Alectinib (CH5424802)

Catalog No.S2762 Batch:S276203

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Technical Data

Formula

C30H34N4O2
Molecular Weight 482.62 CAS No. 1256580-46-7
Solubility (25°C)* In vitro DMSO 6 mg/mL (12.43 mM)
Water Insoluble
Ethanol Insoluble
In Vivo (Add solvents to the product individually and in order.)
Homogeneous suspension
CMC-NA
≥5mg/ml Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
* <1 mg/ml means slightly soluble or insoluble.
* Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations.
* Room temperature shipping (Stability testing shows this product can be shipped without any cooling measures.)

Preparing Stock Solutions

Biological Activity

Description Alectinib is a potent ALK inhibitor with IC50 of 1.9 nM in cell-free assays, sensitive to L1196M mutation and higher selectivity for ALK than PF-02341066, NVP-TAE684 and PHA-E429.
Targets
ALK (F1174L)
(Cell-free assay)		 ALK
(Cell-free assay)		 ALK (R1275Q)
(Cell-free assay)
1 nM 1.9 nM 3.5 nM
In vitro The dissociation constant (KD) value of CH5424802 for ALK in an ATP-competitive manner is 2.4 nM. CH5424802 has substantial inhibitory potency against both native ALK and L1196M with Ki of 0.83 nM and 1.56 nM, respectively. CH5424802 prevents autophosphorylation of ALK in NCI-H2228 NSCLC cells expressing EML4-ALK. CH5424802 also suppresses the phosphorylation of STAT3 and AKT, but not of ERK1/2. CH5424802 completely inhibits the phosphorylation of STAT3 at Tyr705. CH5424802 is preferentially efficacious against NCI-H2228 cells expressing EML4-ALK, but not ALK fusion-negative NSCLC cell lines, including HCC827 cells (EGFR exon 19 deletion), A549 cells (KRAS mutant), or NCI-H522 cells (EGFR wild-type, KRAS wild-type, and ALK wild-type) in monolayer culture. CH5424802 elicits an apoptotic marker—caspase-3/7-like activation—in NCI-H2228 spheroid cells. CH5424802 blocks the growth of two lymphoma lines, KARPAS-299 and SR, with NPM-ALK fusion protein but does not influence the growth of an HDLM-2 lymphoma line without ALK fusion. CH5424802 displays high target selectivity and the stronger anti-proliferative activity against KARPAS-299. CH5424802 inhibits KAPRAS-299 with an IC50 of 3 nM, and KDR with IC50 of 1.4 μM. The metabolic stability of CH5424802 is very high.
In Vivo Oral administration of CH5424802 dose-dependently inhibits tumour growth with an ED50 of 0.46 mg/kg and tumour regression. Treatment of 20 mg/kg CH5424802 reveals rapid tumour regression by 168%, the tumour volume in any mouse is <30 mm3 after 11 days of treatment (at day 28), a potent anti-tumour effect is maintained, and tumour regrowth does not occur throughout the 4-week drug-free period. The half-life and the oral bioavailability of CH5424802 in mice are 8.6 hours and 70.8%, respectively. At a repeated dose of 6 mg/kg, the mean plasma levels reached 1.7, 1.5, and 0.3 nM at 2, 7, and 24 hours post-dose, respectively. Administration of CH5424802 leads to tumour growth prevention and tumour regression. Tumour growth inhibition at 20 mg/kg is 119% for KARPAS-299 and 104% for NB-1 on day 20. CH5424802 inhibits the phosphorylation of STAT3 in a dose-dependent manner (2–20 mg/kg). A partial decrease in AKT phosphorylation is also observed in CH5424802-treated xenograft tumours.

Protocol (from reference)

Kinase Assay:[1]
  • Kinase inhibitory assays in vitro

    The inhibitory ability against each kinase except for MEK1 and Raf-1 is evaluated by examining their ability to phosphorylate various substrate peptides in the presence of CH5424802 using time-resolved fluorescence resonance energy transfer (TR-FRET) assay or fluorescence polarization (FP) assay. The inhibitory activity against MEK1 is evaluated by quantitative analysis of the phosphorylation of a substrate peptide by a recombinant ERK2 protein in the presence of CH5424802. The inhibitory activity against Raf-1 is evaluated by examining the ability of the kinases to phosphorylate MEK1 in the presence of CH5424802.
Cell Assay:[1]
  • Cell Lines

