ALK (IC50 <0.4 nM); MET (IC50 = 0.74 nM)

Ensartinib (X-396) diHClide 具有双重 ALK/MET 抑制活性，有效 IC50 分别低于 0.4 nM 和 0.74 nM。在表达 EML4-ALK E13;A20 的 H3122 肺癌细胞中，二盐酸恩替尼表现出效力（IC50：15 nM）。在表达 EML4-ALK E6a/b 的 H2228 肺癌细胞中； A20，二盐酸依沙替尼同样有效（IC50：45 nM）。此外，X-376 对含有 NPM-ALK 的 SUDHL-1 淋巴瘤细胞具有高效力 (IC50：9 nM)。

根据药代动力学分析，恩沙替尼具有显着的生物利用度和中等的体内半衰期。检查Ensartinib (X-396)对H3122异种移植物的体内作用。 Ensartinib，25 mg/kg bid，给予含有 H3122 异种移植物的裸鼠。与单独的载体相比，恩沙替尼显着减缓肿瘤的生长。在异种移植实验中，恩沙替尼似乎在体内具有良好的耐受性。恩沙替尼治疗对小鼠体重没有影响。给予药物的小鼠看起来很健康，并且没有表现出任何化合物毒性症状。在 Sprague Dawley (SD) 大鼠中进行了额外的全身毒性和毒代动力学研究，以进一步评估 ensartinib 的潜在副作用。在 SD 大鼠中以 20、40 和 80 mg/kg 的剂量重复口服 Ensartinib 十天后，每只动物都存活到研究结束。已发现 Ensartinib 的无显着毒性 (NST) 水平为 80 mg/kg。 Ensartinib 在 NST 水平下的 AUC 为 66 μMHhr，C_max 为 7.19 μM[1]。

在 96 孔板中，细胞以 25%–33% 汇合度接种，并暴露于药物中进行活力实验。 Ensartinib（10、30、100、300 和 1000 nM）用于治疗人肺腺癌细胞系 H3122 和 H2228。 Ensartinib（5、10、30、100 和 300 nM）用于治疗 SUDHL-1 淋巴瘤细胞。 Ensartinib（30、100、300 和 1000 nM）用于治疗 SY5Y 神经母细胞瘤细胞。加入Ensartinib 72小时后，加入Cell Titer Blue Reagent，使用Spectramax分光光度计测量荧光。每个实验点重复设置3次，并且至少独立运行两次。 Windows 版 GraphPad Prism 版本 5 用于计算 IC50。通过对数（抑制剂）与响应公式，使用非线性回归模型来拟合曲线[1]。

Mice: H3122 cells are injected into naked mice (nu/nu). Following the tumors' average volume of 450 mm³, 27 athymic mice with H3122 tumors are randomly assigned to receive either the control vehicle or 25 mg/kg of ensartinib orally via gavage. Mice are killed two, five, and fifteen hours after the single treatment (3 tumors/timepoint/group), and serum is taken for an LC-MS based bioanalytical method to determine the drug concentration[1].

Absorption
The mean Cmax of orally administered 25 mg and 100 mg ensartnib is 292 ng/mL, with the Tmax being between 2 hours and 8 hours and a median Tmax of 3 hours. Ensartinib at approved recommended dosages has an AUC of 4,920 ng\*h/ml. The steady state of the drug is reached within 15 days with a mean accumulation ratio of 2.7. Ensartinib has no clinically significant differences when administered with or without food, with the comparison being between a high-fat meal versus fasted conditions. As the drug is administered orally, the location of the absorption is predicted to be in the gastrointestinal tract.

Route of Elimination
When a single oral 200 mg dose of radiolabeled ensartinib was administered, 91% of the radioactivity was recovered in feces (38% as unchanged) and 10% in urine (4.4% as unchanged).

Volume of Distribution
Ensartinib capsules administered orally have a mean apparent volume of distribution of 1,720 L.

Protein Binding
Ensartinib oral capsules have been found to be 91.6% bound to human plasma protein.

Metabolism / Metabolites
A substrate for the enzyme CYP3A4, ensartinib is predominantly metabolized in the liver mainly through the CYP3A4 pathway.

Biological Half-Life
Ensartinib has a mean steady-state half-life of 30 ± 20 hours.

Hepatotoxicity
In the prelicensure trials of ensartinib as therapy of NSCLC, liver test abnormalities were frequent, with elevations in ALT levels in 59%, AST in 58%, Alk P in 51%, and bilirubin in 12% of treated participants. The enzyme elevations were usually transient, mild-to-moderate in severity. Pruritus was seen in 26%. ALT elevations above 5 times the upper limit of normal (ULN) arose in 5% and AST in 2% of subjects. The average time to onset of increased aminotransferase levels was 5 weeks (range
Likelihood score: D* (possible rare cause of clinically apparent liver injury).

