ALK/EGFR-IN-4
ALK/EGFR-IN-4 is an EGFR and ALK inhibitor (EGFRL858R/T790M/C797S (IC50 1.76 nM), EGFRL858R/T790M (2.97 nM), wild-type EGFR (59.35 nM) and ALK (1.95 nM)). ALK/EGFR-IN-4 induces G0/G1 phase arrest and Apoptosis, inhibits DNA synthesis, downregulates Cyclin D1, and inhibits AKT/ERK/STAT3 phosphorylation. ALK/EGFR-IN-4 shows antitumor efficacy in an Osimertinib (HY-15772)-resistant EGFR triple-mutant non-small cell lung cancer xenograft model.
For research use only. We do not sell to patients.
- CAS No.: 3081622-36-5
- Formula: C24H29ClN7O4P
- Molecular Weight:545.96
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Storage:
Please store the product under the recommended conditions in the Certificate of Analysis.
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Biological Activity
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Biological Activity
Description
ALK/EGFR-IN-4 is an EGFR and ALK inhibitor (EGFRL858R/T790M/C797S (IC50 1.76 nM), EGFRL858R/T790M (2.97 nM), wild-type EGFR (59.35 nM) and ALK (1.95 nM)). ALK/EGFR-IN-4 induces G0/G1 phase arrest and Apoptosis, inhibits DNA synthesis, downregulates Cyclin D1, and inhibits AKT/ERK/STAT3 phosphorylation. ALK/EGFR-IN-4 shows antitumor efficacy in an Osimertinib (HY-15772)-resistant EGFR triple-mutant non-small cell lung cancer xenograft model[1].
IC50 & Target
[1]|
STAT3 |
EGFRL858R/T790M/C797S 1.76 nM (IC50) |
EGFRL858R/T790M 2.97 nM (IC50) |
EGFR (WT) 59.35 nM (IC50) |
ALK 1.95 nM (IC50) |
Cellular Effect
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Cell Line
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Type | Value | Description | References |
|---|---|---|---|---|
| NCI-H1975 | IC50 |
0.080 μM
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Antiproliferative activity against human NCI-H1975 cells assessed as viability reduction incubated 72 hrs by MTT assay.
Antiproliferative activity against human NCI-H1975 cells assessed as viability reduction incubated 72 hrs by MTT assay.
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42868668 |
| HCC827 | IC50 |
0.11 μM
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Antiproliferative activity against human HCC827 cells assessed as viability reduction incubated 72 hrs by MTT assay.
Antiproliferative activity against human HCC827 cells assessed as viability reduction incubated 72 hrs by MTT assay.
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42868668 |
| A549 | IC50 |
1.91 μM
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Antiproliferative activity against human A549 cells assessed as viability reduction incubated 72 hrs by MTT assay.
Antiproliferative activity against human A549 cells assessed as viability reduction incubated 72 hrs by MTT assay.
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42868668 |
| BEAS-2B | IC50 |
3.87 μM
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Cytotoxicity against human BEAS-2B normal lung epithelial cells assessed as viability reduction incubated 72 hrs by MTT assay.
Cytotoxicity against human BEAS-2B normal lung epithelial cells assessed as viability reduction incubated 72 hrs by MTT assay.
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42868668 |
In Vitro
Anti-cancerApoptosisCell Cycle RegulationOthers
Anti-cancer
ALK/EGFR-IN-4 (Compound 6a) (72 h) potently inhibits the proliferation of EGFR-mutant cells, with an IC50 of 0.035 μM in reagent-resistant H1975-M3 cells[1].
ALK/EGFR-IN-4 (starting concentration 50 μM, 3.16-fold serial dilution; 72 h) inhibits the viability of BaF3 cells expressing EGFR L858R/T790M/C797S, with an IC50 of 0.22 μM[1].
ALK/EGFR-IN-4 (0.1-0.5 μM; 14 days) inhibits the long-term clonogenic growth of NCI-H1975 and H1975-M3 cells in a dose-dependent manner[1].
