TWI611806B - A use of an extract of asplenium australasicum (j. sm.) hook. - Google Patents

A use of an extract of asplenium australasicum (j. sm.) hook. Download PDF

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TWI611806B
TWI611806B TW105137326A TW105137326A TWI611806B TW I611806 B TWI611806 B TW I611806B TW 105137326 A TW105137326 A TW 105137326A TW 105137326 A TW105137326 A TW 105137326A TW I611806 B TWI611806 B TW I611806B
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hepatitis
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nanyangshansu
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TW201818951A (en
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黃昆暄
陳景川
黃瑞齡
何淑禎
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景鑫生物科技股份有限公司
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    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
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    • A61P1/16Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine

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Abstract

一種南洋山蘇水萃物的用途,係用以製備C型肝炎治療用的藥物,其中,該南洋山蘇水萃物係以水作為一萃取溶劑,於115~125℃之溫度、0.35~0.5Kg/cm2G之壓力下萃取一南洋山蘇樣品1.5~3小時所獲得。 The utility model relates to a water extract of Nanyang Mountain Soda extract, which is used for preparing a medicine for treating hepatitis C. The water extract of Nanyang Mountain Sophora is used as an extraction solvent at a temperature of 115 to 125 ° C and a temperature of 0.35 to 0.5. It was obtained by extracting a sample of Nanyangshansu under the pressure of Kg / cm 2 G for 1.5 ~ 3 hours.

Description

南洋山蘇水萃物的用途 Uses of Soyang Extract from Nanyang Mountain

本發明係關於一種南洋山蘇水萃物的用途,特別關於一種南洋山蘇水萃物應用於製備C型肝炎治療用藥物的用途。 The present invention relates to the use of a water extract from Nanyang Mountain Perilla, in particular to the use of a water extract from Nanyang Mountain Perilla to prepare a medicine for treating hepatitis C.

C型肝炎(hepatitis C)係為由C型肝炎病毒(hepatitis C virus)所造成的感染性疾病(infectious disease),在感染初期並沒有任何症狀,偶爾會有發燒(fever)、小便黃赤(dark urine)、腹痛(abdominal pain)或黃疸(yellow tinged skin)等症狀,並且在多年後演變為肝病(liver disease)甚至肝硬化(cirrhosis),或其他的併發症(complication),例如肝功能衰竭(liver failure)、肝癌(liver cancer)、食道靜脈曲張或胃底靜脈曲張(esophageal and gastric varices)。 Hepatitis C is an infectious disease caused by the Hepatitis C virus. It has no symptoms in the early stages of infection, and occasionally has fever and yellow urine. symptoms such as dark urine, abdominal pain, or yellow tinged skin, and evolved to liver disease or even cirrhosis, or other complications, such as liver failure, over many years (liver failure), liver cancer, esophageal varices, or esophageal and gastric varices.

習用常會以干擾素(peginterferon)搭配利巴韋林(ribavirin)的合併療法,治癒率(cure rate)大約僅50%且有強烈的副作用,是以,確實仍有必要發展新的抗C型肝炎藥物。 The combination therapy of interferon (peginterferon) and ribavirin often has a cure rate of about 50% and has strong side effects. Therefore, it is still necessary to develop new anti-hepatitis C drug.

為解決上述問題,本發明提供一種南洋山蘇水萃物的用途,係將萃取自南洋山蘇之活性成分,應用於製備C型肝炎治療用的藥物者。 In order to solve the above problems, the present invention provides the use of a water extract from Nanyang Mountain Perilla, which is an active ingredient extracted from Nanyang Mountain Perilla, and is used to prepare a medicine for treating hepatitis C.

本發明之南洋山蘇水萃物的用途,係應用於製備C型肝炎治療用的藥物,其中,該南洋山蘇水萃物係水於115~125℃之溫度、0.35~0.5Kg/cm2G之壓力下萃取1.5~3小時所獲得;如此藉由萃取自南洋山蘇 之活性成分,可以抑制C型肝炎病毒之複製,因而可以應用於製備C型肝炎治療用的藥物,為本發明之功效。 The use of the Nanyang Mountain Soda extract is used for preparing medicine for treating hepatitis C, wherein the Nanyang Mountain Souce extract is water at a temperature of 115-125 ° C and 0.35 ~ 0.5Kg / cm 2 It is obtained by extracting for 1.5 ~ 3 hours under the pressure of G. In this way, by extracting the active ingredients from Nanyangshansu, it can inhibit the replication of hepatitis C virus, so it can be applied to the preparation of drugs for the treatment of hepatitis C. efficacy.

