CDK4 (IC50 = 1.2 nM); CDK6 (IC50 = 1.3 nM)

Abemaciclib 的代谢物 M2 (LSN2839567) 与人类、狗和金的束缚蛋白结合率为 83-92%，而 Abemaciclib 的结合率为 95-99% [2]。人类中最显着（活跃且重要）的转运蛋白是 Abelacil 代谢物 M2 [2]。

[1]. Abstract 2830: The major human metabolites of abemaciclib are inhibitors of CDK4 and CDK6. Cancer Research. July 2016, 76 (14).

[2]. CHMP. Assessment report Verzenios. 26 July 2018 EMA/551438/2018.

Abemaciclib (LY2835219) is an ATP-competitive inhibitor of cyclin dependent kinases 4 and 6 (CDK4 and CDK6) which is currently undergoing clinical evaluation for the treatment of breast and lung cancers. A radiolabeled disposition study following a single 150-mg oral dose of [14C]LY2835219 in healthy subjects indicated that in plasma, in addition to parent drug, the presence of 5 metabolites denoted as M1, M2, M18, M20 and M22. Abemaciclib (34%), M20 (26%), M2 (13%), and M18 (5%) constituted the majority of the plasma exposure. This study investigated the in vitro biological activity of these human circulating metabolites, with the exception of the trace metabolite, M1, and compared their potencies with the parent drug abemaciclib. Specifically non-small cell lung cancer (NSCLC) cells, colorectal cancer (CRC) cells and breast cancer cell lines were evaluated for growth inhibition, cell cycle inhibition and biomarker expression following treatment with abemaciclib and the metabolites. The metabolites were also profiled and compared to abemaciclib for inhibition of CDK4, CDK6, CDK1, and CDK9 in cell-free biochemical kinase assays. The IC50 values for the inhibition of CDK4 and CDK6, for metabolites M2, M18, and M20, but not M22, were between 1 and 3 nM and were nearly equivalent in potency to abemaciclib. Likewise, metabolites M2, M20, and M18 inhibited cell growth and cell cycle progression in a concentration-dependent manner that was consistent with the inhibition of CDK4 and CDK6 since these outcomes correlated with the concentration-dependent inhibition of various biomarkers such a phospho-serine 780-Rb (pRb), topoisomerase II-alpha (Topo IIα), and phospho-serine 10-histone H3 (pHH3). In this regard, metabolites M2 and M20 showed potencies nearly identical with abemaciclib in the cancer cell lines evaluated, whereas depending on the endpoint measured, the potency of M18 was approximately 3-20-fold lower than abemaciclib. M22 showed the least potency for growth inhibition and little or no inhibition of biomarker expression or cell cycle progression at concentrations below 2 μM. Although the cell-free kinase assays showed that like abemaciclib, M2, M18, and M20 had potential to inhibit CDK9, no measurable inhibition of CDK9 by any of these compounds was observed in cancer cells, indicating that the primary targets driving cell cycle inhibition for these metabolites in cancer cells were CDK4 and CDK6 and not CDK9. Studies with abemaciclib, M2 or M20 in breast cancer cells showed that all 3 compounds induced senescence in addition to growth inhibition following 6-8 days of treatment at 200 and 500 nM. In total the results indicated that the major human metabolites of abemaciclib, M2 and M20, are effective inhibitors of CDK4 and CDK6 that are remarkably similar to abemaciclib in regards to their effects in cancer cells on growth, senescence, and other phenotypic responses.[1]

C25H28F2N8

478.540230751038

478.24

1231930-57-6

Abemaciclib metabolite M2-d6

59376686

Light yellow to yellow solid powder

3

83.8

2

9

6

35

680

0

FC1=CC(C2C(=CN=C(N=2)NC2=CC=C(C=N2)CN2CCNCC2)F)=CC2=C1N=C(C)N2C(C)C

IXGZDCRFGCEEBU-UHFFFAOYSA-N

InChI=1S/C25H28F2N8/c1-15(2)35-16(3)31-24-19(26)10-18(11-21(24)35)23-20(27)13-30-25(33-23)32-22-5-4-17(12-29-22)14-34-8-6-28-7-9-34/h4-5,10-13,15,28H,6-9,14H2,1-3H3,(H,29,30,32,33)

5-fluoro-4-(7-fluoro-2-methyl-3-propan-2-ylbenzimidazol-5-yl)-N-[5-(piperazin-1-ylmethyl)pyridin-2-yl]pyrimidin-2-amine

1231930-57-6; Abemaciclib Metabolites M2; Abemaciclib metabolite M2; 5-Fluoro-4-(4-fluoro-1-isopropyl-2-methyl-1H-benzo[d]imidazol-6-yl)-N-[5-(1-piperazinylmethyl)-2-pyridyl]pyrimidin-2-amine; 5-fluoro-4-(7-fluoro-2-methyl-3-propan-2-ylbenzimidazol-5-yl)-N-[5-(piperazin-1-ylmethyl)pyridin-2-yl]pyrimidin-2-amine; Des-Et-Abemaciclib; MFCD32067988; LSN2839567;

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)

DMSO : ~2 mg/mL (~4.18 mM)

配方 1 中的溶解度: ≥ 2.5 mg/mL (5.22 mM) (饱和度未知) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
例如，若需制备1 mL的工作液，可将100 μL 25.0 mg/mL澄清DMSO储备液加入到400 μL PEG300中，混匀；然后向上述溶液中加入50 μL Tween-80，混匀；加入450 μL生理盐水定容至1 mL。
*生理盐水的制备：将 0.9 g 氯化钠溶解在 100 mL ddH₂O中，得到澄清溶液。

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配方 2 中的溶解度: ≥ 2.5 mg/mL (5.22 mM) (饱和度未知) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
例如，若需制备1 mL的工作液，可将 100 μL 25.0 mg/mL澄清DMSO储备液加入900 μL 20% SBE-β-CD生理盐水溶液中，混匀。
*20% SBE-β-CD 生理盐水溶液的制备（4°C，1 周）：将 2 g SBE-β-CD 溶解于 10 mL 生理盐水中，得到澄清溶液。

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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：
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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；
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