Engineered Ribosyl-1-Kinase Enables Concise Synthesis of Molnupiravir, an Antiviral for COVID-19

McIntosh, Jason S.; Benkovics, Tamas; Silverman, Steven M.; Huffman, Mark A.; Kong, Jongrock; Maligres, Peter E.; Itoh, Tetsuji; Yang, Hao; Verma, Deeptak; Pan, Weilan; Ho, Hsing‐I; Vroom, Jonathan; Knight, Anders M.; Hurtak, Jessica A.; Klapars, Artis; Fryszkowska, Anna; Morris, William; Strotman, Neil A.; Murphy, Grant S.; Maloney, Kevin M.; Fier, Patrick S.

doi:10.1021/acscentsci.1c00608

Cited by 112 publications

(143 citation statements)

References 25 publications

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“…Indeed, such methods are already commonly used for screening libraries. 2,4,9,15,28,46 Notably, because variants are identified based on relative changes in product distribution, SUMS data can be analyzed without quantitation of absolute product concentration. Last, the substrate mixtures used in this work featured compounds with which the parent enzyme already had detectable baseline activity.…”

Section: Discussionmentioning

confidence: 99%

Substrate multiplexed protein engineering facilitates promiscuous biocatalytic synthesis

McDonald

Higgins

Buller

2021

Preprint

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Enzymes with high activity are readily produced through protein engineering, but intentionally and efficiently engineering enzymes for an expanded scope is a contemporary challenge. Measuring reaction outcomes on mixtures of substrates, called here SUbstrate Multiplexed Screening (SUMS), has long been used to rigorously quantitate enzyme specificity. Despite the potential utility of SUMS to guide engineering of promiscuous enzymes, this approach has not found widespread adoption in biocatalysis. Here, we develop principles of how to design robust SUMS methods that, rather than assess absolute specificity, use heuristic readouts of substrate promiscuity to identify hits for further investigation. This rich information enables engineering of activity for multiple substrates simultaneously and identifies enzyme variants with altered promiscuity, even when overall activity is lower. We demonstrate the effectiveness of SUMS by engineering two enzymes to produce pharmacologically active tryptamines from simple indole precursors in a biocatalytic cascade. These advances leverage common laboratory equipment and represent a highly accessible and customizable method for enzyme engineering.

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Section: Discussionmentioning

confidence: 99%

Substrate multiplexed protein engineering facilitates promiscuous biocatalytic synthesis

McDonald

Higgins

Buller

2021

Preprint

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“…NHC-TP is incorporated into viral RNA by the RdRp, resulting in viral error induction (i.e. accumulation of errors in the viral genome ultimately leading to inhibition of replication) [ 7 , 16 , 17 ]. Consistent with this mechanism of action, molnupiravir treatment was associated with numerically higher SARS-CoV-2 viral RNA error rates versus placebo in patients with COVID-19 in clinical trials [ 18 – 20 ].…”

Section: Scientific Summarymentioning

confidence: 99%

Molnupiravir: First Approval

Syed

2022

Drugs

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Molnupiravir (Lagevrio ® ) is an orally-administered antiviral prodrug that inhibits replication of RNA viruses through viral error induction. It is being developed by Merck and Ridgeback Biotherapeutics for the prevention and treatment of Coronavirus disease 2019 (COVID-19). Molnupiravir received its first approval on 4 November 2021 in the UK for the treatment of mild to moderate COVID-19 in adults with a positive severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) diagnostic test and who have at least one risk factor for developing severe illness. Molnupiravir is filed for approval and has emergency use authorization for the treatment of COVID-19 in several countries, including the USA, Japan and those in the EU. This article summarizes the milestones in the development of molnupiravir leading to this first approval for COVID-19. Supplementary Information The online version contains supplementary material available at 10.1007/s40265-022-01684-5.

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“…Merck published a route starting from commodity chemicals, ribose and uracil, which are converted to molnupiravir in high yields in three biocatalytic steps; reactions use designer enzymes (Codexis) derived by directed evolution, and these enzymes are not currently commercially available to other manufacturers 15 .…”

Section: Introductionmentioning

confidence: 99%

Cost of Goods Analysis Facilitates an Integrated Approach to Identifying Alternative Synthesis Methodologies for Lower Cost Manufacturing of the COVID-19 Antiviral Molnupiravir

Peterson

Paria

Deshpande³

et al. 2022

Gates Open Res

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Orally delivered drugs offer significant benefits in the fight against viral infections, and cost-effective production is critical to their impact on pandemic response in low- and middle-income countries. One example, molnupiravir, a COVID-19 therapy developed by Emory, Ridgeback, and Merck & Co., had potential to benefit from significant cost of goods (COGs) reductions for its active pharmaceutical ingredient (API), including starting materials. A holistic approach to identifying, developing, and evaluating optimized synthetic routes, which includes detailed COGs modeling, provides a rapid means to increase the availability, uptake and application of molnupiravir and other antivirals in global markets. Identification and development of alternate processes for the synthesis of molnupiravir has been conducted by the Medicines for All Institute at Virginia Commonwealth University (M4ALL) and the Green and Turner Labs at the University of Manchester. Both groups developed innovative processes based on synthetic route design and biocatalysis aimed at lowering costs and improving global access. The authors then performed COGs modeling to assess cost saving opportunities. This included a focus on manufacturing environments and facilities amenable to global public health and the identification of key parameters using sensitivity analyses. While all of the evaluated routes provide efficiency benefits, the best options yielded 3-6 fold API COGs reductions leading to treatment COGs as low as <$3/regimen. Additionally, key starting materials and cost drivers were quantified to evaluate the robustness of the savings. Finally, COGs models can continue to inform the focus of future development efforts on the most promising routes for additional cost savings. While the full price of a treatment course includes other factors, these alternative API synthetic approaches have significant potential to help facilitate broader access in low- and middle-income countries. As other promising therapeutics are developed, a similar process could enable rapid cost reductions while enhancing global access.

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Engineered Ribosyl-1-Kinase Enables Concise Synthesis of Molnupiravir, an Antiviral for COVID-19

Cited by 112 publications

References 25 publications

Substrate multiplexed protein engineering facilitates promiscuous biocatalytic synthesis

Substrate multiplexed protein engineering facilitates promiscuous biocatalytic synthesis

Molnupiravir: First Approval

Cost of Goods Analysis Facilitates an Integrated Approach to Identifying Alternative Synthesis Methodologies for Lower Cost Manufacturing of the COVID-19 Antiviral Molnupiravir

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