Creation of cell-internalizing antibody for novel tumor endothelial missile therapy

2009 Fiscal Year Final Research Report

• Project/Area Number: 20790134
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Single-year Grants
• Research Field: Medical pharmacy
• Research Institution: Osaka University
• Principal Investigator: MUKAI Yohei Osaka University, 大学院・薬学研究科, 助教 (60444501)
• Project Period (FY): 2008 – 2009
• Keywords: 医療薬剤学

Research Abstract

We rapidly isolated "cell-internalizing antibody" against tumor endothelial markers (VEGFR2, Robo4 etc.) using optimized phage display system. Obtained cell-internalizing antibody, anti-VEGFR2 scFv fused with PSIF, significantly suppressed the tumor growth on experimental mouse model. This result indicated that the rapid isolation of cell-internalizing antibody contributes the development DDS medicine, such as immunoconjugate against cancer.

Research Products

• [Journal Article] Simple PEG Conjugation of SPIO via an Au-S Bond Improves Its Tumor Targeting Potency as a Novel MR Tumor Imaging Agent (2010)
  • Author(s): Kojima H, Mukai Y, Yoshikawa M, Kamei K, Yoshikawa T, Morita M, Inubushi T, Yamamoto TA, Yoshioka Y, Okada N, Seino S, Nakagawa S.
  • Journal Title: Bioconjug Chem. (in press)
  • Peer Reviewed
• [Journal Article] Therapeutic effect of PEGylated TNFR1-selective antagonistic mutant TNF in experimental autoimmune encephalomyelitis mice. (2010)
  • Author(s): Nomura T, Abe Y, Kamada H, Shibata H, Kayamuro H, Inoue M, Kawara T, Arita S, Furuya T, Yamashita T, Nagano K, Yoshikawa T, Yoshioka Y, Mukai Y, Nakagawa S, Taniai M, Ohta T, Serada S, Naka T, Tsunoda SI, Tsutsumi Y.
  • Journal Title: J Control Release (in press)
  • Peer Reviewed
• [Journal Article] Creation of a LIGHT mutant with the capacity to evade the decoy receptor for cancer therapy (2010)
  • Author(s): Morishige T, Yoshioka Y, Inakura H, Tanabe A, Yao X, Tsunoda S, Tsutsumi Y, Mukai Y, Okada N, Nakagawa S.
  • Journal Title: Biomaterials 31(12) Pages: 3357-63
  • Peer Reviewed
• [Journal Article] Creation of a lysine-deficient LIGHT mutant with the capacity for site-specific PEGylation and low affinity for a decoy receptor (2010)
  • Author(s): Morishige T, Yoshioka Y, Inakura H, Tanabe A, Yao X, Tsunoda S, Tsutsumi Y, Mukai Y, Okada N, Nakagawa S.
  • Journal Title: Biochem Biophys Res Commun 393(4) Pages: 888-93
  • Peer Reviewed
• [Journal Article] Tumor-targeting CTL expressing a single-chain Fv specific for VEGFR2 (2010)
  • Author(s): Kanagawa N, Yanagawa T, Mukai Y, Yoshioka Y, Okada N, Nakagawa S.
  • Journal Title: Biochem Biophys Res Commun. 394(1) Pages: 54-8
  • Peer Reviewed
• [Journal Article] Creation of lysine-deficient mutant lymphotoxin-alpha with receptor selectivity by using a phage display system. (2010)
  • Author(s): Yoshioka Y, Watanabe H, Morishige T, Yao X, Ikemizu S, Nagao C, Ahmad S, Mizuguchi K, Tsunoda S, Tsutsumi Y, Mukai Y, Okada N, Nakagawa S.
  • Journal Title: Biomaterials 31(7) Pages: 1935-43
  • Peer Reviewed
• [Journal Article] Comparison of the anti-tumor activity of native, secreted, and membrane-bound LIGHT in mouse tumor models. (2010)
  • Author(s): Morishige T, Yoshioka Y, Tanabe A, Yao X, Mizuguchi H, Tsunoda S, Tsutsumi Y, Mukai Y, Okada N, Nakagawa S.
