PF-4989216 is a novel, potent, orally bioactive and selective small molecule inhibitor of PI3K (phosphatidylinositol 3-kinase) with potential anticancer activity. It blocks the PI3K isoforms p110α, p110β, p110γ, p110δ, and VPS34 with respective IC50 values of 2 nM, 142 nM, 65 nM, 1 nM, and 110 nM. In SCLCs with a PIK3CA mutation, PF-4989216 reduced PI3K downstream signaling, which induced apoptosis and inhibited cell viability, transformation, and the growth of xenograft tumors.
Physicochemical Properties
| Molecular Formula | C18H13N6OFS |
| Molecular Weight | 380.399 |
| Exact Mass | 380.086 |
| Elemental Analysis | C, 56.83; H, 3.44; F, 4.99; N, 22.09; O, 4.21; S, 8.43 |
| CAS # | 1276553-09-3 |
| Related CAS # | 1276553-09-3 |
| PubChem CID | 51033720 |
| Appearance | White to off-white solid powder |
| Density | 1.5±0.1 g/cm3 |
| Boiling Point | 693.7±65.0 °C at 760 mmHg |
| Flash Point | 373.4±34.3 °C |
| Vapour Pressure | 0.0±2.2 mmHg at 25°C |
| Index of Refraction | 1.696 |
| LogP | 0.48 |
| Hydrogen Bond Donor Count | 1 |
| Hydrogen Bond Acceptor Count | 8 |
| Rotatable Bond Count | 3 |
| Heavy Atom Count | 27 |
| Complexity | 628 |
| Defined Atom Stereocenter Count | 0 |
| SMILES | N#CC1=C(N2CCOCC2)SC(C3=NC=NN3)=C1C4=CC=C(C#N)C=C4F |
| InChi Key | MUENOTXSRZEFJV-UHFFFAOYSA-N |
| InChi Code | InChI=1S/C18H13FN6OS/c19-14-7-11(8-20)1-2-12(14)15-13(9-21)18(25-3-5-26-6-4-25)27-16(15)17-22-10-23-24-17/h1-2,7,10H,3-6H2,(H,22,23,24) |
| Chemical Name | 4-(4-cyano-2-fluorophenyl)-2-morpholino-5-(1H-1,2,4-triazol-5-yl)thiophene-3-carbonitrile |
| Synonyms | PF 4989216; PF-4989216; PF4989216 |
| 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 | PI3Kα (IC50 = 0.6 nM); mTOR (IC50 = 1440 nM) |
| ln Vitro | PF-4989216 (Compound 10) has good cellular potency (S473 IC50=79 nM), good selectivity against mTOR (mTOR Ki=1440 nM), and excellent PI3K Ki (0.6 nM). Both PI3Kα and mTOR Ki are greater than or equal to 1 nM in PF-4989216. Additionally, PF-4989216 has excellent selectivity against 40 additional kinases and exhibits no significant CYP inhibition. CYP enzymes 1A2, 2C9, 2D6, and 3A4 exhibit less than 30% inhibition at 3 μM[1]. The drug PF-4989216 is tested for toxicity in a number of drug-sensitive and MDR cancer cell lines, including cancer cells that overexpress ABCB1 or ABCG2 and HEK293 cells that have been transfected with human ABCB1 or ABCG2. With IC50 values of 1.11 0.09 and 6.79 1.00 uM, respectively, PF-4989216 inhibits the ABCG2-overexpressing subline S1-M1-80 of human colon carcinoma S1 cells. The ABCG2-overexpressing sublines MCF7-FLV1000 and MCF7-AdVp3000 of human breast carcinoma MCF-7 are inhibited by PF-4989216, with IC50 values of 2.300.68, 23.262.94, and 62.575.46 uM, respectively. The inhibitory concentrations (IC50s) of PF-4989216 are 0.440.05, 0.380.06, and 5.050.89 uM, respectively, for pcDNA-HEK293, ABCB1-transfected MDR19-HEK293, and ABCG2-tranfected R482-HEK293 cells[2]. |
| ln Vivo | PF-4989216 (Compound 10) is dosed orally in our in vivo antitumor model, PI3K driven NCI-H1975 xenograft tumors. PF-4989216 demonstrates dose responsive tumor growth inhibitory activity from 25 to 200 mg/kg in QD oral dosing[1]. |
| Cell Assay | MTT and CCK-8 assays are performed to determine the general sensitivities of cells to the tested drugs. PF-4989216 (0.1, 1 and 10 M) is used to treat the human colon carcinoma S1 cell line and ABCG2-overexpressing subline S1-M1-80. PF-4989216 (0.1, 1, 10, and 100 M) is used to treat the human breast cancer subtype MCF-7, which overexpresses ABCG2, and the subtypes MCF7-FLV1000 and MCF7-AdVp3000. PF-4989216 (0.01, 0.1, 1 and 10 M) is used to treat the parental HEK293 and ABCG2-tranfected R482-HEK293 cells. When used to reverse cytotoxicity assays, PF-4989216, Ko143, or Lapatinib is added at a nontoxic concentration to the assay, and the degree of reversal is then calculated[2]. |
| Animal Protocol | Mice: For animal studies, 6-8 week old nu/nu athymic female mice are used. When 2 106 cells suspended in a 1:1 (v/v) suspension of reconstituted basement membrane are injected, tumors are created. Mice with established tumors of less than 150 mm3 are randomly assigned for studies on the inhibition of tumor growth. A mouse PI3K-driven NCI-H1975 xenograft tumor model is treated with oral doses of PF-4989216 (Compound 10) of 25, 50, 100, and 200 mg/kg. Vernier calipers are used to measure the tumor's dimensions while also calculating the tumor's volume. The percentage that prevents tumor growth is calculated (TGI%). |
| References |
[1]. Highly Selective and Potent Thiophenes as PI3K Inhibitors with Oral Antitumor Activity. ACS Med Chem Lett. 2011 Sep 19;2(11):809-813. |
Solubility Data
| Solubility (In Vitro) |
DMSO: ~76 mg/mL (199.8 mM) Water: <1 mg/mL Ethanol: 7 mg/mL warming (18.4 mM) |
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (6.57 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. Solubility in Formulation 2: ≥ 2.5 mg/mL (6.57 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 3: Suspended in 0.5% methylcellulose: 1mg/mL (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.6288 mL | 13.1441 mL | 26.2881 mL | |
| 5 mM | 0.5258 mL | 2.6288 mL | 5.2576 mL | |
| 10 mM | 0.2629 mL | 1.3144 mL | 2.6288 mL |