TWI602563B - Aurantiamide dipeptide derivatives for treatment or prevention of angiogenesis-related diseases - Google Patents

Description

橙黃醯胺雙肽衍生物用於治療或預防血管新生相關疾病 Anoxin dipeptide derivative for treating or preventing angiogenesis-related diseases

本發明係有關治療或預防血管新生相關疾病。特定言之，本發明係關於橙黃醯胺雙肽衍生物在治療或預防血管新生相關疾病中之用途。 The present invention relates to the treatment or prevention of diseases associated with angiogenesis. In particular, the present invention relates to the use of an orange indamine dipeptide derivative for the treatment or prevention of an angiogenesis-related disease.

血管新生在生理性病況(諸如胚胎發育、生殖、組織修復及骨重構)中起到重要作用。相比之下，血管新生被公認為構成多種致命及致衰弱人類疾病(包括癌症、年齡相關之黃斑變性(AMD)及各種發炎疾病)基礎之共同點。腫瘤血管新生已展示出會促進微環境中之癌症進展及癌轉移。病理性血管新生亦為AMD之主要病因，其為老年人群體中視覺嚴重損失的最常見不可逆病因之一。新生血管性AMD之特徵為侵入視網膜下腔之脈絡膜新生血管(CNV)，通常引起泌出及出血。 Angiogenesis plays an important role in physiological conditions such as embryonic development, reproduction, tissue repair, and bone remodeling. In contrast, angiogenesis is recognized as a commonality that forms the basis of a variety of fatal and debilitating human diseases, including cancer, age-related macular degeneration (AMD), and various inflammatory diseases. Tumor angiogenesis has been shown to promote cancer progression and cancer metastasis in the microenvironment. Pathological angiogenesis is also a major cause of AMD, one of the most common irreversible causes of severe visual loss in the elderly population. Neovascular AMD is characterized by choroidal neovascularization (CNV) that invades the subretinal space, often causing secretion and bleeding.

新血管生成涉及循環內皮祖細胞(EPC)自骨髓募集至缺氧部位以及先存在的內皮細胞之出芽。EPC為循環中之細胞群體，且執行血管新生及血管重構，因為其能夠分化成內皮細胞且形成血管。晚期EPC具有更致力於內皮系分化且促進血管新生之優點。此外，已提出早期EPC會經由製造可能會活化鄰近內皮細胞之血管生成細胞激素(例如VEGF及IL-8)來改進血管新生/血小管生成。近來，據報導EPC藉由干 預血管生成開關來介導早期腫瘤生長及晚期轉移性進展。累積的證據指示EPC在AMD期間促進缺血缺氧組織中之新血管生成。此等發現建立EPC在病理性血管新生中之作用且支持EPC靶向療法可為阻斷血管新生相關疾病之有前景的策略。 Neovascularization involves the recruitment of circulating endothelial progenitor cells (EPC) from the bone marrow to the site of hypoxia and the sprouting of pre-existing endothelial cells. EPC is a cell population in the circulation and performs angiogenesis and vascular remodeling because it is capable of differentiating into endothelial cells and forming blood vessels. Advanced EPC has the advantage of being more committed to endothelial differentiation and promoting angiogenesis. In addition, it has been suggested that early EPC may improve angiogenesis/tubule production by making angiogenic cytokines (eg, VEGF and IL-8) that may activate adjacent endothelial cells. Recently, it has been reported that EPC is doing Pre-angiogenesis switches to mediate early tumor growth and advanced metastatic progression. Cumulative evidence indicates that EPC promotes neovascularization in ischemic and hypoxic tissue during AMD. These findings establish a role for EPC in pathological angiogenesis and support EPC-targeted therapies as a promising strategy for blocking angiogenesis-related diseases.

雙肽衍生物Fmoc-10a(9H-茀-9-基)甲基(S)-1-((R)-1-羥基-3-苯基丙-2-基胺基)-3-甲基-1-側氧基丁-2-基胺基甲酸酯)為橙黃醯胺衍生物，其由***醇及***酸構成，且為蜆殼花椒(Zanthoxylum dissitum)、天葵(Semiaquilegia adoxoides)及火炭母(Polygonum chinensis)之主要組分。橙黃醯胺展現抗細菌、消炎(Chiao-Ting Yen等人，European Journal of Medicinal Chemistry 44(2009)1933-1940)、抗氧化及抗HIV作用。近來，許多雙肽展示出對抗癌細胞之細胞毒性作用(Chiao-Ting Yen等人，European Journal of Medicinal Chemistry 45(2010)2494-2502)。舉例而言，***酸芥肽(Peptichemio，PTC)及硼替佐米(Bortezomib，VELCADE^®)已經開發及銷售用於治療癌症。 Bipeptide derivative Fmoc-10a(9 H -茀-9-yl)methyl( S )-1-(( R )-1-hydroxy-3-phenylpropan-2-ylamino)-3-methyl The base-1-yloxybutan-2-ylcarbamate is an orange xanthine derivative composed of amphetamine and phenylalanine, and is Zanthoxylum dissitum , Semiaquilegia adoxoides and The main component of Polygonum chinensis . The orange yellow amine exhibits antibacterial and anti-inflammatory effects (Chiao-Ting Yen et al., European Journal of Medicinal Chemistry 44 (2009) 1933-1940), antioxidant and anti-HIV effects. Recently, many dipeptides have exhibited cytotoxic effects against cancer cells (Chiao-Ting Yen et al., European Journal of Medicinal Chemistry 45 (2010) 2494-2502). For example, Peptichemio (PTC) and Bortezomib (VELCADE ^® ) have been developed and marketed for the treatment of cancer.

本發明提供一種抑制、改善、預防或治療血管新生相關疾病之方法，其包含向個體投與有效量之本文所述之式(I)化合物或其互變異構體、立體異構體或對映異構體、或其溶劑合物、前藥或醫藥學上可接受之鹽。 The present invention provides a method for inhibiting, ameliorating, preventing or treating an angiogenesis-related disease comprising administering to an individual an effective amount of a compound of the formula (I) described herein or a tautomer, stereoisomer or enantiomer thereof. Isomer, or a solvate, prodrug or pharmaceutically acceptable salt thereof.

在一些實施例中，該式(I)化合物係選自(S)-1-((S)-1-羥基-3-苯基丙-2-基胺基)-1-側氧基-3-苯基丙-2-基胺基甲酸(9H-茀-9-基)甲酯；(S)-1-((R)-1-羥基-3-苯基丙-2-基胺基)-1-側氧基-3-苯基丙-2-基胺基甲酸(9H-茀-9-基)甲酯；(S)-1-((S)-1-羥基-3-苯基丙-2-基胺基)-3-(1H-吲哚-3-基)-1-側氧基丙-2-基胺基甲酸(9H-茀-9-基)甲酯；(S)-1-((R)-1-羥基-3-苯基丙-2-基胺基)-3-(1H-吲哚-3-基)-1-側氧基丙-2-基胺基甲酸(9H-茀-9-基)甲酯；(S)-1-((S)-1-羥基-3-苯基丙-2-基胺基)-3-(萘-2-基)- 1-側氧基丙-2-基胺基甲酸(9H-茀-9-基)甲酯；(S)-1-((R)-1-羥基-3-苯基丙-2-基胺基)-3-(萘-2-基)-1-側氧基丙-2-基胺基甲酸(9H-茀-9-基)甲酯；(S)-1-((S)-1-羥基-3-苯基丙-2-基胺基)-3-甲基-1-側氧基丁-2-基胺基甲酸(9H-茀-9-基)甲酯；及(S)-1-((R)-1-羥基-3-苯基丙-2-基胺基)-3-甲基-1-側氧基丁-2-基胺基甲酸(9H-茀-9-基)甲酯；或其互變異構體、立體異構體或對映異構體、或其溶劑合物、前藥或醫藥學上可接受之鹽。在另一實施例中，該化合物為(S)-1-((R)-1-羥基-3-苯基丙-2-基胺基)-3-甲基-1-側氧基丁-2-基胺基甲酸(9H-茀-9-基)甲酯。 In some embodiments, the compound of formula (I) is selected from the group consisting of (S)-1-((S)-1-hydroxy-3-phenylpropan-2-ylamino)-1-oxooxy-3 -Phenylpropan-2-ylaminocarboxylic acid (9H-fluoren-9-yl)methyl ester; (S)-1-((R)-1-hydroxy-3-phenylpropan-2-ylamino) -1-oxooxy-3-phenylpropan-2-ylaminocarboxylic acid (9H-fluoren-9-yl)methyl ester; (S)-1-((S)-1-hydroxy-3-phenyl Propan-2-ylamino)-3-(1H-indol-3-yl)-1-oxoylpropan-2-ylaminocarboxylic acid (9H-fluoren-9-yl)methyl ester; (S) 1-((R)-1-hydroxy-3-phenylpropan-2-ylamino)-3-(1H-indol-3-yl)-1-yloxypropan-2-ylamino (9H-fluoren-9-yl)methyl formate; (S)-1-((S)-1-hydroxy-3-phenylpropan-2-ylamino)-3-(naphthalen-2-yl) 1- 1-oxopropan-2-ylaminocarboxylic acid (9H-fluoren-9-yl)methyl ester; (S)-1-((R)-1-hydroxy-3-phenylpropan-2-yl Amino)-3-(naphthalen-2-yl)-1-oxoylpropan-2-ylaminocarboxylic acid (9H-fluoren-9-yl)methyl ester; (S)-1-((S)- 1-hydroxy-3-phenylpropan-2-ylamino)-3-methyl-1-oxobutan-2-ylaminocarboxylic acid (9H-fluoren-9-yl)methyl ester; and (S )-1-((R)-1-hydroxy-3-phenylpropan-2-ylamino)-3-methyl-1-oxobutan-2-ylaminocarboxylic acid (9H-茀-9) -yl)methyl ester; or its tautomers, stereoisomers Of acceptable enantiomer, or a solvate, pharmaceutically acceptable prodrug, or salt thereof. In another embodiment, the compound is ( S )-1-(( R )-1-hydroxy-3-phenylpropan-2-ylamino)-3-methyl-1-oxetoxy- yl carbamic acid 2- (9 H - fluorenyl-9-yl) methyl ester.

