Y15   中文名称: 1,2,4,5-苯四胺四盐酸盐

FAK

Y15 体外处理会导致结肠癌细胞、乳腺癌细胞和黑色素瘤细胞活力降低、脱离增加和细胞凋亡增加。 Y15 剂量依赖性地抑制 TPC1、BCPAP、K1 和 TT 细胞系中总 FAK 和 pY397 的表达。甲状腺癌的所有细胞系都会经历有效的剂量依赖性分离。所有髓样和乳头状甲状腺癌细胞系都表现出坏死增加，并且当暴露于 Y15 时，所有这些细胞系都表现出集落形成的剂量依赖性减少[1]。 Y15 不靶向同源 Pyk-2、c-Src、c-RAF、EGFR、IGFR、PDGFR、PI3K、VEGFR-3 和 c-Met [3]。

Y15 在体内抑制神经母细胞瘤、胰腺肿瘤和乳腺肿瘤的生长[2]。根据对小鼠进行的药代动力学研究，Y15在这些动物体内吸收非常快，腹腔注射30mg/kg剂量后4.8分钟内达到最大血浆浓度。 Y15 的半衰期分别为 6.9 分钟和 11.6 分钟，在小鼠和人肝微粒体中代谢迅速。在为期 7 天的研究中，Y15 单次口服给药的最大耐受剂量为 200 mg/kg，多次口服给药的最大耐受剂量为 100 mg/kg。在 28 天的研究中，腹腔注射 30 毫克/公斤，在 7 天的研究中，口服 100 毫克/公斤，Y15 不会导致任何死亡或体重的统计学显着变化。在小鼠的各个器官中，在28天内腹腔注射30mg/kg剂量和在7天内口服100mg/kg剂量时，没有出现临床、化学、血液学或组织病理学变化[3]。

含有 10 μCi [γ-³²P]-ATP 的激酶缓冲液 将含有 10 μCi [γ-³²P]-ATP 的激酶缓冲液与 0.1 μg纯化的 FAK 蛋白和 20 mM HEPES，pH 7.4、5 mM MgCl₂、5 mM MnCl₂、0.1 mM Na₃VO _4..激酶反应在室温下运行五分钟后，添加 2× Laemmli 缓冲液以停止反应。使用 Ready SDS-10% PAGE 凝胶分离蛋白质，并使用放射自显影术显示磷酸化烯醇酶。

将每孔一万个细胞接种到 96 孔培养皿中，加入 100 μL 含有 10% FBS 和 1% 青霉素/链霉素的培养基。抑制剂处理 24 小时后，每孔加入 20 μL Cell Titer 96 Aqueous One Solution Cell Proliferation Assay。在两小时的试剂孵育期后，在 490 nm 处读取板的读数。

Mice: Female, naked mice six weeks of age are employed. Athalic nude mice are given a subcutaneous injection of 5×10⁶ Panc si5-IGF-1R cells, which have been mixed with matrigel, in their flank on day zero. By day seven, the animals are split into two groups at random. 30 mg/kg of Y15 was administered to one group (n = 5), while PBS was given to the other group (n = 5). Using 5×10⁶ Panc si-ctrl cells combined with matrigel, subcutaneous injections are made into the flanks of naked mice to create Panc si-ctrl xenografts. On day 7, these animals are also split into two groups at random; five animals each group received TAE226 (30 mg/kg), while the other five animals received PBS as a control. Intraperitoneal injections of 0.1 mL in total volume are used to administer the medications and PBS. Beginning on day 10, tumor sizes are measured every three or four days in terms of length (mm) and width (mm). The formula to calculate the volume of a tumor is volume (cm3) = 1/2×length (cm)×width (cm)².

[1]. Oncotarget . 2014 Sep 15;5(17):7945-59.

[2]. Carcinogenesis, Volume 33, Issue 5, May 2012, Pages 1004–1013

[3]. Arch Toxicol . 2015 Jul;89(7):1095-101.

