WARNING: This product is for research use only, not for human or veterinary use.
MedKoo CAT#: 202228
CAS#: 939791-38-5 (besylate)
Description: PF-562271 Besylate, also known as PF-562,271 and PF-271, is an orally bioavailable small molecule and ATP-competitive focal adhesion kinase (FAK) inhibitor with potential antineoplastic and antiangiogenic activities. PF-562271 inhibits the tyrosine kinase FAK, and to a lesser extent, proline-rich tyrosine kinase (PYK2), which may inhibit tumor cell migration, proliferation, and survival. Note: PF-562271 benzesulfonate is also called PF-562271 besylate.
MedKoo Cat#: 202228
Name: PF-562271 Besylate
CAS#: 939791-38-5 (besylate)
Chemical Formula: C27H26F3N7O6S2
Molecular Weight: 665.66
Elemental Analysis: C, 48.72; H, 3.94; F, 8.56; N, 14.73; O, 14.42; S, 9.63
Related CAS #: 939791-38-5 (besylate) 717907-75-0 (free base) 939791-39-6 (mesylate) 939791-41-0 (HCl) 939791-40-9 (tosylate)
Synonym: PF562271 benzesulfonate salt; PF-271; PF562271 besylate; PF562271; PF562271; PF-562271; PF562,271; PF562,271; PF-562,271; PF-00562271; PF00562271; PF 00562271; PF271, PF-271, PF 271; VS-6062; VS-6062; VS 6062;
IUPAC/Chemical Name: N-methyl-N-(3-(((2-((2-oxoindolin-5-yl)amino)-5-(trifluoromethyl)pyrimidin-4-yl)amino)methyl)pyridin-2-yl)methanesulfonamide benzenesulfonate
InChi Key: LKLWTLXTOVZFAE-UHFFFAOYSA-N
InChi Code: InChI=1S/C21H20F3N7O3S.C6H6O3S/c1-31(35(2,33)34)19-12(4-3-7-25-19)10-26-18-15(21(22,23)24)11-27-20(30-18)28-14-5-6-16-13(8-14)9-17(32)29-16;7-10(8,9)6-4-2-1-3-5-6/h3-8,11H,9-10H2,1-2H3,(H,29,32)(H2,26,27,28,30);1-5H,(H,7,8,9)
SMILES Code: CS(=O)(N(C)C1=NC=CC=C1CNC2=NC(NC3=CC4=C(NC(C4)=O)C=C3)=NC=C2C(F)(F)F)=O.O=S(C5=CC=CC=C5)(O)=O
Appearance: white solid powder
Purity: >98% (or refer to the Certificate of Analysis)
Shipping Condition: Shipped under ambient temperature as non-hazardous chemical. This product is stable enough for a few weeks during ordinary shipping and time spent in Customs.
Storage Condition: Dry, dark and at 0 - 4 C for short term (days to weeks) or -20 C for long term (months to years).
Solubility: Soluble in DMSO, not in water
Shelf Life: >2 years if stored properly
Drug Formulation: This drug may be formulated in 50% DMSO and 50% PEG400.
Stock Solution Storage: 0 - 4 C for short term (days to weeks), or -20 C for long term (months).