在一個實施例中，該血管新生相關疾病為侵襲性及轉移性血管新生相關癌症或眼新血管生成。 In one embodiment, the angiogenesis-related disease is an invasive and metastatic angiogenesis-related cancer or ocular neovascularization.

在一些實施例中，該侵襲性及轉移性血管新生相關癌症包括(但不限於)口腔癌、腦癌、乳癌、腎癌、卵巢癌、肺癌、結腸癌、肝癌(諸如肝細胞癌)及黑素瘤。在另一實施例中，該侵襲性及轉移性血管新生相關癌症為黑素瘤。在一些實施例中，對於侵襲性及轉移性血管新生相關癌症，該投藥可與手術、放射線療法或化學療法組合、在其之前或在其之後使用。 In some embodiments, the invasive and metastatic angiogenesis-related cancer includes, but is not limited to, oral cancer, brain cancer, breast cancer, kidney cancer, ovarian cancer, lung cancer, colon cancer, liver cancer (such as hepatocellular carcinoma), and black Prime tumor. In another embodiment, the invasive and metastatic angiogenesis-related cancer is melanoma. In some embodiments, for invasive and metastatic angiogenesis-related cancers, the administration can be combined with, before, or after surgery, radiation therapy, or chemotherapy.

在一個實施例中，眼新血管生成為黃斑變性。黃斑變性較佳為濕性黃斑變性。濕性黃斑變性更佳地為AMD。在一個實施例中，對於黃斑變性，該投藥係以玻璃體內、眼內或眼周途徑進行。在另一實施例中，該化合物調配成注射劑或點眼劑。 In one embodiment, ocular neovascularization is macular degeneration. Macular degeneration is preferably wet macular degeneration. The wet macular degeneration is more preferably AMD. In one embodiment, for macular degeneration, the administration is by intravitreal, intraocular or periocular routes. In another embodiment, the compound is formulated as an injection or eye drop.

圖1 A至E展示出Fmoc-10a抑制晚期EPC之血管生成作用。A.Fmoc-10a之化學結構。B.使用MTT分析，Fmoc-10a以濃度依賴性方式抑制晚期EPC之細胞生長。C.使用LDH分析，Fmoc-10a不誘導晚期EPC中之LDH釋放。D.使用Transwell遷移分析，Fmoc-10a抑制晚期EPC之細胞遷移。E.使用管形成分析，Fmoc-10a消除晚期EPC之毛 細管狀結構。資料表示為五次獨立實驗之平均值±SEM。*，p<0.01，與對照組相比。 Figures 1A to E show that Fmoc-10a inhibits the angiogenesis of advanced EPC. A. The chemical structure of Fmoc-10a. B. Using MTT assay, Fmoc-10a inhibited cell growth of late EPC in a concentration dependent manner. C. Using LDH analysis, Fmoc-10a does not induce LDH release in late EPC. D. Using Transwell migration assay, Fmoc-10a inhibits cell migration in advanced EPC. E. Using tube formation analysis, Fmoc-10a eliminates the capillary structure of late EPC. Data are expressed as the mean ± SEM of five independent experiments. *, p < 0.01 compared to the control group.

圖2 A及B展示出Fmoc-10a抑制早期EPC之血管生成潛能。A及B.使用CFU-希爾分析，Fmoc-10a抑制早期EPC之群落形成單位(CFU)。C.使用趨化運動分析，Fmoc-10a抑制早期EPC之趨化性。資料表示為四次獨立實驗之平均值±SEM。 Figures 2A and B show that Fmoc-10a inhibits the angiogenic potential of early EPC. A and B. Using CFU-Hill analysis, Fmoc-10a inhibited early EPC community forming units (CFU). C. Using chemotactic analysis, Fmoc-10a inhibits the chemotaxis of early EPC. Data are expressed as the mean ± SEM of four independent experiments.

圖3 A及B展示出Fmoc-10a離體及在活體內抑制血管新生。A.使用主動脈環出芽分析，Fmoc-10a消除VEGF誘導之血管出芽。B.使用活體內小鼠DIVAA模型，Fmoc-10a減弱VEGF誘導之微血管形成。資料表示為三次獨立實驗之平均值±SEM。*，p<0.01，及**，p<0.001，與媒劑組相比。 Figures 3A and B show that Fmoc-10a is isolated and inhibits angiogenesis in vivo. A. Using aortic ring budding assay, Fmoc-10a abolishes VEGF-induced vascular sprouting. B. Fmoc-10a attenuates VEGF-induced microvascular formation using an in vivo mouse DIVAA model. Data are expressed as mean ± SEM of three independent experiments. *, p < 0.01, and **, p < 0.001, compared to the vehicle group.

圖4 A至D顯示Fmoc-10a抑制黑素瘤癌症之細胞侵襲、VEGF釋放、腫瘤生長及轉移。A.Fmoc-10a顯著抑制B16F10黑素瘤細胞之細胞侵襲。B.Fmoc-10a濃度依賴性抑制VEGF自B16F10黑素瘤細胞釋放。C.使用腫瘤異種移植模型，Fmoc-10a誘導B16F10黑素瘤生長之劑量依賴性抑制。D.使用實驗癌轉移模型，Fmoc-10a抑制B16F10細胞之肺群落數目。資料表示為三個獨立實驗之平均值±S.E.M.。*，p<0.05，**，p<0.01，及***，P<0.001，與媒劑組相比。 Figure 4 A to D show that Fmoc-10a inhibits cell invasion, VEGF release, tumor growth and metastasis of melanoma cancer. A. Fmoc-10a significantly inhibited cell invasion of B16F10 melanoma cells. B. Fmoc-10a concentration-dependently inhibits the release of VEGF from B16F10 melanoma cells. C. Using a tumor xenograft model, Fmoc-10a induced dose-dependent inhibition of B16F10 melanoma growth. D. Using the experimental cancer metastasis model, Fmoc-10a inhibits the number of lung communities in B16F10 cells. Data are expressed as the mean ± SEM of three independent experiments. *, p < 0.05, **, p < 0.01, and ***, P < 0.001, compared to the vehicle group.

圖5 A及B顯示Fmoc-10a抑制AMD動物模型中之脈絡膜新血管生成(CNV)。A.執行氧誘導視網膜病變(OIR)之小鼠模型以在氧過多條件下誘導7天大小鼠之視網膜新血管生成。視網膜新血管生成之整體裝片(whole-mount)分析顯示氧過多顯著增加同工凝集素(isolectin)之螢光。Fmoc-10a顯著阻斷小鼠之氧過多誘導之病理性新血管生成，且癌思停(Avastin)(10μg)用作陽性對照。B.視網膜中新血管生成之定量偵測係自四個獨立實驗呈現。資料表示為平均值±S.E.。*，p<0.001，與常氧組相比；#，p<0.05，與在氧過多下之媒劑處理組相 比。 Figure 5 A and B show that Fmoc-10a inhibits choroidal neovascularization (CNV) in an animal model of AMD. A. A mouse model of oxygen-induced retinopathy (OIR) was performed to induce retinal neovascularization in 7-day-old mice under hyperoxic conditions. Whole-mount analysis of retinal neovascularization showed that hyperoxia significantly increased the fluorescence of isolectin. Fmoc-10a significantly blocked hyperoxia-induced pathological neovascularization in mice, and Avastin (10 μg) was used as a positive control. B. Quantitative detection of neovascularization in the retina is presented in four independent experiments. Data are expressed as mean ± SE. *, p < 0.001, compared with the normoxic group; #, p < 0.05, compared to the vehicle treated group under hyperoxic conditions.

本發明至少部分基於發現橙黃醯胺雙肽衍生物抑制血管新生。因此，橙黃醯胺雙肽衍生物可以用作血管新生抑制劑，藉此預防或治療侵襲性及轉移性血管新生相關癌症(特別是黑素瘤之轉移)及眼新血管生成(特別是黃斑變性，諸如年齡相關之黃斑變性(AMD)之病理性新血管生成)。 The present invention is based, at least in part, on the discovery that an orange xanthine dipeptide derivative inhibits angiogenesis. Therefore, the orange-ylidene dipeptide derivative can be used as an angiogenesis inhibitor to prevent or treat invasive and metastatic angiogenesis-related cancers (especially melanoma metastasis) and ocular neovascularization (especially macular degeneration). , such as age-related macular degeneration (AMD) pathological neovascularization).

為方便起見，此處收集在本發明之情形下使用之某些術語。除非另外定義，否則本文中所使用之所有技術及科學術語具有如本發明所屬之一般技術者通常理解之相同含義。 For convenience, certain terms used in the context of the present invention are collected herein. All technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art, unless otherwise defined.

除非上下文以其他方式明確規定，否則單數形式「一(a/an)」及「該(the)」在本文中用以包括複數參照物。 The singular forms "a", "the", "the" and "the" are used to include the plural referents.

如本文所用之術語「治療(treatment或treating)」欲意謂獲得所需藥理學及/或生理作用，例如改善與疾病相關之症狀。該作用可在完全或部分預防疾病或其症狀方面為預防性的及/或在部分或完全治癒疾病及/或可歸因於該疾病之不良影響方面為治療性的。如本文所用之「治療」包括對哺乳動物(特定言之人類)之疾病的防治性(例如預防性)、治癒性或緩解性治療；且包括：(1)對在易患疾病但尚未診斷為患有該疾病之個體中存在的該疾病或病況之防治性(例如預防性)、治癒性或緩解性治療；(2)抑制疾病(例如藉由遏制其進展來抑制)；或(3)緩解疾病(例如減輕與疾病相關的症狀)。 The term "treatment" or "treating" as used herein is intended to mean obtaining the desired pharmacological and/or physiological effect, for example to ameliorate the symptoms associated with the disease. This effect may be therapeutic in terms of preventing, in whole or in part, the disease or its symptoms and/or in the partial or complete cure of the disease and/or attributable to the adverse effects of the disease. "Treatment," as used herein, includes prophylactic (eg, prophylactic), curative or palliative treatment of a disease in a mammal (specifically, a human); and includes: (1) being susceptible to a disease but not yet diagnosed Preventive (eg, prophylactic), curative or palliative treatment of the disease or condition present in an individual having the disease; (2) inhibition of the disease (eg, inhibition by inhibition of its progression); or (3) alleviation of the disease (eg, alleviating symptoms associated with the disease).

