VEGFR2 (IC50 = 1.5 nM); PDGFRα (IC50 = 1.1 nM); FGFR1 (IC50 = 2.2 nM); c-Kit (IC50 = 12.5 nM)

AP24534 (2.5 mg/kg and 5 mg/kg) increases mice median survival in a mouse xenograft model of Ba/F3 cells expressing native Bcr-Abl. AP24534 (10 mg/kg–50 mg/kg) dramatically inhibits tumor growth in the Ba/F3 Bcr-AblT315I xenograft model. Phosphorylated Bcr-Abl and Phosphorylated CrkL are reduced in the tumors by AP24534 (30 mg/kg).[2]

The impact of AP24534 (0-320 nM) on the activity of GST-Abl kinase is measured with a synthetic peptide substrate (Abltide: EAIYAAPFAKKK). In 25 μL reaction mixture, assays are run for 15 minutes at 30 °C. 8 mM MOPS (pH 7), 0.2 mM EDTA, 50 μM Abltide, 30 mM MgCl₂, 10 mM β-glycerol phosphate, 1 mM EGTA, 0.002% Brij-35, 0.4 mM DTT, 0.2 mg/mL BSA, 0.4 mM sodium orthovanadate, 10 nM WT or mutant GST-Abl kinase, and 100 µM ATP/γ-³²[P]ATP (5000 cpm/pmol). An immersion in 0.75% phosphoric acid is required to stop a reaction after part of the reaction mixture has been transferred onto a p81 phosphocellulose filter. Phosphate incorporation is measured using scintillation counting; filters are air dried after three rounds of washing in 0.75% phosphoric acid and rinsing in acetone. By removing the peptide substrate from the kinase reaction, background binding to the filters is taken into account for all results. Kinase assays come before time course experiments to determine the linear range of enzymatic activity.

Ba/F3 cell lines are arranged in 96-well plates (4 × 10³ cells/well) and given a 72-hour incubation period with AP24534. A methanethiosulfonate (MTS)-based viability assay (CellTiter96 Aqueous One Solution) is used to measure proliferation. Every value is compared to the drug-free control wells. The mean of three separate, quadruplicat experiments is used to report IC50 values.

Mice: In the Ba/F3 survival model, 100 μL of a 1×10⁷ cells/mL suspension in serum-free medium is injected into the tail vein of female SCID mice expressing native BCR-ABL or BCR-ABL^T315I. Mice are treated with vehicle (25 mM citrate buffer, pH 2.75), dasatinib, or ponatinib once daily for up to 19 days in a row starting 72 hours later. IACUC guidelines are followed when sacrificing morbid animals. Mice with evident splenomegaly from tumor cell infiltration were found during necropsy. The Kaplan-Meier method is utilized to analyze the survival data, and a Log-rank test is employed to assess statistical significance by comparing the survival time of each treatment group with that of the vehicle group. Ba/F3 BCR-ABL^T315I cells, 100 μL of a 1×10⁷ cells/mL cell suspension in serum-free medium, are subcutaneously inserted into the right flank of female nude mice for the Ba/F3 Tumor Model. The mice are assigned to treatment groups at random once the tumor volume averages around 500 mm³. For a maximum of 19 days, mice are given oral gavage once a day with either vehicle (25 mM citrate buffer, pH 2.75) or ponatinib. It computes the tumor volume (mm³). At the final measurement, mean tumor volume for treatment group/mean tumor volume for control group (%T/C) is calculated to determine tumor growth inhibition when the treatment period is over.

Absorption, Distribution and Excretion
The absolute bioavailability of ponatinib is unknown. Peak concentrations of ponatinib are observed within 6 hours after Iclusig oral administration. Food does not affect absorption of food. The aqueous solubility of ponatinib is pH dependent, with higher pH resulting in lower solubility. When 45 mg of ponatinib is given to cancer patients, the pharmacokinetic parameters are as follows: Cmax = 73 ng/mL; AUC = 1253 ng•hr/mL;
Ponatinib is mainly eliminated via feces. Following a single oral dose of [14C]-labeled ponatinib, approximately 87% of the radioactive dose is recovered in the feces and approximately 5% in the urine.
After oral administration of 45 mg ponatinib once daily for 28 days in cancer patients, the steady state volume of distribution is 1223 L. Ponatinib is a weak substrate for P-gp and ABCG2.
Ponatinib is greater than 99% bound to plasma proteins in vitro. The geometric mean (CV%) apparent steady state volume of distribution is 1223 liters (102%) following oral administration of Iclusig 45 mg once daily for 28 days in patients with cancer. Ponatinib is a weak substrate for both P-gp and ABCG2 [also known as BCRP] in vitro. Ponatinib is not a substrate for organic anion transporting polypeptides (OATP1B1, OATP1B3) and organic cation transporter 1 (OCT1) in vitro.
Exposure increased by approximately 90% (median) (range: 20% to 440%) between the first dose and presumed steady state. Ponatinib is mainly eliminated via feces. Following a single oral dose of (14)C-labeled ponatinib, approximately 87% of the radioactive dose is recovered in the feces and approximately 5% in the urine.
The absolute bioavailability of ponatinib is unknown. Peak concentrations of ponatinib are observed within 6 hours after Iclusig oral administration. Following ingestion of either a high-fat or low-fat meal by 22 healthy volunteers, plasma ponatinib exposures (AUC and Cmax) were not different when compared to fasting conditions. The aqueous solubility of ponatinib is pH dependent, with higher pH resulting in lower solubility.
Ponatinib was equally distributed into RBCs and plasma, and did not show preferential partitioning into red blood cells in mouse, rat, monkey, or human blood. Drug derived radioactivity was found in the brain, the Tmax being 48 hours.
For more Absorption, Distribution and Excretion (Complete) data for Ponatinib (6 total), please visit the HSDB record page.

