The New LC-MS/MS Method for Determination of Unbound Tacrolimus in Plasma

NCT04657562 | Unknown FDA Drug

• 1 other identifier: 2019/33/N/NZ7/01631
• Study Type: observational
• Target: 380
• Locations: 1 country
• Sites: 1

Brief Summary

Tacrolimus (TAC) is characterized by a narrow therapeutic window, as well as high inter- and intra-individual variability in pharmacokinetics. Both under- and overexposure may lead to severe adverse effects. Therapeutic drug monitoring (TDM) is an essential element of post-transplant patient care. Most transplantation centers use C0 to adjust TAC dosage. Some controversies remain about relationship between C0 and clinical outcome. It is generally accepted that only protein-unbound drug molecules can cross cellular membranes, which imply that TDM of free tacrolimus fraction may be of paramount importance and improve clinical management of organ recipients. Whole blood TAC concentrations and dose requirements are strongly associated with CYP3A5 polymorphism. Routine CYP3A5 genotyping on the waiting lists might be useful to guide tacrolimus dosing. This interdisciplinary project tackles the research problem from three angles - biochemistry, genetics and clinical observation. The primary goal of the study is to evaluate clinical usefulness of different TDM protocols in patients after kidney and liver transplantation.

Conditions

Immunosuppression; Kidney Transplant Rejection; Kidney Transplant; Complications; Liver Transplant Rejection; Liver Transplant; Complications; Metabolism Medication Toxicity; Genetic Predisposition

Keywords

Kidney transplant; liver transplant; CYP3A; unbound tacrolimus; therapeutic drug monitoring

Outcome Measures

Primary Outcomes (4)

• Development and validation of the new LC-MS/MS measurement method

  Development and validation of the extremely sensitive method for unbound tacrolimus determination using the EMA and FDA guidelines

  1 year

• A comparison of different TAC TDM protocols depending on the matrix

  1. concentration of unbound tacrolimus in plasma ultrafiltrate 2. concentration of tacrolimus in plasma 3. concentration of tacrolimus in whole blood The measures will be obtained with the use of LC-MS/MS method.

  1 year

• Equation to calculate unbound TAC concentration

  Development of the equation (using the concentration of free TAC, plasma and whole blood, and blood components) by statistical methods.

  1 year

• A correlation between free TAC and symptoms of underimmunosuppresion

  Biopsy proven acute rejection

  1 year

Secondary Outcomes (3)

• A correlation between free TAC and nephrotoxicity

  1 year

• CYP3A expression

  1 year

• Blood components

  1 year

Study Arms (3)

De novo renal/liver transplant recipients

The group of 40 consecutive adult (age \> 18 years) male and female recipients of deceased kidney or liver transplant from the Regional Qualification Center (Warsaw, Poland).

Drug: Tacrolimus; Diagnostic Test: Unbound tacrolimus measurement; Diagnostic Test: CYP3A4 and CYP3A5 genotyping

Random renal transplant recipients

The group of 300 random adult (age \> 18 years) male and female recipients of deceased kidney attending the local outpatient clinic.

Drug: Tacrolimus; Diagnostic Test: Unbound tacrolimus measurement; Diagnostic Test: CYP3A4 and CYP3A5 genotyping

Renal transplant recipients experiencing graft rejection

The group of 40 consecutive adult (age \> 18 years) male and female recipients of deceased kidney experiencing acute rejection of the renal allograft.

Drug: Tacrolimus; Diagnostic Test: Unbound tacrolimus measurement; Diagnostic Test: CYP3A4 and CYP3A5 genotyping

Interventions

Tacrolimus DRUG

Prevention of rejection in kidney or liver transplant: a standard immunosuppressive therapy according to international protocols.

Also known as: Prograf, Advagraf

De novo renal/liver transplant recipients; Random renal transplant recipients; Renal transplant recipients experiencing graft rejection

Unbound tacrolimus measurement DIAGNOSTIC_TEST

Unbound tacrolimus measurement in plasma ultrafiltrate.