  • Journal Title: Int Immunopharmacol 10(1) Pages: 26-33
  • Peer Reviewed
• [Journal Article] 腫瘍MRイメージングにおける造影剤開発 (2010)
  • Author(s): 向洋平、中川晋作
  • Journal Title: JSMI Report. 3(1) Pages: 14-17
  • Peer Reviewed
• [Journal Article] The treatment of established murine collagen-induced arthritis with a TNFR1-selective antagonistic mutant TNF (2009)
  • Author(s): Shibata H, Yoshioka Y, Abe Y, Ohkawa A, Nomura T, Minowa K, Mukai Y, Nakagawa S, Taniai M, Ohta T, Kamada H, Tsunoda S, Tsutsumi Y.
  • Journal Title: Biomaterials 30(34) Pages: 6638-47
  • Peer Reviewed
• [Journal Article] LIGHT protein suppresses tumor growth by augmentation of immune response (2009)
  • Author(s): Morishige T, Yoshioka Y, Inakura H, Tanabe A, Watanabe H, Yao X, Tsunoda S, Tsutsumi Y, Mukai Y, Okada N, Nakagawa S.
  • Journal Title: Immunol Lett. 127(1) Pages: 33-8
  • Peer Reviewed
• [Journal Article] Novel protein engineering strategy for creating highly receptor-selective mutant TNFs (2009)
  • Author(s): Nomura T, Abe Y, Kamada H, Inoue M, Kawara T, Arita S, Furuya T, Yoshioka Y, Shibata H, Kayamuro H, Yamashita T, Nagano K, Yoshikawa T, Mukai Y, Nakagawa S, Taniai M, Ohta T, Tsunoda SI, Tsutsumi Y.
  • Journal Title: Biochem Biophys Res Commun. 388(4) Pages: 667-71
  • Peer Reviewed
• [Journal Article] Fast binding kinetics and conserved 3D structure underlie the antagonistic activity of mutant TNF: useful information for designing artificial proteo-antagonists (2009)
  • Author(s): Mukai Y, Nakamura T, Yoshioka Y, Shibata H, Abe Y, Nomura T, Taniai M, Ohta T, Nakagawa S, Tsunoda S, Kamada H, Yamagata Y, Tsutsumi Y.
  • Journal Title: J Biochem. 146(2)(表紙掲載) Pages: 167-72
  • Peer Reviewed
• [Journal Article] The augmentation of intracellular delivery of peptide therapeutics by artificial protein transduction domains (2009)
  • Author(s): Yoshikawa T, Sugita T, Mukai Y, Yamanada N, Nagano K, Nabeshi H, Shibata H, Yoshioka Y, Nakagawa S, Kamada H, Tsunoda S, Tsutsumi Y.
  • Journal Title: Biomaterials 30(19) Pages: 3318-23
  • Peer Reviewed
• [Journal Article] Crystallization and preliminary X-ray analysis of the tumour necrosis factorα-tumour necrosis factor receptor type 2 complex (2009)
  • Author(s): Mukai Y, Nakamura T, Yoshioka Y, Tsunoda S, Kamada H, Nakagawa S, Yamagata Y, Tsutsumi Y.
  • Journal Title: Acta Cryst F65(Pt3) Pages: 295-8
  • Peer Reviewed
• [Journal Article] Direct cell-entry of gold/iron-oxide magnetic nanoparticles in adenovirus mediated gene delivery (2009)
  • Author(s): Kamei K, Mukai Y, Kojima H, Yoshikawa T, Yoshikawa M, Kiyohara G, Yamamoto TA, Yoshioka Y, Okada N, Seino S, Nakagawa S.
  • Journal Title: Biomaterials 30(9) Pages: 1809-14
  • Peer Reviewed
• [Journal Article] Structure-function relationship of tumor necrosis factor (TNF) and its receptor interaction based on 3D structural analysis of a fully active TNFR1-selective TNF mutant (2009)
  • Author(s): Mukai Y, Shibata H, Nakamura T, Yoshioka Y, Abe Y, Nomura T, Taniai M, Ohta T, Ikemizu S, Nakagawa S, Tsunoda S, Kamada H, Yamagata Y, Tsutsumi Y.
  • Journal Title: J Mol Biol. 385(4) Pages: 1221-9
  • Peer Reviewed
• [Journal Article] Ligand-independent assembly of purified soluble magic roundabout (Robo4), a tumor-specific endothelial marker. (2008)
  • Author(s): Yoshikawa M, Mukai Y, Okada Y, Yoshioka Y, Tsunoda SI, Tsutsumi Y, Okada N, Aird WC, Doi T, Nakagawa S.
