We use cookies to improve your experience. By continuing to browse this site, you accept our cookie policy.×
Skip main navigation
Open Drawer MenuClose Drawer Menu
Bioanalysis
BioTechniques
Future Drug Discovery
Future Medicinal Chemistry
Future Science OA
International Journal of Pharmacokinetics
Pharmaceutical Patent Analyst
Therapeutic Delivery
Research Article

Engineering ‘Enzymelink’ for screening lead compounds to inhibit mPGES-1 while maintaining prostacyclin synthase activity

    Diana T Ruan

    Columbia University Vagelos College of Physicians & Surgeons, Columbia University Irving Medical Center, New York, NY, USA

    Search for more papers by this author

    ,
    Nanhong Tang

    The Center for Experimental Therapeutics & Pharmacoinformatics, Department of Pharmacological & Pharmaceutical Sciences, College of Pharmacy, University of Houston, Houston, TX, USA

    Visiting Scholar from Department of Hepatobiliary Surgery & Fujian Institute of Hepatobiliary Surgery, Cancer Center of Fujian Medical University, Fujian Medical University Union Hospital, Fuzhou, China

    Search for more papers by this author

    ,
    Hironari Akasaka

    The Center for Experimental Therapeutics & Pharmacoinformatics, Department of Pharmacological & Pharmaceutical Sciences, College of Pharmacy, University of Houston, Houston, TX, USA

    Search for more papers by this author

    ,
    Renzhong Lu

    The Center for Experimental Therapeutics & Pharmacoinformatics, Department of Pharmacological & Pharmaceutical Sciences, College of Pharmacy, University of Houston, Houston, TX, USA

    Search for more papers by this author

    &
    Ke-He Ruan

    *Author for correspondence: Tel.: +1 713 743 1771;

    E-mail Address: khruan@uh.edu

    The Center for Experimental Therapeutics & Pharmacoinformatics, Department of Pharmacological & Pharmaceutical Sciences, College of Pharmacy, University of Houston, Houston, TX, USA

    Search for more papers by this author

    Published Online:3 Jun 2021https://doi.org/10.4155/fmc-2021-0056

    Abstract

    Aim: This study investigated our Enzymelinks, COX-2-10aa-mPGES-1 and COX-2-10aa-PGIS, as cellular cross-screening targets for quick identification of lead compounds to inhibit inflammatory PGE2 biosynthesis while maintaining prostacyclin synthesis. Methods: We integrated virtual and wet cross-screening using Enzymelinks to rapidly identify lead compounds from a large compound library. Results: From 380,000 compounds virtually cross-screened with the Enzymelinks, 1576 compounds were identified and used for wet cross-screening using HEK293 cells that overexpressed individual Enzymelinks as targets. The top 15 lead compounds that inhibited mPGES-1 activity were identified. The top compound that specifically inhibited inflammatory PGE2 biosynthesis alone without affecting COX-2 coupled to PGI2 synthase (PGIS) for PGI2 biosynthesis was obtained. Conclusion: Enzymelink technology could advance cyclooxygenase pathway-targeted drug discovery to a significant degree.

