CN103623076B - The application of the extract of nasturtium in the preparation of medicine for treating viral diseases - Google Patents
The application of the extract of nasturtium in the preparation of medicine for treating viral diseases Download PDFInfo
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Abstract
Description
技术领域 technical field
本发明涉及医药技术领域，涉及金莲花提取物在制药领域的新用途，具体涉及金莲花提取物在制备治疗病毒性疾病药物中的应用。 The present invention relates to the field of medical technology, and relates to the new application of the extract of Trollius chinensis in the field of pharmacy, in particular to the application of the extract of Trollius chinensis in the preparation of drugs for treating viral diseases.
背景技术 Background technique
从2003年“SARS病毒”横行肆虐，到2009年“甲型H1N1流感”汹汹来袭，再到今年“H7N9禽流感”余威尚存，短短十年间，我们已一而再、再而三地遭到病毒的攻击。虽然历史的记载告诉我们，病毒作为一类物种的存在时间远远超过人类，在人类的发展史上，它对人类带来的生存威胁也远超其他物种。 From the rampant "SARS virus" in 2003, to the fierce attack of "H1N1 influenza" in 2009, and to the lingering power of "H7N9 bird flu" this year, in just ten years, we have repeatedly and repeatedly Attacked by a virus. Although historical records tell us that viruses, as a species, have existed far longer than humans, and in the history of human development, they have posed far more threats to human survival than other species.
由各种呼吸道病毒所引起的病毒感染性疾病是一组常见病，其发病率在全球呈逐步上升趋势，而且不断出现新的致病病毒。如2003年初在我国部分省市流行的非典型肺炎（SARS）就是由冠状病毒的变异病毒所引起的一种严重的病毒性传染病。H5N1病毒及目前出现的H1N1、H7N9等病毒感染的疾病，都属于病毒感染性疾病。 Viral infectious diseases caused by various respiratory viruses are a group of common diseases, the incidence of which is gradually increasing in the world, and new pathogenic viruses are constantly emerging. For example, atypical pneumonia (SARS), which was prevalent in some provinces and cities in my country in early 2003, is a serious viral infectious disease caused by a mutant virus of coronavirus. H5N1 virus and currently emerging H1N1, H7N9 and other virus-infected diseases are all viral infectious diseases.
病毒感染性疾病的危害不仅仅在于所引起的各种感染，更主要是其大多继发或合并细菌感染，故致使临床症状较重，病程较长，表现多样化，一直是危害人类健康的主要疾病之一。 The hazards of viral infectious diseases are not only the various infections caused by them, but more importantly, most of them are secondary or combined with bacterial infections, so the clinical symptoms are severe, the course of the disease is long, and the manifestations are diverse. It has always been the main cause of harm to human health. one of the diseases.
目前临床治疗病毒性疾病以化学合成和半合成的药物为主。但是，由于半个多世纪以来化学合成和半合成抗菌、抗病毒药物的广泛使用，已诱发病菌产生了耐药性菌株，且日渐增多条件性致病菌。加之其毒副作用较大等缺点，致使该类药物尽管作为一线治疗药、但已存在诸多不足。因此，抗病毒中药具有副作用小而具有广阔的应用前景，此新药的开发成功将产生重大的社会效益。 At present, the clinical treatment of viral diseases is mainly based on chemically synthesized and semi-synthesized drugs. However, due to the widespread use of chemically synthesized and semi-synthesized antibacterial and antiviral drugs for more than half a century, drug-resistant strains of pathogens have been induced, and opportunistic pathogenic bacteria are increasing day by day. Coupled with its disadvantages such as relatively large toxic and side effects, this type of drug has many shortcomings even though it is used as a first-line treatment drug. Therefore, the antiviral traditional Chinese medicine has few side effects and has broad application prospects, and the successful development of this new medicine will produce significant social benefits.
