VEGFR2 4.2 nM (IC50) PDGFRβ 6.6 nM (IC50) c-kit 2.3 nM (IC50)

苹果酸法米替尼抑制置于基质胶中的大鼠主动脉环的微血管喷出，以及 VEGF 诱导的人脐静脉内皮细胞的增殖、迁移和小管形成[1]。在胃癌细胞系中，苹果酸夫米替尼（1.8 和 3.6 μM；48 小时）会诱导细胞周期停滞在 G2/M 期，从而减少细胞生长，并导致剂量依赖性细胞凋亡[2]。苹果酸夫米替尼（0.6-20.0 µM；24-72 小时）以剂量依赖性方式抑制胃癌细胞的生长[2]。

许多由人类肿瘤细胞系制成的异种移植物在接触具有广泛而强大的抗肿瘤作用的苹果酸夫米替尼时会出现消退或生长停滞[1]。通过抑制血管生成，苹果酸米替尼（50 和 100 mg/kg；口服，每日一次，持续 3 周）可减少体内肿瘤的生长[2]。

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在体内，famitinib表现出广泛而有效的抗肿瘤活性，导致各种来源于人类肿瘤细胞系的已建立异种移植物的退化或生长停滞。此外，当联合使用奥沙利铂或5-氟尿嘧啶时，法米替尼显著增强了其疗效。综上所述，famitinib具有强大的临床前抗肿瘤活性，支持其在临床上的进一步评估。Famitinib目前正在中国进行I期临床试验[1]。

Famitinib抑制c-kit、VEGFR-2、PDGFRα和PDGFRβ的活性，IC50值分别为2.3 nM、4.7 nM和6.6 nM。此外，Famitinib抑制了VEGF诱导的人脐静脉内皮细胞的增殖、迁移和小管形成，以及从基质凝胶包埋的大鼠主动脉环中喷出的微血管。[1]

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末端脱氧核苷酸转移酶dUTP缺口末端标记（TUNEL）法根据制造商的方案通过TUNEL法测量细胞凋亡。用famitinib处理细胞48小时后，用磷酸缓冲盐水（PBS）洗涤细胞，并在室温下用4%多聚甲醛固定10分钟。然后用TUNEL检测试剂盒中提供的相应试剂对细胞进行染色。在覆盖盖玻片后，在荧光显微镜下对载玻片进行成像。阳性细胞显示绿色荧光，并从三个随机显微镜视野中计数[2]。

细胞增殖测定[2]
细胞类型：人胃癌细胞BGC-823和MGC-803
测试浓度：0、0.6、1.25、2.5 、5.0、10.0 和 20.0 µM
孵育时间：24、48 和 72 小时
实验结果：以剂量依赖性方式抑制细胞生长BGC-823 和 MGC -803 细胞的 IC50 值分别为 3.6 和 3.1 µM。

Animal/Disease Models: 18-20 g female BALB/c athymic nu/nu (nude) mice (age, 6–8 weeks) bearing BGC-823 xenografts[2]
Doses: 50 and 100 mg/kg
Route of Administration: po (oral gavage); 50 and 100 mg/kg; one time/day for 3 weeks
Experimental Results: Inhibited BGC-823 xenograft growth (tumor volume, 395.2 vs. 2,690.5 mm3), and animal weights were similar between groups (21.6 vs. 18.7 g).

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In vivo xenograft model experiments BGC-823 cells were suspended in PBS (1×107 cells/ml), and 100 µl of the cell suspension was subcutaneously injected into the right axillary area of 18-20-g female BALB/c athymic nu/nu mice (n=40; age, 6–8 weeks). The temperature of the housing conditions was maintained at 23–25°C with a humidity of 50–60% and a 10/14 h light/dark cycle. Food and water were changed 3 times a week. When the tumor volume reached ~100 mm3, mice were randomized into treatment groups. Tumors and animal weights were measured twice weekly, and tumor volume was calculated using the following formula: V=LxW2x1/2 (where V represents tumor volume, L is the length of the tumor and W is the width of the tumor). To measure famitinib, three groups were randomized (n=5 mice/group) as follows: Control group (gavage, physiological saline, once daily for 3 weeks); low-dose famitinib group (gavage, 50 mg/kg, once daily for 3 weeks); and high-dose famitinib group (gavage, 100 mg/kg, once for 3 weeks). A dose of 50 mg/kg was used for the following experiments. To compare famitinib with other drugs, animals were randomized (n=5 mice/group) as follows: Control group (gavage, physiological saline, once daily for 3 weeks); famitinib group (gavage, 50 mg/kg, once daily for 3 weeks); 5-FU group [10 mg/kg, intraperitoneal (ip), once every 2 days for 3 weeks]; DDP group (3 mg/kg, ip, once weekly for 3 weeks); and PTX group (10 mg/kg, ip, once a week for 3 weeks). Then, tumors and weight were quantified[2].

