Vamotinib (PF-114) is a potent, selective, orally bioactive tyrosine kinase inhibitor (TKI). Vamotinib inhibits the autophosphorylation of BCR/ABL and BCR/ABL-T315I. Vamotinib causes apoptosis. Vamotinib displays antiproliferation and anti-tumor effects. Vamotinib has potential in the research/study of drug-resistant Philadelphia chromosome-positive (Ph+) leukemias. The IC50 of Vamotinib inhibiting ABL series kinases are 0.49 nM (ABL), 0.78 nM (ABLT315I), 9.5 nM (ABLE255K), 2.0 nM (ABLF317I), 7.4 nM (ABLG250E), 1.0 nM (ABLH396P), 2.8 nM (ABLM351T). , 12 nM (ABLQ252H) and 4.1 nM (ABLY253F). Vamotinib is a click chemical agent. It has Alkyne groups and could undergo CuAAc (copper-catalyzed azide-alkyne cycloaddition reaction) with compounds bearing Azide groups.

Physicochemical Properties

Molecular Formula	C29H27F3N6O
Molecular Weight	532.559496164322
Exact Mass	532.219
CAS #	1416241-23-0
Related CAS #	2141996-20-3 (mesylate);1416241-23-0;
PubChem CID	71475839
Appearance	Off-white to light yellow solid powder
LogP	5
Hydrogen Bond Donor Count	1
Hydrogen Bond Acceptor Count	8
Rotatable Bond Count	6
Heavy Atom Count	39
Complexity	910
Defined Atom Stereocenter Count	0
SMILES	C(NC1=CC=C(CN2CCN(C)CC2)C(C(F)(F)F)=C1)(=O)C1=CC=C(C)C(C#CC2N3C(=NN=2)C=CC=C3)=C1
InChi Key	SLIVDYMORZGPLW-UHFFFAOYSA-N
InChi Code	InChI=1S/C29H27F3N6O/c1-20-6-7-22(17-21(20)9-11-27-35-34-26-5-3-4-12-38(26)27)28(39)33-24-10-8-23(25(18-24)29(30,31)32)19-37-15-13-36(2)14-16-37/h3-8,10,12,17-18H,13-16,19H2,1-2H3,(H,33,39)
Chemical Name	4-methyl-N-[4-[(4-methylpiperazin-1-yl)methyl]-3-(trifluoromethyl)phenyl]-3-[2-([1,2,4]triazolo[4,3-a]pyridin-3-yl)ethynyl]benzamide
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 Note: Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture and light.
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

