳台湾红藜的50E-H 2O layer-only partitioned EA萃物)等多种萃取方法的去壳台湾红藜萃取物是否具降低细胞内脂质堆积的功效，并辅以silymarin作为一对照组进行测试。其中前述50E是指50％ethanol，前述EA是指ethyl acetate。其中于图7、图8与图9，CTL为控制组，Vehicle为空白组，前述空白组 系以体细胞3T3-L1细胞株同样以胰岛素、地塞米松和罗格列酮诱导分化与脂质生成。
进一步，如图5，前述评估测试方法的实验步骤如下所述：(1)将3T3-L1小鼠胚胎成纤维细胞株种于黑孔透明底的微量培养盘96孔，每一微孔以4000个细胞均匀分散于多孔盘，以10％NBCS(new born calf serum)的DMEM在培养箱温度为37℃及5％CO 2环境中培养，当细胞生长至微量培养盘每一孔约8～9成满，即可配置3T3-L1细胞分化套组(differentiation cocktail，#K579-100,BioVision,USA)于10％胎牛血清蛋白的DMEM/F12(1:1)中，加入前述每一微孔的细胞中并继续培养72小时以诱导细胞分化；(2)另将胰岛素(1.5ug/mL)配置于10％胎牛血清蛋白的DMEM/F12(1:1)而成为一Maintenance kit以诱导细胞脂质堆积，并分别加入前述10μg/mL、30μg/mL、100μg/mL等不同剂量的多种萃取方法的去壳台湾红藜萃取物，培养48小时后，更换培养液并进而再次培养48小时；(3)藉磷酸盐缓冲液(PBS)清洗，加入含10％福尔马林(formalin)固定细胞1小时；(4)移除10％福尔马林，以二次水清洗后加入10μg/mL尼罗红(Nile Red，#72485,Sigma,USA)，染色10分钟；(5)移除尼罗红，以二次水清洗后加入10μg/mL Hoechst 33342(H3570,Invitrogen,USA)，染色10分钟；(6)移除Hoechst 33342，以二次水清洗后检测数值(Nile red：RFU485/580nm；Hoechst：RFU350/461nm)，并以荧光显微镜进行细胞脂质拍照纪录(如图6A～图6Q所示)与进一步分析。
如图7、图8与图9，前述实验结果显示，尽管PDC-2747与PDC-2898并没有随着施予剂量增加而抑制细胞脂肪生成的结果，然而PDC-2749、 PDC-2899、PDC-2900、PDC-2948与silymarin组的细胞内脂肪显著降低，显示具有缓解细胞脂质堆积的功效。此外，PDC-2899与PDC-2948抑制细胞脂质堆积的功效较PDC-2749为佳。
上述实施例仅为说明本发明的原理及其功效，其目的在使熟习前述技术者能了解本发明的内容并据以实施，并非限制本发明。因此习于此技术的人士对上述实施例进行等效修饰、修改及变化仍不脱本发明的精神。本发明的权利范围应如前述的申请专利范围所列。

Claims (10)

  1. 一种制备含去壳台湾红藜萃取物而可治疗非酒精性脂肪肝的医药组合物的应用，其特征是，所述医药组合物包含台湾红藜萃取物。
  2. 如权利要求1所述的应用，其特征是，所述台湾红藜萃取物是以水为溶剂进行萃取获得的萃取物。
  3. 如权利要求2所述的应用，其特征是，所述台湾红藜萃取物为去壳台湾红藜萃取物。
  4. 如权利要求1所述的应用，其特征是，所述治疗非酒精性脂肪肝为改善肝脏中脂肪累积、油滴累积。
  5. 如权利要求1所述的应用，其特征是，所述医药组合物能有效恢复脂肪肝变性程度为35％～60％。
  6. 如权利要求1所述的应用，其特征是，所述去壳台湾红藜萃取物是以有效剂量施予所需生物个体或生物细胞，所述医药组合物每次施予所述所需生物个体的有效剂量为800～1200毫克/公斤体重，所述医药组合物每次施予所述所需生物 细胞的有效剂量浓度为10～100μg/mL。
  7. 如权利要求6所述的应用，其特征是，所述生物选自哺乳动物、伴侣动物。
  8. 如权利要求6所述的应用，其特征是，所述所需生物个体是为肥胖个体，所述肥胖个体的身体质量指数为24.0～27.9或/及大于或等于28.0。
  9. 如权利要求8所述的应用，其特征是，所述医药组合物施予所述肥胖个体能抑制其体脂肪累积。
  10. 如权利要求9所述的应用，其特征是，所述体脂肪包括皮下脂肪与内脏脂肪。
PCT/CN2020/117553 2019-10-17 2020-09-24 制备含去壳台湾红藜萃取物而可治疗非酒精性脂肪肝的医药组合物及其应用 WO2021073397A1 (zh)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US201962916457P 2019-10-17 2019-10-17
US62/916,457 2019-10-17

Publications (1)

Publication Number Publication Date
WO2021073397A1 true WO2021073397A1 (zh) 2021-04-22