術語「投與(administered)」、「投與(administering)」或「投與(administration)」在本文中可互換使用以指傳遞模式，包括(但不限於)靜脈內、肌內、腹膜內、動脈內、皮下或經皮投與本發明之試劑(例如化合物或組合物)。 The terms "administered", "administering" or "administration" are used interchangeably herein to refer to modes of delivery, including but not limited to intravenous, intramuscular, intraperitoneal, The agents (e.g., compounds or compositions) of the invention are administered intra-arterially, subcutaneously or transdermally.

如本文中所用之術語「預防(prevent)」、「預防(preventing)」及 「預防(prevention)」係指預防疾病或病症或其一或多種症狀的發作、復發或擴散。在某些實施例中，該術語係指在存在或不存在一或多種其他額外活性劑之情況下，在症狀發作之前，尤其對處於本文提供之疾病或病症風險下的患者，用本文提供之化合物或抗體或劑型治療或投與本文提供之化合物或抗體或劑型。該等術語涵蓋具體疾病之症狀之抑制或減少。就此而言，術語「預防」可與術語「預防性治療」互換使用。 As used herein, the terms "prevent", "preventing" and "Prevention" means preventing the onset, recurrence or spread of a disease or condition or one or more of its symptoms. In certain embodiments, the term refers to a patient provided prior to the onset of symptoms, particularly at the risk of the disease or condition provided herein, in the presence or absence of one or more additional additional active agents. The compound or antibody or dosage form is treated or administered a compound or antibody or dosage form provided herein. These terms encompass the inhibition or reduction of the symptoms of a particular disease. In this regard, the term "prevention" can be used interchangeably with the term "preventive treatment."

除非另外指明，否則如本文所用之術語「共投與」及「與……組合」包括同步、同時或在無特定時限內依序投與兩種或兩種以上治療劑。 The terms "co-administered" and "in combination with" as used herein, unless otherwise indicated, include the simultaneous or simultaneous administration of two or more therapeutic agents within a specified time period.

如本文所用之術語「有效量」係指在特定劑量及時間段下有效達成關於疾病治療所需結果之量。有效量之試劑不需要治癒疾病或病況，但將提供疾病或病況之治療，使得該疾病或病況之發作得以延遲、受阻或預防，或改善該疾病或病況症狀。有效量可為單一劑量或分成呈適合形式的兩個或兩個以上劑量，其欲在整個指定時間段中以一次、兩次或大於兩次投與。 The term "effective amount" as used herein refers to an amount effective to achieve a desired result for the treatment of a disease at a particular dosage and time period. An effective amount of the agent does not require a cure for the disease or condition, but will provide a treatment for the disease or condition such that the onset of the disease or condition is delayed, blocked or prevented, or the symptoms of the disease or condition are ameliorated. An effective amount can be a single dose or divided into two or more doses in a suitable form, which are intended to be administered once, twice or more than twice throughout the specified period of time.

如本文所用之術語「個體」或「患者」係指可用本發明之方法治療之動物，包括人類物種。除非特定地指明一種性別，否則術語「個體」或「患者」意欲指男性及女性兩種性別。因此，術語「個體」或「患者」包含可得益於本發明之治療方法的任何哺乳動物。 The term "individual" or "patient" as used herein refers to an animal, including a human species, that can be treated by the methods of the invention. Unless specifically specifying a gender, the terms "individual" or "patient" are intended to mean both male and female genders. Thus, the term "individual" or "patient" encompasses any mammal that can benefit from the methods of treatment of the present invention.

如本文所用之術語「血管新生相關疾病」係指特徵為病理性血管新生之病症。特徵為病理性血管新生之病症係指其中單獨或與其他組合之不正常或異常血管新生促成病症之起因、起源或症狀的病症。 The term "angiogenesis-related disease" as used herein refers to a condition characterized by pathological angiogenesis. A condition characterized by pathological angiogenesis refers to a condition in which the abnormal or abnormal angiogenesis alone or in combination with other causes contributes to the cause, origin or symptom of the condition.

如本文中所用，術語「組合療法」係指兩種或兩種以上不同醫藥劑以重疊方案投與以使個體同時暴露於兩種藥劑之情形。 As used herein, the term "combination therapy" refers to the situation in which two or more different pharmaceutical agents are administered in an overlapping regimen to expose the individual to both agents simultaneously.

如本文所用，術語「抑制」意謂防止某事發生，延遲某事發生 出現及/或降低某事發生之程度或可能性。因此，「抑制血管新生」及「抑制新血管系(neovasculature)形成」意欲涵蓋預防、延遲及/或降低血管新生發生之可能性以及降低新血管之數目、生長速率、大小等。 As used herein, the term "inhibiting" means preventing something from happening and delaying something to happen. Appear and/or reduce the extent or likelihood of something happening. Therefore, "inhibiting angiogenesis" and "inhibiting the formation of neovasculature" are intended to cover the prevention, delay, and/or reduction of the likelihood of angiogenesis and the reduction in the number, growth rate, size, and the like of new blood vessels.

如本文所用，術語「黃斑變性」係指由於視網膜之損壞而引起視野中心(黃斑)視覺喪失之醫學病況。「濕性黃斑變性」(亦稱為新血管型或滲出型)係指涉及來自視網膜後方脈絡膜之血管生長的黃斑變性。在濕性黃斑變性中，視網膜有時可能剝離。「年齡相關之黃斑變性」(AMD)係指最常見形式之黃斑變性，其通常在晚年開始，黃斑中有特徵黃色沈積物。 As used herein, the term "macular degeneration" refers to a medical condition that causes visual loss of the center of the field (macular) due to damage to the retina. "Welken macular degeneration" (also known as neovascular or exudative) refers to macular degeneration involving the growth of blood vessels from the choroid of the posterior retina. In wet macular degeneration, the retina may sometimes peel off. "Age-related macular degeneration" (AMD) refers to the most common form of macular degeneration, which usually begins in later years and has characteristic yellow deposits in the macula.

如本文所用，術語「轉移(metastasis)」(有時縮寫為「mets」；複數「metastases」)係指腫瘤細胞自一個器官或組織擴散至另一部位。該術語亦指由於轉移而在新部位形成之腫瘤組織。「轉移性癌症」為自其原始或原發部位擴散之癌症，且亦可稱為「繼發性癌症」或「繼發性腫瘤」。一般而言，轉移性腫瘤係以其起源之原發性腫瘤之組織命名。 As used herein, the term "metastasis" (sometimes abbreviated as "mets"; plural "metastases") refers to the spread of tumor cells from one organ or tissue to another. The term also refers to tumor tissue formed at a new site due to metastasis. "metastatic cancer" is a cancer that spreads from its original or primary site and can also be called "secondary cancer" or "secondary tumor." In general, metastatic tumors are named after the tissue of the primary tumor from which they originated.

術語「醫藥學上可接受」在本文中用於指在醫學判斷範疇內，適用於與人類及動物之組織接觸而無過度毒性、刺激、過敏反應或其他問題或併發症、且與合理益處/風險比相匹配之化合物、物質、組合物及/或劑型。 The term "pharmaceutically acceptable" is used herein to mean in the context of medical judgment, for use in contact with human and animal tissues without excessive toxicity, irritation, allergic reactions or other problems or complications, and with reasonable benefits/ A compound, substance, composition, and/or dosage form that matches the risk ratio.

如本文所使用，「醫藥學上可接受之鹽」係指所揭示之化合物的衍生物，其中母體化合物藉由製得其酸鹽或鹼鹽而改質。醫藥學上可接受之鹽之實例包括(但不限於)鹼性殘基(諸如胺、吡啶、嘧啶及喹唑啉)之礦物質或有機酸鹽；酸性殘基(諸如羧酸)之鹼金屬或有機鹽；及其類似物。 As used herein, "pharmaceutically acceptable salts" refers to derivatives of the disclosed compounds wherein the parent compound is modified by the preparation of its acid or base salt. Examples of pharmaceutically acceptable salts include, but are not limited to, mineral or organic acid salts of basic residues such as amines, pyridines, pyrimidines, and quinazolines; alkali metals of acidic residues such as carboxylic acids. Or an organic salt; and the like.

在一個態樣中，本發明提供一種抑制、改善、預防或治療血管 新生相關疾病之方法，其包含向個體投與有效量之以下式(I)化合物， 其中R為-(CH₂)_1-4苯基，其中苯基未經取代或經一至三個羥基、NH₂、NO₂或烷基取代；-(CH₂)_1-4吲哚基；-(CH₂)_1-4萘基；或異丙基， 或其互變異構體、立體異構體或對映異構體、或其溶劑合物、前藥或醫藥學上可接受之鹽。 In one aspect, the invention provides a method of inhibiting, ameliorating, preventing or treating an angiogenesis-related disease, comprising administering to an individual an effective amount of a compound of formula (I) below, Wherein R is -(CH ₂ ) _1-4 phenyl, wherein phenyl is unsubstituted or substituted with one to three hydroxy, NH ₂ , NO ₂ or alkyl; -(CH ₂ ) _1-4 fluorenyl; (CH ₂ ) _1-4 naphthyl; or isopropyl, or a tautomer, stereoisomer or enantiomer thereof, or a solvate, prodrug or pharmaceutically acceptable salt thereof.

在一些實施例中，R為-CH₂苯基、-CH₂吲哚基、-CH₂萘基或異丙基。在另一實施例中，R為異丙基。 In some embodiments, R is -CH ₂ phenyl, -CH ₂ fluorenyl, -CH ₂ naphthyl or isopropyl. In another embodiment, R is isopropyl.

