ALK (IC50 = 1.9 nM); ALK^F1174L (IC50 = 1 nM); ALK^R1275Q (IC50 = 3.5 nM); ALK (Kd = 2.4 nM)

体外活性：CH5424802 以 ATP 竞争方式对 ALK 的解离常数 (KD) 值为 2.4 nM。 CH5424802 对天然 ALK 和 L1196M 具有显着的抑制效力，Ki 分别为 0.83 nM 和 1.56 nM。 CH5424802 可防止表达 EML4-ALK 的 NCI-H2228 NSCLC 细胞中 ALK 的自身磷酸化。 CH5424802 还抑制 STAT3 和 AKT 的磷酸化，但不抑制 ERK1/2 的磷酸化。 CH5424802 完全抑制 STAT3 Tyr705 的磷酸化。 CH5424802 优先对表达 EML4-ALK 的 NCI-H2228 细胞有效，但对 ALK 融合阴性 NSCLC 细胞系无效，包括 HCC827 细胞（EGFR 外显子 19 缺失）、A549 细胞（KRAS 突变体）或 NCI-H522 细胞（EGFR 野生型）单层培养中的（KRAS 野生型和 ALK 野生型）。 CH5424802 在 NCI-H2228 球状细胞中引发凋亡标记物 - caspase-3/7 样激活。 CH5424802 使用 NPM-ALK 融合蛋白阻断两种淋巴瘤系 KARPAS-299 和 SR 的生长，但不影响无 ALK 融合的 HDLM-2 淋巴瘤系的生长。 CH5424802对KARPAS-299表现出高靶点选择性和更强的抗增殖活性。 CH5424802 抑制 KAPRAS-299，IC50 为 3 nM，抑制 KDR，IC50 为 1.4 μM。 CH5424802的代谢稳定性非常高。激酶测定：通过使用时间分辨荧光共振能量转移 (TR-FRET) 测定或荧光偏振 (TR-FRET) 测定在 CH5424802 存在下检查其磷酸化各种底物肽的能力，评估对除 MEK1 和 Raf-1 之外的每种激酶的抑制能力。 FP）测定。在 CH5424802 存在的情况下，通过重组 ERK2 蛋白对底物肽的磷酸化进行定量分析来评估针对 MEK1 的抑制活性。通过检查激酶在 CH5424802 存在的情况下磷酸化 MEK1 的能力来评估对 Raf-1 的抑制活性。细胞测定：将细胞（NSCLC、A549 和 HCC827）接种在 96 孔板中过夜，并与不同浓度的 CH5424802 一起孵育指定时间。对于球状细胞生长抑制测定，将细胞接种在球状体板上，孵育过夜，然后用化合物处理指定的时间。通过发光细胞活力测定来测量活细胞。使用 Caspase-Glo 3/7 检测试剂盒评估 Caspase-3/7 检测。

口服 CH5424802 剂量依赖性地抑制肿瘤生长（ED50 为 0.46 mg/kg）并抑制肿瘤消退。 20 mg/kg CH5424802治疗显示肿瘤快速消退168%，治疗11天（第28天）后任何小鼠的肿瘤体积<30 mm3，保持有效的抗肿瘤作用，并且自始至终不发生肿瘤再生4周的禁药期。 CH5424802在小鼠体内的半衰期和口服生物利用度分别为8.6小时和70.8%。重复剂量为 6 mg/kg 时，给药后 2、7 和 24 小时的平均血浆水平分别达到 1.7、1.5 和 0.3 nM。 CH5424802的施用导致肿瘤生长预防和肿瘤消退。第 20 天，20 mg/kg 剂量时，KARPAS-299 的肿瘤生长抑制率为 119%，NB-1 的肿瘤生长抑制率为 104%。CH5424802 以剂量依赖性方式（2-20 mg/kg）抑制 STAT3 的磷酸化。在 CH5424802 处理的异种移植肿瘤中也观察到 AKT 磷酸化部分降低。

通过使用时间分辨荧光共振能量转移 (TR-FRET) 测定或荧光偏振 (FP) 测定，通过观察每种激酶（MEK1 和 Raf-1 除外）磷酸化不同激酶的能力来评估其抑制能力。 CH542480 存在下的底物肽。在 CH5424802 存在的情况下，定量分析重组 ERK2 蛋白对底物肽的磷酸化，以确定对 MEK1 的抑制活性。当 CH5424802 存在时，激酶磷酸化 MEK1 的能力可用于测量其对 Raf-1 的抑制活性。

在 96 孔板中，将 NSCLC、A549 和 HCC827 等细胞接种过夜，然后与不同浓度的 CH5424802 一起孵育指定的时间。在球状细胞生长抑制测定中，将化合物添加到已接种在球状体平板上的细胞中，孵育一整晚，然后处理指定的持续时间。发光细胞活力测定用于确定活细胞的数量。 Caspase-Glo 3/7 检测试剂盒用于评估 Caspase-3/7 检测。