ln Vitro	At IC50 values of 0.49, 0.78, and 1.0 µM for ABL, ABL(T315l), and ABL(H396P), respectively, vamotinib (0-1 µM) inhibits ABL kinase and its mutations [1]. In a dose-dependent way, vamotinib (0-1000 nM) suppresses BCR/ABL and BCR/ABL-T315I autophosphorylation [1]. When Ba/F3 cells expressing natural BCR/ABL are exposed to vamotinib (0-2000 nM), it shows antiproliferative action [1]. Ba/F3 cells that express BCR/ABL and BCR/ABL-T315I undergo apoptosis when exposed to vamotinib (0-100 nM) [1]. Ph+ patient-derived cell lines grown in k562, kcl-22, SupB15, Tom-1, and BV-173 cells are inhibited by vamotinib (0-1000 nM) [1]. Ph+ PD-LTC with non-mutational resistance and the T315I mutation is inhibited in growth by vamotinib (0-1000 nM) [1].
ln Vivo	Vamotinib (25, 40 mg/kg; ir; once daily for 14 days) demonstrates anticancer efficacy [1]. Vamotinib (50 mg/kg; oral; once daily for 20 days) prolong survival in mice with BCR/ABL and BCR/ABL-T315I driven CML-like illness [1].
Cell Assay	Western Blot Analysis[1] Cell Types: Ba/F3 Cell Tested Concentrations: 0, 10, 25, 50, 100, 500, 1000 nM Incubation Duration: Experimental Results: A single dose inhibits the autophosphorylation of BCR/ABL and BCR/ABL-T315I dependent manner and inhibited substrate phosphorylation, as shown by BCR/ABL and BCR/ABL-T315I diminished Crk1 phosphorylation and downstream Stat5 activation. Cell proliferation assay[1] Cell Types: Ba/F3 Cell Tested Concentrations: 0, 50, 500, 2000 nM Incubation Duration: Experimental Results: Effectively inhibited the proliferation of Ba/F3 cells expressing native BCR/ABL in a dose-dependent manner and did not The effect on Ba/F3 cells transduced with empty vector in the presence of IL-3 (10 ng/ml) is shown. Apoptosis analysis [1] Cell Types: Ba/F3 Cell Tested Concentrations: 0-100 nM Incubation Duration: Experimental Results: Apoptosis was induced in Ba/F3 cells expressing BCR/ABL and BCR/ABL-T315I in a dose-dependent manner.
Animal Protocol	Animal/Disease Models: Female BALB/cAnNRj-Foxn1nu (nude) mice (K562 nude mouse xenograft model) [1] Doses: 25, 40 mg/kg Route of Administration: po (oral gavage); one time/day for 14 days Experimental Results: Average tumor within 4 weeks Volume diminished by 100%. Animal/Disease Models: 8-12 weeks, C57BL/6N mice (mouse model of CML-like disease) [1] Doses: 50 mg/kg Route of Administration: Po; one time/day for 20 days Experimental Results: Median survival from 28 days was Dramatically extended to 39 days.
References	[1]. PF-114, a potent and selective inhibitor of native and mutated BCR/ABL is active against Philadelphia chromosome-positive (Ph+) leukemias harboring the T315I mutation. Leukemia. 2015 May;29(5):1104-14. [2]. PF-114, a potent and selective inhibitor of native and mutated BCR/ABL is active against Philadelphia chromosome-positive (Ph+) leukemias harboring the T315I mutation. Leukemia. 2015 May;29(5):1104-14. [3]. PF‑114, a novel selective inhibitor of BCR‑ABL tyrosine kinase, is a potent inducer of apoptosis in chronic myelogenous leukemia cells. Int J Oncol. 2019 Jul;55(1):289-297.
Additional Infomation	Vamotinib (PF-114) is under investigation in clinical trial NCT02885766 (Study to Evaluate Tolerability, Safety, Pharmacokinetics and Preliminary Efficacy of PF-114 for Oral Administration in Adults With Ph+ Chronic Myeloid Leukemia, Which is Resistant to the 2-nd Generation Bcr-Abl Inhibitors or Has T315I Mutation in the BCR-ABL Gene). Vamotinib is an orally bioavailable, Bcr-Abl tyrosine kinase inhibitor, with potential antineoplastic activity. Designed to overcome resistance of tumor cells to second generation Bcr-Abl inhibitors, vamotinib targets and binds to the Bcr-Abl fusion oncoprotein, including those fusion proteins with the 'gatekeeper' resistance mutation T315I, an amino acid substitution at position 315 in Bcr-Abl from a threonine (T) to an isoleucine (I). This inhibits Bcr-Abl-mediated proliferation of, and enhances apoptosis in, Philadelphia chromosome-positive (Ph+) hematologic malignancies. The Bcr-Abl fusion protein is an aberrantly activated tyrosine kinase produced by leukemia cells that contain the Philadelphia chromosome.

Solubility Data

Solubility (In Vitro)

DMSO : ~50 mg/mL (~93.89 mM)

Solubility (In Vivo)

Solubility in Formulation 1: 2.5 mg/mL (4.69 mM) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), suspension solution; with sonication. 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 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 2: ≥ 2.5 mg/mL (4.69 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in 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 25.0 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	1.8777 mL	9.3886 mL	18.7772 mL
	5 mM	0.3755 mL	1.8777 mL	3.7554 mL
	10 mM	0.1878 mL	0.9389 mL	1.8777 mL

*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.