MedKoo Cat#: 202228 Name: PF-562271 (benzesulfonate salt) CAS#: 939791-38-5 (besylate) Chemical Formula: C27H26F3N7O6S2 Exact Mass: 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; PF562271; PF-562271; PF-00562271; PF00562271; PF 00562271; PF562271 PhSO3H; PF562271 benzesulfonate salt; PF-271; PF562271 besylate.

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/h4-8,11H,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

Technical Data

Appearance: white solid powder

Purity: >98% (or refer to the Certificate of Analysis)

Certificate of Analysis: View CoA: current batch, Lot#BBC50325 View CoA: previous batch, Lot#BBC40612 View CoA: previous batch, Lot#SSC20130322 View QC data: current batch, Lot#BBC50325 View QC data: previous batch, Lot#BBC40612 View QC data:  previous batch, Lot#SSC20130322

Safety Data Sheet (MSDS): View Material Safety Data Sheet (MSDS)

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).

Harmonized System Code: 293490

Additional Information

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.

Related CAS# CAS#939791-38-5 (PF-562271 benzenesulfonate salt); CAS#717907-75-0 (PF-562271 free base).

References

1: Fang Y, Wang D, Xu X, Liu J, Wu A, Li X, Xue Q, Wang H, Wang H, Zhang H. Synthesis, biological evaluation, and molecular dynamics (MD) simulation studies of three novel F-18 labeled and focal adhesion kinase (FAK) targeted 5-bromo pyrimidines as radiotracers for tumor. Eur J Med Chem. 2017 Feb 15;127:493-508. doi: 10.1016/j.ejmech.2017.01.015. Epub 2017 Jan 11. PubMed PMID: 28109944.

2: Kessler BE, Sharma V, Zhou Q, Jing X, Pike LA, Kerege AA, Sams SB, Schweppe RE. FAK Expression, Not Kinase Activity, Is a Key Mediator of Thyroid Tumorigenesis and Protumorigenic Processes. Mol Cancer Res. 2016 Sep;14(9):869-82. doi: 10.1158/1541-7786.MCR-16-0007. Epub 2016 Jun 3. PubMed PMID: 27259715; PubMed Central PMCID: PMC5025360.

3: Howe GA, Xiao B, Zhao H, Al-Zahrani KN, Hasim MS, Villeneuve J, Sekhon HS, Goss GD, Sabourin LA, Dimitroulakos J, Addison CL. Focal Adhesion Kinase Inhibitors in Combination with Erlotinib Demonstrate Enhanced Anti-Tumor Activity in Non-Small Cell Lung Cancer. PLoS One. 2016 Mar 10;11(3):e0150567. doi: 10.1371/journal.pone.0150567. eCollection 2016 Mar 10. PubMed PMID: 26962872; PubMed Central PMCID: PMC4786279.

4: Fan GP, Wang W, Zhao H, Cai L, Zhang PD, Yang ZH, Zhang J, Wang X. Pharmacological Inhibition of Focal Adhesion Kinase Attenuates Cardiac Fibrosis in Mice Cardiac Fibroblast and Post-Myocardial-Infarction Models. Cell Physiol Biochem. 2015;37(2):515-26. PubMed PMID: 26330161.

5: Rolón-Reyes K, Kucheryavykh YV, Cubano LA, Inyushin M, Skatchkov SN, Eaton MJ, Harrison JK, Kucheryavykh LY. Microglia Activate Migration of Glioma Cells through a Pyk2 Intracellular Pathway. PLoS One. 2015 Jun 22;10(6):e0131059. doi: 10.1371/journal.pone.0131059. eCollection 2015 Jun 22. PubMed PMID: 26098895; PubMed Central PMCID: PMC4476590.

6: Shelef MA, Bennin DA, Yasmin N, Warner TF, Ludwig T, Beggs HE, Huttenlocher A. Focal adhesion kinase is required for synovial fibroblast invasion, but not murine inflammatory arthritis. Arthritis Res Ther. 2014 Oct 4;16(5):464. doi: 10.1186/s13075-014-0464-6. PubMed PMID: 25280866; PubMed Central PMCID: PMC4203874.

7: Stone RL, Baggerly KA, Armaiz-Pena GN, Kang Y, Sanguino AM, Thanapprapasr D, Dalton HJ, Bottsford-Miller J, Zand B, Akbani R, Diao L, Nick AM, DeGeest K, Lopez-Berestein G, Coleman RL, Lutgendorf S, Sood AK. Focal adhesion kinase: an alternative focus for anti-angiogenesis therapy in ovarian cancer. Cancer Biol Ther. 2014 Jul;15(7):919-29. doi: 10.4161/cbt.28882. Epub 2014 Apr 23. PubMed PMID: 24755674; PubMed Central PMCID: PMC4100993.

8: Yoon H, Choi YL, Song JY, Do I, Kang SY, Ko YH, Song S, Kim BG. Targeted inhibition of FAK, PYK2 and BCL-XL synergistically enhances apoptosis in ovarian clear cell carcinoma cell lines. PLoS One. 2014 Feb 11;9(2):e88587. doi: 10.1371/journal.pone.0088587. eCollection 2014 Feb 11. PubMed PMID: 24523919; PubMed Central PMCID: PMC3921183.