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Metabolism / Metabolites
At least 64% of a ponatinib dose undergoes phase I and phase II metabolism. CYP3A4 and to a lesser extent CYP2C8, CYP2D6 and CYP3A5 are involved in the phase I metabolism of ponatinib in vitro. Ponatinib is also metabolized by esterases and/or amidases.
In vivo, ponatinib was hydrolysed by non-specific esterases or amidases at the amide bond to an acid and aniline. AP24600 was the major metabolite in rat and human plasma but was a trace level metabolite in monkey plasma. In rat, monkey and human plasma, the amide hydrolysis metabolite AP24600 was 263%, < 1% and 58.4% of the ponatinib levels. In rats, the metabolism of ponatinib was mainly to the N-desmethyl metabolite AP24567, which was excreted in feces, and AP24600 (and its downstream metabolites) which was excreted in urine. In monkey feces drug-related radioactivity was present mostly as the parent compound or as N-desmethyl ponatinib (M42), hydroxy ponatinib (M31), a double lactam at piperazine moiety (M35) and N-oxide ponatinib (M36). In human feces, ponatinib accounted for 23.7% of the radioactivity and there was extensive metabolism of ponatinib. Other metabolites identified in human feces were hydroxy ponatinib, N-desmethyl ponatinib, and several minor metabolites resulting from two or more modifications.
At least 64% of a ponatinib dose undergoes phase I and phase II metabolism. CYP3A4 and to a lesser extent CYP2C8, CYP2D6 and CYP3A5 are involved in the phase I metabolism of ponatinib in vitro. Ponatinib is also metabolized by esterases and/or amidases.

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Biological Half-Life
After oral administration of 45 mg ponatinib once daily for 28 days in cancer patients, the terminal elimination half-life is 24 hours (range of 12 - 66 hours).
The terminal half-life of ponatinib in plasma after an IV dose was 9.7 hr in the rat and 5.3 hr in the monkey.
The geometric mean (range) terminal elimination half-life of ponatinib was approximately 24 (12 to 66) hours following Iclusig 45 mg oral administration once daily for 28 days in patients with cancer.

Hepatotoxicity
In large clinical trials, elevations in serum aminotransferase levels during ponatinib therapy occurred in up to 56% of patients and were above 5 times upper limit of normal (ULN) in 8% of patients. While these abnormalities were reversible in most patients, they were prolonged or severe in some. Instances of clinically apparent liver disease and progressive hepatic failure and death were reported in clinical trials of ponatinib, although the clinical features of the liver injury have not been well described. The latency until onset can be rapid and most cases have had a hepatocellular pattern of serum enzyme elevations. Because of the potential for serious hepatotoxicity, routine monitoring of liver tests is recommended during ponatinib therapy and dose modification or discontinuation recommended for ALT or AST elevations above 3 times ULN. Thus, ponatinib therapy is associated with a high rate of transient serum aminotransferase elevations and is reported to cause rare instances of severe hepatic injury, but there have been no cases described in the literature.
Reactivation of hepatitis B has been reported with imatinib and nilotinib therapy of CML, but not with ponatinib. Reactivation typically occurs in an HBsAg positive person treated with the tyrosine kinase inhibitor for 3 to 6 months, presenting with jaundice, marked serum aminotransferase elevations and an increase in HBV DNA levels. Reactivation of hepatitis B can be severe, and fatal instances have been reported after imatinib and nilotinib therapy. Screening of patients for HBsAg and anti-HBc is sometimes recommended before starting cancer chemotherapy and those with HBsAg offered prophylaxis with oral antiviral agents, such as lamivudine, tenofovir or entecavir. Whether reactivation occurs with ponatinib therapy is unknown.
Likelihood score: E* (unproven but suspected cause of clinically apparent liver injury).