第1圖:投予本發明南洋山蘇水萃物對含有HCV複製子的人類肝癌細胞株之存活率的影響結果折線圖。 FIG. 1 is a line chart showing the results of the effect of administering the extract of Nanyang Mountain Solanum nigra on the survival rate of human liver cancer cell lines containing HCV replicons.

第2a圖:投予本發明南洋山蘇水萃物對含有HCV複製子的人類肝癌細胞株之非結構蛋白NS5B的蛋白質含量的影響結果影像圖。 Fig. 2a is an image showing the results of the effect of administering the extract of Nanyang Mountain Solanum nigrum L. to the protein content of non-structural protein NS5B of a human liver cancer cell line containing HCV replicon.

第2b圖:投予本發明南洋山蘇水萃物對含有HCV複製子的人類肝癌細胞株之非結構蛋白NS5B的RNA含量的影響結果柱狀圖。 Fig. 2b: Histogram of the results of the effect of administering the extract of Nanyang Mountain Solanum officinalis on the RNA content of nonstructural protein NS5B of a human liver cancer cell line containing HCV replicon.

第3圖:投予本發明南洋山蘇水萃物對含有HCV複製子的人類肝癌細胞株之非結構蛋白NS5B的蛋白質含量的影響結果影像圖。 FIG. 3 is an image showing the result of the effect of administering the extract of Nanyang Mountain Solanum nigrum L. to the protein content of non-structural protein NS5B of a human liver cancer cell line containing HCV replicon.

第4圖:投予本發明南洋山蘇水萃物對受C型肝炎感染之人類肝癌細胞株之非結構蛋白NS5B的RNA含量的影響結果柱狀圖。 Figure 4: Histogram of the effect of the administration of the extract of Nanyang Mountain Solanum nigrum L. on the RNA content of the non-structural protein NS5B of a human liver cancer cell line infected with hepatitis C.

為讓本發明之上述及其他目的、特徵及優點能更明顯易懂,下文特舉本發明之較佳實施例,並配合所附圖式,作詳細說明如下:本發明所述之「南洋山蘇(Asplenium australasicum(J.Sm.)Hook.)」,係指鐵角蕨科(Aspleniaceae)鐵角蕨屬(Asplenium)的植物,其新生嫩葉(young emerging frond)常被作為食用菜葉。 In order to make the above and other objects, features, and advantages of the present invention more comprehensible, the following describes the preferred embodiments of the present invention and the accompanying drawings in detail, as follows: "Su ( Asplenium australasicum (J.Sm.) Hook.)" Refers to a plant of the genus Asplenium of the Aspleniaceae family, and its young emerging frond is often used as a vegetable leaf.

本發明之南洋山蘇水萃物,係可以抑制C型肝炎病毒之複製,因而可以應用於製備C型肝炎治療用的藥物,該南洋山蘇水萃物與醫藥學上可以接受之載劑或賦形劑組合形成一醫藥組合物,其中,該南洋山蘇水萃物係可以製備成任何方便食用之型式,如錠劑、膠囊、粉劑、粒劑或液劑等,或者將該南洋山蘇水萃物與其他食品或飲料組合,以適於食用 之樣態供生物體以口服方式服用。 The water extract of Nanyang Mountain Sophora according to the present invention can inhibit the replication of Hepatitis C virus, so it can be applied to the preparation of drugs for the treatment of Hepatitis C. The excipients are combined to form a pharmaceutical composition, wherein the Nanyangshansu water extract can be prepared into any convenient form, such as tablets, capsules, powders, granules or liquids, etc. Water extracts are combined with other foods or beverages to make them suitable for consumption This state is for the organism to take orally.

其中,該南洋山蘇水萃物較佳係可以藉由一包含以下步驟之方法所製得:提供一南洋山蘇樣品;以水作為一萃取溶劑萃取該南洋山蘇樣品;及將南洋山蘇粗萃液進行濃縮,以獲得該南洋山蘇水萃物。 Among them, the Nanyangshansu water extract is preferably prepared by a method including the following steps: providing a Nanyangshansu sample; using water as an extraction solvent to extract the Nanyangshansu sample; and The crude extract was concentrated to obtain the Nanyangshansu extract.

