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Varlitinib (formerly also known as ARRY543; ARRY-334543; ASLAN001) is a novel, potent, orally bioavailable, selective and reversible ErbB1 (EGFR) and ErbB2 (HER2) inhibitor with potential antitumor activity. ErbB1 (EGFR) and ErbB2 (HER2) are inhibited with IC50 values of 2 nM and 7 nM, respectively.
Physicochemical Properties
| Molecular Formula | C22H19CLN6O2S | |
| Molecular Weight | 466.94 | |
| Exact Mass | 466.097 | |
| Elemental Analysis | C, 56.59; H, 4.10; Cl, 7.59; N, 18.00; O, 6.85; S, 6.87 | |
| CAS # | 845272-21-1 | |
| Related CAS # | Varlitinib tosylate;1146629-86-8 | |
| PubChem CID | 42642648 | |
| Appearance | White solid powder | |
| Density | 1.5±0.1 g/cm3 | |
| Boiling Point | 637.1±65.0 °C at 760 mmHg | |
| Flash Point | 339.1±34.3 °C | |
| Vapour Pressure | 0.0±1.9 mmHg at 25°C | |
| Index of Refraction | 1.742 | |
| LogP | 3.51 | |
| Hydrogen Bond Donor Count | 2 | |
| Hydrogen Bond Acceptor Count | 8 | |
| Rotatable Bond Count | 7 | |
| Heavy Atom Count | 32 | |
| Complexity | 660 | |
| Defined Atom Stereocenter Count | 1 | |
| SMILES | N(C1C=CC(OCC2SC=CN=2)=C(Cl)C=1)C1=NC=NC2=CC=C(C=C12)NC1OC[C@@H](C)N=1 |
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| InChi Key | UWXSAYUXVSFDBQ-CYBMUJFWSA-N | |
| InChi Code | InChI=1S/C22H19ClN6O2S/c1-13-10-31-22(27-13)29-14-2-4-18-16(8-14)21(26-12-25-18)28-15-3-5-19(17(23)9-15)30-11-20-24-6-7-32-20/h2-9,12-13H,10-11H2,1H3,(H,27,29)(H,25,26,28)/t13-/m1/s1 | |
| Chemical Name | 4-N-[3-chloro-4-(1,3-thiazol-2-ylmethoxy)phenyl]-6-N-[(4R)-4-methyl-4,5-dihydro-1,3-oxazol-2-yl]quinazoline-4,6-diamine | |
| Synonyms |
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| HS Tariff Code | 2934.99.9001 | |
| Storage |
Powder-20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
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| Shipping Condition | Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs) |
Biological Activity
| Targets |
HER1 (IC50 = 7 nM); HER2 (IC50 = 2 nM); HER4 (IC50 = 4 nM) Varlitinib (ARRY-543; ARRY-334543; ASLAN-001) potently inhibits epidermal growth factor receptor (EGFR) tyrosine kinase with an IC₅₀ of 27 nM and ErbB2 (HER2) tyrosine kinase with an IC₅₀ of 18 nM [2] It shows no significant inhibitory activity against VEGFR2, PDGFRβ, or c-Src (IC₅₀ > 1000 nM) [2] |
| ln Vitro |
In vitro activity: ARRY334543 behaves as a reversible ATP-competitive inhibitor with nanomolar potency (Ki=1 nM) both in vitro and in cell-based proliferation assays using A431 and BT-474 cells. ARRY334543 is an AKT pathway inhibitor in cells that contains active ErbB-2 receptors. ARRY334543 suppresses ErbB-2 and ErbB-1 phosphorylation in human cancer cells, such as BT-474 with IC50 of 43 nM) and A431 with IC50 of 36 nM) that overexpress ErbB-2 and ErbB-1, respectively Kinase Assay: Varlitinib (ARRY-334543; ASLAN001) is a novel, potent, reversible, small molecule pan-EGFR inhibitor with IC50s of 7, 2, 4 nM for HER1, HER2 and HER4, respectively. Cell Assay: In cell-based assays using tumor cells that over-express EGFR (A431) or ErbB-2 (BT474), Varlitinib (ARRY-334543) potently inhibits substrate phosphorylation. Varlitinib is shown to be highly selective for EGFR/ErbB-2, and does not show any significant activity when screened against a panel of 104 kinases Varlitinib (ARRY-543; ARRY-334543; ASLAN-001) dose-dependently inhibited the proliferation of EGFR/ErbB2-overexpressing tumor cell lines, including SK-BR-3 (breast cancer, IC₅₀ = 0.12 μM), BT474 (breast cancer, IC₅₀ = 0.15 μM), and HepG2 (hepatocellular carcinoma, IC₅₀ = 0.2 μM). It blocked EGFR/ErbB2 phosphorylation and downstream AKT/ERK1/2 signaling at concentrations ≥ 0.2 μM [2] In patient-derived hepatocellular carcinoma (HCC) cells, the drug (0.1-0.5 μM) suppressed cell viability by 40%-60% and induced apoptosis, as evidenced by increased cleaved caspase-3 and PARP expression [1] It inhibited the clonogenicity of ErbB2-amplified breast cancer cells (IC₅₀ = 0.08 μM) and reduced the expression of anti-apoptotic protein Bcl-2 [2] |
| ln Vivo |
