Reutilization of Tacrolimus Extracted from Expired Prograf® Capsules: Physical, Chemical, and Pharmacological Assessment

In this study, we investigated whether tacrolimus extracted and purified from the commercial capsules (Prograf® 5 mg) have retained its original quality and activity beyond the capsules expiration date in order to be reused for research purposes after extraction. High-performance liquid chromatography (HPLC) assay method was developed and validated for the quantification of tacrolimus, using cyclosporine A as an internal standard (IS). Moreover, a combination of analytical methods, including nuclear magnetic resonance (NMR), gas chromatography–mass spectrometry (GC-MS), Fourier transform-infrared (FT-IR) spectroscopy, X-ray diffraction (XRD), and differential scanning calorimetry (DSC) were used to assess the quality of extracted/purified tacrolimus. Suppression of murine peripheral-blood mononuclear cells (PBMC) proliferation and the levels of interleukin-2 (IL-2) and interferon gamma (IFN-γ) were also assessed. The data obtained showed no detectable differences in the quality profile between the authentic sample and extracted drug. Also, the results showed that the extracted/purified tacrolimus was able to suppress T cell proliferation, induced by concanavalin A, indicating the retained pharmacological activity. We proved that tacrolimus extracted/purified from expired Prograf® capsuled retains its purity and immunosuppressive activity and can be reused for research and possibly in pharmaceutical manufacturing.

[1]  M. D’Agostino,et al.  Tacrolimus 0.1% ointment in nail psoriasis: a randomized controlled open‐label study , 2013, Journal of the European Academy of Dermatology and Venereology : JEADV.

[2]  H. Patel,et al.  Self micro-emulsifying drug delivery system of tacrolimus: Formulation, in vitro evaluation and stability studies , 2013, International journal of pharmaceutical investigation.

[3]  J. Zwischenberger,et al.  Design and physicochemical characterization of advanced spray-dried tacrolimus multifunctional particles for inhalation , 2013, Drug design, development and therapy.

[4]  T. Mehta,et al.  Improvement of dissolution rate of tacrolimus by solid dispersion technique , 2013, Journal of Pharmaceutical Investigation.

[5]  R. O. Macêdo,et al.  Correlation of thermal analysis and pyrolysis coupled to GC-MS in the characterization of tacrolimus. , 2013, Journal of pharmaceutical and biomedical analysis.

[6]  T. Rath.  Tacrolimus in transplant rejection , 2013, Expert opinion on pharmacotherapy.

[7]  Fang Liu,et al.  SAHA, an HDAC inhibitor, synergizes with tacrolimus to prevent murine cardiac allograft rejection , 2012, Cellular and Molecular Immunology.

[8]  Koichiro Nakamura,et al.  A Randomized, Open-Label, Multicenter Trial of Topical Tacrolimus for the Treatment of Pruritis in Patients with Atopic Dermatitis , 2012, Annals of dermatology.

[9]  D. Colombo,et al.  Evaluation, synthesis and characterization of tacrolimus impurities , 2012, The Journal of Antibiotics.

[10]  Jian-xin Lin,et al.  IL-2 family cytokines: new insights into the complex roles of IL-2 as a broad regulator of T helper cell differentiation. , 2011, Current opinion in immunology.

[11]  Ramon Arens,et al.  Autocrine IL-2 is required for secondary population expansion of CD8+ memory T cells , 2011, Nature Immunology.

[12]  M. Fukuzawa,et al.  Effects of a calcineurin inhibitor, FK506, and a CCR5/CXCR3 antagonist, TAK-779, in a rat small intestinal transplantation model. , 2011, Transplant immunology.

[13]  M. Udompataikul,et al.  Effectiveness of 0.1% topical tacrolimus in adult and children patients with vitiligo , 2011, The Journal of dermatology.

[14]  S. G. Cardoso,et al.  Determination of tacrolimus in pharmaceutical formulations by validated spectrophotometric methods , 2009 .

[15]  T. Oshiro,et al.  Kinetics of IFN-gamma, TNF-alpha, IL-10 and IL-4 production by mononuclear cells stimulated with gp43 peptides, in patients cured of paracoccidioidomycosis. , 2007, Revista da Sociedade Brasileira de Medicina Tropical.