De novo renal/liver transplant recipients; Random renal transplant recipients; Renal transplant recipients experiencing graft rejection

CYP3A4 and CYP3A5 genotyping DIAGNOSTIC_TEST

DNA purification and genotyping

De novo renal/liver transplant recipients; Random renal transplant recipients; Renal transplant recipients experiencing graft rejection

Eligibility Criteria

• Age: 18 Years+
• Sex: all
• Healthy Volunteers: No
• Age Groups: Adult (18-64), Older Adult (65+)
• Sampling Method: Probability Sample

Study Population

1. 40 consecutive de novo adult kidney or liver transplant recipients on TAC-based immunosuppression. 2. 300 adult kidney transplant recipients attending the local outpatient clinic. 3. 40 adult kidney transplant recipients experiencing acute rejection of the renal allograft.

You may qualify if:

• adult (\>18 yo), recipient of kidney or liver transplant from the Regional Qualification Center, tacrolimus-based immunosuppression

You may not qualify if:

• the use of agents significantly influencing TAC metabolism, double organ recipients, HBV, HCV and HIV infection, neurological disorders.

Sponsors & Collaborators

• National Science Centre, Poland: lead
• Medical University of Warsaw: collaborator

Study Sites (1)

Department of General and Transplant Surgery, Medical University of Warsaw

Warsaw, 02-006, Poland

RECRUITING

Related Publications (24)

• Kidney Disease: Improving Global Outcomes (KDIGO) Transplant Work Group. KDIGO clinical practice guideline for the care of kidney transplant recipients. Am J Transplant. 2009 Nov;9 Suppl 3:S1-155. doi: 10.1111/j.1600-6143.2009.02834.x.

  PMID: 19845597 BACKGROUND

• European Association for the Study of the Liver. EASL Clinical Practice Guidelines: Liver transplantation. J Hepatol. 2016 Feb;64(2):433-485. doi: 10.1016/j.jhep.2015.10.006. Epub 2015 Nov 17. No abstract available.

  PMID: 26597456 BACKGROUND

• Bittersohl H, Schniedewind B, Christians U, Luppa PB. A simple and highly sensitive on-line column extraction liquid chromatography-tandem mass spectrometry method for the determination of protein-unbound tacrolimus in human plasma samples. J Chromatogr A. 2018 Apr 27;1547:45-52. doi: 10.1016/j.chroma.2018.03.010. Epub 2018 Mar 7.

  PMID: 29544893 BACKGROUND

• Brunet M, van Gelder T, Asberg A, Haufroid V, Hesselink DA, Langman L, Lemaitre F, Marquet P, Seger C, Shipkova M, Vinks A, Wallemacq P, Wieland E, Woillard JB, Barten MJ, Budde K, Colom H, Dieterlen MT, Elens L, Johnson-Davis KL, Kunicki PK, MacPhee I, Masuda S, Mathew BS, Millan O, Mizuno T, Moes DAR, Monchaud C, Noceti O, Pawinski T, Picard N, van Schaik R, Sommerer C, Vethe NT, de Winter B, Christians U, Bergan S. Therapeutic Drug Monitoring of Tacrolimus-Personalized Therapy: Second Consensus Report. Ther Drug Monit. 2019 Jun;41(3):261-307. doi: 10.1097/FTD.0000000000000640.

  PMID: 31045868 BACKGROUND

• Ekberg H, Tedesco-Silva H, Demirbas A, Vitko S, Nashan B, Gurkan A, Margreiter R, Hugo C, Grinyo JM, Frei U, Vanrenterghem Y, Daloze P, Halloran PF; ELITE-Symphony Study. Reduced exposure to calcineurin inhibitors in renal transplantation. N Engl J Med. 2007 Dec 20;357(25):2562-75. doi: 10.1056/NEJMoa067411.

  PMID: 18094377 BACKGROUND

• Pascual J, Berger SP, Witzke O, Tedesco H, Mulgaonkar S, Qazi Y, Chadban S, Oppenheimer F, Sommerer C, Oberbauer R, Watarai Y, Legendre C, Citterio F, Henry M, Srinivas TR, Luo WL, Marti A, Bernhardt P, Vincenti F; TRANSFORM Investigators. Everolimus with Reduced Calcineurin Inhibitor Exposure in Renal Transplantation. J Am Soc Nephrol. 2018 Jul;29(7):1979-1991. doi: 10.1681/ASN.2018010009. Epub 2018 May 11.

  PMID: 29752413 BACKGROUND

• Stienstra NA, Sikma MA, van Dapperen AL, de Lange DW, van Maarseveen EM. Development of a Simple and Rapid Method to Measure the Free Fraction of Tacrolimus in Plasma Using Ultrafiltration and LC-MS/MS. Ther Drug Monit. 2016 Dec;38(6):722-727. doi: 10.1097/FTD.0000000000000351.

