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

Alectinib (0-1000 nM; 2 hours; NCI-H2228 cells) treatment was able to prevent autophosphorylation of ALK and significantly suppress phosphorylation of STAT3 and AKT, in NCI-H2228 cells expressing EML4-ALK[1].
Alectinib (0-1000 nM; 5 days; HCC827, A549, or NCIH522 cells) treatment reduces cell activity in a dose-dependent manner[1].

Alectinib (0.2-20 mg/kg; oral administration; once daily; for 11 days; SCID or nude mice bearing NCI-H2228 cells), tumor regression and dose-dependent inhibition of tumor growth (EC50 of 0.46 mg/kg) are possible outcomes of treatment. There are no noticeable toxicity symptoms or variations in body weight at any dosage level[1].

Through the use of time-resolved fluorescence resonance energy transfer (TR-FRET) assay or fluorescence polarization (FP) assay, the inhibitory ability against each kinase—apart from MEK1 and Raf-1—is assessed by looking at their capacity to phosphorylate different substrate peptides in the presence of CH542480. The phosphorylation of a substrate peptide by a recombinant ERK2 protein in the presence of CH5424802 is quantitatively analyzed to determine the inhibitory activity against MEK1. When CH5424802 is present, the kinases' capacity to phosphorylate MEK1 is used to gauge their inhibitory activity against Raf-1.

In 96-well plates, cells such as NSCLC, A549, and HCC827 are seeded overnight and then incubated with different concentrations of CH5424802 for the specified amount of time. In the spheroid cell growth inhibition assay, the compound is added to cells that have been seeded on spheroid plates, incubated for a full night, and then treated for the designated durations. The Luminescent Cell Viability Assay is used to determine the number of viable cells. The Caspase-Glo 3/7 Assay Kit is used to evaluate the Caspase-3/7 assay.

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

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)

Solubility in Formulation 1: ≥ 0.38 mg/mL (0.79 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 3.8 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: ≥ 0.38 mg/mL (0.79 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 3.8 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.

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Solubility in Formulation 3: 30% PEG400+0.5% Tween80+5% propylene glycol: 30mg/mL

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Solubility in Formulation 4: 20 mg/mL (41.44 mM) in 0.5% CMC-Na/saline water (add these co-solvents sequentially from left to right, and one by one), suspension solution; with ultrasonication.
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution.

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 (Please use freshly prepared in vivo formulations for optimal results.)