ALK/EGFR-IN-4 (0.1-0.5 μM; 24 h) significantly inhibits DNA synthesis in NCI-H1975 and H1975-M3 cells, reducing EdU-positive cells to approximately 5-6% at 0.5 μM[1].
ALK/EGFR-IN-4 (0.1-0.5 μM; 24 h) inhibits EGFR phosphorylation and downstream PI3K/AKT, MAPK/ERK, and STAT3 signaling in NCI-H1975 and H1975-M3 cells[1].
ALK/EGFR-IN-4 (0.1-0.5 μM; 24 h) inhibits the migration and invasion of NCI-H1975 and H1975-M3 cells in a dose-dependent manner[1].
ALK/EGFR-IN-4 (starting concentration 50 μM, 3.16-fold serial dilution; 72 h) inhibits the viability of BaF3 cells expressing EGFR L858R/T790M/C797S, with an IC50 of 0.22 μM[1].
ALK/EGFR-IN-4 (0.1-0.5 μM; 14 days) inhibits the long-term clonogenic growth of NCI-H1975 and H1975-M3 cells in a dose-dependent manner[1].
ALK/EGFR-IN-4 (0.1-0.5 μM; 24 h) significantly inhibits DNA synthesis in NCI-H1975 and H1975-M3 cells, reducing EdU-positive cells to approximately 5-6% at 0.5 μM[1].
ALK/EGFR-IN-4 (0.1-0.5 μM; 24 h) inhibits EGFR phosphorylation and downstream PI3K/AKT, MAPK/ERK, and STAT3 signaling in NCI-H1975 and H1975-M3 cells[1].
ALK/EGFR-IN-4 (0.1-0.5 μM; 24 h) inhibits the migration and invasion of NCI-H1975 and H1975-M3 cells in a dose-dependent manner[1].
Anti-cancer
Apoptosis
Anti-cancer
Cell Cycle Regulation
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.
Cell Viability Assay[1]
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Cell Line:H1975-M3, NCI-H1975, HCC827, A549, and BEAS-2B cells
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Concentration:concentration series used for IC50 determination
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Incubation Time:72 h (EGFR-IN-222); 4 h (MTT)
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Result:IC50 values were 0.035 μM in H1975-M3, 0.080 μM in NCI-H1975, 0.11 μM in HCC827, 1.91 μM in A549, and 3.87 μM in BEAS-2B.
Cell Viability Assay[1]
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Cell Line:BaF3 cells stably expressing EGFR L858R/T790M/C797S
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Concentration:3.16-fold serial dilutions starting at 50 μM
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Incubation Time:72 h
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Result:Inhibited BaF3-EGFR L858R/T790M/C797S viability with IC50 = 0.22 μM.
Cell Proliferation Assay[1]
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Cell Line:NCI-H1975 and H1975-M3 cells
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Concentration:0.1, 0.25, 0.5 μM
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Incubation Time:14 days
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Result:Inhibited colony formation in a dose-dependent manner in both NCI-H1975 and H1975-M3 cells.
Cell Proliferation Assay[1]
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Cell Line:NCI-H1975 and H1975-M3 cells
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Concentration:0.1, 0.25, 0.5 μM (EGFR-IN-222); 10 μM (EdU)
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Incubation Time:24 h (EGFR-IN-222); 2 h (EdU)
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Result:In vehicle controls, approximately 32% of cells were EdU-positive.
At 0.5 μM, EdU-positive cells were reduced to 5.30% in NCI-H1975 cells and 6.34% in H1975-M3 cells, representing an approximately 80% reduction in proliferating cells; the effect was concentration-dependent.
Cell Cycle Analysis[1]
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Cell Line:NCI-H1975 and H1975-M3 cells
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Concentration:0.1, 0.25, 0.5 μM
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Incubation Time:24 h
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Result:In NCI-H1975 cells, the G0/G1 fraction increased from 39.74% (control) to 52.29% at 0.1 μM, 57.52% at 0.25 μM, and 64.56% at 0.5 μM; corresponding S-phase fractions were 21.94%, 14.26%, and 10.66%, and G2/M fractions were 21.68%, 21.54%, and 18.79%.
