Bioproduction of 3,4-methylenedioxymethamphetamine and derivatives.

Summary

A new method combining yeast fermentation and enzymes produces the recreational drug MDMA (Ecstasy) and related compounds, offering an alternative to chemical synthesis. The process starts with a yeast-based system that converts ring-substituted benzaldehyde precursors into phenylacetylcarbinol derivatives using variants of pyruvate decarboxylase. Among seven wildtype enzymes tested, Candida tropicalis PDC gave the highest yield from piperonal, and a mutant further improved production. Five of sixteen tested benzaldehyde analogs yielded corresponding derivatives with 20–70% efficiency, allowing purification of milligram quantities. A stereoselective transaminase then converted all five derivatives, followed by N-methylation and chemical reduction to produce MDMA and its analog 6-chloro-MDMA. This is the first reported bioproduction method for MDMA and other methamphetamine derivatives.

Study at a glance

Abstract

3,4-Methylenedioxymethamphetamine (MDMA, Ecstasy) is a recreational drug under clinical investigation for the treatment of central nervous system disorders, including post-traumatic stress disorder. Chemical synthesis of MDMA mainly relies on the use of safrole and piperonal as starting materials. We report a novel strategy integrating bioconversion and biocatalysis in the bioproduction of MDMA and other methamphetamine derivatives. For the initial step, a yeast-based bioconversion system was used to produce phenylacetylcarbinol (PAC) derivatives from ring-substituted benzaldehyde precursors, including piperonal, using variants of pyruvate decarboxylase (PDC). Among seven wildtype enzymes tested, Candida tropicalis PDC (CtPDC) showed the highest yield from piperonal, and a CtPDC1-I479A mutant further improved the titer. Five of sixteen ring-substituted benzaldehyde analogs (i.e., piperonal, 6-chloropiperonal, 4-acetylbenzaldehyde, 2-fluoro-4-methoxybenzaldehyde, and 2-fluoro-4-propoxybenzaldehyde) yielded corresponding PAC derivatives with yields between 20 and 70 ​%, which allowed the purification of multiple milligram quantities of each product. Three stereoselective ω-transaminases were evaluated for their ability to catalyze the transamination of PAC derivatives, with the (R)-selective enzyme ATA-117-Rd11 able to convert all five isolated PAC derivatives. Isolated transamination products were subsequently N-methylated using human phenylethanolamine N-methyltransferase. Chemical reduction facilitated the final production of MDMA and its analog 6-chloro-MDMA. Our work represents the first reported bioproduction method leading to MDMA and other methamphetamine derivatives, suitable for future pathway and strain optimization.