中国药典1977年版中规定金莲花为毛茛科植物金莲花Trollius chinensis Bunge的干燥花。夏季花开时采摘，除去杂质，阴干。苦，寒。归肝经。具有清热散风，解毒消肿，平肝明目。用于口疮喉肿，浮热牙宣，耳疼目痛，山岚瘴气，疔疮火毒，上呼吸道感染，急性肠炎，泌尿系统感染及疮疖脓肿等作用。 The 1977 edition of the Chinese Pharmacopoeia stipulates that Trollius is the dried flower of Trollius chinensis Bunge, a plant in the Ranunculaceae family. Pick when the flowers bloom in summer, remove impurities, and dry in the shade. Bitter, cold. Return liver channel. Have heat-clearing and wind-dispelling, detoxification and detumescence, suppress liver and improve eyesight. It is used for aphthous sores and swollen throats, feverish teeth, earaches and eye pains, miasma, furuncles, upper respiratory tract infections, acute enteritis, urinary system infections and sores, furuncles and abscesses.
金莲花提取物是一种由天然植物、中药材或中药饮片金莲花提取出来的具有显著抗病毒作用的提取物。金莲花提取物中含有多种化学成分，现已发现该类提取物主要以黄酮类成分为主，包含各种类型的黄酮成分，如黄酮、黄酮醇、二氢黄酮、二氢黄酮醇、异黄酮、二轻异黄酮、查尔酮等大类成分。 Trollius extract is a kind of extract which is extracted from natural plants, Chinese medicinal materials or Chinese herbal pieces, and has significant antiviral effect. The extract of nasturtium lotus contains a variety of chemical components. It has been found that this type of extract is mainly composed of flavonoids, including various types of flavonoids, such as flavonoids, flavonols, dihydroflavones, dihydroflavonols, iso Flavonoids, dihydroxyisoflavones, chalcones and other major ingredients.
发明内容 Contents of the invention
本发明所要解决的技术问题是提供一种金莲花提取物用于治疗病毒性疾病的新用途。在临床上可以制备成用于治疗病毒性疾病的药物制剂。所述的金莲花提取物，是从毛茛科植物金莲花Trollius chinensis Bunge的干燥花中提取出来的一大类物质。用该金莲花提取物做为药效成分来制备用于治疗病毒性疾病的药物制剂，可单独或与制药工艺中可接受的赋形剂结合，按照常规方法制成口服给药、腔道、黏膜给药、肌肉或静脉给药的药物剂型。所述金莲花提取物总黄酮含量大于50%以上。金莲花提取物在制备治疗流感病毒H1N1的药物中的应用；金莲花提取物在制备治疗呼吸道合胞病毒RSV药物中的应用；金莲花提取物在制备治疗腺病毒Adv3抑制活药物中的应用。 The technical problem to be solved by the present invention is to provide a new application of nasturtium extract for treating viral diseases. Clinically, it can be prepared as a pharmaceutical preparation for treating viral diseases. The Trollius extract is a large class of substances extracted from the dried flowers of the Trollius chinensis Bunge of the Ranunculaceae plant. Use the nasturtium extract as a medicinal ingredient to prepare pharmaceutical preparations for treating viral diseases, which can be prepared alone or in combination with acceptable excipients in the pharmaceutical process, and prepared according to conventional methods for oral administration, cavities, Drug formulations for mucosal, intramuscular or intravenous administration. The total flavonoid content of the Trollica chinensis extract is greater than 50%. The application of the Trollius flower extract in the preparation of the medicine for treating influenza virus H1N1; the application of the Trollius flower extract in the preparation of the medicine for the treatment of respiratory syncytial virus RSV;
本发明优点是，本发明发现了中药金莲花提取物治疗病毒性疾病的药效物质，为其临床应用提供了科学依据，使金莲花提取物可做为一个天然的抗病毒剂，具有无成瘾性、用量小的特点。金莲花提取物采用体外培养法筛选实验，结果表明：在625μg/ml的浓度时对甲型流感病毒A1的抑制率为75.6%、212.5μg/ml的浓度时对呼吸道合胞病毒RSV的抑制率为62.3%、39.07μg/ml的浓度时对腺病毒Adv3的抑制率为67.1%。金莲花提取物为一种从天然植物中提取得到的抗病毒物质，总黄酮含量在50%以上，是一种具有开发前景的抗病毒中药新药制剂。 The advantage of the present invention is that the present invention has discovered the effective substance of the traditional Chinese medicine Trollius flower extract for treating viral diseases, which provides a scientific basis for its clinical application, so that the Trollius flower extract can be used as a natural antiviral agent, and has no effect. The characteristics of addiction and small dosage. The extract of nasturtium lotus was screened by in vitro culture method, and the results showed that the inhibition rate of influenza A virus A1 was 75.6% at the concentration of 625 μg/ml, and the inhibition rate of respiratory syncytial virus RSV at the concentration of 212.5 μg/ml The inhibitory rate to adenovirus Adv3 was 67.1% at the concentration of 62.3% and 39.07μg/ml. Trollius extract is an antiviral substance extracted from natural plants, with a total flavonoid content of more than 50%. It is a new antiviral traditional Chinese medicine preparation with development prospects.