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Effects During Pregnancy and Lactation
◉ Summary of Use during Lactation
No information is available on the clinical use of ponatinib during breastfeeding. Because ponatinib is more than 99% bound to plasma proteins, the amount in milk is likely to be low. However, its half-life is about 24 hours and it might accumulate in the infant. National Comprehensive Cancer Network guidelines recommend avoiding breastfeeding during ponatinib therapy and the manufacturer recommends withholding breastfeeding for 6 days following the last dose.
◉ Effects in Breastfed Infants
Relevant published information was not found as of the revision date.
◉ Effects on Lactation and Breastmilk
Relevant published information was not found as of the revision date.

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Protein Binding
> 99% bound to plasma proteins.

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Interactions
Ponatinib is a BCR-ABL tyrosine kinase inhibitor (TKI) approved for the treatment of chronic myeloid leukemia and Philadelphia chromosome-positive acute lymphoblastic leukemia in patients resistant or intolerant to prior TKIs. In vitro studies suggested that metabolism of ponatinib is partially mediated by CYP3A4. The effects of CYP3A4 inhibition on the pharmacokinetics of ponatinib and its CYP3A4-mediated metabolite, AP24567, were evaluated in a single-center, randomized, two-period, two-sequence crossover study in healthy volunteers. Subjects (N = 22) received two single doses (orally) of ponatinib 15 mg, once given alone and once coadministered with daily (5 days) ketoconazole 400 mg, a CYP3A4 inhibitor. Ponatinib plus ketoconazole increased ponatinib maximum plasma concentration (C(max)) and area under the concentration-time curve (AUC) compared with ponatinib alone. The estimated mean ratios for AUC0-8, AUC0-t, and C(max) indicated increased exposures to ponatinib of 78%, 70%, and 47%, respectively; exposure to AP24567 decreased by 71%. Exposure to AP24567 was marginal after ponatinib alone (no more than 4% of the exposure to ponatinib). These results suggest that caution should be exercised with the concurrent use of ponatinib and strong CYP3A4 inhibitors and that a ponatinib dose decrease to 30 mg daily, from the 45 mg daily starting dose, could be considered.
... Ponatinib at pharmacologically relevant concentrations produced synergistic cytotoxicity with ABCB1 and ABCG2 substrate chemotherapy drugs and enhanced apoptosis induced by these drugs, including daunorubicin, mitoxantrone, topotecan, and flavopiridol, in cells overexpressing these transport proteins. ...

[1]. Cancer Cell . 2009 Nov 6;16(5):401-12.

[2]. J Med Chem . 2010 Jun 24;53(12):4701-19.

[3]. Mol Cancer Ther . 2011 Jun;10(6):1028-35.

[4]. Mol Cancer Ther . 2012 Mar;11(3):690-9.

[5]. Blood . 2004 Oct 15;104(8):2532-9.

Therapeutic Uses
Antineoplastic Agents; Protein Kinase Inhibitors
Iclusig (ponatinib) is a kinase inhibitor indicated for the: Treatment of adult patients with T315I-positive chronic myeloid leukemia (CML) (chronic phase, accelerated phase, or blast phase) and T315I-positive Philadelphia chromosome positive acute lymphoblastic leukemia (Ph+ALL). /Included in US product label/
Iclusig (ponatinib) is a kinase inhibitor indicated for the: Treatment of adult patients with chronic phase, accelerated phase, or blast phase chronic myeloid leukemia or Ph+ ALL for whom no other tyrosine kinase inhibitor (TKI) therapy is indicated. /Included in US product label/