在一些實施例中，式(I)化合物係選自：(S)-1-((S)-1-羥基-3-苯基丙-2-基胺基)-1-側氧基-3-苯基丙-2-基胺基甲酸(9H-茀-9-基)甲酯；(S)-1-((R)-1-羥基-3-苯基丙-2-基胺基)-1-側氧基-3-苯基丙-2-基胺基甲酸(9H-茀-9-基)甲酯；(S)-1-((S)-1-羥基-3-苯基丙-2-基胺基)-3-(1H-吲哚-3-基)-1-側氧基丙-2-基胺基甲酸(9H-茀-9-基)甲酯；(S)-1-((R)-1-羥基-3-苯基丙-2-基胺基)-3-(1H-吲哚-3-基)-1-側氧基丙-2-基胺基甲酸(9H-茀-9-基)甲酯；(S)-1-((S)-1-羥基-3-苯基丙-2-基胺基)-3-(萘-2-基)-1-側氧基丙-2-基胺基甲酸(9H-茀-9-基)甲酯；(S)-1-((R)-1-羥基-3-苯基丙-2-基胺基)-3-(萘-2-基)-1-側氧基丙-2-基胺基甲酸(9H-茀-9-基)甲酯；(S)-1-((S)-1-羥基-3-苯基丙-2-基胺基)-3-甲基-1-側氧基丁-2-基胺 基甲酸(9H-茀-9-基)甲酯；及(S)-1-((R)-1-羥基-3-苯基丙-2-基胺基)-3-甲基-1-側氧基丁-2-基胺基甲酸(9H-茀-9-基)甲酯 In some embodiments, the compound of formula (I) is selected from the group consisting of: (S)-1-((S)-1-hydroxy-3-phenylpropan-2-ylamino)-1-oxooxy-3 -Phenylpropan-2-ylaminocarboxylic acid (9H-fluoren-9-yl)methyl ester; (S)-1-((R)-1-hydroxy-3-phenylpropan-2-ylamino) -1-oxooxy-3-phenylpropan-2-ylaminocarboxylic acid (9H-fluoren-9-yl)methyl ester; (S)-1-((S)-1-hydroxy-3-phenyl Propan-2-ylamino)-3-(1H-indol-3-yl)-1-oxoylpropan-2-ylaminocarboxylic acid (9H-fluoren-9-yl)methyl ester; (S) 1-((R)-1-hydroxy-3-phenylpropan-2-ylamino)-3-(1H-indol-3-yl)-1-yloxypropan-2-ylamino (9H-fluoren-9-yl)methyl formate; (S)-1-((S)-1-hydroxy-3-phenylpropan-2-ylamino)-3-(naphthalen-2-yl) (1-H-oxypropan-2-ylaminocarboxylic acid (9H-fluoren-9-yl)methyl ester; (S)-1-((R)-1-hydroxy-3-phenylpropan-2-yl Amino)-3-(naphthalen-2-yl)-1-oxoylpropan-2-ylaminocarboxylic acid (9H-fluoren-9-yl)methyl ester; (S)-1-((S)- 1-hydroxy-3-phenylpropan-2-ylamino)-3-methyl-1-oxobutan-2-ylamine (9H-fluoren-9-yl)methyl carbamic acid; and (S)-1-((R)-1-hydroxy-3-phenylpropan-2-ylamino)-3-methyl-1- 2-oxet-2-ylaminocarbamic acid (9H-fluoren-9-yl)methyl ester

或其互變異構體、立體異構體或對映異構體、或其溶劑合物、前藥或醫藥學上可接受之鹽。 Or a tautomer, a stereoisomer or an enantiomer thereof, or a solvate, prodrug or pharmaceutically acceptable salt thereof.

在另一實施例中，該化合物為(S)-1-((R)-1-羥基-3-苯基丙-2-基胺基)-3-甲基-1-側氧基丁-2-基胺基甲酸(9H-茀-9-基)甲酯。該化合物之 化學結構為。 In another embodiment, the compound is ( S )-1-(( R )-1-hydroxy-3-phenylpropan-2-ylamino)-3-methyl-1-oxetoxy- yl carbamic acid 2- (9 H - fluorenyl-9-yl) methyl ester. The chemical structure of the compound is .

血管新生涉及新微血管自先存在的血管生長或出芽，且涉及新生血管生長之血小管生成為許多生理性及病理性病況所必需的，該等病況包括胚胎發生、侵襲性及轉移性癌症、類風濕性關節炎、糖尿病性視網膜病變、肥胖症、動脈粥樣硬化、缺血性心臟及肢體疾病以及創傷癒合。超過70種疾病已鑑別為血管新生依賴性(Carmeliet,Nature,438：932-6,2005)。新血管之不正常發展已涉及許多病理生理學過程。舉例而言，血管之不正常生長與腸閉鎖、消化性潰瘍、類風濕性關節炎、全身性紅斑狼瘡、牛皮癬、增殖性視網膜病變及動脈粥樣硬化相關。由於血管新生涉及多種病理性過程，故本發明方法可適用於治療、改善或預防諸如侵襲性及轉移性癌症及眼新血管生成(諸如黃斑變性)之疾病。 Angiogenesis involves the growth or sprouting of blood vessels from pre-existing new microvasculature, and the formation of blood tubules involved in the growth of new blood vessels is essential for many physiological and pathological conditions, including embryogenesis, invasive and metastatic cancer, and Rheumatoid arthritis, diabetic retinopathy, obesity, atherosclerosis, ischemic heart and limb disease, and wound healing. More than 70 diseases have been identified as angiogenesis dependent (Carmeliet, Nature, 438: 932-6, 2005). The abnormal development of new blood vessels has involved many pathophysiological processes. For example, abnormal growth of blood vessels is associated with intestinal atresia, peptic ulcer, rheumatoid arthritis, systemic lupus erythematosus, psoriasis, proliferative retinopathy, and atherosclerosis. Since angiogenesis involves a variety of pathological processes, the methods of the invention are applicable to the treatment, amelioration or prevention of diseases such as invasive and metastatic cancer and ocular neovascularization such as macular degeneration.

在一些實施例中，侵襲性及轉移性癌症為血管新生相關癌症。根據一些實施例，侵襲性及轉移性血管新生相關癌症包括(但不限於)口腔癌、腦癌、乳癌、腎癌、卵巢癌、肺癌、結腸癌、肝癌、肝細胞癌及黑素瘤。 In some embodiments, the aggressive and metastatic cancer is an angiogenesis-related cancer. According to some embodiments, invasive and metastatic angiogenesis-related cancers include, but are not limited to, oral cancer, brain cancer, breast cancer, kidney cancer, ovarian cancer, lung cancer, colon cancer, liver cancer, hepatocellular carcinoma, and melanoma.

在一個實施例中，眼新血管生成為黃斑變性。黃斑變性為65歲及65歲以上人視力損失及失明之主要病因。黃斑變性通常以年齡相關之形式發生(通常稱作AMD)，但亦會發生青少年的黃斑變性。在AMD中，黃斑為造成急劇的中央視力退化之視網膜的一部分。 In one embodiment, ocular neovascularization is macular degeneration. Macular degeneration is the main cause of loss of vision and blindness in people 65 years of age and older. Macular degeneration usually occurs in an age-related form (commonly known as AMD), but adolescent macular degeneration also occurs. In AMD, the macula is part of the retina that causes dramatic central vision deterioration.

如「新生血管性」名稱所表明，濕性黃斑變性之特徵為新血管生長異常，例如在黃斑上生長。此類新血管可在視網膜下生長，血液及體液滲漏。此類滲漏導致感光視網膜細胞永久性損傷，該等細胞死亡且在中央視力中產生盲點。濕性黃斑變性可進一步分成兩類。在濕性黃斑變性之隱性形式中，在視網膜下生長之新血管不顯著且滲漏較不明顯，通常產生較不嚴重之視力下降。在濕性黃斑變性之典型形式中，血管生長及結疤具有極其清楚、可描繪之輪廓，其可在視網膜下觀測到。經典濕性黃斑變性亦稱為經典脈絡膜新生血管且通常導致更嚴重的視力損失。 As indicated by the name "neovascular", wet macular degeneration is characterized by abnormal growth of new blood vessels, such as growth on the macula. These new blood vessels can grow under the retina, leaking blood and body fluids. Such leakage results in permanent damage to the photoreceptor cells, which die and produce blind spots in central vision. Wet macular degeneration can be further divided into two categories. In the recessive form of wet macular degeneration, new blood vessels growing under the retina are less pronounced and less pronounced, often resulting in less severe vision loss. In the typical form of wet macular degeneration, blood vessel growth and scarring have an extremely clear, descriptive profile that can be observed under the retina. Classical wet macular degeneration is also known as classical choroidal neovascularization and usually results in more severe loss of vision.

鑒於血管新生在濕性黃斑變性中之作用，其包含許多AMD情況，本發明方法可適用於治療及/或預防此類病症。濕性黃斑變性之當前療法涉及血管新生抑制劑，視情況與光動力療法(PDT)組合以使藥物靶向特定細胞。亦使用光凝來治療濕性黃斑變性，其中使用較高能量雷射光束以在具有不正常血管之視網膜區域中產生小燒傷。 In view of the role of angiogenesis in wet macular degeneration, which encompasses many AMD conditions, the methods of the invention are applicable to the treatment and/or prevention of such conditions. Current therapies for wet macular degeneration involve angiogenesis inhibitors, optionally combined with photodynamic therapy (PDT) to target drugs to specific cells. Photocoagulation is also used to treat wet macular degeneration, where a higher energy laser beam is used to create a small burn in the area of the retina with abnormal blood vessels.

本發明化合物可利用可有效地將本發明之化合物或組合物傳輸至適當或所需作用部位之任何途徑向哺乳動物、較佳人類投與，該途徑為諸如經口、經鼻、經肺、經皮，諸如被動或離子導入傳遞，或非經腸，例如經直腸、積存、皮下、靜脈內、肌肉內、鼻內、腦內、眼用溶液或軟膏。根據本發明之給藥方案可由單一劑量或經一段時間的複數個劑量組成。投與可為每日、每週(或以一些其他多天間隔)或按間斷的時程一次或多次。欲投與之本文所述之化合物或其醫藥組合物之確切量將隨各個體而變化。 The compounds of the invention may be administered to a mammal, preferably a human, by any route effective to deliver a compound or composition of the invention to the appropriate or desired site of action, such as by oral, nasal, or pulmonary, Transdermal, such as passive or iontophoretic delivery, or parenteral, such as transrectal, accumulative, subcutaneous, intravenous, intramuscular, intranasal, intracerebral, ophthalmic solutions or ointments. The dosage regimen according to the invention may consist of a single dose or multiple doses over a period of time. The vote may be one or more times daily, weekly (or at some other multi-day interval) or intermittent schedule. The exact amount of the compound or pharmaceutical composition thereof to be administered herein will vary with the individual.

