3258-02-4

Usage

Uses

Used in Antiviral Applications:
N(4)-hydroxycytidine is used as a broad-spectrum antiviral agent for inhibiting the replication of various viruses, including SARS-CoV, MERS-CoV, and SARS-CoV-2. It reduces viral titers in infected cells and has potential applications in the treatment of seasonal and pandemic influenza.
Used in Pharmaceutical Industry:
N(4)-hydroxycytidine is used as a pharmaceutical agent for the development of antiviral drugs targeting a wide range of viruses. Its broad-spectrum activity and oral bioavailability make it a promising candidate for the treatment of various viral infections.

References

1) Janion and Glickman (1980),?N4-hydroxycytidine: a mutagen specific for AT to GC transitions;?Mutat. Res.,?72?43 2) Stuyver?et al.?(2003),?Ribonucleotide analogue that blocks replication of bovine viral diarrhea and hepatitis C viruses in culture; Antimicrob. Agents Chemother.,?47?244 3) Costantini?et al.?(2012),?Antiviral activity of nucleoside analogues against norovirus; Antivir. Ther.,?17?981 4) Reynard?et al.?(2015),?Identification of a New Ribonucleoside Inhibitor of Ebola Virus Replication; Viruses,?7?6233 5) Ehteshami?et al.?(2017),?Characterization of ?-D-N4-Hydroxycytidine as a Novel Inhibitor of Chikungunya Virus; Antimicrob. Agents Chemother.,?61?e02395-16 6) Yoon?et al.?(2018),?Orally efficacious broad-spectrum ribonucleoside analog inhibitor of influenza and respiratory syncytial viruses; Antimicrob. Agents Chemother.,?62?e00766-18 7) Barnard?et al.?(2004),?Inhibition of severe acute respiratory syndrome-associated coronavirus (SARSCoV) by calpain inhibitors and beta-D-N4-hydroxycytidine; Antivir. Chem. Chemother., 15 15 8) Pyrc et al. (2006),?Inhibition of human coronavirus NL63 infection at early stages of the replication cycle; Antimicrob. Agents Chemother.,?50?2000 9) Toots et al. (2019),?Characterization of orally efficacious influenza drug with high resistance barrier in ferrets and human airway epithelia; Sci. Transl. Med.,?11?eaax5866

InChI:InChI=1/C9H13N3O6/c13-3-4-6(14)7(15)8(18-4)12-2-1-5(11-17)10-9(12)16/h1-2,4,6-8,13-15,17H,3H2,(H,10,11,16)/t4-,6-,7-,8-/m1/s1

Upstream product

• 82913-19-7 [(2R,3R,4R)-3,4-diacetoxy-5-[2-oxo-4-(1,2,4-triazol-1-yl)pyrimidin-1-yl]tetrahydrofuran-2-yl]methyl acetate 
• 104105-76-2 1-(β-D-erythro-Pentofuranosyl)-4-(1,2,4-triazol-1-yl)-2(1H)-pyrimidinone 
• 2346620-55-9 molnupiravir acetonide 
• 58-96-8 uridine 
• 362-43-6 2',3'-O-isopropylideneuridine 
• 13957-31-8 4-thiouridine 
• 65-46-3 CYTIDINE 
• 800379-06-0 3-benzyloxy-1-(3,4-dihydroxy-5-hydroxymethyltetrahydrofuran-2-yl)-4-imino-3,4-dihydropyrimidin-2(1H)-one 
• 3310-47-2 4-amino-1-(3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-1H-pyrimidin-2-one-3-oxide 
• 800379-05-9 1-(3,4-dihydroxy-5-hydroxymethyltetrahydrofuran-2-yl)pyrimidine-2,4(1H,3H)-dione 4-(O-benzyloxime) 
• 10526-27-9 2',3'-O-(isopropylidene)-5'-O-(triphenylmethyl)uridine 
• 64898-15-3 2',3',5'-tris-O-(tert-butyldimethylsilyl)uridine 
• 15049-50-0 1-(2',3',5'-tri-O-benzoyl-β-D-ribofuranosyl)-4-thiouracil 
• 1748-04-5 tribenzoyl uridine 

