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TW201615184A - 莫諾苯宗藥物應用於癌症治療 - Google Patents

莫諾苯宗藥物應用於癌症治療 Download PDF

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TW201615184A
TW201615184A TW104134950A TW104134950A TW201615184A TW 201615184 A TW201615184 A TW 201615184A TW 104134950 A TW104134950 A TW 104134950A TW 104134950 A TW104134950 A TW 104134950A TW 201615184 A TW201615184 A TW 201615184A
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cancer
monobenzone
cells
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drug
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陳丘泓
莊秀美
魏宗德
蕭乃文
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朗齊生物醫學股份有限公司
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Abstract

本發明提供莫諾苯宗藥物的新臨床之應用。莫諾苯宗藥物是經過衛生署通過核准上市的局部使用的脫色劑藥物。本發明提供莫諾苯宗藥物有效的抑制不同類型的癌細胞。

Description

莫諾苯宗藥物應用於癌症治療
本發明係為莫諾苯宗藥物的新適應症之應用，尤其該莫諾苯宗藥物具有抑制多種癌症之用途。
莫諾苯宗(Monobenzone)是一種局部使用的脫色劑，用於治療色素沉著過度，如各種色斑、老年斑、黑色素瘤等，效果很明顯。它能分解皮膚中的黑色素，阻止皮膚中黑色素的生成，使皮膚恢復健康的色澤，而並不破壞黑色素細胞，毒性很輕，通常製成軟膏或搽劑。莫諾苯宗是早已被FDA所認可的用藥，具有大量藥物機轉及人體研究成果資料。
由於臨床應用上的明顯差異，因此從來沒有人認為莫諾苯宗具有抑制癌症細胞的可能性。
癌症長期高居全球死因之首，且罹患癌症人數更是逐年攀升，因此治療癌症儼然成為重要的課題。一般來說，癌症藥物治療無論是化學治療或標靶治療，目的多是讓癌細胞無法複製、分裂，來阻斷腫瘤的蔓延擴張。根據統計，平均每一萬個新藥約只有五個能夠進入第一期臨床試驗。進一步，許多癌細胞相繼產生抗藥性，使藥物的使用成效大幅降低，最終導致癌症治療失敗。由此可見藥物開發 具有一定的困難程度。
因此，如何能讓癌症藥物的快速的可以被開發出來，並且盡可能的減少臨床失敗的機率，在癌症病學上是一個非常急迫又重要的課題。
為解決上述的問題，本發明係針對莫諾苯宗藥物進行新適應症的研發，而達到老藥新用的目標。
經過實驗設計結果顯示莫諾苯宗藥物對正常細胞沒有或僅有微小的毒性，但莫諾苯宗在腫瘤細胞之間具有選擇性的影響。
本發明提供一種制備抑制癌症之醫藥組合物之方法，其中該醫藥組合物包含一莫諾苯宗藥物(Monobenzone)及一藥物可接受的鹽類所組成。
本發明一實施例中，其中該癌症係選自由胸腔相關癌症、腹腔相關癌症、內分泌相關癌症及消化道相關癌症所組成之群組。
本發明一實施例中，其中該癌症係骨肉瘤相關癌症、皮膚癌相關癌症。
本發明一實施例中，其中該胸腔相關癌症係指肺癌。
本發明一實施例中，其中該腹腔相關癌症係選自由膀胱癌、腎臟癌及及子宮頸癌所組成之群組。
本發明一實施例中，其中該內分泌相關癌症係選自由前列腺癌、乳癌及卵巢癌所組成之群組。
本發明一實施例中，其中該內分泌相關癌症係選自由胃癌、肝癌、大腸癌、胰臟癌及舌癌所組成之群組。
本發明一實施例中，其中該莫諾苯宗藥物的有效劑量濃度為每天20mg/kg~500mg/kg。
第一圖顯示本發明莫諾苯宗用藥應用於抑制癌細胞分析結果
第二圖顯示莫諾苯宗用藥可對腫瘤體積之效果
第三圖顯示高劑量及低劑量莫諾苯宗腫瘤生長之抑制效果
各種癌症細胞株建立
將不同癌症類型的細胞株進行繼代培養，癌症細胞種類包含肺癌、胃癌、肝癌、直腸癌、皮膚癌、子宮頸癌、前列腺癌、膀胱癌、乳癌、血癌、胰腺癌、卵巢癌、舌癌、骨肉瘤、腎臟癌，並且對照組亦使用腎臟(HEK293(Kidney))、纖維母細胞HFW(fibroblast)、肺部上皮細胞BEAS-2B(Lung Epithelial)的正常細胞做測試。(表一)