可能需要降低眼新血管生成疾病中之血管新生程度。在一些實施例中，本文所述之化合物可傳遞至眼。傳遞至眼可例如使用玻璃體內、眼內及/或眼周途徑(諸如玻璃體內注射劑、結膜下注射劑等)達成。將本文所述之化合物局部施加至眼亦可例如使用點眼劑達成。眼投與途徑可尤其適用於治療眼新血管生成疾病，諸如黃斑變性。 It may be necessary to reduce the degree of angiogenesis in ocular neovascularization diseases. In some embodiments, the compounds described herein can be delivered to the eye. Delivery to the eye can be achieved, for example, using intravitreal, intraocular and/or periocular routes (such as intravitreal injections, subconjunctival injections, etc.). Topical application of the compounds described herein to the eye can also be achieved, for example, using an eye drop. The ocular route can be particularly useful for treating ocular neovascular diseases such as macular degeneration.

視投與途徑而定，有效劑量可根據以下計算：體重；體表面積；原發器官/腫瘤大小；及/或待治療之個體之癌轉移之數目、大小及/或類型。最終給藥方案將由主治醫師考慮會改變藥物作用之各種因素來確定，該等因素例如為藥物之比活性；患者之損傷及反應之強度；患者之年齡、健康狀況、體重、性別及飲食；任何存在之感染的嚴重度；投藥時間；其他療法之使用(或不使用)；及其他臨床因素。 Depending on the route of administration, the effective dose can be calculated based on: body weight; body surface area; primary organ/tumor size; and/or number, size and/or type of cancer metastasis of the individual to be treated. The final dosing regimen will be determined by the attending physician considering various factors that alter the action of the drug, such as the specific activity of the drug; the strength of the patient's injury and response; the age, health, weight, sex and diet of the patient; The severity of the infection present; the time of administration; the use of other therapies (or not); and other clinical factors.

本發明之方法可與其他療法組合使用(亦即，根據本發明之治療可與一或多種所需治療劑或醫療程序同時、在其之前或在其之後投與)。用於此類組合方案之療法(治療劑或程序)之具體組合將考慮所需治療劑及/或程序之相容性以及欲達成之所需治療作用。 The methods of the invention can be used in combination with other therapies (i.e., the treatment according to the invention can be administered simultaneously with, prior to, or subsequent to one or more desired therapeutic or medical procedures). The particular combination of therapies (therapeutics or procedures) for such combination regimens will take into account the compatibility of the desired therapeutic agent and/or procedure and the desired therapeutic effect desired.

舉例而言，對於侵襲性及轉移性癌症，本發明之方法可與其他程序一起使用，包括手術、放射線療法(例如γ輻射、神經元波束放射線療法、電子束放射線療法、質子療法、近接療法、全身性放射性同位素)、化學療法、內分泌療法、高溫及超低溫療法，視待治療之腫瘤而定。 For example, for invasive and metastatic cancer, the methods of the invention can be used with other procedures, including surgery, radiation therapy (eg, gamma radiation, neuron beam radiation therapy, electron beam radiation therapy, proton therapy, proximity therapy, Systemic radioisotopes), chemotherapy, endocrine therapy, high temperature and ultra-low temperature therapy, depending on the tumor to be treated.

本發明之方法還可與第二抗癌劑之一或多種其他組合同時、分開或在其之後使用。第二抗癌劑包括(但不限於)抗代謝產物(例如5-氟尿嘧啶、甲胺喋呤、氟達拉賓(fludarabine)、阿糖胞苷(cytarabine)(亦稱為胞嘧啶***糖苷或Ara-C)及高劑量阿糖胞苷)、抗微管劑(例如長春花生物鹼(vinca alkaloid)，諸如長春新鹼(vincristine)及長春鹼(vinblastine)；及紫杉烷(taxane)，諸如太平洋紫杉醇(paclitaxel)及多 西他賽(docetaxel))、烷化劑(例如二氯甲二乙胺(mechlorethamine)、氯芥苯丁酸(chlorambucil)、環磷醯胺(cyclophosphamide)、美法侖(melphalan)、美法侖、異環磷醯胺(ifosfamide)、卡莫司汀(carmustine)、阿紮胞苷(azacitidine)、地西他濱(decitabine)、白消安(busulfan)、環磷醯胺(cyclophosphamide)、達卡巴嗪(dacarbazine)、異環磷醯胺、及亞硝基脲(諸如卡莫司汀(carmustine)、洛莫司汀(lomustine)、雙氯乙基亞硝基脲及羥脲)、鉑試劑(例如順鉑(cisplatin)、卡鉑(carboplatin)、奧沙利鉑(oxaliplatin)、賽特鉑(satraplatin)(JM-216)及CI-973)、蒽環黴素(anthracycline)(例如多柔比星(doxorubicin)及道諾黴素(daunorubicin))、抗腫瘤抗生素(例如絲裂黴素(mitomycin)、博萊黴素(bleomycin)、艾達黴素(idarubicin)、阿黴素(adriamycin)、道諾黴素(daunomycin)(亦稱為道諾黴素(daunorubicin)、紅比黴素(rubidomycin)或柔紅黴素(cerubidine))及米托蒽醌(mitoxantrone))、拓撲異構酶抑制劑(例如依託泊苷(etoposide)及喜樹鹼(camptothecin))、嘌呤拮抗劑或嘧啶拮抗劑(例如6-巰基嘌呤、5-氟尿嘧啶、阿糖胞苷、氯法拉濱(clofarabine)及吉西他濱(gemcitabine))、細胞成熟劑(例如三氧化二砷及維甲酸(tretinoin))、DNA修復酶抑制劑(例如鬼臼毒素(podophyllotoxine)、依託泊苷(etoposide)、伊立替康(irinotecan)、拓撲替康(topotecan)及替尼泊苷(teniposide))、阻止細胞存活之酶(例如天冬醯胺酶及培門冬酶)、組蛋白脫乙醯基酶抑制劑(例如伏立諾他(vorinostat))、任何其他細胞毒性劑(例如磷酸雌莫司汀(estramustine phosphate)、***(dexamethasone)、潑尼氮芥(prednimustine)及丙卡巴肼(procarbazine))、激素(例如***、強的松(prednisone)、甲基潑尼龍(methylprednisolone)、他莫昔芬(tamoxifen)、亮丙立德(leuprolide)、氟他胺(flutamide)及甲地孕酮(megestrol))、單株抗體(例 如吉妥珠單抗奧唑米星(gemtuzumab ozogamicin)、阿侖單抗(alemtuzumab)、利妥昔單抗(rituximab)及釔-90-替伊莫單抗泰澤坦(yttrium-90-ibritumomab tiuxetan))、免疫調節劑(例如沙立度胺(thalidomide)及來那度胺(lenalidomide))、Bcr-Abl激酶抑制劑(例如AP23464、AZD0530、CGP76030、PD180970、SKI-606、伊馬替尼(imatinib)、BMS354825(達沙替尼(dasatinib))、AMN107(尼羅替尼(nilotinib))及VX-680)、激素促效劑或拮抗劑、部分促效劑或部分拮抗劑、激酶抑制劑、手術、放射線療法(例如γ輻射、中子束放射線療法、電子束放射線療法、質子療法、近接療法及全身性放射性同位素)、內分泌療法、生物學反應調節劑(例如干擾素、介白素及腫瘤壞死因子)、高溫及超低溫療法、及減弱任何不良影響之試劑(例如止吐藥)。在一個實施例中，抗癌劑或癌症治療劑為細胞毒性劑、抗代謝物、抗葉酸物、HDAC抑制劑、DNA***劑、DNA交聯劑、DNA烷化劑、DNA裂解劑、拓撲異構酶抑制劑、CDK抑制劑、JAK抑制劑、抗血管生成劑、Bcr-Abl抑制劑、HER2抑制劑、EGFR抑制劑、VEGFR抑制劑、PDGFR抑制劑、HGFR抑制劑、IGFR抑制劑、c-Kit抑制劑、Ras路徑抑制劑、PI3K抑制劑、多靶向激酶抑制劑、mTOR抑制劑、抗***、抗雄激素、芳香酶抑制劑、生長抑素類似物、ER調節劑、抗微管蛋白劑、長春花生物鹼、紫杉烷、HSP抑制劑、Smoothened拮抗劑、端粒酶抑制劑、抗轉移劑、免疫抑制劑、生物製劑(諸如抗體或激素療法)。 The method of the invention may also be used simultaneously, separately or after the one or more other combinations of the second anticancer agent. Second anticancer agents include, but are not limited to, antimetabolites (eg, 5-fluorouracil, methotrexate, fludarabine, cytarabine (also known as cytosine arabinoside or Ara) -C) and high dose cytarabine), anti-microtubule agents (such as vinca alkaloid, such as vincristine and vinblastine; and taxane, such as Pacific paclitaxel and more Cettaxel), alkylating agents (eg mechlorethamine, chlorambucil, cyclophosphamide, melphalan, melphalan) , ifosfamide, carmustine, azacitidine, decitabine, busulfan, cyclophosphamide, Dacarbazine, ifosfamide, and nitrosourea (such as carmustine, lomustine, bischloroethylnitrosourea, and hydroxyurea), platinum reagent (eg cisplatin, carboplatin, oxaliplatin, satraplatin (JM-216) and CI-973), anthracycline (eg more flexible) Doxorubicin and daunorubicin, antitumor antibiotics (eg mitomycin, bleomycin, idarubicin, adriamycin) , daunomycin (also known as daunorubicin, rubidomycin or cerubidine) and mitoxantrone) Topoisomerase inhibitors (eg, etoposide and camptothecin), guanidine antagonists or pyrimidine antagonists (eg 6-mercaptopurine, 5-fluorouracil, cytarabine, clofarabine) Clofarabine) and gemcitabine, cell maturation agents (such as arsenic trioxide and retinoic acid), DNA repair enzyme inhibitors (such as podophyllotoxine, etoposide, irinotecan) , topotecan and teniposide, enzymes that prevent cell survival (such as aspartate and asparaginase), histone deacetylase inhibitors (eg, vorino) He (vorinostat), any other cytotoxic agent (such as estramustine phosphate, dexamethasone, prednimustine and procarbazine), hormones (eg ground Dexamethasone, prednisone, methylprednisolone, tamoxifen, leuprolide, flutamide, and megestrol, Individual antibody (example Such as gemtuzumab ozogamicin, alemtuzumab, rituximab and 钇-90-tenimuzumab zeittan (yttrium-90-ibritumomab Tiuxetan)), immunomodulators (such as thalidomide and lenalidomide), Bcr-Abl kinase inhibitors (eg AP23464, AZD0530, CGP76030, PD180970, SKI-606, imatinib ( Imatinib), BMS354825 (dasatinib), AMN107 (nilotinib) and VX-680), hormonal agonists or antagonists, partial agonists or partial antagonists, kinase inhibitors , surgery, radiation therapy (eg gamma radiation, neutron beam radiotherapy, electron beam radiotherapy, proton therapy, proximity therapy and systemic radioisotopes), endocrine therapy, biological response modifiers (eg interferon, interleukin and Tumor necrosis factor), hyperthermia and ultra-low temperature therapy, and agents that attenuate any adverse effects (eg antiemetics). In one embodiment, the anticancer agent or cancer therapeutic agent is a cytotoxic agent, an antimetabolite, an antifolate, an HDAC inhibitor, a DNA insert, a DNA crosslinker, a DNA alkylating agent, a DNA cleaving agent, a topological difference Enzyme inhibitors, CDK inhibitors, JAK inhibitors, anti-angiogenic agents, Bcr-Abl inhibitors, HER2 inhibitors, EGFR inhibitors, VEGFR inhibitors, PDGFR inhibitors, HGFR inhibitors, IGFR inhibitors, c- Kit inhibitors, Ras pathway inhibitors, PI3K inhibitors, multi-targeted kinase inhibitors, mTOR inhibitors, antiestrogens, antiandrogens, aromatase inhibitors, somatostatin analogues, ER modulators, anti-microtubules Protein agents, vinca alkaloids, taxanes, HSP inhibitors, Smoothened antagonists, telomerase inhibitors, anti-metastatic agents, immunosuppressants, biological agents (such as antibodies or hormonal therapies).