  • Journal Title: Protein Expr Purif 61(1) Pages: 78-82
  • Peer Reviewed
• [Presentation] 抗体-薬物複合体;Antibody-drug conjugateの開発に適う細胞内侵入抗体の効率的探索法 (2010)
  • Author(s): 吉川舞、向洋平、岡田欣晃、吉岡靖雄、岡田直貴、Aird WC、土井健史、堤康央、角田慎一、中川晋作
  • Organizer: 日本薬学会第130年会
  • Place of Presentation: 岡山
  • Year and Date: 2010-03-30
• [Presentation] Antibody Drug Conjugate製剤の開発を目指した細胞内侵入抗体の効率的探索法 (2010)
  • Author(s): 向洋平
  • Organizer: 技術情報協会セミナー「次世代抗体医薬品開発に向けた最新研究動向」
  • Place of Presentation: 東京
  • Year and Date: 2010-01-25
• [Presentation] Ligand independent assembly of purified soluble Magic Roundabout (Robo4), a tumor-specific endothelial marker (2009)
  • Author(s): Mai Yoshikawa, Yohei Mukai, Yoshiaki Okada, Yasuo Yoshioka, Shin-ichi Tsunoda, Yasuo Tsutsumi, Naoki Okada, William C Aird, Takefumi Doi, Shinsaku Nakagawa
  • Organizer: ECCO 15 and 34th ESMO Multidisciplinary Congress
  • Place of Presentation: Berlin (Germany)
  • Year and Date: 20090920-20090924
• [Presentation] Design of tumor targeting magnetic resonance imaging (MRI) agent based on gold/iron-oxide composite nanoparticle (2009)
  • Author(s): Hiroki Kojima, Yohei Mukai, Kazumasa Kamei, Satoshi Seino, Masashi Morita, Toshiro Inubushi, Yasuo Yoshioka, Naoki Okada, Shinsaku Nakagawa
  • Organizer: ECCO 15 and 34th ESMO Multidisciplinary Congress
  • Place of Presentation: Berlin (Germany)
  • Year and Date: 20090920-20090924
• [Presentation] Direct cell entry of gold/iron-oxide magnetic nanoparticles in adenovirus mediated gene delivery (2009)
  • Author(s): Yohei Mukai, Hiroki Kojima, Tomoaki Yoshikawa, Kazumasa Kamei, Mai Yoshikawa, Takao A Yamamoto, Yasuo Yoshioka, Naoki Okada, Satoshi Seino, Shinsaku Nakagawa
  • Organizer: ECCO 15 and 34th ESMO Multidisciplinary Congress
  • Place of Presentation: Berlin (Germany)
  • Year and Date: 20090920-20090924
• [Presentation] Receptor interaction of Tumor Necrosis Factor (TNF) based on 3D structural analysis of a fully active TNFR1-selective TNF mutant (2009)
  • Author(s): Yohei Mukai, Teruya Nakamura, Yasuo Yoshioka, Yasuhiro Abe, Tetsuya Nomura, Madoka Taniai, Tsunetaka Ohta, Shinji Ikemizu, Shinsaku Nakagawa, Shin-ichi Tsunoda, Haruhiko Kamada, Yuriko Yamagata, Yasuo Tsutsumi
  • Organizer: 12th International TNF Conference
  • Place of Presentation: Madrid (Spain)
  • Year and Date: 20090426-20090429
• [Presentation] The development of fully active receptor selective Tumor Necrosis Factor (TNF) mutants (2009)
  • Author(s): Mai Yoshikawa, Yohei Mukai, Hiroko Shibata, Yasuo Yoshioka, Yasuhiro Abe, Tetsuya Nomura, Madoka Taniai, Tsunetaka Ohta, Shin-ichi Tsunoda, Haruhiko Kamada, Shinsaku Nakagawa, Yasuo Tsutsumi