詳而言之,該南洋山蘇樣品係可以為南洋山蘇之成熟老葉(三年以上,背面分布孢子),較佳係可以預先將該南洋山蘇樣品於110~120℃之溫度下進行乾燥(將100克南洋山蘇樣品乾燥至20克),得到一南洋山蘇乾燥物;此外,該南洋山蘇樣品亦可以預先碎成粉粒(粒徑約為1~2mm),以增加該南洋山蘇樣品與該萃取溶劑之接觸表面積,藉此提升後續萃取之萃取效率。 In detail, the Nanyangshansu sample can be the mature old leaves of Nanyangshansu (more than three years, with spores distributed on the back). Preferably, the Nanyangshansu sample can be processed in advance at a temperature of 110 ~ 120 ° C. Drying (drying 100g Nanyangshansu sample to 20g) to obtain a dried Nanyangshansu sample; In addition, the Nanyangshansu sample can also be crushed into powder particles (particle size about 1 ~ 2mm) in advance to increase the The contact surface area of the Nanyang Shansu sample and the extraction solvent improves the extraction efficiency of subsequent extraction.

舉例而言,每4.8kg之南洋山蘇樣品係能夠混合該萃取溶劑,使二者總體積達80L,並於115~125℃之溫度、0.35~0.5Kg/cm2G之壓力下進行萃取1.5~3小時,上述萃取亦可以重複數次,使該南洋山蘇樣品所富含之活性成分可以完整溶出於該萃取溶劑,此為本發明所屬技術領域中具有通常知識者所廣泛應用,在此不加以贅述。 For example, every 4.8kg of Nanyangshansu sample can be mixed with the extraction solvent so that the total volume of the two reaches 80L, and the extraction is performed at a temperature of 115 to 125 ° C and a pressure of 0.35 to 0.5 Kg / cm 2 G. ~ 3 hours, the above extraction can also be repeated several times, so that the active ingredients contained in the Nanyangshansu sample can be completely dissolved in the extraction solvent. This is widely used by those with ordinary knowledge in the technical field to which the present invention belongs. Do not repeat them.

前述之南洋山蘇粗萃液可以經過減壓濃縮及冷凍乾燥,以獲得該南洋山蘇水萃物,藉由此一程序,係可以使該南洋山蘇水萃物之活性成分更加濃縮,是以僅需使用少量之該南洋山蘇水萃物即可以發揮最佳療效。 The aforementioned crude extract of Nanyang Mountain Perilla can be concentrated under reduced pressure and freeze-dried to obtain the Nanyang Mountain Perilla water extract. With this procedure, the active ingredients of the Nanyang Perilla water extract can be more concentrated. Therefore, only a small amount of the Nanyangshan sour water extract can be used to achieve the best effect.

為證實本發明之南洋山蘇水萃物係可以抑制C型肝炎病毒之複製,遂進行以下試驗。 In order to confirm that the water extract of Nanyang Mountain Sorbaria according to the present invention can inhibit the replication of hepatitis C virus, the following tests were performed.

(A)南洋山蘇水萃物對肝癌細胞之細胞毒性 (A) Cytotoxicity of water extract from Nanyang Mountain Sophora to liver cancer cells

請參照第1表所示,本試驗係選用含有HCV複製子的人類肝癌細胞株Ava5進行,係分別以0、10、25、50、75、100、150、200、300及400μL/mL之南洋山蘇水萃物處理該肝癌細胞,於三天後以細胞毒 性測試套組(CellTiter 96 AQueous One Solution Proliferation Assay)觀察該肝癌細胞之細胞存活率,以未處理南洋山蘇水萃物之肝癌細胞的細胞存活率作為100%。 Please refer to Table 1. This test was performed using the human liver cancer cell line Ava5 containing HCV replicons. The test was performed at 0, 10, 25, 50, 75, 100, 150, 200, 300, and 400 μL / mL Nanyang. The hepatocellular carcinoma cells were treated with Sansu water extract, and the cell survival rate of the hepatocellular carcinoma cells was observed with a cytotoxicity test kit (CellTiter 96 AQ ueous One Solution Proliferation Assay) three days later. The cell viability of the cells was taken as 100%.