血管新生在癌症及黃斑變性之進展中起到重要作用。本發明指示橙黃醯胺雙肽化合物深度抑制人類早期及晚期內皮祖細胞之血管生成功能。該等化合物亦展現離體及在活體內兩種情況下之有前景的抗血管生成作用。特定言之，該等化合物為阻斷黑素瘤之腫瘤生長及癌轉移以及AMD之病理性新血管生成的新穎血管新生抑制劑。 Angiogenesis plays an important role in the progression of cancer and macular degeneration. The present invention indicates that the orange flavonoid dipeptide compound deeply inhibits the angiogenic function of human early and late endothelial progenitor cells. These compounds also exhibit promising anti-angiogenic effects in both ex vivo and in vivo. In particular, these compounds are novel angiogenesis inhibitors that block tumor growth and cancer metastasis of melanoma and pathological neovascularization of AMD.

實例Instance

材料與方法：Materials and Methods:

人類CD34陽性內皮祖細胞之分離及培育Isolation and Cultivation of Human CD34 Positive Endothelial Progenitor Cells

此研究符合赫爾辛基宣言(Declaration of Helsinki)中關於人類組織使用中概述之原則。倫理學批准由臺灣臺北的馬偕紀念醫院(Mackay Memorial Hospital,Taipei,Taiwan)之機構審查委員會(Institutional Review Board)授予。在收集周邊血液(80mL)之前自健康供體獲得知情同意書。周邊血液單核細胞(PBMC)藉由離心在Ficoll-PaqueTM plus(GE Healthcare)上根據製造商之說明書而自其他血液組分分級分離。獲得且培養CD34陽性祖細胞。簡言之，細胞使用CD34微珠粒套組及MACSTM細胞分離系統(均來自Miltenyi Biotec)自PBMC分離。CD34陽性EPC維持在MV2完全培養基(PromoCell)中，該培養基由內皮基礎培養基、20%胎牛血清、hEGF、VEGF、hFGF-B、IGF-1、抗壞血酸及肝素組成。將1×10⁶個細胞/cm²接種於經纖維結合蛋白塗佈之培養皿(BD Biosciences)上且在37℃培育箱中在加濕95%空氣及5%CO₂氛圍下維持。 This study is consistent with the principles outlined in the Declaration of Helsinki on the use of human tissue. The ethics approval was granted by the Institutional Review Board of Mackay Memorial Hospital (Taipei, Taiwan). Informed consent was obtained from healthy donors prior to collecting peripheral blood (80 mL). Peripheral blood mononuclear cells (PBMC) were fractionated from other blood components by centrifugation on Ficoll-PaqueTM plus (GE Healthcare) according to the manufacturer's instructions. CD34 positive progenitor cells were obtained and cultured. Briefly, cells were isolated from PBMC using a CD34 microbead set and a MACSTM cell separation system (both from Miltenyi Biotec). CD34-positive EPC was maintained in MV2 complete medium (PromoCell) consisting of endothelial basal medium, 20% fetal bovine serum, hEGF, VEGF, hFGF-B, IGF-1, ascorbic acid and heparin. 1 × 10 ⁶ cells/cm ^{2 was} seeded on a fibronectin-coated petri dish (BD Biosciences) and maintained in a humidified 95% air and 5% CO ₂ atmosphere in a 37 ° C incubator.

細胞增殖分析Cell proliferation analysis

將EPC(5×10³個細胞/孔)接種至96孔盤上。在24小時培育之後，移除培養基，且細胞在不存在或存在Fmoc-10a之情況下與含有2%FBS的新鮮MV2完全培養基一起培育48小時。細胞與MTT(0.5mg/mL)一起培育2小時。甲臢晶體利用二甲亞碸(DMSO)溶解，且在550nm下用ELISA讀取器量測吸光度。 EPC (5 x 10 ³ cells/well) was seeded onto a 96-well plate. After 24 hours of incubation, the medium was removed and cells were incubated with fresh MV2 complete medium containing 2% FBS for 48 hours in the absence or presence of Fmoc-10a. The cells were incubated with MTT (0.5 mg/mL) for 2 hours. The formazan crystals were dissolved using dimethyl hydrazine (DMSO) and the absorbance was measured at 550 nm using an ELISA reader.

細胞毒性分析Cytotoxicity analysis

EPC在不存在或存在Fmoc-10a之情況下用含有2%FBS之MV2完全培養基處理48小時。LDH釋放百分比由在Fmoc-10a處理之後培養基中之LDH活性與細胞溶解物中之LDH活性的比率計算。 EPC was treated with MV2 complete medium containing 2% FBS for 48 hours in the absence or presence of Fmoc-10a. The percentage of LDH release was calculated from the ratio of LDH activity in the medium after treatment with Fmoc-10a to LDH activity in cell lysates.

細胞遷移分析Cell migration analysis

將EPC(5×10⁴個細胞/孔)接種至具有MV2完全培養基之上部腔室上，接著在具有含有2%FBS之具有指定濃度Fmoc-10之MV2完全培養基的底部腔室中培育。在16小時處理之後，機械移除過濾器上側上之細胞，且將遷移在下側上之細胞用4%甲醛固定，接著用0.5%結晶紫染色10分鐘。細胞遷移藉由用倒置相差顯微鏡在10個隨機視野中計數染色細胞數目來定量且進行攝影。 EPC (5 x 10 ⁴ cells/well) was seeded onto the upper chamber with MV2 complete medium, followed by incubation in a bottom chamber with MV2 complete medium containing the indicated concentration of Fmoc-10 containing 2% FBS. After 16 hours of treatment, the cells on the upper side of the filter were mechanically removed, and the cells migrating on the lower side were fixed with 4% formaldehyde, followed by staining with 0.5% crystal violet for 10 minutes. Cell migration was quantified and photographed by counting the number of stained cells in 10 random fields with an inverted phase contrast microscope.

毛細管形成分析Capillary formation analysis

將用於促進EPC分化成毛細管狀結構之基質膠(BD Biosciences,Bedford,MA)添加至48孔盤中。經基質膠塗佈之48孔盤在37℃下培育30分鐘以允許聚合。在凝膠形成之後，每孔接種EPC(6×10⁴個細胞)於含有2%FBS之具有指定濃度Fmoc-10a之MV2完全培養基中的聚合基質膠層上，接著在37℃下培育10-16小時。毛細管形成之顯微照片用倒置相差顯微鏡採集。管形成藉由在每孔3個隨機視野中藉由使用Image-Pro Plus軟體量測每一管之長軸來定量。 Matrigel (BD Biosciences, Bedford, MA) used to promote the differentiation of EPC into a capillary structure was added to a 48-well plate. Matrigel coated 48-well plates were incubated at 37 ° C for 30 minutes to allow for polymerization. After gel formation, EPC (6 x 10 ⁴ cells) was seeded per well on a polymeric matrix gel layer containing 2% FBS in MV2 complete medium with the indicated concentration of Fmoc-10a, followed by incubation at 37 °C. 16 hours. Photomicrographs of capillary formation were collected using an inverted phase contrast microscope. Tube formation was quantified by measuring the long axis of each tube using Image-Pro Plus software in 3 random fields per well.