先將各細胞於培養液培養後，由於各細胞的屬性不同，因此針對不同種的細胞也要用相對應的培養液(表一)，計算細胞數目，回種2X106細胞數，然後加入培養該細胞株之培養液補至體積為10ml，繼續培養2-3天。之後將細胞計數，並分裝至96孔盤，其中每孔的細胞數目固定為3000顆，且體積為100ul。
IC50即半抑制濃度(或稱半抑制率)。在間接競爭ELISA標準曲線中是一個非常重要的資料，標準曲線是一個S型曲線。ICELISA中，不添加藥物的對照組的OD值定為B0，添加了藥物的實驗組的OD值為B，B/B0%就叫做結合率，在結合率為50%時所對應的藥物的濃度就叫做IC50。一般IC50的數值越小表示藥物的抑制效果越強。
細胞存活分析方法
將96孔盤中原有的培養液吸掉，每孔加入濃度10um、體積100ul的市售藥物，放置72小時後，每孔再加入100ul已稀釋的WST-1試劑，該稀釋比例為培養液與WST-1原液的體積比為9：1，最後每個孔盤的總體積為200ul，然後將96孔盤放置於37度30~90分鐘，利用elisa reader於OD450偵測吸光值，並計算各癌症細胞株之存活率。
莫諾苯宗藥對於各種癌細胞分析結果
莫諾苯宗對胸腔相關癌症細胞抑制效果之測試
本分析莫諾苯宗對胸腔相關癌症細胞抑制效果之測試，主要針對兩種肺癌細胞進行測試，癌細胞株分別為A549和H1650兩個細胞株，各做4次的癌細胞抑制實驗結果，並且計算其平均值，結果表列如後。(表二)
莫諾苯宗對腹腔相關癌症細胞抑制效果之測試
本分析莫諾苯宗對腹腔相關癌症細胞抑制效果之測試，主要針對三種腹腔相關癌症種類細胞進行測試，膀胱癌細胞株分別為TSGH和T24兩個細胞株(表三)，子宮頸癌細胞株分別為HeLa細胞株(表四)，腎臟癌細胞株為786-O(表五)抑制實驗結果，並且計算其平均值，結果表列如後。
莫諾苯宗對內分泌相關癌症細胞抑制效果之測試
本分析莫諾苯宗對腹腔相關癌症細胞抑制效果之測試，主要針對三種內分泌相關癌症種類細胞進行測試，前列腺癌細胞株分別為PC-3及LNCap細胞株(表六)，乳癌細胞株分別為MCF7和MDA-MB-231兩個細胞株(表七)，卵巢癌細胞株分別為NIH-OVCAR-3細胞株和TOV-21G(表八)，各做4次的癌細胞抑制實驗結果，並且計算其平均值，結果表列如後。
莫諾苯宗對消化道相關癌症細胞抑制效果之測試
分析莫諾苯宗對消化道相關癌症細胞抑制效果之測試，主要針對五種消化道相關癌症種類細胞進行測試，胃癌細胞株分別為AGS和MKN-45兩個細胞株(表九)，肝癌細胞株分別為HepG2和Hep3B兩個細胞株(表十)，大腸癌細胞株分別為HCT116-wt和LoVo細胞株(表十一)，胰臟癌細胞株分別為AsPC和BxPC細胞株(表十二)，舌癌細胞株為SAS細胞株(表十三)做4次的癌細胞抑制實驗結果，並且計算其平均值，結果表列如後。
莫諾苯宗對其他癌症細胞抑制效果之測試
分析莫諾苯宗對其他類型癌正進行測試，骨肉癌細胞株為U2OS細胞株(表十四)，皮膚癌細胞株分別為A375和BCC兩個細胞株(表十五)，各做4次的癌細胞抑制實驗結果，並且計算其平均值，結果表列如後。
對照組實驗設計
莫諾苯宗對正常細胞抑制效果之測試
分析莫諾苯宗對多種正常細胞進行測試，正常腎臟細胞株為HEK293細胞株(表十六)，正常纖維母細胞細胞株為HFW細胞株(表十七)，正常肺部上皮細胞細胞株為BEAS-2B細胞株(表十八)，各做4次的癌細胞抑制實驗結果，並且計算其平均值，結果表列如後。
針對莫諾苯宗對各種癌症細胞抑制效果彙整於表十九，清楚看出莫諾苯宗對於多項癌症有明顯的抑制效果。經過發明人實驗結果，莫諾苯宗藥物對於不同的癌症細胞有明顯的抑制效果。(圖一)
動物實驗分析
本實驗使用雌性BALB/cAnN.Cg-Foxn1nu/CrlNarl小鼠為樣本(購自國家實驗動物中心)，體重為21±1g，皮下注射胃癌細胞(AGS)後隨機分籠，測試藥物分成三組：正常對照組、低劑量(100mg/kg/day)、高劑量(200mg/kg/day)。腫瘤形成超過100mm3後，每天採取腹腔注射方式給予藥物；每週測量腫瘤大小兩次，腫瘤體積測量公式如下：(L×W2)；L代表腫瘤最長直徑；W代表腫瘤最短直徑。
依據第2圖之結果，低劑量與高劑量之莫諾苯宗均對腫瘤具有良好之抑制效果，且於實驗過程中各組小鼠之重量均未出現明顯降低的現象，因此表示莫諾苯宗不論高低劑量在治療過程均能使受測小鼠具有良好的健康狀態而不死亡。
依據第3圖之結果，高低劑量之莫諾苯宗可有效減緩腫瘤體積增長，並可同時減少腫瘤體積，其中以高劑量之莫諾苯宗具有較佳之效果。
上列詳細說明係針對本發明之一可行實施例之具體說明，惟該實施例並非用以限制本發明之專利範圍，凡未脫離本發明技藝精神所為之等效實施或變更，均應包含於本發明之專利範圍中。