[1]. Insights into ALK-driven cancers revealed through development of novel ALK tyrosine kinaseinhibitors. Cancer Res. 2011 Jul 15;71(14):4920-31.

Non-small cell lung cancer (NSCLC) is one of the most frequent subtypes of lung cancers and encompasses a variety of different lung cancers, with notable ones being adenocarcinoma, squamous cell carcinoma, and large cell carcinoma. Ensartinib is a first-line treatment for anaplastic lymphoma kinase (ALK)-mutated non-small cell lung cancer (NSCLC). Chemically, ensartinib is an aminopyridazine-based, small molecule tyrosine kinase inhibitor (TKI) which inhibits the anaplastic lymphoma kinase (ALK) protein. The drug is 10-fold more potent in the inhibition of the growth of ALK-positive lung cancer cell lines when compared to [crizotinib]. Ensartinib was approved by the FDA for adult patients with anaplastic lymphoma kinase (ALK)-positive locally advanced or metastatic non-small cell lung cancer (NSCLC) who have not previously received an ALK-inhibitor on December 18, 2024. Although there was no statistically significant difference in overall survival when compared to [crizotinib], ensartinib demonstrated a statistically significant progression-free survival improvement when compared to [crizotinib].
Ensartinib is a small molecule inhibitor of anaplastic lymphoma kinase (ALK) and is used to treat adults with locally advanced or metastatic non-small cell lung cancer. Ensartinib is associated with transient elevations in serum aminotransferase levels and bilirubin during therapy and rare instances of drug induced liver injury.

Ensartinib is an orally available small molecule inhibitor of the receptor tyrosine kinase anaplastic lymphoma kinase (ALK), with potential antineoplastic activity. Upon oral administration, ensartinib binds to and inhibits ALK kinase, ALK fusion proteins and ALK point mutation variants. Inhibition of ALK leads to the disruption of ALK-mediated signaling and eventually inhibits tumor cell growth in ALK-expressing tumor cells. In addition, ensartinib inhibits various other kinases, including the receptor tyrosine kinase c-Met (hepatocyte growth factor receptor; HGFR; MET) and the receptor tyrosine kinase C-ros oncogene 1 (ROS1). ALK belongs to the insulin receptor superfamily and plays an important role in nervous system development. ALK is not expressed in healthy adult human tissue but ALK dysregulation and gene rearrangements are associated with a series of tumors; ALK mutations are associated with acquired resistance to small molecule tyrosine kinase inhibitors.
ENSARTINIB is a small molecule drug with a maximum clinical trial phase of IV (across all indications) that was first approved in 2024 and has 3 investigational indications.
anaplastic lymphoma kinase (ALK) inhibitor for the treatment of ALK-positive non-small-cell lung cancer

C26H29CL4FN6O3

634.357265233994

634.1

C, 49.23; H, 4.61; Cl, 22.35; F, 2.99; N, 13.25; O, 7.57

2137030-98-7

Ensartinib;1370651-20-9

138320013

White to light yellow solid powder

123

5

8

6

40

812

3

C[C@@H]1CN(C[C@@H](N1)C)C(=O)C2=CC=C(C=C2)NC(=O)C3=NN=C(C(=C3)O[C@H](C)C4=C(C=CC(=C4Cl)F)Cl)N.Cl.Cl

IERUINQRGJAECT-ISUJJMBGSA-N

InChI=1S/C26H27Cl2FN6O3.2ClH/c1-13-11-35(12-14(2)31-13)26(37)16-4-6-17(7-5-16)32-25(36)20-10-21(24(30)34-33-20)38-15(3)22-18(27)8-9-19(29)23(22)28;;/h4-10,13-15,31H,11-12H2,1-3H3,(H2,30,34)(H,32,36);2*1H/t13-,14+,15-;;/m1../s1

6-amino-5-[(1R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy]-N-[4-[(3R,5S)-3,5-dimethylpiperazine-1-carbonyl]phenyl]pyridazine-3-carboxamide;dihydrochloride

X-396 hydrochloride; X396 hydrochloride; X 396 hydrochloride; Ensartinib dihydrochloride; Ensartinib HCl; Ensartinib hydrochloride

2934.99.9001

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 : ~35.71 mg/mL (~56.29 mM)
H2O : ~33.33 mg/mL (~52.54 mM)

配方 1 中的溶解度: ≥ 2.08 mg/mL (3.28 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 20.8 mg/mL澄清DMSO储备液加入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: ≥ 2.08 mg/mL (3.28 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 20.8 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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配方 3 中的溶解度: ≥ 2.08 mg/mL (3.28 mM) (饱和度未知) in 10% DMSO + 90% Corn Oil (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 20.8 mg/mL 澄清 DMSO 储备液加入到 900 μL 玉米油中并混合均匀。

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配方 4 中的溶解度: 14.29 mg/mL (22.53 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.

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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。