目前临床治疗病毒性疾病以化学合成和半合成的药物为主。但是，由于半个多世纪以来化学合成和半合成抗菌、抗病毒药物的广泛使用，已诱发病菌产生了耐药性菌株，且日渐增多条件性致病菌。加之其毒副作用较大等缺点，致使该类药物尽管作为一线治疗药、但已存在诸多不足。因此，抗病毒中药具有副作用小而具有广阔的应用前景，此新药的开发成功将产生重大的社会效益。 At present, the clinical treatment of viral diseases is mainly based on chemically synthesized and semi-synthesized drugs. However, due to the widespread use of chemically synthesized and semi-synthesized antibacterial and antiviral drugs for more than half a century, drug-resistant strains of pathogens have been induced, and opportunistic pathogenic bacteria are increasing day by day. Coupled with its disadvantages such as relatively large toxic and side effects, this type of drug has many shortcomings even though it is used as a first-line treatment drug. Therefore, the antiviral traditional Chinese medicine has few side effects and has broad application prospects, and the successful development of this new medicine will produce significant social benefits.
具体实施方式 Detailed ways
实施例1：金莲花提取物体外对流感病毒（H1N1）抑制活性检测 Example 1: In vitro detection of anti-influenza virus (H1N1) inhibitory activity of Trollius chinensis extract
材料与方法 Materials and Methods
1.体外宿主细胞：MDCK（狗肾传代细胞）细胞。 1. In vitro host cells: MDCK (dog kidney passage cells) cells.
2.病毒株：流感病毒A型（H1N1）（A3/京科/30/95）。 2. Virus strain: Influenza virus type A (H1N1) (A3/Jingke/30/95).
3. CPE法检测药物对细胞的毒性。 3. CPE method to detect the toxicity of drugs to cells.
4. CPE法(细胞病变)检测药物对流感病毒的抑制作用。 4. CPE method (cytopathic) to detect the inhibitory effect of drugs on influenza virus.
试验过程 Experimental procedure
（1）.药物的细胞毒性检测 (1). Cytotoxicity detection of drugs
MDCK细胞在96孔细胞培养板中培养形成单层后，加入不同浓度的药液，继续培养3天，用CPE法检测药物对MDCK细胞的毒性。 After MDCK cells were cultured in a 96-well cell culture plate to form a monolayer, different concentrations of drug solutions were added, and the culture was continued for 3 days. The toxicity of drugs to MDCK cells was detected by CPE method.
（2）. 药物抗流感病毒作用检测 (2). Detection of drug anti-influenza virus effect
MDCK细胞在96孔细胞培养板中培养形成单层后，用100TCID50的流感病毒感染细胞，再加入无毒浓度下的系列浓度的含药培养液，继续培养3天，经CPE法确定药物对流感病毒的抑制作用。 After the MDCK cells were cultured in a 96-well cell culture plate to form a monolayer, the cells were infected with 100 TCID50 of influenza virus, and then a series of drug-containing culture medium at a non-toxic concentration was added, and the culture was continued for 3 days. The effect of the drug on influenza was determined by the CPE method. Inhibition of the virus. the
实验结果： Experimental results:
实验结果表明金莲花提取物体对MDCK细胞的无毒浓度为625ug/ml，即（TC0=625ug/ml），金莲花提取物在625ug/ml的浓度时对流感病毒A型（H1N1）的抑制率分别为75.6%。 The experimental results show that the non-toxic concentration of Trollius extract to MDCK cells is 625ug/ml, that is (TC0=625ug/ml), the inhibitory rate of Trollius extract to influenza virus type A (H1N1) at the concentration of 625ug/ml 75.6% respectively.
