Journal of Korean Society for Clinical Pharmacology and Therapeutics (임상약리학회지)

Korean Society for Clinical Pharmacology and Therapeutics (대한임상약리학회)
권호 표지

Application of Method to Measure Calcineurin Activity in Healthy Human Volunteers after Cyclosporine Administration

건강한 한국인 지원자에서 싸이클로스포린(Cyclosporine) 투약 후 칼시뉴린(Calcineurin) 활성도 측정법의 적용

  • Lim, Mi-Sun (Clinical Trial Center, Kyungpook National University Hospital, Department of Biomedical Science, Kyungpook National University Graduate School) ;
  • Seong, Sook-Jin (Clinical Trial Center, Kyungpook National University Hospital, Department of Biomedical Science, Kyungpook National University Graduate School) ;
  • Park, Jeong-Hyeon (Clinical Trial Center, Kyungpook National University Hospital, Department of Biomedical Science, Kyungpook National University Graduate School) ;
  • Seo, Jeong-Ju (Clinical Trial Center, Kyungpook National University Hospital, Department of Biomedical Science, Kyungpook National University Graduate School) ;
  • Lee, Joo-Mi (Clinical Trial Center, Kyungpook National University Hospital, Department of Biomedical Science, Kyungpook National University Graduate School) ;
  • Lee, Hae-Won (Clinical Trial Center, Kyungpook National University Hospital, Department of Biomedical Science, Kyungpook National University Graduate School) ;
  • Yoon, Young-Ran (Clinical Trial Center, Kyungpook National University Hospital, Department of Biomedical Science, Kyungpook National University Graduate School)
  • 임미선 (경북대학교 임상시험센터 및 경북대학교 대학원 의과학과) ;
  • 성숙진 (경북대학교 임상시험센터 및 경북대학교 대학원 의과학과) ;
  • 박정현 (경북대학교 임상시험센터 및 경북대학교 대학원 의과학과) ;
  • 서정주 (경북대학교 임상시험센터 및 경북대학교 대학원 의과학과) ;
  • 이주미 (경북대학교 임상시험센터 및 경북대학교 대학원 의과학과) ;
  • 이혜원 (경북대학교 임상시험센터 및 경북대학교 대학원 의과학과) ;
  • 윤영란 (경북대학교 임상시험센터 및 경북대학교 대학원 의과학과)
  • Received : 2011.11.25
  • Accepted : 2011.12.17
  • Published : 2011.12.30
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Abstract

Background: Calcineurin-inhibitors have wide inter-individual variation in drug response. Although therapeutic drug monitoring has been conducted to optimize personalized regimen, toxicity or rejection may occur. In this study, pharmacologic effect was evaluated by measuring calcineurin activity in peripheral blood after administration of a single dose of cyclosporine in healthy volunteers. Methods: 7 healthy Korean male subjects received cyclosporine 200 mg and blood samples were drawn immediately before and at 1, 1.5, 4, 6, 12 h after dosing to measure calcineurin activity. The blood concentrations of cyclosporine were determined for 24 hours. Calcineurin activity assay was done with Calcineurin cellular activity assay kit (Calbiochem, USA). Frozen whole blood samples in liquid $N_2$ were thawed and lysed with lysis buffer. 50 ${\mu}L$ of phosphate standard curve samples were added to each well of a 96-well plate and 10 ${\mu}L$ of diluted lysate were added to the well with RII phosphopeptide substrate. After incubating for 30 min, reaction was terminated by adding 100 ${\mu}L$ $GREEN^{TM}$ reagent. Absorbance was read at 620 nm using spectrophotometer. We evaluated percent change in calcineurin activity from baseline level in relation to the lowest level. Results: Decrease of calcineurin activity was confirmed after cyclosporine administration ($mean{\pm}SD$: $58.9{\pm}48.6$ (%)). Significant correlation was shown between calcineurin activity change and pharmacokinetic parameters ($AUC_{last}$: r = 0.834, p value = 0.01, $C_{max}$: r = 0.774, p value = 0.02). Conclusion: In this study, we confirmed the pharmacologic effect and its correlation with pharmacokinetics after administration of a single dose of cyclosporine by measuring calcineurin activity in peripheral blood in healthy volunteers.

