TWI624264B - 南洋山蘇水萃物的用途 - Google Patents
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- 239000000284 extract Substances 0.000 title claims abstract description 50
- 241001312267 Asplenium australasicum Species 0.000 title description 2
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- 241000725619 Dengue virus Species 0.000 description 16
- 230000000694 effects Effects 0.000 description 11
- 201000007270 liver cancer Diseases 0.000 description 9
- 208000014018 liver neoplasm Diseases 0.000 description 9
- 208000001490 Dengue Diseases 0.000 description 8
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- 230000010076 replication Effects 0.000 description 6
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- 208000009714 Severe Dengue Diseases 0.000 description 4
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- 241000699666 Mus <mouse, genus> Species 0.000 description 3
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- 102100031181 Glyceraldehyde-3-phosphate dehydrogenase Human genes 0.000 description 2
- 238000011529 RT qPCR Methods 0.000 description 2
- 241000700159 Rattus Species 0.000 description 2
- 239000006286 aqueous extract Substances 0.000 description 2
- 201000002950 dengue hemorrhagic fever Diseases 0.000 description 2
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- 108090000623 proteins and genes Proteins 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
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Abstract
一種南洋山蘇水萃物的用途,係用以製備治療登革熱病毒感染之藥物,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。
Description
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治療登革熱病毒感染之藥物的用途。
登革熱病毒感染可能導致登革熱(dengue fever)、出血性登革熱(dengue hemorrhagic fever,簡稱DHF)或登革熱休克症候群(dengue shock syndrome,簡稱DSS);惟,目前尚無任何可以治療登革熱病毒感染之抗病毒藥物,而臨床上僅能夠以支持性療法(supportive therapy)舒緩患者之病徵,故,提供一種可以治療登革熱病毒感染之藥物確實刻不容緩。
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,應用於製備治療登革熱病毒感染之藥物者。
本發明之南洋山蘇水萃物的用途,係應用於製備治療登革熱病毒感染之藥物,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,較佳地,該南洋山蘇水萃物係能夠以口服方式投予一所需個體,且該南洋山蘇水萃物之每天投予該所需個體之劑量為2~6毫升/公斤體重;如此藉由萃取自南洋山蘇之活性成分,可以抑制登革熱病毒之複製,因而可以應用於製備治療登革熱病毒感染之藥物,為本發明之功效。
第1a圖:試驗(A)中,投予本發明南洋山蘇水萃物對經登革熱感染的人類肝癌細胞株之非結構蛋白NS5的RNA含量的影響結果柱狀圖。
第1b圖:試驗(A)中,投予本發明南洋山蘇水萃物對經登革熱感染的人類肝癌細胞株之非結構蛋白NS5的蛋白質含量的影響結果影像圖。
第2圖:試驗(B)中,投予本發明南洋山蘇水萃物對經登革熱感染的人類肝癌細胞株之非結構蛋白NS5的RNA含量的影響結果柱狀圖。
第3a圖:試驗(C)中,投予本發明南洋山蘇水萃物對經登革熱感染的乳鼠之臨床成績的影響結果折線圖。
第3b圖:試驗(C)中,投予本發明南洋山蘇水萃物對經登革熱感染的乳鼠之體重的影響結果折線圖。
第3c圖:試驗(C)中,投予本發明南洋山蘇水萃物對經登革熱感染的乳鼠之存活率的影響結果折線圖。
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。
本發明之南洋山蘇水萃物,係可以抑制登革熱病毒之複製,因而可以應用於製備治療登革熱病毒感染之藥物,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。
又,本發明之南洋山蘇水萃物係可以供投予一所需個體,例如以2~6毫升/公斤體重之劑量投予該所需個體,使該南洋山蘇水萃物係所含有之活性成分得以於該所需個體體內作用,抑制登革熱病毒入侵細胞及於細胞內的複製作用。
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。
