A5467-5mg | Ponatinib (AP24534) [943319-70-8] Clinisciences

Background

BCR-ABL fusion gene forms when the ABL gene from chromosome 9 joins to the BCR gene on chromosome 22. BCR-ABL is translated into a constitutively active tyrosine kinase, which is oncogenic. Depending on the fusion location, multiple protein variants are formed with molecular weight ranging from 185 to 210 kDa. BCR-ABL activates JAK/STAT pathway and MAPK signaling. ^[3] This gene is found in most patients with chronic myelogenous leukemia (CML), and in some patients with acute lymphoblastic leukemia (ALL) or acute myelogenous leukemia (AML).

Ponatinib is the second-generation pan inhibitor of BCR-Abl kinases, which is also effective against the mutant form of BCR-Abl (T315I). ^{[1, 2]} IC50 for WT and mutant form are 0.5 and 11 nM. ^[4] Ponatinib also inhibits several other clinically relevant kinases (RET, FLT3, KIT, PDGFRα, PDGFRβ, and FGFR1) in vitro, with IC50s of 5, 25, 100, 5, 9, and 23) in Ba/F3 cells lines. ^[4]

References:
1. Huang WS, Metcalf CA, Sundaramoorthi R, Wang Y, Zou D, Thomas RM, Zhu X, Cai L, Wen D, Liu S, Romero J, Qi J, Chen I, Banda G, Lentini SP, Das S, Xu Q, Keats J, Wang F, Wardwell S, Ning Y, Snodgrass JT, Broudy MI, Russian K, Zhou T, Commodore L, Narasimhan NI, Mohemmad QK, Iuliucci J, Rivera VM, Dalgarno DC, Sawyer TK, Clackson T, Shakespeare WC (June 2010). Discovery of 3-[2-(imidazo[1,2-b]pyridazin-3-yl)ethynyl]-4-methyl-N-{4-[(4-methylpiperazin-1-yl) methyl]-3-(trifluoromethyl)phenyl}benzamide (AP24534), a potent, orally active pan-inhibitor of breakpoint cluster region-abelson (BCR-ABL) kinase including the T315I gatekeeper mutant. J. Med. Chem. 53 (12): 4701–19.
2. O'Hare T, Pollock R, Stoffregen EP, Keats JA, Abdullah OM, Moseson EM, Rivera VM, Tang H, Metcalf Ca CA, Bohacek RS, Wang Y, Sundaramoorthi R, Shakespeare WC, Dalgarno D, Clackson T, Sawyer TK, Deininger MW, Druker BJ (2004). Inhibition of wild-type and mutant Bcr-Abl by AP23464, a potent ATP-based oncogenic protein kinase inhibitor: implications for CML. Blood 104 (8): 2532–2539.
3. Cilloni D and Saglio G. Molecular pathways: BCR-ABL. Clinical Cancer Res (2011) 18(4):930-937
4. Gozgit JM, Wong MJ, Zhu X, Schrock AB, Chen T, Clackson T and Rivera VM. Ponatinib, a potent pan-BCR-ABL inhibitor, retains activity against gatekeeper mutants of FLT3, RET, KIT, PDGFR α/β and FGFR1. 2012 AACR poster.

Product Citation

1. MOHAMMAD B. HOSSAIN, et al. "TIE2-mediated tyrosine phosphorylation of H4 regulates DNA damage response by recruiting ABL1" SCIENCE ADVANCES01 APR 2016 : E1501290.

Chemical Properties

Physical Appearance	A solid
Storage	Store at -20°C
M.Wt	532.56
Cas No.	943319-70-8
Formula	C₂₉H₂₇F₃N₆O
Solubility	≥53.3 mg/mL in DMSO; insoluble in H2O; insoluble in EtOH
Chemical Name	3-(2-imidazo[1,2-b]pyridazin-3-ylethynyl)-4-methyl-N-[4-[(4-methylpiperazin-1-yl)methyl]-3-(trifluoromethyl)phenyl]benzamide
SDF	Download SDF
Canonical SMILES	CC1=C(C=C(C=C1)C(=O)NC2=CC(=C(C=C2)CN3CCN(CC3)C)C(F)(F)F)C#CC4=CN=C5N4N=CC=C5
Shipping Condition	Small Molecules with Blue Ice, Modified Nucleotides with Dry Ice.
General tips	We do not recommend long-term storage for the solution, please use it up soon.

Protocol

Cell experiment: [1]
Cell lines	BaF3 cells stably expressing ZMYM2-FGFR1 and CEP110-FGFR1 or BCR-FGFR1
Preparation method	The solubility of this compound in DMSO is >10 mM. General tips for obtaining a higher concentration: Please warm the tube at 37 °C for 10 minutes and/or shake it in the ultrasonic bath for a while.Stock solution can be stored below -20°C for several months.
Reaction Conditions	48 hours, 50 nM for BaF3-ZMYM2, BAF3-CEP 100 nM for BaF3-BCR
Applications	Ponatinib treatment reduced phosphorylation FGFR1 levels. The percentage of cells in S-phase was also dramatically decreased, while the percentage of apoptotic cells was increased in the three different chimeric kinase-transformed BaF3 cells which suggested that their survival depended on activated FGFR1.
Animal experiment: [2]
Animal models	Female CB.17 severe combined immunodeficient mice injected with MV4-11 cells
Dosage form	Oral administration, 1–25 mg/kg, once daily for 4 weeks
Applications	Ponatinib potently inhibited tumor growth in a dose-dependent manner. Administration of 1 mg/kg, the lowest dose tested, led to significant inhibition of tumor growth (TGI = 46%, P < 0.01) and doses of 2.5 mg/kg or greater resulted in tumor regression. Notably, dosing with 10 or 25 mg/kg led to complete and durable tumor regression with no palpable tumors detected during a 31-day follow up.
Other notes	Please test the solubility of all compounds indoor, and the actual solubility may slightly differ with the theoretical value. This is caused by an experimental system error and it is normal.
References: [1] Ren M, Qin H, Ren R, Cowell JK. Ponatinib suppresses the development of myeloid and lymphoid malignancies associated with FGFR1 abnormalities. Leukemia. 2013 Jan;27(1):32-40. [2] Gozgit JM, Wong MJ, Wardwell S, Tyner JW, Loriaux MM, Mohemmad QK, Narasimhan NI, Shakespeare WC, Wang F, Druker BJ, Clackson T, Rivera VM. Potent activity of ponatinib (AP24534) in models of FLT3-driven acute myeloid leukemia and other hematologic malignancies. Mol Cancer Ther. 2011 Jun;10(6):1028-35.

Biological Activity

Description	Ponatinib (AP24534) is a novel, potent multi-target inhibitor of Abl, PDGFRα, VEGFR2, FGFR1 and Src with IC50 of 0.37 nM, 1.1 nM, 1.5 nM, 2.2 nM and 5.4 nM, respectively.
Targets	Abl	PDGFRα	VEGFR2	FGFR1	Src
IC50	0.37 nM	1.1 nM	1.5 nM	2.2 nM	5.4 nM