Pemigatinib (formerly INCB054828; INCB-054828; Pemazyre) is a novel, potent, orally bioavailable and selective FGFR inhibitor with IC50 of 0.4 nM, 0.5 nM, 1.2 nM, 30 nM for FGFR1, FGFR2, FGFR3, FGFR4, respectively. For the treatment of bile duct cancer (cholangiocarcinoma), pemigatinib has been approved as a medicine. Inhibiting FGFR1/2/3 through binding to it, INCB054828 may lead to the suppression of signal transduction pathways associated with FGFR1/2/3. When FGFR1/2/3 is overexpressed in tumor cells, this prevents their proliferation.

Physicochemical Properties

Molecular Formula	C24H27F2N5O4
Molecular Weight	487.4991
Exact Mass	487.203
Elemental Analysis	C, 59.13; H, 5.58; F, 7.79; N, 14.37; O, 13.13
CAS #	1513857-77-6
Related CAS #	1513857-77-6;1513857-77-6 (HCl);
PubChem CID	86705695
Appearance	White to light yellow solid powder
Density	1.4±0.1 g/cm3
Index of Refraction	1.620
LogP	0.66
Hydrogen Bond Donor Count	1
Hydrogen Bond Acceptor Count	8
Rotatable Bond Count	6
Heavy Atom Count	35
Complexity	731
Defined Atom Stereocenter Count	0
SMILES	FC1C(=C([H])C(=C(C=1N1C(N(C([H])([H])C([H])([H])[H])C2C(=C([H])N=C3C=2C([H])=C(C([H])([H])N2C([H])([H])C([H])([H])OC([H])([H])C2([H])[H])N3[H])C1([H])[H])=O)F)OC([H])([H])[H])OC([H])([H])[H]
InChi Key	HCDMJFOHIXMBOV-UHFFFAOYSA-N
InChi Code	InChI=1S/C24H27F2N5O4/c1-4-30-21-14(11-27-23-16(21)9-15(28-23)13-29-5-7-35-8-6-29)12-31(24(30)32)22-19(25)17(33-2)10-18(34-3)20(22)26/h9-11H,4-8,12-13H2,1-3H3,(H,27,28)
Chemical Name	11-(2,6-difluoro-3,5-dimethoxyphenyl)-13-ethyl-4-(morpholin-4-ylmethyl)-5,7,11,13-tetrazatricyclo[7.4.0.02,6]trideca-1,3,6,8-tetraen-12-one
Synonyms	INCB054828; INCB54828; INCB-054828; INCB 054828; INCB 54828; INCB-54828
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
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	FGFR1 (IC50 = 0.4 nM); FGFR2 (IC50 = 0.5 nM); FGFR3 (IC50 = 1.2 nM); FGFR4 (IC50 = 30 nM)
ln Vitro	Pemigatinib effectively reduces reactive astrocytes' ability to attract myeloid cells. Pemigatinib's potential to modulate FGFR may hold promise for promoting protective mechanisms in both human and murine systems and suppressing proinflammatory astrocyte responses.[2]
ln Vivo	Pemigatinib does not appear to have any effect on the acute EAE disease course.[2]
Cell Assay	Afatinib (10 μM), UNC2025 (10 μM), or pemigatinib (10 μM) were used for a 24-hour period to stimulate primary mouse astrocytes. ACM or a control medium was used to stimulate N2A neuronal cells for a full day. After being detached, primary mouse astrocytes and N2A neuronal cells were once again washed in cold 1× PBS. It was done using live/dead staining. Furthermore, annexin V–propidium iodide staining was carried out. Prior to being acquired on a 3L Cytek Northern Lights flow cytometer, cells were first rinsed and then resuspended in annexin V binding buffer. The OMIQ platform was used to analyze the flow cytometry data.
Animal Protocol	Female C57Bl/6J mice of experimental autoimmune encephalomyelitis (EAE) model 2.5 mg/kg i.n.
ADME/Pharmacokinetics	Absorption, Distribution and Excretion Following administration of a single oral dose of 13.5 mg pemigatinib, the median Tmax was 1.13 (0.50-6.00) hours. The steady state was reached within 4 days following repeated once daily dosing, with the median drug accumulation ratio of 1.63 (range 0.63 to 3.28). Steady-state concentration of pemigatinib increased in a dose-proportional manner over the dose range of 1 to 20 mg, which is about 0.07 to 1.5 times the recommended dose. The mean steady-state AUC and Cmax were 2620 nM x h (54% CV) and 236 nM, respectively. Following oral administration of a single radiolabeled dose of 11 mg pemigatinib, about 82.4% of the dose was recovered in feces. Of this recovered drug, about 1.4% of the dose was unchanged parent compound. About 12.6% of the dose was recovered in urine, where 1% of the dose