[1]. Abstract 3604: Preclinical antitumor study of famitinib, an orally available multi-targeted kinase inhibitor of VEGFR/PDGFR/c-Kit in phase I clinical trials.

[2]. Famitinib exerted powerful antitumor activity in human gastric cancer cells and xenografts. Oncol Lett. 2016 Sep;12(3):1763-1768.

Famitinib (SHR1020), a novel multi-targeted tyrosine kinase inhibitor, has antitumor activity against several solid tumors via targeting vascular endothelial growth factor receptor 2, c-Kit and platelet-derived growth factor receptor β. The present study investigated famitinib's activity against human gastric cancer cells in vitro and in vivo. Cell viability and apoptosis were measured, and cell cycle analysis was performed following famitinib treatment using 3-(4,5-dimethylthiazol -2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium assay, flow cytometry, terminal deoxynucleotidyl transferase dUTP nick end labeling assay and western blotting. Subsequently, cluster of differentiation 34 staining was used to evaluate microvessel density. BGC-823-derived xenografts in nude mice were established to assess drug efficacy in vivo. Famitinib inhibited cell proliferation by inducing cell cycle arrest at the G2/M phase and caused cell apoptosis in a dose-dependent manner in gastric cancer cell lines. In BGC-823 xenograft models, famitinib significantly slowed tumor growth in vivo via inhibition of angiogenesis. Compared with other chemotherapeutics such as 5-fluorouracil, cisplatin or paclitaxel alone, famitinib exhibited the greatest tumor suppression effect (>85% inhibition). The present study demonstrated for the first time that famitinib has efficacy against human gastric cancer in vitro and in vivo, which may lay the foundations for future clinical trials.[2]

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Angiogenesis plays a key role in tumor progression and anti-angiogenic agents including sunitinib and sorafenib that target VEGF/VEGFR signaling pathway have been proved to an effective therapy for cancer in the clinic. However, severe side effects such as hypertension and bone marrow toxicity limit their clinical application. Thus, development of novel anti-angiogenic agents with fewer side effects is still an unmet challenge. In this study, we characterized the in vitro and in vivo antitumor activity of famitinib, an orally active multi-targeted kinase inhibitor. Famitinib inhibited the activity of c-kit, VEGFR-2, PDGFRα and PDGFRβ with IC50 values of 2.3 nM, 4.7 nM and 6.6 nM, respectively. In addition, Famitinib inhibited the VEGF-induced proliferation, migration and tubule formation of human umbilical vein endothelial cells, and micro-vessel spouting from matrigel-embedded rat aortic rings. In vivo, famitinib exhibited broad and potent anti-tumor activity, leading to regression or growth arrest of various established xenografts derived from human tumor cell lines. Moreover, famitinib significantly enhanced the efficacy of oxaliplatin or 5-fluorouracil when they were combined. In summary, famitinib has potent preclinical antitumor activity which supports its further evaluation in clinic. Famitinib is currently in phase I clinical trials in China.[1]

C27H33FN4O7

544.233

1256377-67-9

Famitinib;1044040-56-3

49840531

Typically exists as solid at room temperature

2.224

166.76

5

9

9

39

824

1

[C@@H](O)(C(=O)O)CC(=O)O.C(C1NC2CCN(CCN(CC)CC)C(=O)C=2C=1C)=C1C(NC2=CC=C(F)C=C12)=O

JNDRBKCNKMZANY-QLTVYZEUSA-N

InChI=1S/C23H27FN4O2.C4H6O5/c1-4-27(5-2)10-11-28-9-8-19-21(23(28)30)14(3)20(25-19)13-17-16-12-15(24)6-7-18(16)26-22(17)29;5-2(4(8)9)1-3(6)7/h6-7,12-13,25H,4-5,8-11H2,1-3H3,(H,26,29);2,5H,1H2,(H,6,7)(H,8,9)/b17-13-;/t;2-/m.0/s1

5-[2-(diethylamino)ethyl]-2-[(Z)-(5-fluoro-2-oxo-1H-indol-3-ylidene)methyl]-3-methyl-6,7-dihydro-1H-pyrrolo[3,2-c]pyridin-4-one;(2S)-2-hydroxybutanedioic acid

Famitinib malate; Famitinib S-malate; 4RST0F28MR; UNII-4RST0F28MR; 1256377-67-9; Famitinib L-Malate; Famitinib malate [WHO-DD]; DTXSID30154828