  PMID: 27805928 BACKGROUND

• Bouamar R, Shuker N, Hesselink DA, Weimar W, Ekberg H, Kaplan B, Bernasconi C, van Gelder T. Tacrolimus predose concentrations do not predict the risk of acute rejection after renal transplantation: a pooled analysis from three randomized-controlled clinical trials(dagger). Am J Transplant. 2013 May;13(5):1253-61. doi: 10.1111/ajt.12191. Epub 2013 Mar 8.

  PMID: 23480233 BACKGROUND

• Israni AK, Riad SM, Leduc R, Oetting WS, Guan W, Schladt D, Matas AJ, Jacobson PA; DeKAF Genomics Investigators. Tacrolimus trough levels after month 3 as a predictor of acute rejection following kidney transplantation: a lesson learned from DeKAF Genomics. Transpl Int. 2013 Oct;26(10):982-9. doi: 10.1111/tri.12155. Epub 2013 Jul 24.

  PMID: 23879408 BACKGROUND

• Kershner RP, Fitzsimmons WE. Relationship of FK506 whole blood concentrations and efficacy and toxicity after liver and kidney transplantation. Transplantation. 1996 Oct 15;62(7):920-6. doi: 10.1097/00007890-199610150-00009.

  PMID: 8878385 BACKGROUND

• Zahir H, McCaughan G, Gleeson M, Nand RA, McLachlan AJ. Changes in tacrolimus distribution in blood and plasma protein binding following liver transplantation. Ther Drug Monit. 2004 Oct;26(5):506-15. doi: 10.1097/00007691-200410000-00008.

  PMID: 15385833 BACKGROUND

• Zahir H, Nand RA, Brown KF, Tattam BN, McLachlan AJ. Validation of methods to study the distribution and protein binding of tacrolimus in human blood. J Pharmacol Toxicol Methods. 2001 Jul-Aug;46(1):27-35. doi: 10.1016/s1056-8719(02)00158-2.

  PMID: 12164257 BACKGROUND

• Hendijani F, Azarpira N, Kaviani M. Effect of CYP3A5*1 expression on tacrolimus required dose for transplant pediatrics: A systematic review and meta-analysis. Pediatr Transplant. 2018 Jun 19:e13248. doi: 10.1111/petr.13248. Online ahead of print.

  PMID: 29920880 BACKGROUND

• Rojas L, Neumann I, Herrero MJ, Boso V, Reig J, Poveda JL, Megias J, Bea S, Alino SF. Effect of CYP3A5*3 on kidney transplant recipients treated with tacrolimus: a systematic review and meta-analysis of observational studies. Pharmacogenomics J. 2015 Feb;15(1):38-48. doi: 10.1038/tpj.2014.38. Epub 2014 Sep 9.

  PMID: 25201288 BACKGROUND

• Haufroid V, Wallemacq P, VanKerckhove V, Elens L, De Meyer M, Eddour DC, Malaise J, Lison D, Mourad M. CYP3A5 and ABCB1 polymorphisms and tacrolimus pharmacokinetics in renal transplant candidates: guidelines from an experimental study. Am J Transplant. 2006 Nov;6(11):2706-13. doi: 10.1111/j.1600-6143.2006.01518.x.

  PMID: 17049058 BACKGROUND

• Venkataramanan R, Swaminathan A, Prasad T, Jain A, Zuckerman S, Warty V, McMichael J, Lever J, Burckart G, Starzl T. Clinical pharmacokinetics of tacrolimus. Clin Pharmacokinet. 1995 Dec;29(6):404-30. doi: 10.2165/00003088-199529060-00003.

  PMID: 8787947 BACKGROUND

• Kuypers DR, Claes K, Evenepoel P, Maes B, Vanrenterghem Y. Clinical efficacy and toxicity profile of tacrolimus and mycophenolic acid in relation to combined long-term pharmacokinetics in de novo renal allograft recipients. Clin Pharmacol Ther. 2004 May;75(5):434-47. doi: 10.1016/j.clpt.2003.12.009.