Figure TWI611806BD00001
Figure TWI611806BD00001

請參照第1圖所示,南洋山蘇水萃物於濃度為50μL/mL以下不具明顯毒性。 Please refer to Figure 1. Nanyang Mountain Soy extract has no obvious toxicity at a concentration of 50 μL / mL or less.

(B)南洋山蘇水萃物對C型肝炎病毒複製作用的影響 (B) Effect of extracts from Nanyang Mountain Soda on replication of hepatitis C virus

如第2表所示,本試驗同樣以含有HCV複製子的人類肝癌細胞株Ava5進行試驗,以未處理該南洋山蘇水萃物之肝癌細胞作為正控制組(第B0組),另分別以10μL/mL(第B1組)、25μL/mL(第B2組)、50μL/mL(第B3組)、75μL/mL(第B4組)之濃度的南洋山蘇水萃物處理該肝癌細胞三天後,將各組細胞裂解處理後,以西方墨點法分析各組細胞中非結構蛋白NS5B之蛋白質含量。 As shown in Table 2, this test was also performed on the human liver cancer cell line Ava5 containing HCV replicon, and the liver cancer cells that were not treated with the Nanyangshan thaliana water extract were used as the positive control group (Group B0). The liver cancer cells were treated with a water extract of Nanyang Mountain Solanum at a concentration of 10 μL / mL (Group B1), 25 μL / mL (Group B2), 50 μL / mL (Group B3), and 75 μL / mL (Group B4) for three days. Then, after lysing the cells of each group, the protein content of non-structural protein NS5B in each group of cells was analyzed by Western blot method.

Figure TWI611806BD00002
Figure TWI611806BD00002

請參照第2a圖所示,第B1~B3組之非結構蛋白NS5B的蛋白質含量均有減少,顯示該南洋山蘇水萃物可以抑制C型肝炎病毒之複製作用。 Please refer to Figure 2a, the protein content of the non-structural proteins NS5B in groups B1 to B3 has been reduced, indicating that the Nanyang Mountain Sophora extract can inhibit the replication of hepatitis C virus.

續以即時聚合酶鏈式反應偵測第B0~B4組細胞中非結構蛋白NS5B之RNA含量,如第2b圖所示,第B1~B4組之非結構蛋白NS5B之RNA含量亦有減少,同樣顯示該南洋山蘇水萃物可以抑制C型肝炎病毒之複製作用。 Continue to use real-time polymerase chain reaction to detect the RNA content of non-structural protein NS5B in cells of groups B0 ~ B4. As shown in Figure 2b, the RNA content of non-structural protein NS5B of groups B1 to B4 also decreased. It has been shown that the Nanyangshansu water extract can inhibit the replication of hepatitis C virus.

(C)長時間處理南洋山蘇水萃物的影響 (C) The effect of long-term treatment of the extracts from Nanyang Mountain

如第3表所示,本試驗以依上述之流程進行,惟改於以不同濃度的南洋山蘇水萃物處理該肝癌細胞六天後,才分析各組細胞中非結構蛋白NS5B之蛋白質含量。 As shown in Table 3, this test was performed according to the above procedure, but the protein content of the non-structural protein NS5B in each group of cells was analyzed after changing the liver cancer cells for six days with different concentrations of Nanyangshansu water extracts. .

Figure TWI611806BD00003
Figure TWI611806BD00003

請參照第3圖所示,第C2組之非結構蛋白NS5B的蛋白質 含量有明顯減少,顯示該南洋山蘇水萃物可以抑制C型肝炎病毒之複製作用。 Please refer to Figure 3, the protein of group C2 non-structural protein NS5B The content was significantly reduced, showing that the Nanyang Mountain Solanum extract can inhibit the replication of hepatitis C virus.

(D)南洋山蘇水萃物對C型肝炎病毒複製作用的影響 (D) Effect of extracts from Nanyang Mountain Soda on replication of hepatitis C virus

請參照第4表所示,本試驗係選用人類肝癌細胞株Huh-7進行,於C型肝炎病毒感染(MOI=0.1)後,分別以0、10、25、50μL/mL之南洋山蘇水萃物處理該肝癌細胞,於三天後以即時聚合酶鏈式反應偵測第D0~D4組細胞中非結構蛋白NS5B之RNA含量。 Please refer to Table 4. This test was performed using human liver cancer cell line Huh-7. After Hepatitis C virus infection (MOI = 0.1), the test was performed at 0, 10, 25, and 50 μL / mL. The liver cancer cells were treated with the extract, and the RNA content of the non-structural protein NS5B in the cells of groups D0 to D4 was detected by real-time polymerase chain reaction after three days.