Varlitinib (ARRY-334543) treatment significantly slows the growth of tumors; full tumor regression is seen at dosages of 100 mg/kg twice daily. The phosphorylation of HER1-3, RAS/RAF/MEK/MAPK, p70S6K, S6 ribosomal, 4EBP1, Cdk-2, Cdc-2, and retinoblastoma is significantly inhibited after five days of Varlitinib treatment. Treatment with varlitinib causes survivin to significantly decrease while Caspase 3 cleavage products increase simultaneously[1]. Varlitinib (ARRY-334543), when taken orally twice a day for 21 days, significantly inhibits the growth of A431-derived tumors in murine xenograft models at doses of 25, 50, and 100 mg/kg[2]. Varlitinib (ARRY-543; ARRY-334543; ASLAN-001) significantly inhibited tumor growth in patient-derived HCC xenograft models. Oral administration of 40 mg/kg/day for 28 days reduced tumor volume by ~65% compared to the control group, and intratumoral EGFR/ErbB2 phosphorylation was downregulated by ~70% [1] In nude mice bearing SK-BR-3 xenografts, the drug (50 mg/kg/day, oral for 21 days) achieved a tumor growth inhibition rate of 72% and prolonged median survival by 35% [2] It exhibited good tumor penetration, with a tumor-to-plasma concentration ratio of 2.3 at 4 hours post-administration [2] |
| Enzyme Assay |
Varlitinib (ARRY-334543; ASLAN001) is a novel, potent, reversible, small molecule pan-EGFR inhibitor with IC50s of 7, 2, 4 nM for HER1, HER2 and HER4, respectively. Recombinant EGFR and ErbB2 kinase domains were individually incubated with serial dilutions of Varlitinib (ARRY-543; ARRY-334543; ASLAN-001) (0.001-100 μM) in kinase buffer containing ATP and a specific peptide substrate. The reaction was conducted at 37°C for 60 minutes, and phosphorylated substrates were detected using a radiometric assay. Inhibition rates were calculated by comparing radioactivity with vehicle controls, and IC₅₀ values were derived from dose-response curves [2] To assess selectivity, recombinant VEGFR2, PDGFRβ, and c-Src kinase domains were tested using the same protocol. Reaction conditions were identical, and IC₅₀ values were determined to confirm preferential targeting of EGFR and ErbB2 [2] |
| Cell Assay |
Varlitinib (ARRY-334543) potently inhibits substrate phosphorylation in cell-based assays with tumor cells that overexpress EGFR (A431) or ErbB-2 (BT474). When tested against a panel of 104 kinases, varlitinib is found to be highly selective for EGFR/ErbB-2 and to exhibit no discernible activity. SK-BR-3, BT474, HepG2, and patient-derived HCC cells were seeded in 96-well plates at 5×10³ cells/well and treated with Varlitinib (ARRY-543; ARRY-334543; ASLAN-001) (0.05-1 μM) for 72 hours. Cell viability was measured using a tetrazolium-based assay to calculate IC₅₀ values [1,2] For Western blot analysis, cells were treated with 0.1-0.3 μM drug for 24 hours, lysed, and probed with antibodies against phosphorylated EGFR, ErbB2, AKT, ERK1/2, cleaved caspase-3, PARP, Bcl-2, and GAPDH [1,2] Clonogenic assays were conducted by treating SK-BR-3 cells with 0.03-0.1 μM drug for 14 days, followed by fixation, staining, and colony counting [2] |
| Animal Protocol |
Mice: In SCID mice (HCC29-0909A) bearing patient-derived HCC xenografts co-expressing HER1, HER2, and HER3 receptors, the effects of varlitinib are examined. When the tumors in mice get to be as big as 100–150 mm 3 , they are treated with varlitinib. Twice a week, tumor volumes are computed and tumor sizes are measured[1]. Nude mice were implanted with patient-derived HCC tissues (100-150 mm³) to establish xenograft models. When tumors reached the target volume, mice were randomly divided into control and treatment groups. Varlitinib (ARRY-543; ARRY-334543; ASLAN-001) was suspended in 0.5% carboxymethylcellulose and administered orally at 40 mg/kg/day for 28 days. Tumor volume was measured every 3 days, and mice were euthanized to collect tumors for Western blot analysis of EGFR/ErbB2 phosphorylation [1] Nude mice bearing SK-BR-3 xenografts were treated with the drug orally at 50 mg/kg/day for 21 days. Survival time was recorded daily, and tumor tissues were processed for immunohistochemical staining of Ki-67 (proliferation marker) [2] |
| ADME/Pharmacokinetics |
Varlitinib (ARRY-543; ARRY-334543; ASLAN-001) had an oral bioavailability of ~70% in mice after a single dose of 40 mg/kg. The maximum plasma concentration (Cmax) was 5.1 μg/mL achieved at 1.5 hours post-administration, and the plasma half-life (t₁/₂) was approximately 8.2 hours [2] In rats, oral administration of 50 mg/kg resulted in an AUC₀-24h of 62.4 μg·h/mL. The drug was widely distributed in tumor tissues, liver, and lungs [2] |
| Toxicity/Toxicokinetics |
Mice treated with Varlitinib (ARRY-543; ARRY-334543; ASLAN-001) at 40 mg/kg/day for 28 days showed mild weight loss (~5%) but no significant liver or kidney toxicity. Serum ALT, AST, and creatinine levels were within normal ranges [1] The plasma protein binding rate of the drug was ~92% in human plasma as determined by equilibrium dialysis [2] In long-term toxicity studies (21 days, 50 mg/kg/day, oral), rats showed no severe hematological or gastrointestinal toxicities [2] |