SCID or nude mice bearing NCI-H2228
20 mg/kg
Oral administration

Absorption, Distribution and Excretion
Alectinib reached maximal concentrations at 4 hours following administration of 600 mg twice daily under fed conditions in patients with ALK-positive NSCLC. The absolute bioavailability was 37% in the fed state. A high-fat, high-calorie meal increased the combined exposure of alectinib and its major metabolite M4 by 3.1-fold following oral administration of a single 600 mg dose.
When radioactively labeled, 98% of radioactivity was found in feces with 84% of that amount excreted as unchanged alectinib and 6% as M4. Less than 0.5% was found to be recovered in urine.
4016 L
The apparent clearance is 81.9L/hr for alectinib and 217 L/hr for M4.

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Metabolism / Metabolites
Alectinib is metabolized by CYP3A4 to its major active metabolite M4. M4 is then further metabolized by CYP3A4. Both alectinib and M4 demonstrate similar in vivo and in vitro activity. In vitro studies suggest that alectinib is not a substrate for P-gp while M4 is.

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Biological Half-Life
The mean elimination half life is 33 hr for alectinib and 31 hr for M4.

Hepatotoxicity
In preregistration trials of alectinib, ALT elevations occurred in up to 50% of patients, but values above 5 times the upper limit of normal (ULN) were found in only 1% to 4%. Alectinib therapy was also associated with frequent elevations in alkaline phosphatase (47%) and bilirubin (39%), but these abnormalities were usually mild-to-moderate in degree, as well as asymptomatic and transient in nature. Clinically apparent liver injury with jaundice was rare, but cases were reported and at least 2% of alectinib treated subjects discontinued therapy early because of severe liver test abnormalities. The clinical features of these episodes were not reported and, since its approval and more widescale use, there have been no published cases of liver injury attributable to alectinib therapy. Use of this agent, however, has been limited. Thus, alectinib has been reported to cause liver injury that can be clinically significant and require drug discontinuation, but the clinical features of the injury have not been well defined and their relationship to treatment not definitively shown.
Likelihood score: D (possible cause of clinically apparent liver injury).

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Effects During Pregnancy and Lactation
◉ Summary of Use during Lactation
No information is available on the clinical use of alectinib during breastfeeding. Because alectinib is more than 99% bound to plasma proteins, the amount in milk is low. However, its half-life is about 33 hours and it might accumulate in the infant. The manufacturer recommends that breastfeeding be discontinued during alectinib therapy and for 1 week after the last dose.
◉ Effects in Breastfed Infants
Relevant published information was not found as of the revision date.
◉ Effects on Lactation and Breastmilk
Relevant published information was not found as of the revision date.

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Protein Binding
Alectinib and its major metabolite M4 are >99% bound to human plasma proteins.

[1]. CH5424802, a selective ALK inhibitor capable of blocking the resistant gatekeeper mutant. Cancer Cell. 2011, 19(5), 679-690.

[2]. Alectinib versus crizotinib in treatment-naive anaplastic lymphoma kinase-positive (ALK+) non-small-cell lung cancer: CNS efficacy results from the ALEX study. Ann Oncol. 2018 Nov 1;29(11):2214-2222.

Alectinib is an organic heterotetracyclic compound that is 6,6-dimethyl-5,6-dihydro-11H-benzo[b]carbazol-11-one carrying additional cyano, 4-(morpholin-4-yl)piperidin-1-yl and ethyl substituents at positions 3, 8 and 9 respectively. Used (as the hydrochloride salt) for the treatment of patients with anaplastic lymphoma kinase-positive, metastatic non-small cell lung cancer. It has a role as an EC 2.7.10.1 (receptor protein-tyrosine kinase) inhibitor and an antineoplastic agent. It is an organic heterotetracyclic compound, a member of morpholines, a member of piperidines, a nitrile and an aromatic ketone. It is a conjugate base of an alectinib(1+).
Alectinib is a second generation oral drug that selectively inhibits the activity of anaplastic lymphoma kinase (ALK) tyrosine kinase. It is specifically used in the treatment of non-small cell lung cancer (NSCLC) expressing the ALK-EML4 (echinoderm microtubule-associated protein-like 4) fusion protein that causes proliferation of NSCLC cells. Inhibition of ALK prevents phosphorylation and subsequent downstream activation of STAT3 and AKT resulting in reduced tumour cell viability. Approved under accelerated approval in 2015, alectinib is indicated for use in patients who have progressed on or were not tolerant of crizotinib, which is associated with the development of resistance.
Alectinib is a Kinase Inhibitor. The mechanism of action of alectinib is as a Kinase Inhibitor.
Alectinib is a tyrosine kinase receptor inhibitor and antineoplastic agent used in the therapy of selected forms of advanced non-small cell lung cancer. Alectinib is associated with a moderate rate of transient elevations in serum aminotransferase levels during therapy and with rare instances of clinically apparent acute liver injury.
Alectinib is an orally available inhibitor of the receptor tyrosine kinase anaplastic lymphoma kinase (ALK) with antineoplastic activity. Upon administration, alectinib binds to and inhibits ALK kinase, ALK fusion proteins as well as the gatekeeper mutation ALKL1196M known as one of the mechanisms of acquired resistance to small-molecule kinase inhibitors. The inhibition leads to disruption of ALK-mediated signaling and eventually inhibits tumor cell growth in ALK-overexpressing tumor cells. ALK belongs to the insulin receptor superfamily and plays an important role in nervous system development. ALK dysregulation and gene rearrangements are associated with a series of tumors.
See also: Alectinib Hydrochloride (active moiety of).

