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. 2024 Aug 2;16(8):1032.
doi: 10.3390/pharmaceutics16081032.

Identification of the Biotransformation Pathways of a Potential Oral Male Contraceptive, 11β-Methyl-19-Nortestosterone (11β-MNT) and Its Prodrugs: An In Vitro Study Highlights the Contribution of Polymorphic Intestinal UGT2B17

Namrata Bachhav  1 , Dilip Kumar Singh  1 , Diana L Blithe  2 , Min S Lee  2 , Bhagwat Prasad  1
Affiliations

Identification of the Biotransformation Pathways of a Potential Oral Male Contraceptive, 11β-Methyl-19-Nortestosterone (11β-MNT) and Its Prodrugs: An In Vitro Study Highlights the Contribution of Polymorphic Intestinal UGT2B17

Namrata Bachhav et al. Pharmaceutics. .

Abstract

11β-Methyl-19-nortestosterone dodecylcarbonate (11β-MNTDC) is a prodrug of 11β-MNT and is being considered as a promising male oral contraceptive candidate in clinical development. However, the oral administration of 11β-MNTDC exhibits an ~200-fold lower serum concentration of 11β-MNT compared to 11β-MNTDC, resulting in the poor bioavailability of 11β-MNT. To elucidate the role of the first-pass metabolism of 11β-MNT in its poor bioavailability, we determined the biotransformation products of 11β-MNT and its prodrugs in human in vitro models. 11β-MNT and its two prodrugs 11β-MNTDC and 11β-MNT undecanoate (11β-MNTU) were incubated in cryopreserved human hepatocytes (HHs) and subjected to liquid chromatography-high resolution tandem mass spectrometry analysis, which identified ten 11β-MNT biotransformation products with dehydrogenated and glucuronidation (11β-MNTG) metabolites being the major metabolites. However, 11β-MNTG formation is highly variable and prevalent in human intestinal S9 fractions. A reaction phenotyping study of 11β-MNT using thirteen recombinant UDP-glucuronosyltransferase (UGT) enzymes confirmed the major role of UGT2B17 in 11β-MNTG formation. This was further supported by a strong correlation (R2 > 0.78) between 11β-MNTG and UGT2B17 abundance in human intestinal microsomes, human liver microsomes, and HH systems. These results suggest that 11β-MNT and its prodrugs are rapidly metabolized to 11β-MNTG by the highly polymorphic intestinal UGT2B17, which may explain the poor and variable bioavailability of the drug.

Keywords: 11β-MNT; 11β-MNTDC; 11β-Methyl-19-nortestosterone; LC-MS; MetID; UGT2B17; biotransformation; metabolite identification; oral male contraceptive.

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Conflict of interest statement

Bhagwat Prasad (Corresponding author) is a co-founder of Precision Quantomics Inc. and a recipient of research funding from AbbVie, Boehringer Ingelheim, Bristol Myers Squibb, Genentech, Generation Bio, Gilead, Merck, Novartis, and Takeda. All other authors declare no conflicts of interest.

Figures

Figure 1
Figure 1
11β-Methyl-19-nortestosterone dodecylcarbonate (11β-MNTDC) and 11β-Methyl-19-nortestosterone undecanoate (11β-MNTU) are prodrugs metabolized by esterases into their active metabolite, 11β-Methyl-19-nortestosterone (11β-MNT) (A). Structural similarities amongst testosterone, dimethandrolone (DMA), and 11β-MNT (B). Red spheres in (B) indicate oxygen atom.
Figure 2
Figure 2
Volcano plot showing fold-differences and p-values of m/z features detected in a 2 h incubation of human hepatocytes with 11β-MNT as compared to the untreated (control) samples (A). P represents the parent active metabolite (11β-MNT); M1 and M3–M7, six identified metabolites; M2, four potential structural isomers. The black dotted line indicates −log10 (p-value, i.e., 1.33). Relative MS intensity of 11β-MNT metabolites in human hepatocyte incubation (B).
Figure 3
Figure 3
Box and Whisker plot representing 11β-MNT and its major metabolites, M4, i.e., 11β-MNTG (A) and M7 (B) showing signal in the treatment (50 µM of 11β-MNT) compared to the untreated group in human hepatocytes.
Figure 4
Figure 4
The levels of dehydrogenated 11β-MNT and 11β-MNTG formed in human hepatocyte incubations of 50 µM 11β-MNT (A) and 10 µM 11β-MNTU (B), and 11β-MNTDC (C). HH1–4: (Average of all four hepatocytes with SD). HH1 and HH2 represent UGT2B17 high expressors, whereas HH3 and HH4 are the null expressors of UGT2B17.
Figure 5
Figure 5
Metabolite/11β-MNT ratio of dehydrogenated 11β-MNT and 11β-MNTG in the pooled human intestinal (A) and liver (B) S9 fraction incubations with 50 µM of 11β-MNT, 11β-MNTDC, and 11β-MNTU.
Figure 6
Figure 6
Normalized 11β-MNTG levels (with internal standard) detected in the incubation of individual recombinant UGT isoforms. SD represents triplicate analysis.
Figure 7
Figure 7
Correlation of UGT2B17 abundance and relative 11β-MNTG levels in HIMs (A), HLMs (B), and human hepatocytes (C).
See this image and copyright information in PMC

References

    1. Bearak J., Popinchalk A., Alkema L., Sedgh G. Global, Regional, and Subregional Trends in Unintended Pregnancy and Its Outcomes from 1990 to 2014: Estimates from a Bayesian Hierarchical Model. Lancet Glob. Health. 2018;6:e380–e389. doi: 10.1016/S2214-109X(18)30029-9. - DOI - PMC - PubMed
    1. Abbe C.R., Page S.T., Thirumalai A. Male Contraception. Yale J. Biol. Med. 2020;93:603–613. - PMC - PubMed
    1. Long J.E., Lee M.S., Blithe D.L. Update on Novel Hormonal and Nonhormonal Male Contraceptive Development. J. Clin. Endocrinol. Metab. 2021;106:e2381–e2392. doi: 10.1210/clinem/dgab034. - DOI - PMC - PubMed
    1. Attardi B.J., Hild S.A., Koduri S., Pham T., Pessaint L., Engbring J., Till B., Gropp D., Semon A., Reel J.R. The Potent Synthetic Androgens, Dimethandrolone (7α,11β-Dimethyl-19-nortestosterone) and 11β-Methyl-19-nortestosterone, Do Not Require 5α-Reduction to Exert Their Maximal Androgenic Effects. J. Steroid Biochem. Mol. Biol. 2010;122:212–218. doi: 10.1016/j.jsbmb.2010.06.009. - DOI - PMC - PubMed
    1. Sharma S., Ahire D., Basit A., Lajoie M., Wang C., Lee M.S., Blithe D.L., Amory J.K., Singh D.K., Heyward S., et al. Dimethandrolone, a Potential Male Contraceptive Pill, Is Primarily Metabolized by the Highly Polymorphic UDP-Glucuronosyltransferase 2B17 Enzyme in Human Intestine and Liver. Drug Metab. Dispos. 2022;50:1493–1500. doi: 10.1124/dmd.122.001041. - DOI - PMC - PubMed

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