群落形成分析Community formation analysis

將經分離之PBMC再懸浮在MV2完全培養基中。在2天之後，收集非附著細胞，且再接種至經纖維結合蛋白塗佈之24孔盤上。五天後，計數每孔早期EPC之群落形成單位(CFU)之數目。 The isolated PBMC were resuspended in MV2 complete medium. After 2 days, non-adherent cells were collected and re-inoculated onto a fibronectin coated 24-well plate. After five days, the number of community forming units (CFU) of early EPC per well was counted.

早期EPC趨化性分析Early EPC chemotaxis analysis

早期EPC之趨化運動使用Transwell評估。下層過濾器表面經明膠塗佈。將存在或不存在MV2完全培養基之Fmoc-10a置放於下層孔中。腔室在37℃下培育4小時。細胞用冷甲醇固定且用0.5%結晶紫染色。 The early EPC chemotaxis exercise was evaluated using Transwell. The lower filter surface is coated with gelatin. Fmoc-10a in the presence or absence of MV2 complete medium was placed in the lower wells. The chamber was incubated at 37 ° C for 4 hours. The cells were fixed with cold methanol and stained with 0.5% crystal violet.

主動脈環出芽分析Aortic ring sprouting analysis

主動脈收集自8至10週齡史泊格多利大白鼠(Sprague-Dawley rat)。在澈底洗滌之後，將主動脈切成1mm環片段。將主動脈環置放 於48孔盤中，該等盤預塗佈130μl基質膠且在37℃下聚合。該等孔隨後再用50μl基質膠覆蓋以用於密封。接著將具有或不具有Fmoc-10a之VEGF(20ng/ml)添加至孔中。每3天更換培養基。在第8天觀測出芽內皮細胞且攝影。出芽血管之面積利用Image-Pro Plus軟體定量地量測。 The aorta was collected from 8 to 10 weeks old Sprague-Dawley rat. After the clear wash, the aorta was cut into 1 mm loop segments. Place the aortic annulus In a 48-well plate, the disks were precoated with 130 μl of Matrigel and polymerized at 37 °C. The wells were then covered with 50 μl of Matrigel for sealing. VEGF (20 ng/ml) with or without Fmoc-10a was then added to the wells. The medium was changed every 3 days. Bud endothelial cells were observed on day 8 and photographed. The area of the budding blood vessels was quantitatively measured using Image-Pro Plus software.

定向活體內血管新生分析(DIVAA)Directed in vivo angiogenesis analysis (DIVAA)

手術矽酮試管在4℃下用含有具有或不具有Fmoc-10a之VEGF的基質膠填充。接著，將經麻醉小鼠(C57BL/6小鼠)之腰部背刮毛且滅菌準備。製得5mm皮膚切口且用無菌止血鉗形成10mm深的皮下凹穴。DIVAA試管在37℃下培育1小時以允許凝膠形成且接著植入至小鼠之背側中。在15天之後，獲取DIVAA試管且攝影。新血管藉由用德拉布金法(Drabkin method)量測栓塞之血紅素來定量。所有涉及動物實驗之程序均由馬偕醫學院(Mackay Medical College)之機構動物護理及使用委員會(Institutional Animal Care and Use Committee)批准。 The surgical ketone tube was filled with Matrigel containing VEGF with or without Fmoc-10a at 4 °C. Next, the waist of the anesthetized mouse (C57BL/6 mouse) was shaved back and sterilized. A 5 mm skin incision was made and a 10 mm deep subcutaneous pocket was formed with a sterile hemostat. DIVAA tubes were incubated for 1 hour at 37 °C to allow gel formation and subsequent implantation into the dorsal side of the mice. After 15 days, DIVAA tubes were taken and photographed. New blood vessels were quantified by measuring the hemoglobin of the embolization using the Drabkin method. All procedures involving animal experiments were approved by the Institutional Animal Care and Use Committee of Mackay Medical College.

黑素瘤細胞之培養Culture of melanoma cells

小鼠黑素瘤細胞株(B16F10)獲自美國典型培養物保藏中心(American Type Culture Collection，ATCC，Manassas,VA)且培養在含有10%FBS、青黴素(100單位/ml)、鏈黴素(100μg/ml)及L-麩醯胺酸(2mM)中之DMEM。將細胞維持在37℃含有5%CO2之加濕空氣中。 The mouse melanoma cell line (B16F10) was obtained from the American Type Culture Collection (ATCC, Manassas, VA) and cultured in 10% FBS, penicillin (100 units/ml), streptomycin ( 100 μg/ml) and DMEM in L-glutamic acid (2 mM). The cells were maintained at 37 ° C in humidified air containing 5% CO 2 .

侵襲分析Invasion analysis

過濾器上側經濃度為125μg/cm²之基質膠(BD Biosciences，Bedford,MA)塗佈。將B16F10細胞(5×10⁴個細胞/孔)接種至具有無血清培養基之上部腔室上，接著在具有含有10%FBS之不存在或存在Fmoc-10a之培養基的底部腔室中培育。在16小時處理之後，機械移除過濾器上側上之細胞，且將遷移在下側上之細胞用4%甲醛固定，接著用0.5%結晶紫染色10分鐘。最後，侵襲之細胞藉由用倒置相差顯微 鏡在10個隨機視野中計數染色細胞數目來定量且進行攝影。 The upper side of the filter was coated with Matrigel (BD Biosciences, Bedford, MA) at a concentration of 125 μg/cm ² . B16F10 cells (5 x 10 ⁴ cells/well) were seeded onto a chamber with serum-free medium above, followed by incubation in a bottom chamber with medium containing 10% FBS in the absence or presence of Fmoc-10a. After 16 hours of treatment, the cells on the upper side of the filter were mechanically removed, and the cells migrating on the lower side were fixed with 4% formaldehyde, followed by staining with 0.5% crystal violet for 10 minutes. Finally, the invading cells were quantified and photographed by counting the number of stained cells in 10 random fields with an inverted phase contrast microscope.

VEGF ELISA分析VEGF ELISA analysis

B16F10細胞用各種濃度之Fmoc-10a或媒劑處理24小時，且接著收集上清液以檢查血管內皮生長因子(VEGF)蛋白之量。VEGF小鼠ELISA套組(Novex,Invitrogen)根據製造商之說明書使用。 B16F10 cells were treated with various concentrations of Fmoc-10a or vehicle for 24 hours, and then the supernatant was collected to examine the amount of vascular endothelial growth factor (VEGF) protein. The VEGF mouse ELISA kit (Novex, Invitrogen) was used according to the manufacturer's instructions.

皮下異種移植模型Subcutaneous xenograft model

將體積為100μL之B16F10細胞(2×10⁶個)皮下注射至C57BL/6小鼠之右側腹中。在9天之後，當腫瘤直徑達至2mm時，小鼠每日藉由腹膜內注射來隨機給予媒劑或指定劑量之Fmoc-10a。每日量測腫瘤大小及體重。腫瘤體積根據下式計算：V=(長度×寬度²/2)。根據動物倫理委員會(Animal Ethics Committee)，當動物中出現不堪忍受之情形時，應停止實驗。 A volume of 100 μL of B16F10 cells (2 × 10 ⁶ cells) was subcutaneously injected into the right abdomen of C57BL/6 mice. After 9 days, when the tumor diameter reached 2 mm, the mice were randomly administered daily with a vehicle or a prescribed dose of Fmoc-10a by intraperitoneal injection. Tumor size and body weight were measured daily. The tumor volume was calculated according to the following formula: V = (length x width ² /2). According to the Animal Ethics Committee, experiments should be stopped when there is an unbearable situation in the animal.

實驗癌轉移模型Experimental cancer metastasis model

將B16F10細胞(2×10⁶個)緩慢注射至C57BL/6小鼠之側尾靜脈中以引發腫瘤癌轉移。自腫瘤細胞注射之前3天至腫瘤細胞注射之後21天(處死之日)每日向小鼠腹膜內投與Fmoc-10a(1.5或4.5mg/kg)。 B16F10 cells (2 x 10 ⁶ cells) were slowly injected into the lateral tail vein of C57BL/6 mice to initiate tumor metastasis. Fmoc-10a (1.5 or 4.5 mg/kg) was administered intraperitoneally to mice daily from 3 days before tumor cell injection to 21 days after tumor cell injection (day of sacrifice).

氧誘導之視網膜病變(OIR)Oxygen-induced retinopathy (OIR)

OIR之小鼠模型已在與視網膜缺血疾病(包括AMD)相關之研究中及在評估抗血管生成化合物之功效的研究中廣泛用於誘導視網膜新血管生成。簡言之，出生後(P7)之BALB/c裸鼠與其哺乳母鼠(nursing dam)一起經歷氧過多(75%氧氣)持續5天(P12)，且接著經歷室內空氣以製造視網膜新血管生成。在返回至常氧後發生新血管生成且在P17達至峰值。OIR模型中之所有小鼠在安樂死時經由左心室穿刺用5ml若丹明(rhodamine)結合之葡聚糖灌注固定。接著，執行視網膜血管之血管造影，且製備視網膜整體裝片以對視網膜病變特徵進行評分。 Mouse models of OIR have been widely used to induce retinal neovascularization in studies related to retinal ischemic diseases (including AMD) and in studies evaluating the efficacy of anti-angiogenic compounds. Briefly, postnatal (P7) BALB/c nude mice, together with their nursing dam, experience hyperoxia (75% oxygen) for 5 days (P12) and then undergo indoor air to create retinal neovascularization . New angiogenesis occurs after returning to normoxia and peaks at P17. All mice in the OIR model were perfused with 5 ml of rhodamine-conjugated dextran via levation by left ventricle during euthanasia. Next, angiography of the retinal blood vessels is performed, and a retinal integral patch is prepared to score the retinopathy features.

實例1 Fmoc-10a抑制人類晚期EPC之細胞增殖、遷移及管形成且無細Example 1 Fmoc-10a inhibits cell proliferation, migration and tube formation in human advanced EPC without fine 胞毒性作用.Cytotoxic effect.