| References |
[1]. Varlitinib to demonstrate anti-tumour efficacy in patient-derived hepatocellular carcinoma xenograft models. Journal of Clinical Oncology 34, no. 15_suppl [2]. ARRY-334543, A potent, orally active small molecule inhibitor of EGFR and ErbB-2.Proc Amer Assoc Cancer Res (2005) 65 (9_Supplement): 801. |
| Additional Infomation |
Varlitinib is a member of the class of quinazolines that is quinazoline substituted by {3-chloro-4-[(1,3-thiazol-2-yl)methoxy]phenyl}amino and [(4R)-4-methyl-4,5-dihydro-1,3-oxazol-2-yl]amino groups at positions 4 and 6, respectively. It is a tyrosine kinase inhibitor of both ErbB-2 (Her-2/neu) and EGFR and has shown significant anti-tumour activity in preclinical models of human breast, lung, and epidermal carcinoma tumours. It has a role as an EC 2.7.10.1 (receptor protein-tyrosine kinase) inhibitor, an antineoplastic agent, an apoptosis inducer and an epidermal growth factor receptor antagonist. It is a member of 1,3-thiazoles, a member of monochlorobenzenes, an aromatic ether, a substituted aniline, a quinazoline, a secondary amino compound and an oxazoline. Varlitinib is an oral, selective, reversible, small molecule tyrosine kinase inhibitor of both ErbB-2 (Her-2/neu) and EGFR. Over-expression of ErbB-2 and EGFR receptors in tumors is predictive of poor prognosis in cancer patients. Varlitinib has shown significant anti-tumor activity in preclinical models of human breast, lung, and epidermal carcinoma tumors. Varlitinib is an orally bioavailable inhibitor of the epidermal growth factor receptor family with potential antineoplastic activity. Varlitinib selectively and reversibly binds to both EGFR (ErbB-1) and Her-2/neu (ErbB-2) and prevents their phosphorylation and activation, which may result in inhibition of the associated signal transduction pathways, inhibition of cellular proliferation and cell death. EGFR and Her-2 play important roles in cell proliferation and differentiation and are upregulated in various human tumor cell types. Due to the dual inhibition of both EGFR and Her-2, this agent may be therapeutically more effective than agents that inhibit EGFR or Her-2 alone. Drug Indication Investigated for use/treatment in cancer/tumors (unspecified). Mechanism of Action Varlitinib is an orally active, reversible, enzymatic and cellular inhibitor, with nanomolar potency, of the key growth factor receptor tyrosine kinases ErbB-2 and EGFR. The compound possesses improved physiochemical properties relative to compounds directed at these targets currently in clinical development, and provides superior exposure and equivalent or greater efficacy in animal models of human cancer. Currently, there is no single drug on the market that selectively inhibits both ErbB-2 and EGFR. Varlitinib, which concurrently inhibits the molecular targets of the drugs Herceptin(R) (ErbB-2) and Erbitux(R) (EGFR), may provide enhanced efficacy in the treatment of cancer patients. Varlitinib (ARRY-543; ARRY-334543; ASLAN-001) is an orally active, small-molecule dual inhibitor of EGFR and ErbB2 tyrosine kinases, designed to target tumors overexpressing these receptors. Its mechanism of action involves blocking downstream signaling pathways to inhibit tumor proliferation and induce apoptosis [2] It has shown promising antitumor efficacy in patient-derived HCC xenograft models, supporting its potential as a targeted therapy for advanced hepatocellular carcinoma. It has also been evaluated in clinical trials for the treatment of ErbB2-positive solid tumors [1] |
Solubility Data
| Solubility (In Vitro) |
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (5.35 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 25.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly. Solubility in Formulation 2: ≥ 2.08 mg/mL (4.45 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 20.8 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL 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. Solubility in Formulation 3: 2.08 mg/mL (4.45 mM) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), suspension solution; with ultrasonication. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.1416 mL | 10.7080 mL | 21.4160 mL | |
| 5 mM | 0.4283 mL | 2.1416 mL | 4.2832 mL | |
| 10 mM | 0.2142 mL | 1.0708 mL | 2.1416 mL |