9: Grädler U, Bomke J, Musil D, Dresing V, Lehmann M, Hölzemann G, Greiner H, Esdar C, Krier M, Heinrich T. Fragment-based discovery of focal adhesion kinase inhibitors. Bioorg Med Chem Lett. 2013 Oct 1;23(19):5401-9. doi: 10.1016/j.bmcl.2013.07.050. Epub 2013 Jul 31. PubMed PMID: 23973211.

10: Wiemer AJ, Wernimont SA, Cung TD, Bennin DA, Beggs HE, Huttenlocher A. The focal adhesion kinase inhibitor PF-562,271 impairs primary CD4+ T cell activation. Biochem Pharmacol. 2013 Sep 15;86(6):770-81. doi: 10.1016/j.bcp.2013.07.024. Epub 2013 Aug 5. PubMed PMID: 23928188; PubMed Central PMCID: PMC3762933.

11: Lagares D, Busnadiego O, García-Fernández RA, Lamas S, Rodríguez-Pascual F. Adenoviral gene transfer of endothelin-1 in the lung induces pulmonary fibrosis through the activation of focal adhesion kinase. Am J Respir Cell Mol Biol. 2012 Dec;47(6):834-42. doi: 10.1165/rcmb.2011-0446OC. Epub 2012 Sep 6. PubMed PMID: 22962065.

12: Lagares D, Busnadiego O, García-Fernández RA, Kapoor M, Liu S, Carter DE, Abraham D, Shi-Wen X, Carreira P, Fontaine BA, Shea BS, Tager AM, Leask A, Lamas S, Rodríguez-Pascual F. Inhibition of focal adhesion kinase prevents experimental lung fibrosis and myofibroblast formation. Arthritis Rheum. 2012 May;64(5):1653-64. doi: 10.1002/art.33482. PubMed PMID: 22492165; PubMed Central PMCID: PMC3338902.

13: Stokes JB, Adair SJ, Slack-Davis JK, Walters DM, Tilghman RW, Hershey ED, Lowrey B, Thomas KS, Bouton AH, Hwang RF, Stelow EB, Parsons JT, Bauer TW. Inhibition of focal adhesion kinase by PF-562,271 inhibits the growth and metastasis of pancreatic cancer concomitant with altering the tumor microenvironment. Mol Cancer Ther. 2011 Nov;10(11):2135-45. doi: 10.1158/1535-7163.MCT-11-0261. Epub 2011 Sep 8. PubMed PMID: 21903606; PubMed Central PMCID: PMC3213273.

14: Serrels A, McLeod K, Canel M, Kinnaird A, Graham K, Frame MC, Brunton VG. The role of focal adhesion kinase catalytic activity on the proliferation and migration of squamous cell carcinoma cells. Int J Cancer. 2012 Jul 15;131(2):287-97. doi: 10.1002/ijc.26351. Epub 2011 Aug 30. PubMed PMID: 21823119.

15: Schultze A, Fiedler W. Clinical importance and potential use of small molecule inhibitors of focal adhesion kinase. Anticancer Agents Med Chem. 2011 Sep;11(7):593-9. Review. PubMed PMID: 21787277.

16: Wendt MK, Smith JA, Schiemann WP. Transforming growth factor-β-induced epithelial-mesenchymal transition facilitates epidermal growth factor-dependent breast cancer progression. Oncogene. 2010 Dec 9;29(49):6485-98. doi: 10.1038/onc.2010.377. Epub 2010 Aug 30. PubMed PMID: 20802523; PubMed Central PMCID: PMC3076082.

17: Sun H, Pisle S, Gardner ER, Figg WD. Bioluminescent imaging study: FAK inhibitor, PF-562,271, preclinical study in PC3M-luc-C6 local implant and metastasis xenograft models. Cancer Biol Ther. 2010 Jul 1;10(1):38-43. Epub 2010 Jul 9. PubMed PMID: 20495381; PubMed Central PMCID: PMC3087944.

18: Hao H, Naomoto Y, Bao X, Watanabe N, Sakurama K, Noma K, Motoki T, Tomono Y, Fukazawa T, Shirakawa Y, Yamatsuji T, Matsuoka J, Wang ZG, Takaoka M. Focal adhesion kinase as potential target for cancer therapy (Review). Oncol Rep. 2009 Nov;22(5):973-9. Review. PubMed PMID: 19787209.

19: Wendt MK, Schiemann WP. Therapeutic targeting of the focal adhesion complex prevents oncogenic TGF-beta signaling and metastasis. Breast Cancer Res. 2009;11(5):R68. doi: 10.1186/bcr2360. PubMed PMID: 19740433; PubMed Central PMCID: PMC2790843.

20: Slack-Davis JK, Hershey ED, Theodorescu D, Frierson HF, Parsons JT. Differential requirement for focal adhesion kinase signaling in cancer progression in the transgenic adenocarcinoma of mouse prostate model. Mol Cancer Ther. 2009 Aug;8(8):2470-7. doi: 10.1158/1535-7163.MCT-09-0262. Epub 2009 Aug 11. PubMed PMID: 19671741; PubMed Central PMCID: PMC2728172.