HS Tariff Code: 2934.99.9001
|Biological target:||PF-562271 besylate is an ATP-competitive, reversible inhibitor of FAK and Pyk2 kinase, with an IC50 of 1.5 nM and 13 nM, respectively.|
|In vitro activity:||Phosphorylated FAK (Y397) was highly expressed in primary human osteosarcoma tumor samples and was associated with osteosarcoma prognosis and lung metastasis. PF562271 greatly suppressed proliferation and colony formation in human osteosarcoma cell lines. In addition, treatment of osteosarcoma cell lines with PF562271 induced apoptosis and downregulated the activity of the protein kinase B/mammalian target of rapamycin pathway. PF562271 also impaired the tube formation ability of endothelial cells in vitro. Reference: Cancer Sci. 2017 Jul;108(7):1347-1356.|
|In vivo activity:||Mouse bone marrow-derived macrophages (BMDMs) were obtained and treated with ATP or nigericin in the presence and absence of the clinical trial-tested Pyk2/FAK dual inhibitor, PF-562271. Consistent with findings in human monocyte-derived macrophages and THP-1 cells (Fig. 1A,B), the IL-1β secretion induced by ATP or nigericin was significantly blocked by pretreatment of BMDMs with PF-562271 (Fig. 6A). Next, this study analyzed the inhibition effect of PF-562271 on MSU-induced peritonitis (Fig. 6B). In vivo, MSU induced the production of IL-1β and the recruitment of cells (e.g., Gr-1 + F4/80− neutrophils and F4/80 + monocytes and macrophages) to the peritoneal cavity. However, pretreatment with PF-562271 significantly reduced the amounts of IL-1β and the numbers of recruited cells, compared to the DMSO control (Fig. 6C,D). These results suggest that the PF-562271-induced blockade of Pyk2 and FAK signaling reduces IL-1β production and the recruitment of inflammatory cells to the peritoneal cavity, thus alleviating the inflammatory symptoms in this in vivo model. Reference: Sci Rep. 2016; 6: 36214. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5087076/|
|Solvent||Max Conc. mg/mL||Max Conc. mM|
The following data is based on the product molecular weight 665.66 Batch specific molecular weights may vary from batch to batch due to the degree of hydration, which will affect the solvent volumes required to prepare stock solutions.
|Concentration / Solvent Volume / Mass||1 mg||5 mg||10 mg|
|1 mM||1.15 mL||5.76 mL||11.51 mL|
|5 mM||0.23 mL||1.15 mL||2.3 mL|
|10 mM||0.12 mL||0.58 mL||1.15 mL|
|50 mM||0.02 mL||0.12 mL||0.23 mL|
|Formulation protocol:||Tips for making stock solution: Due to its chemical nature, pure PF-562271 benzenesulfonate solid powder was found to have low solubility in common organic solvents and buffers. User may use the following information as reference when making stock solution: 1. PF562271 benzenesulfonate solubility data:(25°C): DMSO: ~ 0.4 mg/mL; Water ~ 0.8 mg/mL; Ethanol ~ 0.7 mg/mL. PBS (10 mM, pH 7.4) not soluble 2. Since PF562271 benzenesulfonate is not very soluble, when preparing stock solution, please always try small quantity first. After you get familiar with its solubility properties, you can scale up to make stock solution and do formulation. 3. The following method may help you to make high concentration stock solution: 5 mg PF562271 benzenesulfonate is mixed with 0.9 mL DMSO. The solution is sonicated for a few seconds till all big solid particles were disappeared. A cloudy solution was formed at this point. Then 0.1 mLwater was added and sonicated for a few seconds, which would give you a clear solution. The solution made by this method may be used as a stock solution. This stock solution can be diluted with water without precipitation. However, when it’s diluted with PBS buffer (10 mM, pH 7.4), the solution becomes cloudy, indicating that PF562271 was precipited from the solution. 4. Ward et al. had used 30% 2-hydroxypropyl-beta-cyclodextrin (HPCD) in 3 % dextrose to formulate PF562271. HPCD is a macrocyclic molecule, the inner core is more hydrophobic and would form complex with PF5622271. In other words, the PF562271 is encapsulated into the inner core of HPCD to form a complex, which would be more aqueous soluble. Many water insoluble drugs were formulated in this method. (reference: Ward et al. Clinical experimental metastasis June 2013, vol 30, 5:579-594).|