    References

    • 1. Vane JR. Back to an aspirin a day? Science 296(5567), 474–475 (2002).
      Medline CASGoogle Scholar
    • 2. Funk CD. Prostaglandins and leukotrienes: advances in eicosanoid biology. Science 294(5548), 1871–1875 (2001).
      Medline CASGoogle Scholar
    • 3. Murakami M, Naraba H, Tanioka T et al. Regulation of prostaglandin E2 biosynthesis by inducible membrane-associated prostaglandin E2 synthase that acts in concert with cyclooxygenase-2. J. Biol. Chem. 275(42), 32783–32792 (2000).
      Medline CASGoogle Scholar
    • 4. Makoto M, Nakatani Y, Tanioka T, Kudo I. Prostaglandin E synthase. Prostaglandins Other Lipid Mediat. 68-69, 383–399 (2002).
      MedlineGoogle Scholar
    • 5. Ruan KH. Advance in understanding the biosynthesis of prostacyclin and thromboxane A2 in the endoplasmic reticulum membrane via the cyclooxygenase pathway. Mini. Rev. Med. Chem. 4(6), 639–647 (2004).
      Medline CASGoogle Scholar
    • 6. Dogné JM, de Leval X, Hanson J et al. New developments on thromboxane and prostacyclin modulators part I: thromboxane modulators. Curr. Med. Chem. 11(10), 1223–1241 (2004).
      Medline CASGoogle Scholar
    • 7. Murakami PM, Nakatani Y, Tanioka T, Kudo I. Prostaglandin E synthase. Prostaglandins Other Lipid Mediat. 68-69, 383–399 (2002).
      Medline CASGoogle Scholar
    • 8. Yamada T, Komoto J, Watanabe K et al. Crystal structure and possible catalytic mechanism of microsomal prostaglandin E synthase type 2 (mPGES-2). J. Mol. Biol. 348(5), 1163–1176 (2005).
      Medline CASGoogle Scholar
    • 9. Hironari A, So SP, Ruan KH. Relationship of the topological distances and activities between mPGES-1 and COX-2 versus COX-1: implications of the different post-translational endoplasmic reticulum organizations of COX-1 and COX-2. Biochemistry 54(23), 3707–3715 (2015).
      MedlineGoogle Scholar
    • 10. Zhu L, Xu C, Huo X et al. The cyclooxygenase-1/mPGES-1/endothelial prostaglandin EP4 receptor pathway constrains myocardial ischemia-reperfusion injury. Nat. Commun. 10, 1888 (2019).
      MedlineGoogle Scholar
    • 11. Ruan KH, Cervantes V, So SP. Engineering of a novel hybrid enzyme: an anti-inflammatory drug target with triple catalytic activities directly converting arachidonic acid into the inflammatory prostaglandin E2. Protein Eng. Des. Sel. 22(12), 733–740 (2009).
      Medline CASGoogle Scholar
    • 12. Ruan KH, Deng H, So SP. Engineering of a protein with cyclooxygenase and prostacyclin synthase activities that converts arachidonic acid to prostacyclin. Biochemistry 45(47), 14003–14011 (2006).
      Medline CASGoogle Scholar
    • 13. Ruan K-H, So S-P, Cervantes V et al. An active triple-catalytic hybrid enzyme engineered by linking cyclo-oxygenase isoform-1 to prostacyclin synthase that can constantly biosynthesize prostacyclin, the vascular protector. FEBS J. 275(23), 5820–5829 (2008).
      Medline CASGoogle Scholar
    • 14. Vane JR, Bakhle YS, Botting RM. Cyclooxygenases 1 and 2. Ann. Rev. Pharmacol. Toxicol. 38(January), 97–120; (1998).
      Medline CASGoogle Scholar
    • 15. Ding K, Zhou Z, Hou S et al. Structure-based discovery of mPGES-1 inhibitors suitable for preclinical testing in wild-type mice as a new generation of anti-inflammatory drugs. Sci. Rep. 8, 5205 (2018).
      MedlineGoogle Scholar
    • 16. Lauro G, Cantone V, Potenza M et al. Discovery of 3-hydroxy-3-pyrrolin-2-one-based mPGES-1 inhibitors using a multi-step virtual screening protocol. Medchemcomm. 9(12), 2028–2036 (2018).
      Medline CASGoogle Scholar
    • 17. Zhou S, Zhou Z, Ding K et al. DREAM-in-CDM approach and identification of a new generation of anti-inflammatory drugs targeting mPGES-1. Sci. Rep. 10, 10187 (2020).
      Medline CASGoogle Scholar