[16]  Christopher B Wilson,et al.  Regulation of interferon-gamma during innate and adaptive immune responses. , 2007, Advances in immunology.

[17]  Ajaz S Hussain,et al.  Stability profiles of drug products extended beyond labeled expiration dates. , 2006, Journal of pharmaceutical sciences.

[18]  A. Tyznik,et al.  Interleukin-2 signals during priming are required for secondary expansion of CD8+ memory T cells , 2006, Nature.

[19]  J. Morales,et al.  Ten years of treatment with tacrolimus is related to an excellent renal function, allowing monotherapy in a large proportion of cases: unicentric results of the tacrolimus versus cyclosporine A European Multicentric Study in kidney transplant patients. , 2005, Transplantation proceedings.

[20]  H. Kirchner,et al.  Sensitivity of whole-blood T lymphocytes in individual patients to tacrolimus (FK 506): impact of interleukin-2 mRNA expression as surrogate measure of immunosuppressive effect. , 2004, Clinical chemistry.

[21]  R. Ochs,et al.  Mechanism for the paradoxical inhibition and stimulation of calcineurin by the immunosuppresive drug tacrolimus (FK506). , 2003, Archives of biochemistry and biophysics.

[22]  W. Almawi,et al.  Tacrolimus (FK506) versus cyclosporin A microemulsion (Neoral) maintenance immunosuppression: effects on graft survival and function, infection, and metabolic profile following kidney transplantation (KT). , 2003, Molecular immunology.

[23]  J. Fechner,et al.  Effect of immunosuppressants on T-cell subsets observed in vivo using carboxy-fluorescein diacetate succinimidyl ester labeling1 , 2003, Transplantation.

[24]  J. Sandford-Smith.  Outdated drugs may be useful , 2003, BMJ : British Medical Journal.

[25]  昌彦 杭ﾉ瀬.  Calcineurin Antagonists Inhibit Interferon-gamma Production by Downregulation of Interleukin-18 in Human Mixed Lymphocyte Reactions , 2001 .

[26]  N. Tanaka,et al.  Calcineurin antagonists inhibit interferon-gamma production by downregulation of interleukin-18 in human mixed lymphocyte reactions. , 2000, Acta medica Okayama.

[27]  P. Halloran,et al.  Calcineurin and the biological effect of cyclosporine and tacrolimus. , 1998, Transplantation proceedings.

[28]  Bonanno,et al.  The effect of cyclosporin A, FK506 and rapamycin on the murine contact sensitivity reaction , 1998, Clinical and experimental immunology.

[29]  K. Nakahara,et al.  Discovery and Development of a Novel Immunosuppressant, Tacrolimus Hydrate , 1998 .

[30]  G. Geginat,et al.  Effects of FK-506 on the course of murine salmonellosis. , 1996, Journal of chemotherapy.

[31]  T. Kuno,et al.  Interaction of tacrolimus(FK506) and its metabolites with FKBP and calcineurin. , 1994, Biochemical and biophysical research communications.

[32]  C. Klee,et al.  Calcineurin phosphatase activity in T lymphocytes is inhibited by FK 506 and cyclosporin A. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[33]  W. Li,et al.  Cyclosporin A and FK506 mediate differential effects on T cell activation in vivo. , 1992, Journal of immunology.

[34]  P. Schmieder,et al.  Conformational Analysis of the cis‐ and trans‐Isomers of FK506 by NMR and Molecular Dynamics , 1991 .

[35]  S. Schreiber,et al.  Total synthesis of FK506 and an FKBP probe reagent, [C(8),C(9)-13C2]-FK506 , 1990 .

[36]  R. Carlsson,et al.  Kinetics of IL-2 and interferon-gamma production, expression of IL-2 receptors, and cell proliferation in human mononuclear cells exposed to staphylococcal enterotoxin A. , 1985, Cellular immunology.

[37]  B. Benacerraf,et al.  GENETIC CONTROL OF THYMUS-DERIVED CELL FUNCTION : I. IN VITRO DNA SYNTHETIC RESPONSE OF NORMAL MOUSE SPLEEN CELLS STIMULATED BY THE MITOGENS CONCANAVALIN A AND PHYTOHEMAGGLUTININ , 1972 .