Keywords

References

  1. Klee CB, Crouch TH, Krinks MH. Calcineurin: a calcium- and calmodulin-binding protein of the nervous system. Proc Natl Acad Sci U S A, 1979;76(12):6270-6273. https://doi.org/10.1073/pnas.76.12.6270
  2. Liu J, Farmer JD Jr, Lane WS, Friedman J, Weissman I, Schreiber SL. Calcineurin is a common target of cyclophilin-cyclosporin A and FKBP-FK506 complexes. Cell, 1991;66(4):807-815. https://doi.org/10.1016/0092-8674(91)90124-H
  3. Friedman J, Weissman I. Two cytoplasmic candidates for immunophilin action are revealed by affinity for a new cyclophilin: one in the presence and one in the absence of CsA. Cell, 1991;66(4):799-806. https://doi.org/10.1016/0092-8674(91)90123-G
  4. Liu J, Albers MW, Wandless TJ, Luan S, Alberg DG, Belshaw PJ, Cohen P, MacKintosh C, Klee CB, Schreiber SL. Inhibition of T cell signaling by immunophilin-ligand complexes correlates with loss of calcineurin phosphatase activity. Biochemistry, 1992;31(16):3896-3901. https://doi.org/10.1021/bi00131a002
  5. O'Keefe SJ, Tamura J, Kincaid RL, Tocci MJ, O'Neill EA. FK-506- and CsA-sensitive activation of the interleukin-2 promoter by calcineurin. Nature, 1992;357(6380):692-694. https://doi.org/10.1038/357692a0
  6. Clipstone NA, Crabtree GR. Identification of calcineurin as a key signalling enzyme in T-lymphocyte activation. Nature, 1992;357(6380):695-697. https://doi.org/10.1038/357695a0
  7. Shaw KT, Ho AM, Raghavan A, Kim J, Jain J, Park J, Sharma S, Rao A, Hogan PG. Immunosuppressive drugs prevent a rapid dephosphorylation of transcription factor NFAT1 in stimulated immune cells. Proc Natl Acad Sci USA, 1995;92(24):11205-11209. https://doi.org/10.1073/pnas.92.24.11205
  8. Park J, Yaseen NR, Hogan PG, Rao A, Sharma S. Phosphorylation of the transcription factor NFATp inhibits its DNA binding activity in cyclosporin A-treated human B and T cells. J Biol Chem, 1995;270(35):20653-20659. https://doi.org/10.1074/jbc.270.35.20653
  9. Loh C, Shaw KT, Carew J, Viola JP, Luo C, Perrino BA, Rao A. Calcineurin binds the transcription factor NFAT1 and reversibly regulates its activity. J Biol Chem, 1996;271(18):10884-10891. https://doi.org/10.1074/jbc.271.18.10884
  10. Crabtree GR. Contingent genetic regulatory events in T lymphocyte activation. Science, 1989;243(4889):355-361. https://doi.org/10.1126/science.2783497
  11. Shaw JP, Utz PJ, Durand DB, Toole JJ, Emmel EA, Crabtree GR. Identification of a putative regulator of early T cell activation genes. Science, 1988;241(4862):202-205. https://doi.org/10.1126/science.3260404
  12. Kahan BD, Shaw LM, Holt D, Grevel J, Johnston A. Consensus document: Hawk's Cay meeting on therapeutic drug monitoring of cyclosporine. Clin Chem, 1990;36:1510-1516.
  13. Kahan BD, Grevel J. Optimization of cyclosporine therapy in renal transplantation by a pharmacokinetic strategy. Transplantation, 1988;46(5):631-644. https://doi.org/10.1097/00007890-198811000-00002
  14. Nashan B, Cole E, Levy G, Thervet E. Clinical validation studies of Neoral C(2) monitoring: a review. Transplantation, 2002;73(9 Suppl):S3-S11.
  15. Levy G, Thervet E, Lake J, Uchida K; Consensus on Neoral C(2): Expert Review in Transplantation (CONCERT) Group. Patient management by Neoral C(2) monitoring: an international consensus statement. Transplantation, 2002;73(9 Suppl):S12-S18.