In H1975-M3 cells, the G0/G1 fraction increased from 38.47% (control) to 40.93% at 0.1 μM, 60.68% at 0.25 μM, and 61.53% at 0.5 μM; corresponding S-phase fractions were 15.24%, 15.36%, and 11.44%, and G2/M fractions were 29.69%, 17.53%, and 18.68%.
Western Blot Analysis[1]
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Cell Line:NCI-H1975 and H1975-M3 cells
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Concentration:0.1, 0.25, 0.5 μM
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Incubation Time:24 h
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Result:Induced a marked, dose-dependent downregulation of cyclin D1 protein in both NCI-H1975 and H1975-M3 cells.\nProduced concentration-dependent reductions in p-EGFR (Tyr992), p-AKT (Ser473), p-p44/42 MAPK (Thr202/Tyr204), and p-STAT3 (Ser727) in both NCI-H1975 and H1975-M3 cells, while total EGFR, AKT, p44/42 MAPK, and STAT3 levels remained unchanged.
In H1975-M3 cells, p-EGFR was potently suppressed at 0.5 μM.
Cell Invasion Assay[1]
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Cell Line:NCI-H1975 and H1975-M3 cells
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Concentration:0.1, 0.25, 0.5 μM
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Incubation Time:24 h
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Result:Caused a concentration-dependent reduction in the number of migrated cells through uncoated membranes and invaded cells through Matrigel-coated membranes in both NCI-H1975 and H1975-M3 cells.
Apoptosis Analysis[1]
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Cell Line:NCI-H1975 and H1975-M3 cells
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Concentration:0.1, 0.25, 0.5 μM
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Incubation Time:48 h
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Result:Induced dose-dependent apoptosis in both cell lines.
At 0.5 μM, apoptotic cells reached 52.91% in NCI-H1975 cells and 32.92% in H1975-M3 cells.
Parmacokinetics
In Vivo
Anti-cancer
ALK/EGFR-IN-4 (10-30 mg/kg; intraperitoneal; once daily; 17 days) produces dose-dependent tumor growth inhibition in H1975-M3 xenograft mice, reaching 86.47% tumor growth inhibition at 30 mg/kg[1].
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.
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Animal Model:BALB/c nude (male, five weeks old)[1]
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Dosage:10 mg/kg; 30 mg/kg
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Administration:intraperitoneal; once daily; 17 consecutive days
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Result:At 10 mg/kg, a tumor growth inhibition rate of 42.87% was achieved.
At 30 mg/kg, a tumor growth inhibition rate of 86.47% was achieved.
Tumor growth was suppressed in a dose-dependent manner.
Related Protocols :
- Western Blot
- Annexin V plus membrane-impermeant dye apoptosis staining
- BrdU Incorporation Assay
- Protocol for Cell Cycle
- Kinase activity and phosphorylation assays
- EdU Incorporation Assay (Click Chemistry-Based DNA Synthesis Measurement)
- Detection of Apoptosis
- TUNEL staining for apoptotic DNA fragmentation
- Flow cytometric DNA-content cell-cycle staining
- Patient-Derived Xenograft (PDX)
- Apoptosis Solutions
- Splenic/Portal-Vein Liver Metastasis Xenograft
- Protocol for Kinase activity and phosphorylation assays
- Subcutaneous Cell-Line-Derived Xenograft
- Orthotopic Cell-Line Xenograft
- Intraperitoneal/Peritoneal Dissemination Xenograft
Chemical Information
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CAS No. 3081622-36-5
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Molecular Weight 545.96
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Formula C24H29ClN7O4P
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SMILES
CP(C)(C1=CC=CC=C1NC2=NC(NC3=CC([N+]([O-])=O)=C(N4CCN(C)CC4)C=C3OC)=NC=C2Cl)=O
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Room temperature in continental US; may vary elsewhere.
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Storage
Please store the product under the recommended conditions in the Certificate of Analysis.
Purity & Documentation
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ALK/EGFR-IN-4 Related Classifications
HY-190486 Related Classifications
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