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples

注意: 如下所列的是一些常用的体内动物实验溶解配方，主要用于溶解难溶或不溶于水的产品（水溶度<1 mg/mL）。 建议您先取少量样品进行尝试，如该配方可行，再根据实验需求增加样品量。

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注射用配方1: DMSO : Tween 80： Saline = 10 : 5 : 85 (如： 100 μL DMSO → 50 μL Tween 80 → 850 μL Saline)
*生理盐水/Saline的制备：将0.9g氯化钠/NaCl溶解在100 mL ddH ₂ O中，得到澄清溶液。
注射用配方 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (如： 100 μL DMSO → 400 μL PEG300 → 50 μL Tween 80 → 450 μL Saline)
注射用配方 3: DMSO : Corn oil = 10 : 90 (如： 100 μL DMSO → 900 μL Corn oil)
示例: 以注射用配方 3 (DMSO : Corn oil = 10 : 90) 为例说明, 如果要配制 1 mL 2.5 mg/mL的工作液, 您可以取 100 μL 25 mg/mL 澄清的 DMSO 储备液，加到 900 μL Corn oil/玉米油中, 混合均匀。

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注射用配方 4: DMSO : 20% SBE-β-CD in Saline = 10 : 90 [如：100 μL DMSO → 900 μL (20% SBE-β-CD in Saline)]
*20% SBE-β-CD in Saline的制备（4°C，储存1周）：将2g SBE-β-CD (磺丁基-β-环糊精) 溶解于10mL生理盐水中，得到澄清溶液。
注射用配方 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (如： 500 μL 2-Hydroxypropyl-β-cyclodextrin (羟丙基环胡精)→ 500 μL Saline)
注射用配方 6: DMSO : PEG300 : Castor oil : Saline = 5 : 10 : 20 : 65 (如： 50 μL DMSO → 100 μL PEG300 → 200 μL Castor oil → 650 μL Saline)
注射用配方 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (如： 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline)
注射用配方 8: 溶解于Cremophor/Ethanol (50 : 50), 然后用生理盐水稀释。
注射用配方 9: EtOH : Corn oil = 10 : 90 (如： 100 μL EtOH → 900 μL Corn oil)
注射用配方 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (如： 100 μL EtOH → 400 μL PEG300 → 50 μL Tween 80 → 450 μL Saline)

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口服配方 1: 悬浮于0.5% CMC Na (羧甲基纤维素钠)
口服配方 2: 悬浮于0.5% Carboxymethyl cellulose (羧甲基纤维素)
示例: 以口服配方 1 (悬浮于 0.5% CMC Na)为例说明, 如果要配制 100 mL 2.5 mg/mL 的工作液, 您可以先取0.5g CMC Na并将其溶解于100mL ddH2O中，得到0.5%CMC-Na澄清溶液；然后将250 mg待测化合物加到100 mL前述 0.5%CMC Na溶液中，得到悬浮液。

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口服配方 3: 溶解于 PEG400 (聚乙二醇400)
口服配方 4: 悬浮于0.2% Carboxymethyl cellulose (羧甲基纤维素)
口服配方 5: 溶解于0.25% Tween 80 and 0.5% Carboxymethyl cellulose (羧甲基纤维素)
口服配方 6: 做成粉末与食物混合

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注意: 以上为较为常见方法，仅供参考， InvivoChem并未独立验证这些配方的准确性。具体溶剂的选择首先应参照文献已报道溶解方法、配方或剂型，对于某些尚未有文献报道溶解方法的化合物，需通过前期实验来确定（建议先取少量样品进行尝试），包括产品的溶解情况、梯度设置、动物的耐受性等。

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请根据您的实验动物和给药方式选择适当的溶解配方/方案：
1、请先配制澄清的储备液(如：用DMSO配置50 或 100 mg/mL母液(储备液));
2、取适量母液，按从左到右的顺序依次添加助溶剂，澄清后再加入下一助溶剂。以 下列配方为例说明 (注意此配方只用于说明，并不一定代表此产品 的实际溶解配方):
10% DMSO → 40% PEG300 → 5% Tween-80 → 45% ddH2O (或 saline);
假设最终工作液的体积为 1 mL, 浓度为5 mg/mL: 取 100 μL 50 mg/mL 的澄清 DMSO 储备液加到 400 μL PEG300 中，混合均匀/澄清；向上述体系中加入50 μL Tween-80，混合均匀/澄清；然后继续加入450 μL ddH2O (或 saline)定容至 1 mL；
3、溶剂前显示的百分比是指该溶剂在最终溶液/工作液中的体积所占比例;
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6、如不确定怎么将母液配置成体内动物实验的工作液，请查看说明书或联系我们;
7、 以上所有助溶剂都可在 Invivochem.cn网站购买。