    • 18. Ozen G, Gomez I, Daci A et al. Inhibition of microsomal PGE synthase-1 reduces human vascular tone by increasing PGI2: a safer alternative to COX-2 inhibition. Br. J. Pharmacol. 174(22), 4087–4098 (2017).
      Medline CASGoogle Scholar
    • 19. Wang C, Chen Y, Wang Y et al. Inhibition of COX-2, mPGES-1 and CYP4A by isoliquiritigenin blocks the angiogenic Akt signaling in glioma through ceRNA effect of miR-194-5p and lncRNA NEAT1. J. Exp. Clin. Cancer Res. 38, 371 (2019).
      MedlineGoogle Scholar
    • 20. Kats A, Båge T, Georgsson P, Jönsson J et al. Inhibition of microsomal prostaglandin E synthase-1 by aminothiazoles decreases prostaglandin E2 synthesis in vitro and ameliorates experimental periodontitis in vivo. FASEB J. 27(6), 2328–2341 (2013).
      Medline CASGoogle Scholar
    • 21. Francesco LD, Bruno A, Ricciotti E et al. Pharmacological characterization of the microsomal prostaglandin E2 synthase-1 inhibitor AF3485 in vitro and in vivo. Front Pharmacol. 11, 374 (2020).
      MedlineGoogle Scholar
    • 22. Terracciano S, Lauro G, Strocchia M et al. Structural insights for the optimization of dihydropyrimidin-2(1H)-one based mPGES-1 inhibitors. ACS Med. Chem. Lett. 6(2), 187–191 (2015).
      Medline CASGoogle Scholar
    • 23. Ding K, Zhou Z, Zhou S et al. Design, synthesis, and discovery of 5-((1,3-diphenyl-1H-pyrazol-4-yl) methylene) pyrimidine-2,4,6(1H,3H,5H)-triones and related derivatives as novel inhibitors of mPGES-1. Bioorg. Med. Chem. Lett. 28(5), 858–862 (2018).
      Medline CASGoogle Scholar
    • 24. Waltenberger B, Wiechmann KW, Bauer J et al. Pharmacophore modeling and virtual screening for novel acidic inhibitors of microsomal prostaglandin E2 synthase-1 (mPGES-1). J. Med. Chem. 54(9), 3163–3174 (2011).
      Medline CASGoogle Scholar
    • 25. Frédéric M, Guiral S, Fortin L-J et al. Mark mPGES-1 inhibitor screening: an automated multistep high-throughput screening assay for the identification of lead inhibitors of the inducible enzyme mPGES-1. J. Biomolec. Screen. 10(6), 599–605 (2005).
      MedlineGoogle Scholar
    • 26. Orlando BJ, Lucido MJ, Malkowski MG. The structure of ibuprofen bound to cyclooxygenase-2. J. Struct. Biol. 189, 62–66 (2015).
      Medline CASGoogle Scholar
    • 27. Li Y-C, Chiang C-W, Yeh H-C et al. Structures of prostacyclin synthase and its complexes with substrate analog and inhibitor reveal a ligand-specific heme conformation change. J. Biol. Chem. 283, 2917–2926 (2008).
      Medline CASGoogle Scholar
    • 28. Luz JG, Antonysamy S, Kuklish SL et al. Crystal structures of mPGES-1 inhibitor complexes form a basis for the rational design of potent analgesic and anti-inflammatory therapeutics. J. Med. Chem. 58, 4727–4737 (2015).
      Medline CASGoogle Scholar
    • 29. Joel S. Fluorine – a vital element in the medicine chest. Pharmaceuticals 26–30 (2005).
      Google Scholar
    • 30. Liggett JL, Zhang X, Eling TE, Baek SJ. Anti-tumor activity of non-steroidal anti-inflammatory drugs: cyclooxygenase-independent targets. Cancer Lett. 346(2), 217–224 (2014).
      Medline CASGoogle Scholar
    • 31. Masako N, Rosenberg DW. Multifaceted roles of PGE2 in inflammation and cancer. Sem. Immunopathol. 35(2), 123–137 (2013).
      MedlineGoogle Scholar
    • 32. Masako N, Montrose DC, Clark P, et al. Genetic deletion of mPGES-1 suppresses intestinal tumorigenesis. Cancer Res. 68(9), 3251–3259 (2008).
      MedlineGoogle Scholar
    • 33. Masako N, Gokhale V, Meuillet EJ, Rosenberg DW. MPGES-1 as a target for cancer suppression. a comprehensive invited review ‘phospholipase A2 and lipid mediators’. Biochimie. 92, 660–664 (2010).
      MedlineGoogle Scholar
    • 34. Daisuke K, Murakami M, Nakatani Y et al. Potential role of microsomal prostaglandin E synthase-1 in tumorigenesis. J. Biol. Chem. 278(21), 19396–19405 (2003).
      MedlineGoogle Scholar