PF-03758309 (PF03758309; PF-3758309; PF 03758309; PF3758309), a pyrrolopyrazole analog, is a novel, potent, orally bioavailable, and ATP-competitive small molecule inhibitor of PAK4 (p21-activated kinase 4) with potential antineoplastic activity. It inhibits PAK4 with an IC50 of 1.3 nM. As an inhibitor of PAK4, PF-3758309 has potential antineoplastic activity by binding to PAK4 and inhibiting PAK4 activity and cancer cell growth. PAK4, a serine/threonine kinase belonging to the p21-activated kinase (PAK) family, is often upregulated in a variety of cancer cell types and plays an important role in cancer cell motility, proliferation, and survival.
Physicochemical Properties
| Molecular Formula | C25H30N8OS | |
| Molecular Weight | 490.62 | |
| Exact Mass | 490.226 | |
| CAS # | 898044-15-0 | |
| Related CAS # | PF-3758309 hydrochloride;1279034-84-2;PF-3758309 dihydrochloride | |
| PubChem CID | 25227462 | |
| Appearance | White to light yellow solid powder | |
| Density | 1.3±0.1 g/cm3 | |
| Boiling Point | 647.9±55.0 °C at 760 mmHg | |
| Flash Point | 345.6±31.5 °C | |
| Vapour Pressure | 0.0±1.9 mmHg at 25°C | |
| Index of Refraction | 1.685 | |
| LogP | 3.28 | |
| Hydrogen Bond Donor Count | 3 | |
| Hydrogen Bond Acceptor Count | 7 | |
| Rotatable Bond Count | 6 | |
| Heavy Atom Count | 35 | |
| Complexity | 747 | |
| Defined Atom Stereocenter Count | 1 | |
| SMILES | CC1=NC2=C(C(=N1)NC3=NNC4=C3CN(C4(C)C)C(=O)N[C@H](CN(C)C)C5=CC=CC=C5)SC=C2 |
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| InChi Key | AYCPARAPKDAOEN-LJQANCHMSA-N | |
| InChi Code | InChI=1S/C25H30N8OS/c1-15-26-18-11-12-35-20(18)23(27-15)29-22-17-13-33(25(2,3)21(17)30-31-22)24(34)28-19(14-32(4)5)16-9-7-6-8-10-16/h6-12,19H,13-14H2,1-5H3,(H,28,34)(H2,26,27,29,30,31)/t19-/m1/s1 | |
| Chemical Name | (S)-N-(2-(dimethylamino)-1-phenylethyl)-6,6-dimethyl-3-((2-methylthieno[3,2-d]pyrimidin-4-yl)amino)-4,6-dihydropyrrolo[3,4-c]pyrazole-5(1H)-carboxamide. | |
| Synonyms |
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| 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 |
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| 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 | In comparison to the kinase domains of the other group B PAKs (PAK5, Ki=18.1 nM; PAK6, Ki=17.1 nM) and group A PAK1 (Ki=13.7 nM), PF-3758309 exhibits comparable enzymatic potency. However, it exhibits lower activity against the two other group A PAKs (PAK2, IC50=190 nM; PAK3, IC50=99 nM)[1]. PF-3758309 inhibits the growth of a panel of tumor cell lines that are anchorage-independent (IC50=4.7 nM) as well as the phosphorylation of the PAK4 substrate GEF-H1 (IC50=1.3 nM) in cells[1]. Moreover, PF-3758309 prevents HCT116 cells from accumulating endogenous pGEF-H1. PF-3758309 significantly suppresses A549 cell growth that is anchorage-independent (IC50=27 nM) and proliferation (IC50=20 nM)[1]. |
| ln Vivo | In HCT116 and A549 models, PF-3758309 (7.5-30 mg/kg; po; twice daily for 9–18 days) causes a statistically significant tumor growth inhibition (TGI)[1]. |
| Animal Protocol |
Animal/Disease Models: Female nu/nu, CRL breed 6–8 weeks old mice (bearing HCT116 and A549 tumors)[1] Doses: 7.5-30 mg/kg Route of Administration: Oral administration; twice (two times) daily for 9-18 days Experimental Results: Significant tumor growth inhibition (TGI) in HCT116 and A549 models . |
| References |
[1]. Small-molecule p21-activated kinase inhibitor PF3758309 is a potent inhibitor of oncogenic signaling and tumor growth. Proceedings of the National Academy of Sciences of the United States of America (2010), 107(20), 9446-9451, S94. [2]. Do PAKs make good drug targets? F1000 Biol Rep. 2010 Sep 23;2:70. [3]. PF-3758309, p21-activated kinase 4 inhibitor, suppresses migration and invasion of A549 human lung cancer cells via regulation of CREB, NF-κB, and β-catenin signalings. Mol Cell Biochem. 2014 Apr;389(1-2):69-77. [4]. Association of the epithelial-to-mesenchymal transition phenotype with responsiveness to the p21-activated kinase inhibitor, PF-3758309, in colon cancer models. Front Pharmacol. 2013 Mar 28;4:35. |
| Additional Infomation |
N-[(1S)-2-(dimethylamino)-1-phenylethyl]-6,6-dimethyl-3-[(2-methyl-4-thieno[3,2-d]pyrimidinyl)amino]-1,4-dihydropyrrolo[3,4-c]pyrazole-5-carboxamide is an organic heterobicyclic compound, an organosulfur heterocyclic compound and an organonitrogen heterocyclic compound. PF-03758309 has been used in trials studying the treatment of Advanced Solid Tumors. PAK4 Inhibitor PF-03758309 is an orally bioavailable small-molecule inhibitor of p21-activated kinase 4 (PAK4) with potential antineoplastic activity. PAK4 inhibitor PF-03758309 binds to PAK4, inhibiting PAK4 activity and cancer cell growth. PAK4, a serine/threonine kinase belonging to the p21-activated kinase (PAK) family, is often upregulated in a variety of cancer cell types and plays an important role in cancer cell motility, proliferation, and survival. |
Solubility Data
| Solubility (In Vitro) |
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (5.10 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 (5.10 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. Solubility in Formulation 3: ≥ 2.5 mg/mL (5.10 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. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.0382 mL | 10.1912 mL | 20.3824 mL | |
| 5 mM | 0.4076 mL | 2.0382 mL | 4.0765 mL | |
| 10 mM | 0.2038 mL | 1.0191 mL | 2.0382 mL |