Family

ID=75538323

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2020/117553 WO2021073397A1 (zh) 2019-10-17 2020-09-24 制备含去壳台湾红藜萃取物而可治疗非酒精性脂肪肝的医药组合物及其应用

Country Status (2)

Country Link
TW (1) TW202116338A (zh)
WO (1) WO2021073397A1 (zh)

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102524768A (zh) * 2010-12-29 2012-07-04 屏东科技大学 以台湾藜制备的降血脂的药物或保健食品
US20150265527A1 (en) * 2014-03-20 2015-09-24 Tci Co., Ltd. Method for enhancing collagen secretion and preventing cutaneous aging using chenopodium formosanum extract
CN205700946U (zh) * 2016-03-01 2016-11-23 常春藤生技股份有限公司 一种加工不易形成体脂肪的台湾红藜的设备
CN207545495U (zh) * 2016-02-04 2018-06-29 常春藤生技股份有限公司 加工台湾红藜以提升抗氧化、减缓心血管疾病及提升保肝成分的设备
CN108852924A (zh) * 2017-05-11 2018-11-23 嘉药学校财团法人嘉南药理大学 一种具抗老化、美白、抗过敏及细胞修护功能的红藜萃取物
CN109938250A (zh) * 2017-12-21 2019-06-28 财团法人石材暨资源产业研究发展中心 供调节血脂的复合液

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102524768A (zh) * 2010-12-29 2012-07-04 屏东科技大学 以台湾藜制备的降血脂的药物或保健食品
US20150265527A1 (en) * 2014-03-20 2015-09-24 Tci Co., Ltd. Method for enhancing collagen secretion and preventing cutaneous aging using chenopodium formosanum extract
CN207545495U (zh) * 2016-02-04 2018-06-29 常春藤生技股份有限公司 加工台湾红藜以提升抗氧化、减缓心血管疾病及提升保肝成分的设备
CN205700946U (zh) * 2016-03-01 2016-11-23 常春藤生技股份有限公司 一种加工不易形成体脂肪的台湾红藜的设备
CN108852924A (zh) * 2017-05-11 2018-11-23 嘉药学校财团法人嘉南药理大学 一种具抗老化、美白、抗过敏及细胞修护功能的红藜萃取物
CN109938250A (zh) * 2017-12-21 2019-06-28 财团法人石材暨资源产业研究发展中心 供调节血脂的复合液

Also Published As

Publication number Publication date
TW202116338A (zh) 2021-05-01

Similar Documents

Publication Publication Date Title
EP2298320B1 (en) Use of lanostane and poria extract in treating diabetes
CN100998650A (zh) 肉桂在治疗糖尿病上的用途及产品和其制备方法
KR20080003931A (ko) 인슐린 저항성 증후군의 치료
WO2020073869A1 (zh) 盐肤木果实或其提取物的用途
KR20120002131A (ko) 울금 추출물을 함유하는 비만 치료 또는 예방용 조성물
Hou et al. Beneficial effects of mung bean seed coat on the prevention of high-fat diet-induced obesity and the modulation of gut microbiota in mice
CN105722519A (zh) 包含琉璃苣提取物的代谢性疾病的改善、预防或治疗用组合物
CN102740871A (zh) 包含荔枝核提取物的脂肪肝或肥胖的预防或治疗用组合物
CN111513155B (zh) 一种防治脂代谢异常的组合物
WO2021073397A1 (zh) 制备含去壳台湾红藜萃取物而可治疗非酒精性脂肪肝的医药组合物及其应用
KR20150083622A (ko) 오갈피나무와 뽕나무를 이용한 항비만 조성물
Hassan et al. Genetic and histopathology studies on mice: effect of fenugreek oil on the efficiency of ovarian and liver tissues
CN108926595A (zh) 一种具有保护肝脏和降血脂功能的保健品
JP6467345B2 (ja) 脂肪蓄積抑制剤、脂肪肝の予防剤又は治療剤及び脂肪酸合成酵素抑制剤
KR20150006224A (ko) 혼합 생약 추출물을 이용한 대사성질환 예방 및 치료용 조성물
CN108404088B (zh) 一种治疗ⅱ型糖尿病的中药及其制备方法
CN105816492A (zh) 明月草提取物的提取方法及用途
CN101291679A (zh) 包含猕猴桃属植物的组合物及其使用方法
KR102063897B1 (ko) 발효 글루텐세이프 밀가루
US20180177836A1 (en) Traditional chinese medicine composition for promotion of browning of white adipocytes, preparation method and use thereof
TWI589297B (zh) Use of melon leaf extract for the treatment and / or prevention of alcoholic fatty liver or its complications
CN104940387B (zh) 一种治疗非酒精性脂肪肝的中药组合物及其制备和应用
KR102514847B1 (ko) 병풀, 벌사상자 및 영하구기자 추출물을 포함하는 인지 기능 장애 또는 신경염증의 예방 또는 치료용 조성물
TWI837372B (zh) 葡萄果皮萃取物及其製造方法、食品組成物以及醫藥組成物
CN108714192B (zh) 用于改善大脑神经营养功能的组合物及其应用

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: 20876804

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: 20876804

Country of ref document: EP

Kind code of ref document: A1