|In vitro protocol:||1. Hu C, Chen X, Wen J, Gong L, Liu Z, Wang J, Liang J, Hu F, Zhou Q, Wei L, Shen Y, Zhang W. Antitumor effect of focal adhesion kinase inhibitor PF562271 against human osteosarcoma in vitro and in vivo. Cancer Sci. 2017 Jul;108(7):1347-1356. doi: 10.1111/cas.13256. Epub 2017 Jun 8. Erratum in: Cancer Sci. 2018 Nov;109(11):3663-3664. PMID: 28406574; PMCID: PMC5497929. 2. Crompton BD, Carlton AL, Thorner AR, Christie AL, Du J, Calicchio ML, Rivera MN, Fleming MD, Kohl NE, Kung AL, Stegmaier K. High-throughput tyrosine kinase activity profiling identifies FAK as a candidate therapeutic target in Ewing sarcoma. Cancer Res. 2013 May 1;73(9):2873-83. doi: 10.1158/0008-5472.CAN-12-1944. Epub 2013 Mar 27. PMID: 23536552.|
|In vivo protocol:||1. Chung IC, OuYang CN, Yuan SN, Li HP, Chen JT, Shieh HR, Chen YJ, Ojcius DM, Chu CL, Yu JS, Chang YS, Chen LC. Pyk2 activates the NLRP3 inflammasome by directly phosphorylating ASC and contributes to inflammasome-dependent peritonitis. Sci Rep. 2016 Oct 31;6:36214. doi: 10.1038/srep36214. PMID: 27796369; PMCID: PMC5087076. 2. Shang N, Arteaga M, Zaidi A, Cotler SJ, Breslin P, Ding X, Kuo P, Nishimura M, Zhang J, Qiu W. FAK Kinase Activity Is Required for the Progression of c-MET/β-Catenin-Driven Hepataocellular Carcinoma. Gene Expr. 2016;17(1):79-88. doi: 10.3727/105221616X691604. Epub 2016 May 2. PMID: 27142958; PMCID: PMC5064945.|
1: Ortiz-Rivera J, Nuñez R, Kucheryavykh Y, Kucheryavykh L. The PYK2 inhibitor PF-562271 enhances the effect of temozolomide on tumor growth in a C57Bl/6-Gl261 mouse glioma model. J Neurooncol. 2023 Feb;161(3):593-604. doi: 10.1007/s11060-023-04260-3. Epub 2023 Feb 15. PMID: 36790653; PMCID: PMC9992029.
2: Yu HJ, Shin JA, Cho SD. Inhibition of focal adhesion kinase/paxillin axis by caffeic acid phenethyl ester restrains aggressive behaviors of head and neck squamous cell carcinoma in vitro. Arch Oral Biol. 2023 Feb;146:105611. doi: 10.1016/j.archoralbio.2022.105611. Epub 2022 Dec 24. PMID: 36577313.
3: Zhao T, Liu B, Zhang M, Li S, Zhao C, Cheng L. Assessment of alterations in histone modification function and guidance for death risk prediction in cervical cancer patients. Front Genet. 2022 Sep 19;13:1013571. doi: 10.3389/fgene.2022.1013571. PMID: 36199574; PMCID: PMC9527294.
4: Qin Q, Wang R, Fu Q, Zhang G, Wu T, Liu N, Lv R, Yin W, Sun Y, Sun Y, Zhao D, Cheng M. Design, synthesis, and biological evaluation of potent FAK-degrading PROTACs. J Enzyme Inhib Med Chem. 2022 Dec;37(1):2241-2255. doi: 10.1080/14756366.2022.2100886. PMID: 35978496; PMCID: PMC9455338.
5: Luo W, Han Y, Li X, Liu Z, Meng P, Wang Y. Breast Cancer Prognosis Prediction and Immune Pathway Molecular Analysis Based on Mitochondria-Related Genes. Genet Res (Camb). 2022 May 31;2022:2249909. doi: 10.1155/2022/2249909. PMID: 35707265; PMCID: PMC9174003.
6: Cho H, Shin I, Yoon H, Jeon E, Lee J, Kim Y, Ryu S, Song C, Kwon NH, Moon Y, Kim S, Kim ND, Choi HG, Sim T. Identification of Thieno[3,2-d]pyrimidine Derivatives as Dual Inhibitors of Focal Adhesion Kinase and FMS-like Tyrosine Kinase 3. J Med Chem. 2021 Aug 26;64(16):11934-11957. doi: 10.1021/acs.jmedchem.1c00459. Epub 2021 Jul 29. PMID: 34324343.
7: Tan H, Liu Y, Gong C, Zhang J, Huang J, Zhang Q. Synthesis and evaluation of FAK inhibitors with a 5-fluoro-7H-pyrrolo[2,3-d]pyrimidine scaffold as anti- hepatocellular carcinoma agents. Eur J Med Chem. 2021 Nov 5;223:113670. doi: 10.1016/j.ejmech.2021.113670. Epub 2021 Jun 25. PMID: 34214842.