实施例2：金莲花提取物体外对呼吸道合胞病毒（RSV）抑制活性检测 Example 2: In vitro detection of anti-respiratory syncytial virus (RSV) inhibitory activity of Trollius chinensis extract
材料与方法 Materials and Methods
1.体外宿主细胞：Vero（非洲绿猴肾细胞）细胞。 1. In vitro host cells: Vero (African green monkey kidney cells) cells.
2.病毒株：呼吸道合胞病毒（RSV）。 2. Virus strain: respiratory syncytial virus (RSV).
3. CPE法检测药物对细胞的毒性。 3. CPE method to detect the toxicity of drugs to cells.
4. CPE法(细胞病变)检测药物对呼吸道合胞病毒的抑制作用。 4. CPE method (cytopathic) to detect the inhibitory effect of drugs on respiratory syncytial virus.
试验过程 Experimental procedure
（1）.药物的细胞毒性检测 (1). Cytotoxicity detection of drugs
Vero细胞在96孔细胞培养板中培养形成单层后，加入不同浓度的药液，继续培养3天，用CPE法检测药物对Vero细胞的毒性。 After Vero cells were cultured in a 96-well cell culture plate to form a monolayer, drug solutions of different concentrations were added and cultured for 3 days, and the toxicity of drugs to Vero cells was detected by CPE method.
（2）. 药物抗呼吸道合胞病毒作用检测 (2). Detection of anti-respiratory syncytial virus effect of drugs
Vero细胞在96孔细胞培养板中培养形成单层后，用100TCID50的呼吸道合胞病毒（RSV）感染细胞，再加入无毒浓度下的系列浓度的含药培养液，继续培养3天，经CPE法确定药物对呼吸道合胞病毒（RSV）的抑制作用。 After Vero cells were cultured in a 96-well cell culture plate to form a monolayer, the cells were infected with 100 TCID50 of respiratory syncytial virus (RSV), and then a series of non-toxic concentration of drug-containing culture solution was added, and the culture was continued for 3 days, and then the cells were treated with CPE. method to determine the inhibitory effect of drugs on respiratory syncytial virus (RSV).
实验结果： Experimental results:
实验结果表明金莲花提取物体对Vero细胞的无毒浓度为212.5ug/ml，即（TC0=625ug/ml），金莲花提取物在212.5ug/ml的浓度时对呼吸道合胞病毒（RSV）的抑制率分别为62.3%。 The experimental results show that the non-toxic concentration of Trollius extract to Vero cells is 212.5ug/ml, that is (TC0=625ug/ml), and the concentration of Trollius extract on Respiratory Syncytial Virus (RSV) is 212.5ug/ml. The inhibition rates were 62.3%.
实施例3：金莲花提取物体外对腺病毒（Adv3）抑制活性检测 Example 3: In vitro detection of the inhibitory activity of Trollius chinensis extracts against adenovirus (Adv3)
材料与方法 Materials and Methods
1.体外宿主细胞：Vero（非洲绿猴肾细胞）细胞。 1. In vitro host cells: Vero (African green monkey kidney cells) cells.
2.病毒株：腺病毒（Adv3）。 2. Virus strain: adenovirus (Adv 3 ).
3. CPE法检测药物对细胞的毒性。 3. CPE method to detect the toxicity of drugs to cells.
4. CPE法(细胞病变)检测药物对腺病毒的抑制作用。 4. CPE method (cytopathic) to detect the inhibitory effect of drugs on adenovirus.
试验过程 Experimental procedure
（1）.药物的细胞毒性检测 (1). Cytotoxicity detection of drugs
Vero细胞在96孔细胞培养板中培养形成单层后，加入不同浓度的药液，继续培养3天，用CPE法检测药物对Vero细胞的毒性。 After Vero cells were cultured in a 96-well cell culture plate to form a monolayer, drug solutions of different concentrations were added and cultured for 3 days, and the toxicity of drugs to Vero cells was detected by CPE method.
（2）. 药物抗腺病毒作用检测 (2). Drug anti-adenovirus detection
Vero细胞在96孔细胞培养板中培养形成单层后，用100TCID50的腺病毒（Adv3）感染细胞，再加入无毒浓度下的系列浓度的含药培养液，继续培养3天，经CPE法确定药物对腺病毒（Adv3）的抑制作用。 After Vero cells were cultured in a 96-well cell culture plate to form a monolayer, the cells were infected with 100 TCID50 of adenovirus (Adv 3 ), and then a series of drug-containing culture medium at a non-toxic concentration was added, and the culture was continued for 3 days. To determine the inhibitory effect of drugs on adenovirus (Adv 3 ).
