Relationship between physicochemical properties of protein aggregates and complement activation on safety of biopharmaceuticals

• Project/Area Number: 17K15537
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Medical pharmacy
• Research Institution: National Institute of Health Sciences
• Principal Investigator: Shibata Hiroko 国立医薬品食品衛生研究所, 生物薬品部, 室長 (60462769)
• Project Period (FY): 2017-04-01 – 2020-03-31
• Project Status: Completed (Fiscal Year 2019)
• Budget Amount: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000); Fiscal Year 2019: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2018: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2017: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
• Keywords: バイオ医薬品 / 補体 / 凝集体 / 補体活性化

Outline of Final Research Achievements

In this study, we attempted to establish ①preparative method of protein aggregates with various physicochemical attributes, ②complement activation assay, and to elucidate the relationship between physicochemical attributes of protein aggregates and their complement activation potency. A preparative method that can constantly provide protein aggregate with various attributes was developed by varying temperature and agitation rate on IVIG and human albumin. Complement activation potency was evaluated by measuring SC5b9, which is an ultimate product of complement pathway. It was suggested that denatured IVIG and aggregated albumin could activate a complement pathway. In the future, improvement of complement activation assay will be needed to get a consistent relationship between attributes of protein aggregates and complement activation potency.

Academic Significance and Societal Importance of the Research Achievements

抗体以外のタンパク質やFc部分が変性した抗体においても，凝集体が形成されることで補体経路が活性化される可能性が示され，バイオ医薬品における凝集体の管理の重要性を支持する成果である．また，抗体以外のタンパク質の凝集体の補体活性化能に関する報告は極めて少なく，学術的な意義もあると考えられる．引き続き研究を進め，補体活性化の強度や形状による活性化能の違いについて一貫した傾向が得られれば，バイオ医薬品製剤の適切なリスク管理戦略を構築するのに有益な情報となり，バイオ医薬品の品質管理の近代化および安全性向上につながるものと期待される．

Research Products

• [Journal Article] Recent Achievements and Current Interests in Research on the Characterization and Quality Control of Biopharmaceuticals in Japan (2020)
  • Author(s): Ishii-Watabe Akiko、Shibata Hiroko、Suetomo Hiroyuki、Ikeda Yosuke、Telikepalli Srivalli、Kiyoshi Masato、Hayashi Yu、Muto Takashi、Tanaka Yukako、Ueda Satomi、Iwura Takafumi、Saitoh Satoshi、Aoyama Michihiko、Harazono Akira、Hyuga Masashi、Goda Yukihiro、Torisu Tetsuo、Uchiyama Susumu
  • Journal Title: Journal of Pharmaceutical Sciences Volume: 109 Issue: 5 Pages: 1652-1661
  • DOI: 10.1016/j.xphs.2020.01.001
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] 日本薬局方の動向と分析担当者の実務対応 タンパク質医薬品注射剤の不溶性微粒子試験法. (2020)
  • Author(s): 柴田寛子，原園景，石井明子
  • Journal Title: PHARM STAGE Volume: 19(10) Pages: 9-12
• [Journal Article] Collaborative Study for Analysis of Subvisible Particles Using Flow Imaging and Light Obscuration: Experiences in Japanese Biopharmaceutical Consortium (2019)
  • Author(s): Kiyoshi Masato、Shibata Hiroko、Uchiyama Susumu、Ishii-Watabe Akiko
  • Journal Title: Journal of Pharmaceutical Sciences Volume: 108 Issue: 2 Pages: 832-841
  • DOI: 10.1016/j.xphs.2018.08.006
  • Peer Reviewed / Open Access
• [Journal Article] バイオ医薬品における凝集体及び不溶性微粒子評価法について (2018)
  • Author(s): 柴田寛子，木吉真人，原園景，石井明子
  • Journal Title: 医療機器レギュラトリーサイエンス Volume: 49(11) Pages: 747-753
  • NAID: 40021739521
• [Presentation] Current status and issues of biopharmaceuticals from regulatory perspective (2019)
  • Author(s): Shibata H, Kiyoshi M, Harazono A, Ishii-Watabe A
  • Organizer: 第19回日本蛋白質科学会年会・第71回日本細胞生物学会大会 合同年次大会
  • Invited
• [Presentation] 注射剤の不溶性微粒子評価に用いられる光遮蔽法とフローイメージング法の分析性能の比較：市販タンパク質医薬品注射剤の測定結果 (2019)
  • Author(s): 柴田 寛子，木吉 真人，原園 景，石井 明子
  • Organizer: 第9回レギュラトリーサイエンス学会学術大会
• [Presentation] フローイメージング法によるシリコーン油滴とタンパク質凝集体／不溶性微粒子の分類における測定条件の影響 (2019)
  • Author(s): 柴田寛子，原園景，木吉真人，石井明子
  • Organizer: 日本薬学会第140年会
• [Presentation] Evaluation of Protein Aggregates/Subvisible Particles in Therapeutic Protein Injections. (2018)
  • Author(s): Shibata H
  • Organizer: CMC Strategy Forum Japan 2018
  • Int'l Joint Research
• [Presentation] Collaborative study for analysis of subvisible particles using flow imaging and light obscuration: experience in Japanese biopharmaceutical consortium (2018)
  • Author(s): Shibata H, Kiyoshi M, Harazono A, Torisu T, Maruno T, Akimaru M, Asano Y, Hirokawa M, Ikemoto K, Itakura Y, Iwura T, Kikitsu A, Kumagai T, Mori N, Murase H, Nishimura H, Oda A, Ogawa T, Ojima T, Okabe S, Saito S, Saitoh S, Suetomo H, Takegami K, Takeuchi M, Yasukawa H, Uchiyama S, Ishii-Watabe A
  • Organizer: 2018 Workshop on Protein Aggregation and Immunogenicity
  • Int'l Joint Research
• [Presentation] Usefulness and Issues of Flow Imaging Analysis for Evaluating Aggregates in Therapeutic Protein Injections. (2017)
  • Author(s): Hiroko S, Kiyoshi M, Harazono A, Ishii-Watabe A
  • Organizer: USP Workshop on Control and Determination of Visible and Sub-visible Particulate Matter in Biologics
  • Int'l Joint Research