ERK2 (Ki = 2 nM); GSK3 (Ki = 395 ); AURA (Ki = 540 ); CDK2 (Ki = 852 )

VX-11e 的 IC50 为 48 nM，有效抑制 HT29 细胞的细胞增殖。

VX-11e 在大鼠和小鼠中均表现出良好的口服生物利用度。[1] VX-11e（50 mg/kg，口服）可强烈抑制 pRSK，并减缓携带人黑色素瘤 RPDX 肿瘤的 NSG 小鼠的肿瘤生长。与 BKM120 联合使用时，VX-11e 显着增强对肿瘤生长的抑制作用。 [2]

通过分光光度偶联酶测定来测定化合物对 ERK2 的抑制作用。在此测定中，固定浓度的活化 ERK2 (10 nM) 与不同浓度的化合物在 DMSO (2.5%) 中孵育 10 分钟。 30°C，0.1 mol/L HEPES 缓冲液，pH=7.5，含有 10 mM MgCl2、2.5 mM 磷酸烯醇丙酮酸、200 μM NADH、150 μg/mL 丙酮酸激酶、50 μg/mL 乳酸脱氢酶和 200 μM erktide 肽。添加 65 μM ATP 开始反应。监测 340 nM 处的吸光度损失。

3H-胸苷的掺入用于测量细胞增殖。在使用含有 10% FBS 的 RPMI 1640 生长培养基的 96 孔板中，细胞以每孔 10,000 个细胞的密度铺板。以连续稀释的量添加化合物。细胞和化合物在 37°C 下孵育 48 小时。 48小时后，向每个孔中添加0.4Ci的3H-胸苷，持续8小时，然后将孔放回37℃培养箱中。使用 Tomtec 96 孔细胞收获机收获细胞后，使用 Wallac 1205 BETAPLATE 液体闪烁计数器计算 CPM。

Prior to the therapy experiments, NSG mice are used to expand in vivo human melanoma RPDX tumors. 50 NSG mice (1:10) are implanted with a collection of tumor fragments that were banked from early mouse passages. These tumors are removed once they have reached the protocol's maximum permitted volume (1,000 mm3), digested, and stored as live cells. To be used in the therapy experiments, a smaller portion of this stock is implanted at a 1:5 ratio into NSG mice while the larger portion is kept as a master bank. With PLX4720 200 ppm chemical additive diet at approximately clinical plasma levels, the expansion phase is continuously under drug pressure. The plasma concentrations of PLX4720 (103.7 μg/mL ±3.2 after 7 days) are comparable to the steady-state concentrations observed in patients receiving vemurafenib 960 mg twice daily (130.6 μg/mL±71.78). Animals are randomly assigned to treatment groups when tumors measure 200 mm3 by caliper measurement, and then there is a 3-day ishout phase. Two times per week, caliper measurements are used to determine tumor size. After two weeks of treatment or as needed for animal welfare, mice are sacrificed. When tumor control is achieved as indicated, dosing is extended. In order to extract proteins, tumor tissue is snap-frozen in liquid N2 and preserved in formalin (for FFPE). 4 hours after the final dose, the treatment groups are sacrificed.

[1]. J Med Chem . 2009 Oct 22;52(20):6362-8.

[2]. Clin Cancer Res . 2016 Apr 1;22(7):1592-602.

C24H20CL2FN5O2

500.35

499.10

C, 57.61; H, 4.03; Cl, 14.17; F, 3.80; N, 14.00; O, 6.40

896720-20-0

(R)-VX-11e;1680187-43-2

Solid powder

CC1=CN=C(N=C1C2=CNC(=C2)C(=O)N[C@H](CO)C3=CC(=CC=C3)Cl)NC4=C(C=C(C=C4)F)Cl

WUTVMXLIGHTZJC-OAQYLSRUSA-N

InChI=1S/C24H20Cl2FN5O2/c1-13-10-29-24(31-19-6-5-17(27)9-18(19)26)32-22(13)15-8-20(28-11-15)23(34)30-21(12-33)14-3-2-4-16(25)7-14/h2-11,21,28,33H,12H2,1H3,(H,30,34)(H,29,31,32)/t21-/m1/s1

4-[2-(2-chloro-4-fluoroanilino)-5-methylpyrimidin-4-yl]-N-[(1S)-1-(3-chlorophenyl)-2-hydroxyethyl]-1H-pyrrole-2-carboxamide

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)

Solubility in Formulation 1: ≥ 3.25 mg/mL (6.50 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 32.5 mg/mL clear DMSO stock solution to 400 μL of PEG300 and mix evenly; then add 50 μL of Tween-80 to the above solution and mix evenly; then add 450 μL of normal saline to adjust the volume to 1 mL.
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution.

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Solubility in Formulation 2: ≥ 3.25 mg/mL (6.50 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 32.5 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.

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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。