was unchanged. The apparent volume of distribution was 235 L (60.8% CV) following a single oral dose of 13.5 mg pemigatinib. Following administration of a single oral dose of 13.5 mg pemigatinib, the geometric mean apparent clearance (CL/F) was 10.6 L/h (54% CV). Metabolism / Metabolites Pemigatinib is predominantly metabolized by the CYP3A4 enzyme _in vitro_. Its specific metabolic pathway and resulting metabolites have not yet been characterized. Biological Half-Life Following administration of a single oral dose of 13.5 mg pemigatinib, the geometric mean elimination half-life (t½) of pemigatinib was 15.4 (51.6% CV) hours.
Toxicity/Toxicokinetics	Hepatotoxicity In preregistration clinical trials of pemigatinib, ALT elevations arose in 43% to 50% of patients and were above 5 times ULN in 4% to 12%. Elevations in serum bilirubin were also common, but usually in the context of cholangiocarcinoma with partial or complete biliary obstruction. There were no cases of clinically apparent liver injury with jaundice and no deaths from liver failure attributable to pemigatinib therapy. The elevations were typically self-limited and resolved rapidly with or without dose adjustments. Since its approval, there have been no reports clinically apparent liver injury attributed to pemigatinib. However, the total clinical experience with its use has been limited and the frequency of serum aminotransferase elevations during therapy suggest that clinically significant liver injury may occur. Likelihood score: E* (unproven but possible rare cause of clinically apparent liver injury). Effects During Pregnancy and Lactation ◉ Summary of Use during Lactation No information is available on the clinical use of pemigatinib during breastfeeding. The manufacturer recommends that breastfeeding be discontinued during pemigatinib therapy and for 1 week after 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. Protein Binding The _in vitro_ serum protein binding of pemigatinib was 90.6% at drug concentrations ranging from 1 to 10 µM.
References	[1]. Pharmacol Res . 2020 Jan:151:104567. [2]. JCI Insight . 2022 Apr 8;7(7):e154824.
Additional Infomation	Pharmacodynamics Pemigatinib is a small molecule kinase inhibitor that exerts anti-tumour activity through inhibition of fibroblast growth factor receptors (FGFRs). With an IC50 of less than 2 nM, pemigatinib displays potent inhibition of FGFR1, FGFR2, and FGFR3. In mouse xenograft models of human tumours with FGFR1, FGFR2, or FGFR3 alterations, pemigatinib exhibited potent anti-tumour activity by suppressing the growth of xenografted tumour models. It also showed efficacy against a patient-derived xenograft model of cholangiocarcinoma that expressed an oncogenic FGFR2­ Transformer-2 beta homolog (TRA2b) fusion protein. Pemigatinib also inhibited FGFR4 _in vitro_, however at a concentration approximately 100 times higher than those that inhibit FGFR1, 2, and 3.

Solubility Data

Solubility (In Vitro)

DMSO: 25~40 mg/mL (51.3~82.1 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.75 mg/mL (5.64 mM) (saturation unknown) in 5% DMSO + 95% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution. 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. Solubility in Formulation 2: 2.08 mg/mL (4.27 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 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 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 (4.27 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 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. Solubility in Formulation 4: (saturation unknown) in (add these co-solvents sequentially from left to right, and one by one),  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	2.0513 mL	10.2564 mL	20.5128 mL
	5 mM	0.4103 mL	2.0513 mL	4.1026 mL
	10 mM	0.2051 mL	1.0256 mL	2.0513 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.