Downstream Products

• 65-46-3 CYTIDINE 

Relevant academic research and scientific papers

Orally efficacious broad-spectrum ribonucleoside analog inhibitor of influenza and respiratory syncytial viruses

Morbidity and mortality resulting from influenza-like disease are a threat, especially for older adults. To improve case management, next-generation broad-spectrum antiviral therapeutics that are efficacious against major drivers of influenza-like disease, including influenza viruses and respiratory syncytial virus (RSV), are urgently needed. Using a dual-pathogen high-throughput screening protocol for influenza A virus (IAV) and RSV inhibitors, we have identified N4-hydroxycytidine (NHC) as a potent inhibitor of RSV, influenza B viruses, and IAVs of human, avian, and swine origins. Biochemical in vitro polymerase assays and viral RNA sequencing revealed that the ribonucleotide analog is incorporated into nascent viral RNAs in place of cytidine, increasing the frequency of viral mutagenesis. Viral passaging in cell culture in the presence of an inhibitor did not induce robust resistance. Pharmacokinetic profiling demonstrated dose-dependent oral bioavailability of 36 to 56%, sustained levels of the active 5=-triphosphate anabolite in primary human airway cells and mouse lung tissue, and good tolerability after extended dosing at 800 mg/kg of body weight/day. The compound was orally efficacious against RSV and both seasonal and highly pathogenic avian IAVs in mouse models, reducing lung virus loads and alleviating disease biomarkers. Oral dosing reduced IAV burdens in a Guinea pig transmission model and suppressed virus spread to uninfected contact animals through direct transmission. Based on its broad-spectrum efficacy and pharmacokinetic properties, NHC is a promising candidate for future clinical development as a treatment option for influenza-like diseases.

Novel cytidine-based orotidine-5′-monophosphate decarboxylase inhibitors with an unusual twist

Orotidine-5′-monophosphate decarboxylase (ODCase) is an interesting enzyme with an unusual catalytic activity and a potential drug target in Plasmodium falciparum, which causes malaria. ODCase has been shown to exhibit unusual and interesting interactions with a variety of nucleotide ligands. Cytidine-5′-monophosphate (CMP) is a poor ligand of ODCase, and CMP binds to the active site of ODCase with an unusual orientation and conformation. We designed N3- and N4-modified CMP derivatives as novel ligands to ODCase. These novel CMP derivatives and their corresponding nucleosides were evaluated against Plasmodium falciparum ODCase and parasitic cultures, respectively. These derivatives exhibited improved inhibition of the enzyme catalytic activity, displayed interesting binding conformations and unusual molecular rearrangements of the ligands. These findings with the modified CMP nucleotides underscored the potential of transformation of poor ligands to ODCase into novel inhibitors of this drug target.

An Engineered Cytidine Deaminase for Biocatalytic Production of a Key Intermediate of the Covid-19 Antiviral Molnupiravir

The Covid-19 pandemic highlights the urgent need for cost-effective processes to rapidly manufacture antiviral drugs at scale. Here we report a concise biocatalytic process for Molnupiravir, a nucleoside analogue recently approved as an orally available treatment for SARS-CoV-2. Key to the success of this process was the development of an efficient biocatalyst for the production of N-hydroxy-cytidine through evolutionary adaption of the hydrolytic enzyme cytidine deaminase. This engineered biocatalyst performs >85 000 turnovers in less than 3 h, operates at 180 g/L substrate loading, and benefits from in situ crystallization of the N-hydroxy-cytidine product (85% yield), which can be converted to Molnupiravir by a selective 5′-acylation using Novozym 435.