  PMID: 15116056 BACKGROUND

• Undre NA, van Hooff J, Christiaans M, Vanrenterghem Y, Donck J, Heeman U, Kohnle M, Zanker B, Land W, Morales JM, Andres A, Schafer A, Stevenson P. Low systemic exposure to tacrolimus correlates with acute rejection. Transplant Proc. 1999 Feb-Mar;31(1-2):296-8. doi: 10.1016/s0041-1345(98)01633-9. No abstract available.

  PMID: 10083114 BACKGROUND

• Zong YP, Wang ZJ, Zhou WL, Zhou WM, Ma TL, Huang ZK, Zhao CC, Xu Z, Tan RY, Gu M. Effects of CYP3A5 polymorphisms on tacrolimus pharmacokinetics in pediatric kidney transplantation: a systematic review and meta-analysis of observational studies. World J Pediatr. 2017 Oct;13(5):421-426. doi: 10.1007/s12519-017-0035-4. Epub 2017 May 24.

  PMID: 28540692 BACKGROUND

• Nankivell BJ, Alexander SI. Rejection of the kidney allograft. N Engl J Med. 2010 Oct 7;363(15):1451-62. doi: 10.1056/NEJMra0902927. No abstract available.

  PMID: 20925547 BACKGROUND

• Randomised trial comparing tacrolimus (FK506) and cyclosporin in prevention of liver allograft rejection. European FK506 Multicentre Liver Study Group. Lancet. 1994 Aug 13;344(8920):423-8.

  PMID: 7520105 BACKGROUND

• U.S. Multicenter FK506 Liver Study Group. A comparison of tacrolimus (FK 506) and cyclosporine for immunosuppression in liver transplantation. N Engl J Med. 1994 Oct 27;331(17):1110-5. doi: 10.1056/NEJM199410273311702.

  PMID: 7523946 BACKGROUND

• de Mattos AM, Olyaei AJ, Bennett WM. Nephrotoxicity of immunosuppressive drugs: long-term consequences and challenges for the future. Am J Kidney Dis. 2000 Feb;35(2):333-46. doi: 10.1016/s0272-6386(00)70348-9.

  PMID: 10676738 BACKGROUND

• Elble R, Comella C, Fahn S, Hallett M, Jankovic J, Juncos JL, Lewitt P, Lyons K, Ondo W, Pahwa R, Sethi K, Stover N, Tarsy D, Testa C, Tintner R, Watts R, Zesiewicz T. Reliability of a new scale for essential tremor. Mov Disord. 2012 Oct;27(12):1567-9. doi: 10.1002/mds.25162. Epub 2012 Oct 2.

  PMID: 23032792 BACKGROUND

Biospecimen

Retention: SAMPLES WITH DNA

Blood samples (2 ml and 10 ml test-tubes with EDTA) will be collected to measure 1. unbound tacrolimus concentration in the human plasma ultrafiltrate 2. plasma tacrolimus concentration in the human plasma 3. whole blood tacrolimus through concentration 4. the DNA will be purified from whole blood and analyzed (RT-PCR).

MeSH Terms

Conditions

Genetic Predisposition to Disease

Interventions

Tacrolimus

Condition Hierarchy (Ancestors)

Disease Susceptibility; Disease Attributes; Pathologic Processes; Pathological Conditions, Signs and Symptoms

Intervention Hierarchy (Ancestors)

Macrolides; Lactones; Organic Chemicals

Study Officials

• Karola Warzyszyńska, MD

  Department of General and Transplant Surgery, Medical University of Warsaw

  PRINCIPAL INVESTIGATOR

Central Study Contacts

Karola Warzyszyńska, MD

CONTACT

+48225021783; karola.warzyszynska@gmail.com

Study Design

• Study Type: observational
• Observational Model: COHORT
• Time Perspective: PROSPECTIVE
• Target Duration: 2 Weeks
• Sponsor Type: OTHER GOV
• Responsible Party: PRINCIPAL INVESTIGATOR
• PI Title: Medical Doctor

Study Record Dates

• First Submitted: November 24, 2020
• First Posted: December 8, 2020
• Study Start: August 1, 2020
• Primary Completion: December 1, 2022
• Study Completion: May 1, 2023
• Last Updated: April 22, 2022

Record last verified: 2022-04

Data Sharing

• IPD Sharing: Will share
• Shared Documents: STUDY PROTOCOL, SAP, ICF
• Time Frame: After publication of the study results (about 2023).
• Access Criteria: Non-comercial, after request

Locations

PL (1)