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Drug Warnings
/BOXED WARNING/ WARNING: VASCULAR OCCLUSION, HEART FAILURE, and HEPATOTOXICITY. Vascular Occlusion: Arterial and venous thrombosis and occlusions have occurred in at least 27% of Iclusig treated patients, including fatal myocardial infarction, stroke, stenosis of large arterial vessels of the brain, severe peripheral vascular disease, and the need for urgent revascularization procedures. Patients with and without cardiovascular risk factors, including patients age 50 years or younger, experienced these events. Monitor for evidence of thromboembolism and vascular occlusion. Interrupt or stop Iclusig immediately for vascular occlusion. A benefit-risk consideration should guide a decision to restart Iclusig therapy. Heart Failure: Heart failure, including fatalities, occurred in 8% of Iclusig-treated patients. Monitor cardiac function. Interrupt or stop Iclusig for new or worsening heart failure. Hepatotoxicity: Hepatotoxicity, liver failure and death have occurred in Iclusig-treated patients. Monitor hepatic function. Interrupt Iclusig if hepatotoxicity is suspected.
Hepatotoxicity and acute hepatic failure with fatal outcome have been reported in patients receiving ponatinib; liver biopsies predominantly showed hepatocellular necrosis.6 Fulminant hepatic failure resulting in death following 1 week of therapy was reported in one ponatinib-treated patient in the premarketing clinical trial. Elevations in serum aminotransferase (ALT or AST) concentrations were reported in 56% of patients receiving ponatinib in the premarketing clinical trial; median time to onset of these elevations was 46 days.6 Severe hepatotoxicity has been reported in all the disease cohorts; however, fatal cases have only been reported in patients with blast phase chronic myelogenous leukemia (CML) or Philadelphia chromosome-positive (Ph+) acute lymphocytic (lymphoblastic) leukemia (ALL).
Arterial thromboembolic events, sometimes serious or fatal, have been reported in patients receiving ponatinib. Specifically, cardiovascular, cerebrovascular, and peripheral vascular thrombosis, including fatal myocardial infarction and stroke, have occurred during ponatinib therapy. In the premarketing clinical trial, arterial thromboembolic events of any grade were reported in 11% of patients receiving ponatinib. Serious arterial thrombosis occurred in 8% of ponatinib-treated patients, and approximately 4.7% of patients required a revascularization procedure. The most common arterial thromboembolic event was myocardial infarction (MI) or worsening coronary artery disease; approximately half of these patients developed congestive heart failure concurrent with or subsequent to the myocardial ischemic event. Serious cerebrovascular events, including hemorrhagic conversion of the initial ischemic event and stenosis of large arterial vessels of the brain (e.g., carotid, vertebral, middle cerebral artery), have been reported. In the premarketing clinical trial, 3 of 449 ponatinib-treated patients developed digital or distal extremity necrosis; amputation was required in 2 patients with concomitant diabetes mellitus and peripheral arterial disease. Most (88%) patients who experienced a serious arterial thromboembolic event had one or more cardiovascular risk factors (e.g., MI, coronary artery disease, angina, stroke, transient ischemic attack (TIA), hypertension, diabetes mellitus, hyperlipidemia, cigarette smoking). If an arterial thromboembolic event occurs in a patient receiving ponatinib, interruption or discontinuance of therapy may be necessary. Venous thromboembolic events, including deep-vein thrombosis, pulmonary embolism, portal vein thrombosis, and retinal vein thrombosis, also have been reported in patients receiving ponatinib. Dosage modification or discontinuance of ponatinib therapy should be considered in patients who develop serious venous thromboembolism.
Ponatinib may cause fetal harm; the drug has been shown to be embryotoxic and fetotoxic in animals. There are no adequate and well-controlled studies in pregnant women. Pregnancy should be avoided during therapy. If used during pregnancy or if the patient becomes pregnant while receiving ponatinib, the patient should be apprised of the potential fetal hazard.
For more Drug Warnings (Complete) data for Ponatinib (22 total), please visit the HSDB record page.

C29H27F3N6O

532.56

532.219

C, 65.40; H, 5.11; F, 10.70; N, 15.78; O, 3.00

943319-70-8

Ponatinib hydrochloride;1114544-31-8;Ponatinib-d8;1562993-37-6

24826799

white solid powder

1.3±0.1 g/cm3

>160 °C

1.622

3.79

1

8

6

39

910

0

O=C(C1C=C(C#CC2N3C(C=CC=N3)=NC=2)C(C)=CC=1)NC1C=C(C(F)(F)F)C(CN2CCN(C)CC2)=CC=1

PHXJVRSECIGDHY-UHFFFAOYSA-N

InChI=1S/C29H27F3N6O/c1-20-5-6-22(16-21(20)8-10-25-18-33-27-4-3-11-34-38(25)27)28(39)35-24-9-7-23(26(17-24)29(30,31)32)19-37-14-12-36(2)13-15-37/h3-7,9,11,16-18H,12-15,19H2,1-2H3,(H,35,39)

3-(2-imidazo[1,2-b]pyridazin-3-ylethynyl)-4-methyl-N-[4-[(4-methylpiperazin-1-yl)methyl]-3-(trifluoromethyl)phenyl]benzamide

AP-24534; AP24534; Ponatinib; AP 24534; Trade name: Iclusig

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

Solubility in Formulation 1: ≥ 2.5 mg/mL (4.69 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 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.

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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.

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Solubility in Formulation 3: 30% PEG400+0.5% Tween80+5% propylene glycol: 30mg/mL

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 (Please use freshly prepared in vivo formulations for optimal results.)