Figure TWI611806BD00004
Figure TWI611806BD00004

請參照第4圖所示,第D2~D4組之非結構蛋白NS5B之RNA含量亦有減少,顯示該南洋山蘇水萃物可以抑制C型肝炎病毒之複製作用。 Please refer to Figure 4, the RNA content of the non-structural protein NS5B of groups D2 to D4 also decreased, indicating that the Nanyang Mountain Solan extract can inhibit the replication of hepatitis C virus.

綜合上述試驗結果,南洋山蘇水萃物可以有效抑制C型肝炎病毒株之複製,因而可以清除生物體內之C型肝炎病毒,以達到治療C型肝炎之功效。 Based on the results of the above tests, the extract of Nanyangshan sour water can effectively inhibit the replication of the hepatitis C virus strain, and thus can eliminate the hepatitis C virus in the body to achieve the effect of treating hepatitis C.

此外,1名罹患良性攝護腺肥大,且同時擁有C型肝炎15年病史之患者,於服用本發明之南洋山蘇水萃物3個月後,肝指數GPT已 成功降至20KU/mL以下;其他10名C型肝炎患者(包含男、女)於服用本發明之南洋山蘇水萃物4個月後,肝指數GPT已成功降至正常值。 In addition, a patient suffering from benign prostatic hypertrophy who also had a 15-year history of hepatitis C had a liver index GPT of 3 months after taking the Nanyangshan Sushui extract of the present invention. Successfully reduced to below 20KU / mL; 10 other patients with hepatitis C (including male and female) took the Nanyangshan Sushui extract of the present invention for 4 months, and the liver index GPT had been successfully reduced to normal value.

綜合上述,本發明之南洋山蘇水萃物的用途係藉由萃取自南洋山蘇之活性成分,可以抑制C型肝炎病毒之複製,清除體內之C型肝炎病毒,因而可以應用於製備C型肝炎治療用的藥物,為本發明之功效。 In summary, the use of the water extract of Nanyang Mountain Perilla in the present invention is to extract the active ingredients of Nanyang Mountain Perilla, which can inhibit the replication of Hepatitis C virus and clear the hepatitis C virus in the body, so it can be applied to the preparation of Type C The medicine for treating hepatitis is the effect of the present invention.

雖然本發明已利用上述較佳實施例揭示,然其並非用以限定本發明,任何熟習此技藝者在不脫離本發明之精神和範圍之內,相對上述實施例進行各種更動與修改仍屬本發明所保護之技術範疇,因此本發明之保護範圍當視後附之申請專利範圍所界定者為準。 Although the present invention has been disclosed using the above-mentioned preferred embodiments, it is not intended to limit the present invention. Anyone skilled in the art can make various changes and modifications to the above embodiments without departing from the spirit and scope of the present invention. The technical scope protected by the invention, so the scope of protection of the present invention shall be determined by the scope of the appended patent application.

Claims (1)

一種南洋山蘇水萃物的用途,係應用於製備C型肝炎治療用藥物,其中,該南洋山蘇水萃物係以水作為一萃取溶劑,於115~125℃之溫度、0.35~0.5Kg/cm2G之壓力下萃取一南洋山蘇樣品1.5~3小時所獲得。 The utility model relates to a water extract of Nanyang Mountain Perilla, which is used for preparing a medicine for treating hepatitis C. The water extract of Nanyang Mountain Perilla uses water as an extraction solvent at a temperature of 115-125 ° C and 0.35 ~ 0.5Kg. It was obtained by extracting a sample of Nanyangshansu under a pressure of 1.5 g / cm 2 G for 1.5 ~ 3 hours.
TW105137326A 2016-11-15 2016-11-15 A use of an extract of asplenium australasicum (j. sm.) hook. TWI611806B (en)

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TWI595881B (en) * 2016-11-15 2017-08-21 景鑫生物科技股份有限公司 A use of an extract of asplenium australasicum (j. sm.) hook.

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Publication number Priority date Publication date Assignee Title
CN110869087A (en) * 2017-06-06 2020-03-06 景鑫生物科技股份有限公司 Method for preventing and treating chemical liver injury

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