实验结果： Experimental results:
实验结果表明金莲花提取物体对Vero细胞的无毒浓度为39.07ug/ml，即（TC0=39.07ug/ml），金莲花提取物在39.07ug/ml的浓度时对腺病毒的抑制率分别为67.1%。 The experimental results show that the non-toxic concentration of Trollius extract to Vero cells is 39.07ug/ml, that is (TC0=39.07ug/ml), and the inhibition rate of Trollius extract on adenovirus at the concentration of 39.07ug/ml is respectively 67.1%.
实施例4：Example 4:
取金莲花提取物25g，加入淀粉细粉200g，用等量递增法混匀，加入滑石粉5g，硬脂酸镁2g，混匀，用85%乙醇制粒，干燥，整粒，压制成1000片，每日3次，每次2片。 Take 25g of nasturtium extract, add 200g of starch fine powder, mix evenly by equal increment method, add 5g of talcum powder, 2g of magnesium stearate, mix evenly, granulate with 85% ethanol, dry, granulate, press into 1000 Tablets, 3 times a day, 2 tablets each time.
实施例5： Example 5:
取金莲花提取物25g，加入淀粉细粉150g，滑石粉10g，硬脂酸镁5g，混匀，制粒，干燥，装入胶囊，制成1000粒，每日3次，每次2片。 Get 25g of nasturtium extract, add 150g of fine starch powder, 10g of talcum powder, 5g of magnesium stearate, mix well, granulate, dry, pack into capsules, make 1000 capsules, 3 times a day, 2 tablets each time.
实施例6： Embodiment 6:
取金莲花提取物50g，加入微晶纤维素250g，交联聚乙烯吡咯烷酮50g，混匀，加入90乙醇制粒，干燥，整粒，加入羧甲基淀粉钠10g，硬脂酸镁5g，混匀，压制成1000片，每日服用3次，每次1片。 Take 50g of nasturtium extract, add 250g of microcrystalline cellulose, 50g of cross-linked polyvinylpyrrolidone, mix well, add 90% ethanol to granulate, dry, granulate, add 10g of sodium carboxymethyl starch, 5g of magnesium stearate, mix Evenly, compressed into 1000 tablets, take 3 times a day, 1 tablet each time.
实施例7： Embodiment 7:
取金莲花提取物50g，甘露醇150g，加注射用水2000ml，搅拌使溶解，用1M NaOH溶液调pH至9.60～10.0，搅拌使溶解，滤过，滤液按1％溶液体积量加入活性炭，在70～80℃下加热30分钟，粗滤过，管道及容器用400mL注射用水洗涤，加水到2500ml，药液经0.22μm无菌过滤器过滤，灌装，冻干，压塞，轧盖，制成1000瓶，即得。 Take 50g of nasturtium extract, 150g of mannitol, add 2000ml of water for injection, stir to dissolve, adjust the pH to 9.60-10.0 with 1M NaOH solution, stir to dissolve, filter, add activated carbon to the filtrate according to the volume of 1% solution, and dissolve it at 70 Heat at ~80°C for 30 minutes, coarsely filter, wash the pipeline and container with 400mL water for injection, add water to 2500ml, filter the liquid medicine through a 0.22μm sterile filter, fill, freeze-dry, press stopper, and cap it. 1000 bottles, that is.
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| CN1660864A (en) * | 2004-12-16 | 2005-08-31 | 中国人民解放军第二军医大学 | The preparation process of total flavonoids and several high-purity medicinal substances from Trollius breviflora |
| CN101234147A (en) * | 2008-03-06 | 2008-08-06 | 江西汇仁药业有限公司 | Preparation method of total flavonoids from Trollus chinensis for injection |
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| CN101234147A (en) * | 2008-03-06 | 2008-08-06 | 江西汇仁药业有限公司 | Preparation method of total flavonoids from Trollus chinensis for injection |
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| Title |
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| 金莲花中荭草苷抗病毒活性及作用机理研究;张宏坡;《中国优秀硕士学位论文全文数据库 医药卫生科技辑》;20110715;第6页1.1节、第14页2.1.2节、第21页2.3节以及第31页表4.5 * |
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