  16. Koefoed-Nielsen PB, Karamperis N, Jorgensen KA. 24-h monitoring of calcineurin phosphatase activity in healthy subjects. Scand J Immunol, 2005;62(3):309-311. https://doi.org/10.1111/j.1365-3083.2005.01660.x
  17. Koefoed-Nielsen PB, Gesualdo MB, Poulsen JH, Jorgensen KA. Blood tacrolimus levels and calcineurin phosphatase activity early after renal transplantation. Am J Transplant, 2002;2(2):173-178. https://doi.org/10.1034/j.1600-6143.2002.020209.x
  18. Yano I. Pharmacodynamic monitoring of calcineurin phosphatase activity in transplant patients treated with calcineurin inhibitors. Drug Metab Pharmacokinet, 2008;23(3):150-157. https://doi.org/10.2133/dmpk.23.150
  19. Sommerer C, Giese T, Meuer S, Zeier M. Pharmacodynamic monitoring of calcineurin inhibitor therapy: is there a clinical benefit? Nephrol Dial Transplant, 2009;24(1):21-27.
  20. van Rossum HH, de Fijter JW, van Pelt J. Pharmacodynamic monitoring of calcineurin inhibition therapy: principles, performance, and perspectives. Ther Drug Monit, 2010;32(1):3-10. https://doi.org/10.1097/FTD.0b013e3181c0eecb
  21. Chang DM, Ding YA, Kuo SY, Chang ML, Wei J. Cytokines and cell surface markers in prediction of cardiac allograft rejection. Immunol Invest, 1996;25(1-2):13-21. https://doi.org/10.3109/08820139609059287
  22. Speiser DE, Loliger CC, Siren MK, Jeannet M. Pretransplant cytotoxic donor T-cell activity specific to patient HLA class I antigens correlating with mortality after unrelated BMT. Br J Haematol, 1996;93(4):935-939.
  23. Halloran PF, Helms LM, Kung L, Noujaim J. The temporal profile of calcineurin inhibition by cyclosporine in vivo. Transplantation, 1999;68(9):1356-1361. https://doi.org/10.1097/00007890-199911150-00023
  24. Fruman DA, Klee CB, Bierer BE, Burakoff SJ. Calcineurin phosphatase activity in T lymphocytes is inhibited by FK 506 and cyclosporin A. Proc Natl Acad Sci USA, 1992;89(9):3686-3690. https://doi.org/10.1073/pnas.89.9.3686
  25. Piccinini G, Gaspari F, Signorini O, Remuzzi G, Perico N. Recovery of blood mononuclear cell calcineurin activity segregates two populations of renal transplant patients with different sensitivities to cyclosporine inhibition. Transplantation, 1996;61(10):1526-1531. https://doi.org/10.1097/00007890-199605270-00020
  26. Koefoed-Nielsen PB, Gesualdo MB, Poulsen JH, Jorgensen KA. Blood tacrolimus levels and calcineurin phosphatase activity early after renal transplantation. Am J Transplant, 2002;2(2):173-178. https://doi.org/10.1034/j.1600-6143.2002.020209.x
  27. Pai SY, Fruman DA, Leong T, Neuberg D, Rosano TG, McGarigle C, Antin JH, Bierer BE. Inhibition of calcineurin phosphatase activity in adult bone marrow transplant patients treated with cyclosporine A. Blood, 1994;84(11):3974-3979.
  28. Sanquer S, Schwarzinger M, Maury S, Yakouben K, Rafi H, Pautas C, Kuentz M, Barouki R, Cordonnier C. Calcineurin activity as a functional index of immunosuppression after allogeneic stem-cell transplantation. Transplantation, 2004;77(6):854-858. https://doi.org/10.1097/01.TP.0000114612.55925.22
  29. Fukudo M, Yano I, Masuda S, Fukatsu S, Katsura T, Ogura Y, Oike F, Takada Y, Tanaka K, Inui K. Pharmacodynamic analysis of tacrolimus and cyclosporine in living-donor liver transplant patients. Clin Pharmacol Ther, 2005;78(2):168-181. https://doi.org/10.1016/j.clpt.2005.04.008