8: Tien (田婷怡) TY, Wu (吳懿哲) YJ, Su (蘇正煌) CH, Wang (王學孝) HH, Hsieh (謝金玲) CL, Wang (王博正) BJ, Su (蘇瑀) Y, Yeh (葉宏一) HI. Reduction of Connexin 43 Attenuates Angiogenic Effects of Human Smooth Muscle Progenitor Cells via Inactivation of Akt and NF-κB Pathway. Arterioscler Thromb Vasc Biol. 2021 Feb;41(2):915-930. doi: 10.1161/ATVBAHA.120.315650. Epub 2020 Dec 24. PMID: 33356390.
9: Shi Y, Bray W, Smith AJ, Zhou W, Calaoagan J, Lagisetti C, Sambucetti L, Crews P, Lokey RS, Webb TR. An exon skipping screen identifies antitumor drugs that are potent modulators of pre-mRNA splicing, suggesting new therapeutic applications. PLoS One. 2020 May 29;15(5):e0233672. doi: 10.1371/journal.pone.0233672. PMID: 32469945; PMCID: PMC7259758.
10: Fenelon JC, Xu B, Baltz JM. Focal adhesion kinase PTK2 autophosphorylation is not required for the activation of sodium-hydrogen exchange by decreased cell volume in the preimplantation mouse embryo. Zygote. 2019 Jun;27(3):173-179. doi: 10.1017/S0967199419000212. Epub 2019 Jun 7. PMID: 31171046.
11: Hong KO, Ahn CH, Yang IH, Han JM, Shin JA, Cho SD, Hong SD. Norcantharidin Suppresses YD-15 Cell Invasion Through Inhibition of FAK/Paxillin and F-Actin Reorganization. Molecules. 2019 May 19;24(10):1928. doi: 10.3390/molecules24101928. PMID: 31109130; PMCID: PMC6572169.
12: Al-Ghabkari A, Qasrawi DO, Alshehri M, Narendran A. Focal adhesion kinase (FAK) phosphorylation is a key regulator of embryonal rhabdomyosarcoma (ERMS) cell viability and migration. J Cancer Res Clin Oncol. 2019 Jun;145(6):1461-1469. doi: 10.1007/s00432-019-02913-3. Epub 2019 Apr 21. PMID: 31006845.
13: Wang S, Hwang EE, Guha R, O'Neill AF, Melong N, Veinotte CJ, Conway Saur A, Wuerthele K, Shen M, McKnight C, Alexe G, Lemieux ME, Wang A, Hughes E, Xu X, Boxer MB, Hall MD, Kung A, Berman JN, Davis MI, Stegmaier K, Crompton BD. High- throughput Chemical Screening Identifies Focal Adhesion Kinase and Aurora Kinase B Inhibition as a Synergistic Treatment Combination in Ewing Sarcoma. Clin Cancer Res. 2019 Jul 15;25(14):4552-4566. doi: 10.1158/1078-0432.CCR-17-0375. Epub 2019 Apr 12. PMID: 30979745; PMCID: PMC6634997.
14: Chi Q, Wang L, Xie D, Wang X. Characterization of in vitro metabolism of focal adhesion kinase inhibitors by LC/MS/MS. J Pharm Biomed Anal. 2019 May 10;168:163-173. doi: 10.1016/j.jpba.2019.02.028. Epub 2019 Feb 19. PMID: 30807921.
15: Xiang M, Luo H, Wu J, Ren L, Ding X, Wu C, Chen J, Chen S, Zhang H, Yu L, Zou Y, Xu H, Ye P, Chen M, Xia J. ADAM23 in Cardiomyocyte Inhibits Cardiac Hypertrophy by Targeting FAK - AKT Signaling. J Am Heart Assoc. 2018 Sep 18;7(18):e008604. doi: 10.1161/JAHA.118.008604. PMID: 30371220; PMCID: PMC6222933.
16: Wang Y, Zheng J, Han Y, Zhang Y, Su L, Hu D, Fu X. JAM-A knockdown accelerates the proliferation and migration of human keratinocytes, and improves wound healing in rats via FAK/Erk signaling. Cell Death Dis. 2018 Aug 28;9(9):848. doi: 10.1038/s41419-018-0941-y. PMID: 30154481; PMCID: PMC6113279.