在48小時處理之後，Fmoc-10a以濃度依賴性方式抑制晚期EPC之細胞增殖(圖1A及B)。另外，Fmoc-10a甚至在高濃度(30μM)下亦不誘導晚期EPC之LDH釋放(圖1C)。Fmoc-10a在16小時處理之後顯著抑制晚期EPC之細胞遷移及管形成(圖1D及E)。此等結果指示Fmoc-10a具有在不以細胞毒性方式起作用的情況下阻斷活體外血管新生之能力。 After 48 hours of treatment, Fmoc-10a inhibited cell proliferation of late EPC in a concentration dependent manner (Figures 1A and B). In addition, Fmoc-10a did not induce LDH release from late EPC even at high concentrations (30 μM) (Fig. 1C). Fmoc-10a significantly inhibited cell migration and tube formation of advanced EPC after 16 hours of treatment (Figures 1D and E). These results indicate that Fmoc-10a has the ability to block angiogenesis in vitro without acting in a cytotoxic manner.

實例2 Fmoc-10a抑制人類早期EPC之群落形成及趨化運動Example 2 Fmoc-10a inhibits community formation and chemotaxis of early human EPC

出現證據展現早期EPC藉由介導血管生成開關來促進血管新生及血小管生成。Fmoc-10a顯著抑制早期EPC之周邊處的CFU-希爾(CFU-Hill)群落及伸長之紡錘狀細胞(圖2A及B)。Fmoc-10a亦顯著抑制早期EPC之趨化性(圖2C)。此等結果指示Fmoc-10a具有阻斷早期EPC之血管生成開關之藥理學功能。 Evidence suggests that early EPC promotes angiogenesis and tubule formation by mediating angiogenesis switches. Fmoc-10a significantly inhibited the CFU-Hill community and elongated spindle cells at the periphery of the early EPC (Fig. 2A and B). Fmoc-10a also significantly inhibited the chemotaxis of early EPC (Fig. 2C). These results indicate that Fmoc-10a has a pharmacological function that blocks the angiogenic switch of early EPC.

實例3 Fmoc-10a離體及在活體內抑制血管新生.Example 3 Fmoc-10a is isolated and inhibits angiogenesis in vivo.

血管內皮生長因子(VEGF)為調節血管新生之最強力的血管生成因子。Fmoc-10a顯著消除來自主動脈環之VEGF誘導之血管出芽(圖3A)。此外，藉由分析血紅素含量，Fmoc-10a濃度依賴性地減弱基質膠栓塞中之微血管形成。此等結果指示Fmoc-10a離體及在活體內阻斷VEGF誘導之血管新生(圖3B)。 Vascular endothelial growth factor (VEGF) is the most potent angiogenic factor that regulates angiogenesis. Fmoc-10a significantly abolished VEGF-induced vascular sprouting from the aortic rings (Fig. 3A). Furthermore, by analyzing the heme content, Fmoc-10a dose-dependently attenuated microvascular formation in matrigel embolization. These results indicate that Fmoc-10a blocks VEGF-induced angiogenesis in vitro and in vivo (Fig. 3B).

實例4 Fmoc-10a在活體外及活體內抑制黑素瘤之腫瘤生長及癌轉移.Example 4 Fmoc-10a inhibits tumor growth and cancer metastasis of melanoma in vitro and in vivo.

黑素瘤為侵襲性皮膚癌且亦幾乎為皮膚癌相關之死亡原因。儘管黑素瘤為相對罕見的癌症，但黑素瘤之臭名昭著的侵襲性係歸因於其顯著的轉移性傾向。惡性黑素瘤已充分記錄為高血管生成腫瘤類型，清楚地展示新血管形成作為非典型黑素細胞之疾病進展中之重要步驟。血管生成生長因子(諸如VEGF)在黑素瘤中過度表現且已發現與疾病進展及預後相關。吾人展現Fmoc-10a顯著抑制黑素瘤細胞中之細胞侵襲及VEGF製造(圖4A及B)。Fmoc-10a以劑量依賴性方式強力 阻斷黑素瘤細胞之腫瘤生長(圖4C)。此外，Fmoc-10a實質上抑制黑素瘤癌症之肺部癌轉移(圖4D)。此等結果指示Fmoc-10a經由抑制黑素瘤細胞中之VEGF製造及侵襲特性以及黑素瘤微環境中之血管新生抑制展現出新穎抗腫瘤及抗癌轉移活性。 Melanoma is an invasive skin cancer and is also a cause of death associated with skin cancer. Although melanoma is a relatively rare cancer, the notorious invasive line of melanoma is attributable to its significant metastatic tendency. Malignant melanoma has been well documented as a type of hyperangiogenic tumor that clearly demonstrates that neovascularization is an important step in the progression of disease as atypical melanocytes. Angiogenic growth factors, such as VEGF, are overexpressed in melanoma and have been found to be associated with disease progression and prognosis. We have shown that Fmoc-10a significantly inhibits cell invasion and VEGF production in melanoma cells (Fig. 4A and B). Fmoc-10a is strong in a dose-dependent manner Tumor growth of melanoma cells was blocked (Fig. 4C). Furthermore, Fmoc-10a substantially inhibits lung cancer metastasis of melanoma cancer (Fig. 4D). These results indicate that Fmoc-10a exhibits novel anti-tumor and anti-cancer metastatic activity via inhibition of VEGF production and invasion properties in melanoma cells and angiogenesis inhibition in the melanoma microenvironment.

實例5 Fmoc-10a抑制AMD動物模型中之脈絡膜新生血管(CNV).Example 5 Fmoc-10a inhibits choroidal neovascularization (CNV) in an animal model of AMD.

新血管生成在AMD中起重要作用。眼新生血管性疾病之治療利用靶向VEGF之玻璃體內療法而得到澈底改變。Fmoc-10a如癌思停般顯著阻斷小鼠之氧過多誘導之病理性新血管生成(圖5)。此等結果指示Fmoc-10a為具有治療血管新生相關疾病(諸如AMD)之潛能的有前景的血管新生抑制劑。 New angiogenesis plays an important role in AMD. The treatment of ocular neovascular diseases is altered by intravitreal treatment with VEGF. Fmoc-10a significantly blocked hyperoxia-induced pathological neovascularization in mice as in the case of cancer (Fig. 5). These results indicate that Fmoc-10a is a promising angiogenesis inhibitor with the potential to treat angiogenesis-related diseases such as AMD.

靶向EPC以阻斷血管新生為用於血管新生相關疾病之目前有吸引力的治療方法。本文中，本發明人已展示Fmoc-10a(橙黃醯胺雙肽化合物)深度抑制人類早期及晚期EPC之血管生成功能。Fmoc-10a亦展現離體及在活體內兩種情況下之有前景的抗血管生成作用。Fmoc-10a為阻斷黑素瘤之腫瘤生長及癌轉移以及AMD之病理性新血管生成的新穎血管新生抑制劑。因此，本發明將具有應用於治療血管新生相關疾病(尤其黑素瘤癌症及AMD)之極大潛能。 Targeting EPC to block angiogenesis is currently an attractive treatment for angiogenesis-related diseases. Herein, the present inventors have shown that Fmoc-10a (orange indole dipeptide compound) deeply inhibits the angiogenic function of human early and late EPC. Fmoc-10a also exhibits promising anti-angiogenic effects in both ex vivo and in vivo conditions. Fmoc-10a is a novel angiogenesis inhibitor that blocks tumor growth and cancer metastasis of melanoma and pathological neovascularization of AMD. Thus, the present invention will have great potential for the treatment of angiogenesis-related diseases, particularly melanoma cancer and AMD.

Claims (9)

一種化合物(S)-1-((R)-1-羥基-3-苯基丙-2-基胺基)-3-甲基-1-側氧基丁-2-基胺基甲酸(9H-茀-9-基)甲酯、其互變異構體、立體異構體、對映異構體或醫藥學上可接受之鹽之用途，其中該化合物具有下式： 其用於製備抑制、改善、預防或治療侵襲性及轉移性癌症或腫瘤之血管新生或眼新血管生成之藥劑，其中該侵襲性及轉移性癌症或腫瘤係選自腦癌、乳癌、腎癌、卵巢癌、肺癌、結腸癌及黑素瘤中之任一者。 A compound ( S )-1-(( R )-1-hydroxy-3-phenylpropan-2-ylamino)-3-methyl-1-oxobutan-2-ylaminocarboxylic acid (9) Use of a H -fluoren-9-yl)methyl ester, a tautomer, a stereoisomer, an enantiomer or a pharmaceutically acceptable salt thereof, wherein the compound has the formula: It is used for the preparation of an agent for inhibiting, ameliorating, preventing or treating angiogenesis or ocular neovascularization in invasive and metastatic cancer or tumor, wherein the invasive and metastatic cancer or tumor is selected from the group consisting of brain cancer, breast cancer, and kidney cancer. Any of ovarian cancer, lung cancer, colon cancer, and melanoma. 如請求項1之用途，其中該侵襲性及轉移性癌症或腫瘤為黑素瘤。 The use of claim 1, wherein the invasive and metastatic cancer or tumor is melanoma. 如請求項1之用途，其中該藥劑與手術、放射線療法或化學療法組合、在其之前或在其之後使用。 The use of claim 1, wherein the agent is used in combination with, before, or after surgery, radiation therapy or chemotherapy. 如請求項1之用途，其中該藥劑可進一步包含第二抗癌劑或與第二抗癌劑同時、分開或隨後投與。 The use of claim 1, wherein the agent may further comprise or be administered simultaneously, separately or subsequently with the second anticancer agent. 如請求項1之用途，其中該眼新血管生成為黃斑變性。 The use of claim 1, wherein the neovascularization of the eye is macular degeneration. 如請求項5之用途，其中該黃斑變性為濕性黃斑變性。 The use of claim 5, wherein the macular degeneration is wet macular degeneration. 如請求項6之用途，其中該濕性黃斑變性為年齡相關之黃斑變性(AMD)。 The use of claim 6, wherein the wet macular degeneration is age-related macular degeneration (AMD). 如請求項5之用途，其中該藥劑係以玻璃體內、眼內或眼周途徑投與。 The use of claim 5, wherein the agent is administered intravitreally, intraocularly or periocularly. 如請求項5之用途，其中該化合物係調配成注射劑或點眼劑。 The use of claim 5, wherein the compound is formulated as an injection or an eye drop.