4-(2,5-Dimethyl-1H-pyrrol-1-yl)-N-(2,5-dioxopyrrolidin-1-yl) benzamide improves monoclonal antibody production in a Chinese hamster ovary cell culture

• Published in: PloS one 2021, Vol.16 (4), p.e0250416-e0250416
• Main Authors: Aki, Yuichi, Katsumata, Yuta, Kakihara, Hirofumi, Nonaka, Koichi, Fujiwara, Kenshu
• Format: Article
• Language: English
• Subjects: Adenosine triphosphate; Antioxidants; Batch culture; Benzamide; Biological products; Biology and Life Sciences; Breeding; Butyric acid; Cell culture; Chemical compounds; Chromatography; Culture; Culture media; Cyclin-dependent kinase 4; Dimethyl sulfoxide; Genetic engineering; Glucose; Glutamine; Immunoglobulin G; Laboratories; Libraries; Life sciences; Lithium; Lithium chloride; Manufacturers; Monoclonal antibodies; Optimization; Ovaries; Phenols; Physical Sciences; Productivity; R&D; Research & development; Research and Analysis Methods; Thymidine; Valeric acid; Valproic acid
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0250416

There is a continuous demand to improve monoclonal antibody production for medication supply and medical cost reduction. For over 20 years, recombinant Chinese hamster ovary cells have been used as a host in monoclonal antibody production due to robustness, high productivity and ability to produce proteins with ideal glycans. Chemical compounds, such as dimethyl sulfoxide, lithium chloride, and butyric acid, have been shown to improve monoclonal antibody production in mammalian cell cultures. In this study, we aimed to discover new chemical compounds that can improve cell-specific antibody production in recombinant Chinese hamster ovary cells. Out of the 23,227 chemicals screened in this study, 4-(2,5-dimethyl-1H-pyrrol-1-yl)-N-(2,5-dioxopyrrolidin-1-yl) benzamide was found to increase monoclonal antibody production. The compound suppressed cell growth and increased both cell-specific glucose uptake rate and the amount of intracellular adenosine triphosphate during monoclonal antibody production. In addition, the compound also suppressed the galactosylation on a monoclonal antibody, which is a critical quality attribute of therapeutic monoclonal antibodies. Therefore, the compound might also be used to control the level of the galactosylation for the N-linked glycans. Further, the structure-activity relationship study revealed that 2,5-dimethylpyrrole was the most effective partial structure of 4-(2,5-dimethyl-1H-pyrrol-1-yl)-N-(2,5-dioxopyrrolidin-1-yl) benzamide on monoclonal antibody production. Further structural optimization of 2,5-dimethylpyrrole derivatives could lead to improved production and quality control of monoclonal antibodies.