    NSCLC, A549 and HCC827 cell lines

  • Concentrations

    0-1 μM

  • Incubation Time

    5 days

  • Method

    Cells including NSCLC, A549 and HCC827 are seeded in 96-well plates overnight and incubated with various concentrations of CH5424802 for the indicated time. For spheroid cell growth inhibition assay, cells are seeded on spheroid plates, incubated overnight, and then treated with compound for the indicated times. The viable cells are measured by the Luminescent Cell Viability Assay. Caspase-3/7 assay is evaluated using the Caspase-Glo 3/7 Assay Kit.
Animal Study:[1]
  • Animal Models

    SCID or nude mice bearing NCI-H2228

  • Dosages

    20 mg/kg

  • Administration

    Oral administration

References

  • https://pubmed.ncbi.nlm.nih.gov/21575866/
  • https://pubmed.ncbi.nlm.nih.gov/22225917/

Customer Product Validation

Sensitivity of the H3122 cell line to ALK inhibitors (crizotinib and alectinib). To examine the sensitivity of ALK inhibitors, used an MTT assay. The experiment was performed in triplicate. (A) Growth inhibitory effect of crizotinib. The H3122 cell line was sensitive to crizotinib under a normoxic state, compared with a hypoxic state. The graphs, mean of independent triplicate experiments; error bars, SD. (B) IC50 of ALK inhibitors. The IC50 values of both inhibitors in the H3122 cell lines were significantly higher under hypoxia than under normoxia (crizotinib, *P=0.028 and alectinib, *P=0.0035). *P<0.05.

Data from [ Int J Oncol , 2014 , 45(4), 1430-6 ]

<p>Western blotting data of NPM/ALK-transformed BaF3 cells, both wild-type and L1196M-mutant, treated with CH5424802 for 4 hours. The blot shows phospho-ALK signal (Y1604), as an indicator of NPM/ALK activation.</p>

, , Prof. Gambacorti from Università degli Studi di Milano Bicocc

<p>Cell growth data, performed by tritiated thymidine uptake assay, using NPM/ALK wild-type -transformed BaF3 cells.CH5424802 demonstrated a very good ability to block NPM/ALK-transformed BaF3 cells proliferation with an IC50 of 3 nM.</p>

, , Prof. Gambacorti from Università degli Studi di Milano Bicocc

Immunoblot analysis of full-length DCTN1-ALK proteins. An expression vector encoding DCTN1-ALK cDNA was introduced into H1299 lung cancer cells, which do not express endogenous ALK. The transfectants were then exposed to crizotinib and alectinib. Levels of phosphorylation at tyrosine 1604 were determined 24 hours after treatment with 0, 0.2, 1.0, or 5.0 μM of each drug.

Data from [ , , Oncologist, 2017, 22(2):158-164 ]

Sellecks Alectinib (CH5424802) Has Been Cited by 114 Publications

RNase1-driven ALK-activation is an oncogenic driver and therapeutic target in non-small cell lung cancer [ Signal Transduct Target Ther, 2025, 10(1):124] PubMed: 40246819
Targeting proteostasis in multiple myeloma through inhibition of LTK [ Leukemia, 2025, 10.1038/s41375-025-02682-8] PubMed: 40634511
Novel selective strategies targeting the BCL-2 family to enhance clinical efficacy in ALK-rearranged non-small cell lung cancer [ Cell Death Dis, 2025, 16(1):194] PubMed: 40113795
MED12 mutation induces RTK inhibitor resistance in NSCLC via MEK/ERK pathway activation by inflammatory cytokines [ Cell Mol Life Sci, 2025, 82(1):314] PubMed: 40833497
Evaluation and modification of tumor cell isolation techniques from malignant effusions for rapid drug sensitivity testing [ Mol Oncol, 2025, 19(9):2474-2490] PubMed: 40525275
Evaluation and modification of tumor cell isolation techniques from malignant effusions for rapid drug sensitivity testing [ Mol Oncol, 2025, 10.1002/1878-0261.70072] PubMed: 40525275
Ferristatin II protects nucleus pulposus against degeneration through inhibiting ferroptosis and activating HIF-1α pathway mediated mitophagy [ Int Immunopharmacol, 2025, 147:113895] PubMed: 39752759
Efficacy of amivantamab, a bi-specific antibody targeting EGFR and MET, in ALK-rearranged non-small-cell lung cancer cell lines [ Lung Cancer, 2025, 201:108415] PubMed: 39922174
A high-throughput screening platform to identify MYCN expression inhibitors for liver cancer therapy [ Front Oncol, 2025, 15:1486671] PubMed: 40027135
Brigatinib activates inflammasomes: Implication for immune-related adverse events [ Toxicol Appl Pharmacol, 2025, 498:117310] PubMed: 40122348

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