Microbial transformation of immunosuppressive compounds. I. Desmethylation of FK 506 and immunomycin (FR 900520) by Actinoplanes sp. ATCC 53771.

Chen, Tom S.; Arison, Byron H.; Wicker, Linda S.; Inamine, Edward; Monaghan, Richard L.

doi:10.7164/antibiotics.45.118

Cited by 21 publications

(12 citation statements)

References 8 publications

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“…Structures of FK506 and its analogues. a,b,c,d Known compounds described in refs , , , and , respectively.…”

Section: Resultsmentioning

confidence: 99%

“…Deletion of either tcsD or tcsB in the fkbM deletion mutant (ΔtcsD – ΔfkbM strain and ΔtcsB – ΔfkbM strain), respectively, produced the new analogue 31- O -demethyl-36,37-dihydroFK506 ( 9 ) (Figure e; Table S2; Figures S10 and S11) and 31- O -demethylFK520 ( 10 ) (Figure f; Table S2; Figures S12 and S13), which has been reported from the biotransformation of FK506 by Actinoplanes sp. ATCC 53771 . Again, a trace amount of 31- O -demethylFK523 was produced from the ΔtcsB – ΔfkbM strain, which was sufficient only for UPLC-qTOF-HR-MS detection (Figure f; Figure S12).…”

Section: Resultsmentioning

confidence: 99%

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Biosynthesis of Nonimmunosuppressive FK506 Analogues with Antifungal Activity

et al. 2019

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A reduction in the strong immunosuppressive activity of FK506 ( 1) is essential for developing this compound as an antifungal agent. Seven new FK506 analogues modified at both the FK506-binding protein 12and the calcineurin-binding regions were biosynthesized. 9-DeoxoFK520 (7) exhibited a >900-fold reduction in the in vitro immunosuppressive activity but maintained significant antifungal activity, indicating that the C-9 and C-21 positions are critical for separation of immunosuppressive and antifungal activities. 7 exhibited robust synergistic antifungal activity with fluconazole. FK506 ( 1) is a 23-membered macrolide produced by several Streptomyces species and is used as an immunosuppressive drug to prevent the rejection of transplanted organs. FK506 has also exhibited antifungal, neuroprotective, and neuroregenerative activities. In humans, FK506 binds to FK506binding protein (FKBP) 12, and the resulting FKBP12−FK506 complex interacts with a Ca 2+ -calmodulin-dependent phosphatase, calcineurin (CaN). Inactivation of CaN by forming the FKBP12−FK506−CaN ternary complex prevents the activation of nuclear factor of activated T cells (NF-AT), inhibiting the production of interleukin-2 and subsequent T-cell proliferation. This CaN signaling pathway also plays a critical role in the growth and pathogenesis of major fungal pathogens such as Cryptococcus neoformans, Candida albicans, and Aspergillus f umigatus. Therefore, the synthesis of FK506 analogues that can discriminate human FKBP12/CaN from its fungal counterparts may separate antifungal activity from the immunosuppressive activity, thereby allowing the development of a novel antifungal agent.

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“…Structures of FK506 and its analogues. a,b,c,d Known compounds described in refs , , , and , respectively.…”

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Biosynthesis of Nonimmunosuppressive FK506 Analogues with Antifungal Activity

et al. 2019

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“…To measure tacrolimus metabolite crossreactivity, metabolites M-I (13-O-demethyltacrolimus), M-II (31-O-demethyltacrolimus), M-III (15-O-demethyltacrolimus), and M-IV(12-hydroxytacrolimus) were prepared in vitro by incubation of tacrolimus with liver microsomes prepared from phenobarbital-treated rats in the presence of nicotinamide adenine dinucleotide phosphate (NADPH) generating system under aerobic condition or bioconverted by incubating tacrolimus with an actinomycete. [2][3][4] Oxidative metabolites formed in the reaction medium were isolated and identified. Purified samples were analyzed by high-performance liquid chromatography with mass spectrometric detection and by nuclear magnetic resonance spectroscopy.…”