Focal adhesion kinase (FAK) is up-regulated in thyroid cancer and small molecule FAK scaffolding inhibitor, Y15, was shown to decrease cancer growth in vitro and in vivo. We sought to test the effectiveness of Y15 in thyroid cancer cell lines, profile gene expression with Y15 compared with clinical trial FAK inhibitor PF-04554878, and use Y15 in novel drug combinations. Cell viability was decreased in a dose dependent manner in four thyroid cancer cell lines with Y15 and with higher doses in PF-04554878. Y397 FAK and total FAK were decreased with Y15 and decreased less with PF-04554878. Detachment and necrosis were increased in a dose-dependent manner in all cell lines with Y15. Clonogenicity was decreased in a dose-dependent manner for both Y15 and PF-04554878. We compared gene profiles between papillary thyroid cell lines, TPC1, BCPAP and K1, and 380, 109, and 74 genes were significantly >2-fold changed with Y15 treatment, respectively. Common up-regulated genes were involved in apoptosis, cell cycle, transcription and heat shock; down-regulated genes were involved in cell cycle, cell-to-cell interactions, and cancer stem cell markers. We also compared gene profiles of TT cells treated with Y15 versus PF-04554878. Y15 caused 144 genes to change over 4 fold and PF-04554878 caused 208 gene changes >4-fold (p<0.05). Among genes changed 4 fold, 11 were shared between the treatments, including those involved in metabolism, cell cycle, migration and transcription. Y15 demonstrated synergy with PF-04554878 in TT cells and also synergy with Cabozantinib, Sorafenib, Pazopanib, and strong synergy with Sunitinib in resistant K1 cells. This report revealed the biological effect of Y15 inhibitor, detected the unique and common gene signature profiles in response to Y15 in 4 different thyroid cancer cell lines, demonstrated differential response changes with Y15 and PF-04554878 treatment, and showed the synergy of Y15 with PF-04554878, Cabozantinib, Sorafenib, Pazopanib, and Sunitinib.[1]

C6H14CL4N4

284.0142

281.997

C, 25.37; H, 4.97; Cl, 49.93; N, 19.73

4506-66-5

4506-66-5 (HCl)

78260

Light green to green solid powder

400.9ºC at 760mmHg

≥300ºC(lit.)

233.6ºC

1.827

5.548

104.08

8

4

0

14

90.3

0

Cl[H].Cl[H].Cl[H].Cl[H].N([H])([H])C1C([H])=C(C(=C([H])C=1N([H])[H])N([H])[H])N([H])[H]

BZDGCIJWPWHAOF-UHFFFAOYSA-N

InChI=1S/C6H10N4.4ClH/c7-3-1-4(8)6(10)2-5(3)9;;;;/h1-2H,7-10H2;4*1H

benzene-1,2,4,5-tetramine;tetrahydrochloride

FAK Inhibitor 14; FAK Inhibitor Y15; Y15 hydrochloride; Y15 tetrahydrochloride; 1,2,4,5-Benzenetetramine tetrahydrochloride; Benzene-1,2,4,5-tetraamine tetrahydrochloride; FAK Inhibitor 14; Y15; Benzene-1,2,4,5-tetramine 4HCl; 1,2,4,5-Tetraaminobenzene tetrahydrochloride; MFCD00012970; Y 15; Y-15.

292159

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

注意： 请将本产品存放在密封且受保护的环境中，避免吸湿/受潮。

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

DMSO: 25~56 mg/mL (197.2~88.0 mM)
Water: ~56 mg/mL (~197.2 mM)

配方 1 中的溶解度: 10 mg/mL (35.21 mM) in PBS (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液; 超声助溶。 (<60°C).

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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;
4、 如产品在配制过程中出现沉淀/析出，可通过加热(≤50℃)或超声的方式助溶;
5、为保证最佳实验结果，工作液请现配现用！
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。