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Drug Indication
Alectinib is a kinase inhibitor indicated for the treatment of patients with anaplastic lymphoma kinase (ALK)-positive, metastatic non-small cell lung cancer (NSCLC) who have progressed on or are intolerant to crizotinib. This indication is approved under accelerated approval based on tumor response rate and duration of response. Continued approval for this indication may be contingent upon verification and description of clinical benefit in confirmatory trials.
FDA Label
Alecensa as monotherapy is indicated for the first-line treatment of adult patients with anaplastic lymphoma kinase (ALK)-positive advanced non-small cell lung cancer (NSCLC). Alecensa as monotherapy is indicated for the treatment of adult patients with ALK‑positive advanced NSCLC previously treated with crizotinib.
Treatment of non-small cell lung carcinoma (NSCLC)

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Mechanism of Action
Alectinib is a second generation oral drug that selectively inhibits the activity of anaplastic lymphoma kinase (ALK) tyrosine kinase. It is specifically used in the treatment of non-small cell lung cancer (NSCLC) expressing the ALK-EML4 (echinoderm microtubule-associated protein-like 4) fusion protein that causes proliferation of NSCLC cells. Inhibition of ALK prevents phosphorylation and subsequent downstream activation of STAT3 and AKT resulting in reduced tumour cell viability. Both alectinib and its major active metabolite M4 demonstrate similar in vivo and in vitro activity against multiple mutant forms of ALK.

C30H34N4O2

482.62

482.268

C, 74.66; H, 7.10; N, 11.61; O, 6.63

1256580-46-7

Alectinib Hydrochloride;1256589-74-8;Alectinib-d8;1256585-15-5;Alectinib-d6;1616374-19-6

49806720

White to off-white solidw powder

1.3±0.1 g/cm3

722.5±60.0 °C at 760 mmHg

390.7±32.9 °C

0.0±2.3 mmHg at 25°C

1.673

5.48

72.36

1

5

3

36

867

0

O1C([H])([H])C([H])([H])N(C([H])([H])C1([H])[H])C1([H])C([H])([H])C([H])([H])N(C2C(C([H])([H])C([H])([H])[H])=C([H])C3C(C4C5C([H])=C([H])C(C#N)=C([H])C=5N([H])C=4C(C([H])([H])[H])(C([H])([H])[H])C=3C=2[H])=O)C([H])([H])C1([H])[H]

KDGFLJKFZUIJMX-UHFFFAOYSA-N

InChI=1S/C30H34N4O2/c1-4-20-16-23-24(17-26(20)34-9-7-21(8-10-34)33-11-13-36-14-12-33)30(2,3)29-27(28(23)35)22-6-5-19(18-31)15-25(22)32-29/h5-6,15-17,21,32H,4,7-14H2,1-3H3

9-ethyl-6,6-dimethyl-8-(4-morpholin-4-ylpiperidin-1-yl)-11-oxo-5H-benzo[b]carbazole-3-carbonitrile

Alectinib; CH5424802; CH 5424802; RO 5424802; AF802; CH-5424802; RO5424802; AF 802; AF-802; RO-5424802; brand name: Alecensa

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)

配方 1 中的溶解度: ≥ 0.38 mg/mL (0.79 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 3.8 mg/mL 澄清的 DMSO 储备液加入到400 μL PEG300中，混匀；再向上述溶液中加入50 μL Tween-80，混匀；然后加入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: ≥ 0.38 mg/mL (0.79 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 3.8 mg/mL 澄清 DMSO 储备液添加到 900 μL 玉米油中并混合均匀。

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配方 3 中的溶解度: 30% PEG400+0.5% Tween80+5% propylene glycol: 30mg/mL

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配方 4 中的溶解度: 20 mg/mL (41.44 mM) in 0.5% CMC-Na/saline water (这些助溶剂从左到右依次添加，逐一添加), 悬浊液; 超声助溶。
*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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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。