Section: Antibody Specificitymentioning

confidence: 99%

Multi-site Analytical Evaluation of the Abbott ARCHITECT Tacrolimus Assay

Wallemacq

Goffinet²,

O'Morchoe³

et al. 2009

Therapeutic Drug Monitoring

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The objective of this study was to evaluate the analytical performance of the Abbott ARCHITECT Tacrolimus immunoassay. Proficiency panels and specimens from a population of organ transplant recipients were analyzed in 6 clinical laboratories in Europe and the United States, and the results were compared with other methods. The ARCHITECT assay requires a whole blood specimen pretreatment step with methanol/zinc sulfate to precipitate protein and extract the drug, followed by a 30-minute immunoassay using anti-tacrolimus antibody-coated paramagnetic microparticles and an acridinium-tacrolimus tracer. The assay was free from hematocrit interference in the range 25%-55% and from interference by extremes of cholesterol, triglycerides, bilirubin, total protein, and uric acid. The total percent of coefficient of variations of the assay were 4.9%-7.6% at 3 ng/mL, 2.9%-4.6% at 8.6 ng/mL, and 3.1%-8.2% at 15.5 ng/mL. Limit of detection was < or =0.5 ng/mL and limit of quantification (LOQ) ranged from 0.69 to 1.07 ng/mL across the 6 sites (based on the upper 95% confidence interval concentrations). The 2007 European Consensus Conference on Tacrolimus Optimization recommended the use of assay methods with an LOQ around 1 ng/mL, based upon the need to measure trough tacrolimus blood concentrations precisely down to 3 ng/mL during low-dose tacrolimus regimens. Tacrolimus International Proficiency Testing Scheme samples were measured by the ARCHITECT immunoassay at 5 sites and showed an average bias of -0.28 to +0.85 ng/mL versus IMx Tacrolimus II immunoassay historical values and -0.21 to +0.68 ng/mL versus liquid chromatography/tandem mass spectrometry (LC-MSMS) Tacrolimus historical values. Method comparison studies were performed with the ARCHITECT Tacrolimus immunoassay on patient specimens with the following results: ARCHITECT Tacrolimus assay versus the Abbott IMx Tacrolimus II immunoassay (4 sites) yielded average biases between -0.94 and +0.26 ng/mL; ARCHITECT assay versus the Dade Dimension Tacrolimus immunoassay (2 sites) yielded average biases of -0.46 and +0.11 ng/mL; and ARCHITECT assay versus LC-MSMS methods at 2 sites yielded average biases of +0.51 and +1.63 ng/mL. Spearman correlation coefficients were >/=0.90 on all method comparisons. The ARCHITECT Tacrolimus assay is a semiautomated, robust, and highly sensitive immunoassay, representing an alternative approach for laboratories not equipped with LC-MSMS, and meets the 1 ng/mL recommendation of LOQ by the European Consensus Conference on Tacrolimus Optimization.

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“…31 -0-DesmethylFK-506 was prepared by microbial demethylation of the FK-506 according to previously reported procedures [14]. 31-0-DesmethylFK-506 was also isolated from a mutant strain of Streptomyces sp.…”

Section: Preparation Of 31-0-desmethylfk-506mentioning

confidence: 99%

Enzymology of FK‐506 biosynthesis

Shafiee

Motamedi

Chen

1994

European Journal of Biochemistry

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FK-506 is a macrolide antibiotic with immunosuppressant activity. Structurally, this compound contains three methylated hydroxyl groups at C13, C15 and C31. Previous biosynthetic studies using stable isotope-feeding experiments have established methionine as the source of the methyl for these methylated hydroxyl groups. Based on this information and also the availability of the 31-0-desmethylFK-506, a metabolic precursor for the biosynthesis of FK-506, a S-adenosyl-L-methionine-dependent enzyme assay was developed and the enzyme 3 1 -0-desmethylFK-506 0 :methyltransferase was isolated from an extract of Streptomyces sp. MA 6858 and purified to near homogeneity. 31-0-DesmethylFK-506 0 :methyltransferase is a monomeric protein with an apparent molecular mass of 30000 Da and a PI of 4.4. The first 38 N-terminal amino acids have been sequenced and are H,N-SDVVETLRLPNGATVAHVNAGEAQFIYREIFTDRXYLRH. Functionally, this enzyme has a requirement for Mg" with an optimum temperature of 34°C and a pH of 7.4 for full activity. Moreover, it catalyses the methylation of 31-0-desmethylimmunomycin as efficiently as its own natural substrate, 31 -0-desmethylFK-506. Additionally, FKMT catalyzes the C31 transmethylation reaction of 13,31-0-bis-desmethyl-, 15,31-O-bisdesmethyl-, 13,15,31-0-trisdesmethyl-and 31 -0-19,22-cyclic-hemiketalimmunomycins, which are all structural analogues of FK-506. The reaction is, however, completely blocked if the vicinal hydroxyl which is present at the C-32 position of the 31-0-desmethylFK-506 structure is replaced with azide, phosphate or other substituents. Finally, evidence is presented indicating the close similarity of FKMT and DIMT, a 31-0-desmethylimmunomycin : 0 methyltransferase, previously isolated from a cell-free extract of Streptomyces hygroscopicus var ascomyceticus, an immunomycin (ascomycin/FK-520) producer. in numerous laboratories searching for compounds having novel structures with useful pharmacological activities. In the mean time, extensive biochemical and genetic studies are being pursued in order to unravel the biosynthetic pathways for the formation of these complex structures, hoping, through gene manipulations [6], to improve the yield [7], biosynthesize modified [8] and hybrid [9] structures andor develop mutants [lo]. To these ends, a program was initiated in our laboratories to carry out biochemical and genetic studies on a newly discovered immunosuppressant agent, FK-506. This compound, which is currently undergoing clinical trial for the prevention of organ transplant rejection [ll], is a macrocyclic polyketide produced by Streptomyces tsukubaensis, first reported in 1987 [12]. We have also discovered an FK-506-producing strain of Streptomyces, identified as MA 6858 (ATCC No. 55098). Precursor-feeding studies of this strain by Byrne et al. [13] have established that acetate, propionate, butyrate, pipecolic acid, shikimic acid and methionine participate in the biosynthesis of FK-506. The methionine has been shown to be the source of the methyl groups for the methylation ...

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Microbial transformation of immunosuppressive compounds. I. Desmethylation of FK 506 and immunomycin (FR 900520) by Actinoplanes sp. ATCC 53771.

Cited by 21 publications

References 8 publications

Biosynthesis of Nonimmunosuppressive FK506 Analogues with Antifungal Activity

Biosynthesis of Nonimmunosuppressive FK506 Analogues with Antifungal Activity

Multi-site Analytical Evaluation of the Abbott ARCHITECT Tacrolimus Assay

Enzymology of FK‐506 biosynthesis

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