2011-05-012024-05-13https://scholars.lib.ntu.edu.tw/handle/123456789/648060摘要：臨床前期驗證標的胜肽對肝癌及胰臟癌之療效我們的實驗室從幾年前開始利用噬菌體展示胜肽庫(phage-displayed randompeptide library)與鼻咽癌細胞(NPC cells)作用而找出一特殊胜肽,將之用來做標的化療,當作治療鼻咽癌的另一療法.在此研究中我們發現一新穎的12 個胺基酸胜肽,可用來做導航藥物到NPC 腫瘤組織.在體外觀察,此胜月太能很特殊的與大多數NPC 的細胞膜結合,同樣地也可與NPC 檢體的癌細胞結合.若與帶螢光物質之微脂體連結,則此胜肽結合體可與NPC 細胞膜結合而被吞噬到胞漿中.在長有NPC 異體腫瘤(xenograft)之免疫失全小鼠(SCID mice)中,此胜肽也能特殊的與腫瘤細胞結合,但不會與正常細胞結合.同樣的若對SCID mice 帶有NPC xenograft 而打入胜肽---微脂體(帶平常藥量的五分之一的化療藥物doxorubicin)時,則可見小鼠之xenograft 變成很小.與對照組比起來可減少80-88% 體積,而此變小之腫瘤在顯微鏡下竟是大部份為凋亡及壞死的死細胞及很小部份的活癌細胞; 同時小鼠體重及所有器官都很正常.這個研究結果非常令人鼓舞,因為用所發現之新穎胜月太所連結裝帶有1/5 量之化療藥物之微脂體既可抑制腫瘤生長,又沒有副作用,這對晚期NPC 病人而言是一大福音.也因此, 這種新的治療法立即被美國Cancer Res.接受發表(2004),而英國Lancet Oncology 也特地為此代為介紹.為此,主持人邀請數位專家共同提出群體研究計畫. 在過去三年中此計畫已有具體成果. 但是在過去, 當我們的實驗室在研究對其他癌症有特殊結合力的新的胜肽時,發現其中有些特殊的胜肽有與L-胜肽相同的片段. 因此, 我們懷疑L-peptide 也可能會與其他癌症結合. 經過我們的先驅實驗, 發現L-peptide 確能與數種癌細胞結合. 因此, 本計畫(第四子計畫)之主要的目的是以動物模型去試LP-LD 是否真能治療肝癌及胰臟癌.這包括觀察L-胜肽能否與此兩種癌細胞結合, 能否與腫瘤的外科檢體之癌細胞結合,能否在動物實驗上證明L-P-Lipo-Dox 能殺死此兩種癌之異體腫瘤, 並觀察其最佳劑量及施藥時間表. 此外, 本計畫也要用L-P 連結氧化鐵之奈米顆粒去做MRI,以觀察在體內腫瘤的影像. 由於在之前的研究中, 我們也發現另一種特殊的胜肽SP94 可專與肝癌細胞結合, 同時, 由第三子計畫(Dr. 游)的研究發現有些L-胜肽的修飾(optimizedL-peptide)胜肽似乎比原來的L 胜肽對鼻咽癌的結合力更強, 因此, 本計畫也同時要用SP94 及修飾胜肽(P13)以相同的方法去觀察是否對肝癌及胰臟癌的細胞結合能力, 療效及MRI 的影像分析有更好的結果. 總之,希望在三年後此子計畫能如期完成,加上東洋藥廠的贊助,我們能很順利的將此計畫推進第一期臨床試驗,去證明以胜肽之標的療法對肝癌及胰臟癌病人有用且有很好的療效而甚少有副作用.到時這些癌症將可能變成一可控制的疾病.<br> Abstract: Recently, in our laboratory we have demonstrated that tumor-specific peptide-targetedchemotherapy is superior to conventional chemotherapy for the treatment of NPC in the animalmodel. In this study, using a random peptide phage- displayed library we have identified a novel12-mer peptide from a NPC cell line that has met several criteria for targeted drug delivery intoNPC. In vitro this peptide can bind specifically to the cell surfaces of most NPC cell lines and somebiopsy specimens; the peptide-linked liposome containing fluorescent substance is capable ofbinding to and translocation across cell membranes; in vivo, this specific peptide is able to bind andaccumulate in the xenograft in SCID mice, but not in normal organs; similarly, the L-peptide-linkedliposome carried doxorubicin (L-P-Lipo-Dox) not only causes marked cytotoxicity of NPC cells invitro, in vivo, it also suppresses markedly the xenograft growth (suppress more than 80-88%tumor size) in SCID mice without systemic side effect. These findings suggest that the novelpeptide-targeted chemotherapy has a great potential for diagnostic imaging and targeted therapy ofNPC, while minimizing systemic side effects. These results have been published in CancerResearch journal (2004) and a patent has been obtained from Taiwan and U.S.A. To pave the wayfor clinical application, the PI has been collaborating with several investigators of diverse expertise(including a drug company) on 3 different inter-related aspects which form the basis for a programproject.This program project will be ended in the near future.In our previous other experiment to identify the specific peptide binding to other cancer cells,we also found that the binding LPY domain in the L-peptide also presented in the other high affinitybinding phage clone specific to other cancer cells. Therefore, in the present grant proposal in thesubproject IV, the major effort will focus on the verification of the efficacy of L-P-Lipo-Dox fortreatment of hepatoma and pancreatic cancer including an observation of the localization of itstargeted protein in their surgical specimens, and of investigation of the possibility to use MRI toanalyze the tumor cell image in the xenografts in SCID mice. Since we have also identified anotherspecific peptide SP94 which can bind hepatoma cells, and in one of our collaborator subproject III(Dr. Yu) who used computer analysis of the target protein structure had identified an optimizedL-peptide sequence (p13) which has higher binding activity than that of the original L-peptide,therefore, in this subproject IV, the PI will also include the optimized L-peptide (p13) and thehepatoma specific SP94 peptide to investigate and compare their activity with L-peptide in theabove mentioned 3 aspects: localization of target proteins in the surgical tissue, efficacy of cancertreatment and MRI analysis of tumor image in vivo.It is hopefully that results from this preclinical research can push this research project into phase Iclinical trial after 3 years.肝癌胰臟癌L-胜肽標的療法L-胜肽之核磁共振影像分析微脂體化療藥物DoxorubicinL-胜肽的修飾型胜肽之標的療效肝癌特殊胜肽之標的療效表L-胜肽組織化學法.