17: Liu NG, Yu JN, Hu B, Guo Y, Guo CQ. [Phosphorylated Focal Adhesion Kinase, Phosphinositides 3 Kinase and Aggrecan Genes and Proteins in Cartilage Cells Are Probably Involved in Needle Knife Intervention Induced Improvement of Knee Osteoarthritis in Rabbits]. Zhen Ci Yan Jiu. 2018 Apr 25;43(4):221-5. Chinese. doi: 10.13702/j.1000-0607.170890. PMID: 29888574.
18: Fang Y, Wang D, Xu X, Dava G, Liu J, Li X, Xue Q, Wang H, Zhang J, Zhang H. Preparation, in vitro and in vivo evaluation, and molecular dynamics (MD) simulation studies of novel F-18 labeled tumor imaging agents targeting focal adhesion kinase (FAK). RSC Adv. 2018 Mar 14;8(19):10333-10345. doi: 10.1039/c8ra00652k. PMID: 35540451; PMCID: PMC9078890.
19: Ali D, Abuelreich S, Alkeraishan N, Shwish NB, Hamam R, Kassem M, Alfayez M, Aldahmash A, Alajez NM. Multiple intracellular signaling pathways orchestrate adipocytic differentiation of human bone marrow stromal stem cells. Biosci Rep. 2018 Jan 30;38(1):BSR20171252. doi: 10.1042/BSR20171252. PMID: 29298881; PMCID: PMC5789155.
20: Mazzu YZ, Hu Y, Soni RK, Mojica KM, Qin LX, Agius P, Waxman ZM, Mihailovic A, Socci ND, Hendrickson RC, Tuschl T, Singer S. miR-193b-Regulated Signaling Networks Serve as Tumor Suppressors in Liposarcoma and Promote Adipogenesis in Adipose-Derived Stem Cells. Cancer Res. 2017 Nov 1;77(21):5728-5740. doi: 10.1158/0008-5472.CAN-16-2253. Epub 2017 Sep 7. PMID: 28882999.
Tips for making stock solution:
Due to its chemical nature, pure PF-562271 benzenesulfonate solid powder was found to have low solubility in common organic solvents and buffers. User may use the following information as reference when making stock solution:
1. PF562271 benzenesulfonate solubility data:(25°C): DMSO: ~ 0.4 mg/mL; Water ~ 0.8 mg/mL; Ethanol ~ 0.7 mg/mL. PBS (10 mM, pH 7.4) not soluble
2. Since PF562271 benzenesulfonate is not very soluble, when preparing stock solution, please always try small quantity first. After you get familiar with its solubility properties, you can scale up to make stock solution and do formulation.
3. The following method may help you to make high concentration stock solution: 5 mg PF562271 benzenesulfonate is mixed with 0.9 mL DMSO. The solution is sonicated for a few seconds till all big solid particles were disappeared. A cloudy solution was formed at this point. Then 0.1 mLwater was added and sonicated for a few seconds, which would give you a clear solution. The solution made by this method may be used as a stock solution. This stock solution can be diluted with water without precipitation. However, when it’s diluted with PBS buffer (10 mM, pH 7.4), the solution becomes cloudy, indicating that PF562271 was precipited from the solution.
4. Ward et al. had used 30% 2-hydroxypropyl-beta-cyclodextrin (HPCD) in 3 % dextrose to formulate PF562271. HPCD is a macrocyclic molecule, the inner core is more hydrophobic and would form complex with PF5622271. In other words, the PF562271 is encapsulated into the inner core of HPCD to form a complex, which would be more aqueous soluble. Many water insoluble drugs were formulated in this method. (reference: Ward et al. Clinical experimental metastasis June 2013, vol 30, 5:579-594).
5. The stock solution (12.5 mg/ml) was made by dissolving the drug in a solution of 50% DMSO, 50% PEG-400 (http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3087944.
6. PF562271 benzenesulfonate is less soluble in buffers with high pH (pH > 7). Therefore, we suggest to use water or acidic buffer for dilution.
7. Pure PF562271 has very low solubility, because of its chemical property, not because of the poor quality of our product. Molecule of PF562271 contains several basic nitrogen atoms, which will be porotonated in acdic media, and made the molecule more water soluble.
CAS#939791-38-5 (PF-562271 benzenesulfonate salt);
CAS#717907-75-0 (PF-562271 free base).