Claims (8)

  1. 一種制備抑制癌症之醫藥組合物之方法，其中該醫藥組合物包含一莫諾苯宗藥物(Monobenzone)及一藥物可接受的鹽類所組成。
  2. 如申請專利範圍第1項所述之方法，其中該癌症係選自由胸腔相關癌症、腹腔相關癌症、內分泌相關癌症及消化道相關癌症所組成之群組。
  3. 如申請專利範圍第1項所述之方法，其中該癌症係選自由骨肉瘤相關癌症及皮膚癌相關癌症相關癌症所組成之群組。
  4. 如申請專利範圍第2項所述之方法，其中該胸腔相關癌症係指肺癌。
  5. 如申請專利範圍第2項所述之方法，其中該腹腔相關癌症係選自由膀胱癌、子宮頸癌及腎臟癌所組成之群組。
  6. 如申請專利範圍第2項所述之方法，其中該內分泌相關癌症係選自由前列腺癌、乳癌及卵巢癌所組成之群組。
  7. 如申請專利範圍第2項所述之方法，其中該消化道相關癌症係選自由胃癌、肝癌、大腸癌、胰臟癌及舌癌所組成之群組。
  8. 如申請專利範圍第1項所述之方法，其中該有效劑量濃度為每日20mg/kg~500mg/kg。
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