Toward a Practical, Two-Step Process for Molnupiravir: Direct Hydroxamination of Cytidine Followed by Selective Esterification

A two-step synthesis of molnupiravir (1) is presented. This work focuses on the development of practical reaction and purification conditions toward a manufacturing route. The sequence commences from highly available cytidine (2), and molnupiravir is formed through direct hydroxamination of the cytosine ring and esterification of the sugar's primary alcohol without use of protecting or activating groups. A highly crystalline hydrate of N-hydroxycytidine (3) resulted in an easily purified intermediate, and a practical, off-the-shelf enzyme was selected for the acylation. The yield was increased through a chemically promoted, selective ester cleavage, which converted a byproduct, molnupiravir isobutyryl oxime ester (4), into the final API. Both reactions proceed in >90% assay yield, and crystallization procedures are used to afford intermediates and active pharmaceutical ingredients in purities above 99% with an overall yield of 60%. Excellent throughput and sustainability are achieved by limiting the total concentration to 7 volumes of solvent in the course of the two reactions with an overall PMI of 26 including work-up and isolation. Environmentally friendly solvents, water and 2-methyl tetrahydrofuran, enhance sustainability of the operation.

Development of a Robust Manufacturing Route for Molnupiravir, an Antiviral for the Treatment of COVID-19

Herein is described the development of a large-scale manufacturing process for molnupiravir, an orally dosed antiviral that was recently demonstrated to be efficacious for the treatment of patients with COVID-19. The yield, robustness, and efficiency of each of the five steps were improved, ultimately culminating in a 1.6-fold improvement in overall yield and a dramatic increase in the overall throughput compared to the baseline process.

Preparation method of N4-hydroxycytidine

The invention discloses a preparation method of N4-hydroxycytidine capable of preventing and treating various virus infection including COVID-19. The preparation method comprises the following steps: taking cytosine and tetraacetyl ribose as initial raw materials, and carrying out hydroxyamination, condensation and hydrolysis reaction to prepare the N4-hydroxycytidine. The preparation method is easily available in raw materials, simple in process, economical, environment-friendly and suitable for industrial production.

N4-HYDROXYCYTIDINE AND DERIVATIVES AND ANTI-VIRAL USES RELATED THERETO

This disclosure relates to certain N4-hydroxycytidine derivatives, pharmaceutical compositions, and methods related thereto. In certain embodiments, the disclosure relates to the treatment or prophylaxis of human coronavirus 2019-nCoV.

A concise route to MK-4482 (EIDD-2801) from cytidine

A two-step route to MK-4482 (EIDD-2801, 1) was developed consisting of an esterification and hydroxamination of cytidine. The selective acylation and direct amination eliminate the need for protecting and activating groups and proceed in overall yield of 75%, a significant advancement over the reported yield of 17%. The step count is reduced from five transformations to two, and expensive uridine is replaced with the more available cytidine. This journal is

A High-Yielding Synthesis of EIDD-2801 from Uridine**

A simple reordering of the reaction sequence allowed the improved synthesis of EIDD-2801, an antiviral drug with promising activity against the SARS-CoV-2 virus, starting from uridine. Compared to the original route, the yield was enhanced from 17 % to 61 %, and fewer isolation/purification steps were needed. In addition, a continuous flow procedure for the final acetonide deprotection was developed, which proved to be favorable toward selectivity and reproducibility.

N4-HYDROXYCYTIDINE AND DERIVATIVES AND ANTI-VIRAL USES RELATED THERETO

This disclosure relates to certain N4-hydroxycytidine derivatives, pharmaceutical compositions, and methods related thereto. In certain embodiments, the disclosure relates to the treatment or prophylaxis of viral infections, such as Eastern, Western, and Venezuelan Equine Encephalitis (EEE, WEE and VEE, respectively), Chikungunya fever (CHIK), Ebola, Influenza, RSV, and Zika virus infection with the disclosed compounds.