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。
舉例而言,每4.8公斤之南洋山蘇樣品係能夠混合該萃取溶劑,使二者總體積達80公升,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。
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。
為證實本發明之南洋山蘇水萃物係可以抑制登革熱病毒入侵細胞及於細胞內的複製作用,遂將每4.8公斤之南洋山蘇樣品萃取所得之南洋山蘇粗萃液,經過減壓濃縮及冷凍乾燥獲得共800克之南洋山蘇水萃物粉末,另將5克之南洋山蘇水萃物粉末溶於20毫升之水後,進行以下試驗。
(A)南洋山蘇水萃物對登革熱病毒之複製作用的影響
請參照第1表所示,本試驗係以經登革熱病毒感染之人類肝癌細胞株Huh-7進行試驗,以未處理該南洋山蘇水萃物之肝癌細胞作為控制組(第A0組),另分別以5μL/mL(第A1組)、10μL/mL(第A2組)、25μL/mL(第A3組)、50μL/mL(第A4組)之濃度的南洋山蘇水萃物處理該肝癌細胞三天後,分別收取總RNA及總蛋白質,以反轉錄即時定量聚合酶連鎖反應(RT-qPCR)偵測非結構蛋白NS5之RNA含量,另以西方墨點法偵測非結構蛋白NS2B之蛋白質含量,並以GAPDH之RNA含量及蛋白質含量作為內控制組。
請參照第1a及1b圖所示,第A1~A4組之非結構蛋白NS5的RNA及蛋白質含量均有減少,顯示該南洋山蘇水萃物可以抑制登革熱病毒於細胞內的複製作用。
(B)南洋山蘇水萃物對登革熱病毒之入侵的影響
請參照第2表所示,本試驗同以經登革熱病毒感染之人類肝癌細胞株Huh-7進行試驗,以未處理該南洋山蘇水萃物之肝癌細胞作為控制組(第B0組),另分別以5μL/mL(第B1組)、10μL/mL(第B2組)、25μL/mL(第B3組)、50μL/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,收取總RNA,以反轉錄即時定量聚合酶連鎖反應(RT-qPCR)偵測非結構蛋白NS5之RNA含量,並以GAPDH之RNA含量作為內控制組。
請參照第2圖所示,第B1~B4組之非結構蛋白NS5的RNA含量均有減少,顯示該南洋山蘇水萃物可以抑制登革熱病毒之進入細胞階段,防止登革熱病毒入侵細胞。
(C)南洋山蘇水萃物對受登革熱病毒感染之乳鼠的影響
請參照第3表所示,本試驗係以經登革熱病毒感染之ICR乳鼠進行試驗,以腦內注射之方式使乳鼠感染登革熱病毒,並於感染後的第1天以腹腔注射之方式投予該南洋山蘇水萃物;於試驗過程中每天觀察該乳鼠之外觀、活動行為,評估其臨床成績,另紀錄該乳鼠之體重增長,以及該乳鼠之存活率。
第3表、本試驗各組條件。
請參照第3a圖所示,感染登革熱病毒之第C1組乳鼠的臨床成績呈上升勢,顯示其生理狀態漸漸惡化,而同時投予該南洋山蘇水萃物之第C2、C3組乳鼠的臨床成績的上升趨勢則較為緩和,顯示該南洋山蘇水萃物可以改善感染登革熱病毒之症狀;續請參照第3b圖所示,該南洋山蘇水萃物之投予亦可以改善感染登革熱病毒後的體重下降現象;另請參照第3c圖所示,同時投予該南洋山蘇水萃物更可以有效改善感染登革熱病毒所造成之死亡現象。
綜合上述,本發明之南洋山蘇水萃物途係藉由萃取自南洋山蘇之活性成分,可以有效抑制登革熱病毒入侵細胞及於細胞內的複製作用,清除體內之登革熱病毒,因而可以應用於製備治療登革熱病毒感染之藥物,為本發明之功效。
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。
Claims (3)
- 一種南洋山蘇水萃物的用途,係應用於製備治療登革熱病毒感染之藥物,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。
- 如申請專利範圍第1項所述之南洋山蘇水萃物的用途,其中,該南洋山蘇水萃物係以口服方式投予一所需個體。
- 如申請專利範圍第2項所述之南洋山蘇水萃物的用途,其中,該南洋山蘇水萃物之每天投予該所需個體之劑量為2~6毫升/公斤體重。
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Application Number | Priority Date | Filing Date | Title |
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TW106127365A TWI624264B (zh) | 2017-08-11 | 2017-08-11 | 南洋山蘇水萃物的用途 |
PCT/US2018/046194 WO2019032951A1 (en) | 2017-08-11 | 2018-08-10 | METHOD OF TREATING INFECTION WITH DENGUE VIRUS |
CN201810909633.2A CN109381495A (zh) | 2017-08-11 | 2018-08-10 | 南洋山苏水萃物的用途 |
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Lih-Shiuh Lai et al,"Chemical compositions and some physical properties of the water and alkali-extracted mucilage from the young fronds of Asplenium australasicum (J. Sm.) Hook", Food Hydrocolloids, 2012, 26(2): 344-349. * |
Lih-Shiuh Lai et al,"Chemical compositions and some physical properties of the water and alkali-extracted mucilage from the young fronds of Asplenium australasicum (J. Sm.) Hook", Food Hydrocolloids, 2012, 26(2): 344-349.。 |
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