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

PLoS One. 2021 Apr 22;16(4):e0250416. doi: 10.1371/journal.pone.0250416. eCollection 2021.


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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Triphosphate / metabolism
  • Animals
  • Antibodies, Monoclonal / biosynthesis*
  • Antibodies, Monoclonal / genetics
  • Benzamides / pharmacology*
  • CHO Cells
  • Cell Culture Techniques
  • Cell Proliferation / drug effects
  • Cell Survival / drug effects
  • Cricetulus
  • Culture Media / chemistry
  • Galactose / metabolism
  • Glucose / metabolism
  • Metabolic Engineering / methods*
  • Polysaccharides / metabolism
  • Pyrroles / pharmacology*
  • Signal Transduction / drug effects*
  • Structure-Activity Relationship


  • Antibodies, Monoclonal
  • Benzamides
  • Culture Media
  • Polysaccharides
  • Pyrroles
  • benzamide
  • Adenosine Triphosphate
  • Glucose
  • 2,5-dimethylpyrrole
  • Galactose

Grants and funding

1. The chemical compound library used in this study was provided by the Platform Project for Supporting Drug Discovery and Life Science Research (the Basis for Supporting Innovative Drug Discovery and Life Science Research [BINDS] program) from the Japan Agency for Medical Research and Development (AMED) under grant numbers JP19am0101086, JP19am0101084, and JP19am0101095 (support numbers 1009, 0847, and 1011, respectively). The funders had no role in the study design, data collection, data analysis, decision to publish, or preparation of the manuscript. 2. Daiichi Sankyo Co., Ltd., provided support in the form of salaries for the authors and supporters (YA, YK, HK, KN, AK, and MK), but the company did not have any additional role in the study design, data collection, data analysis, decision to publish, or preparation of the manuscript. The specific roles of these authors are articulated in the Author Contributions section.