  • Organizer: 12th International TNF Conference
  • Place of Presentation: Madrid (Spain)
  • Year and Date: 20090426-20090429
• [Presentation] Antibody-drug conjugateの開発に適う細胞内侵入抗体の効率的探索法 (2009)
  • Author(s): 吉川舞、向洋平、吉岡靖雄、岡田直貴、堤康央、角田慎一、中川晋作
  • Organizer: 第8回ファーマ・バイオフォーラム
  • Place of Presentation: 名古屋
  • Year and Date: 2009-11-14
• [Presentation] High-throughput screening method of "Cell-internalizing monoclonal antibodies", potent anti-tumor drug delivery carriers (2009)
  • Author(s): 吉川舞、向洋平、吉岡靖雄、岡田直貴、堤康央、角田慎一、中川晋作
  • Organizer: 第68回日本癌学会学術総会
  • Place of Presentation: 横浜
  • Year and Date: 2009-10-02
• [Presentation] 金磁性複合ナノ粒子のバイオ・DDS分野への展開 (2009)
  • Author(s): 向洋平
  • Organizer: 第3回「ナノメディスン・アプリケーション研究会」
  • Place of Presentation: 大阪
  • Year and Date: 2009-09-16
• [Presentation] 金磁性複合ナノ粒子のバイオ分野への応用 (2009)
  • Author(s): 向洋平
  • Organizer: 第31回ナノバイオ磁気工学専門研究会
  • Place of Presentation: 大阪
  • Year and Date: 2009-07-23
• [Presentation] immuno-conjugate製剤の開発に適う細胞内侵入抗体の効率的創製法の確立 (2009)
  • Author(s): 吉川舞、向洋平、久保貴、吉岡靖雄、岡田直貴、堤康央、角田慎一、中川晋作
  • Organizer: 日本薬剤学会第24年会
  • Place of Presentation: 静岡
  • Year and Date: 2009-05-22
• [Presentation] 微細超常磁性酸化鉄粒子(USPIO)をコアとしたPEG化金磁性ナノ粒子の創製とそのMRI腫瘍造影剤としての有用性評価 (2009)
  • Author(s): 小島拓記、向洋平、森田将史、犬伏俊郎、山本孝夫、吉岡靖雄、岡田直貴、清野智史、中川晋作
  • Organizer: 日本薬剤学会第24年会
  • Place of Presentation: 静岡
  • Year and Date: 2009-05-22
• [Presentation] TNFR1選択的阻害剤の開発を目指したTNFR1選択的変異体の創製とそのレセプター結合における構造-活性相関研究 (2009)
  • Author(s): 向洋平、柴田寛子、中村照也、吉岡靖雄、阿部康弘、野村鉄也、谷合まどか、太田恒考、池水信二、鎌田春彦、角田慎一、中川晋作、山縣ゆり子、堤康央
  • Organizer: 第9回蛋白質科学会年会
  • Place of Presentation: 熊本
  • Year and Date: 2009-05-21
• [Presentation] STLUCTURE-FUNCTION RELATIONSHIP OF TNF BASED ON 3D STRUCTURAL ANALYSIS OF FULLY ACTIVE TNFR1-SELECTIVE TNF MUTANT (2008)
  • Author(s): Yohei Mukai, Hiroko Shibata, Teruya Nakamura, Yasuo Yoshioka, Yasuhiro Abe, Tetsuya Nomura, Madoka Taniai, Tsunetaka Ohta, Shinji Ikemizu, Shinsaku Nakagawa, Shin-ichi Tsunoda, Haruhiko Kamada, Yuriko Yamagata, Yasuo Tsutsumi
  • Organizer: Cytokine 2008
  • Place of Presentation: Montreal, CANADA
  • Year and Date: 20081000
• [Presentation] Direct Cell-entry of Gold/iron-oxide Magnetic Nanoparticles Enhanced Adenovirus Mediated Gene Expression in Adenovirus-resistant cells (2008)
  • Author(s): Kamei K, Yoshikawa T, Kojima H, Seino S, Yamamoto T A, Yoshioka Y, Okada N, Mukai Y, Nakagawa S
  • Organizer: 7th international conference on the scientific and clinical applications of magnetic carriers
  • Place of Presentation: Vancouver, CANADA
  • Year and Date: 20080500
• [Remarks] 大阪大学研究者総覧(向洋平)
  • URL: http://www.dma.jim.osaka-u.ac.jp/kg-portal/aspI/RX0011D.asp?UNO=15207&page=1