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1999 Fiscal Year Final Research Report Summary

Molecular targeting against growth factor receptors by a novel drug compord of human fusion proteins

Research Project

Project/Area Number 09470258
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field General surgery
Research InstitutionKeio University

Principal Investigator

UEDA Masakazu  Keio Univ., Surgery, Assistant Professor, 医学部, 講師 (50142419)

Co-Investigator(Kenkyū-buntansha) KAMEYAMA Noriaki  Keio Univ., Surgery, Assistant, 医学部, 助手 (50276268)
WATANABE Yasuo  Keio Univ., Surgery, Assistant, 医学部, 助手 (20265846)
OZAWA Soji  Keio Univ., Surgery, Assistant Professor, 医学部, 講師 (10169287)
Project Period (FY) 1997 – 1999
KeywordsMissils therapy / growth factor / growth factor receplor / IL-2 / EGF / IL-2 receptor / fusion protein
Research Abstract

To develop a new Missile therapy targeted at growth factor receptor, fusion proteins with a human Rnase and monoclonal antibody against growth factor receptors or growth factor. A hybrid human protein was engineered by fusing the genes encoding human pancreatic Rnase 1 and human IL-2, was isolated and refolded from E.coli inclusion bodies, and was purified to homogeneity. The hpRNase1-hIL-2 inhibited protein synthesis in HTLV-1 infected, malignant T lymphocytes, hyperproducing high affinity IL-2 receptors, with an IC50 of 2x10-8M, whereas no inhibition was detectable for receptor deficient control cells or hpRNase 1 alone had an IL50 of almost 10-3M.. A molar excess of hIL-2 locked the protein synthesis inhibition dose dependently. In a human mixed lymphocyte culture, hpRNase 1-hIL-2 inhibited the proliferation of responder cells with potency comparable to that of cyclosporine, while minimal doses of FK506 importantly improved its potency. Concentrations comparable to the IC50 in vitro could be safely achieved in animals, and could be escalated without any toxic side effects. The murine monoclonal antibody (528) against the human epidermal growth factor receptor (EGFR) was conjugated with mammalian pancreatic Rnase via SPDP and 2-IT. The conjugate showed dose-dependent cytotoxicity against EGFR-producing squamous cancer cells and no detectable cytotoxicity against EGFR-deficient small cell lung cancer cells. The cytotoxicity of the conjugate was positively correlated with the EGFR numbers of each cell line. The addition of excess 528 antibody to the medium rotected A431 cells from the conjugate cytotoxicity. This immunoconjugate might be useful for targeted treatment of squamous cell carcinomas hyperexpressing EGFR.

  • Research Products

    (15 results)

All Other

All Publications (15 results)

  • [Publications] Psarras K.: "Targeting activated lymphocytes with an entirely human immunotoxin analog: Human pancreatci RNase1-human IL-2 fusion"Cytokine. (in press).

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Suwa T.: "Epidermal growth factor receptor-dependent cytotoxic effect of anti-EGFR antibody-ribonuclease conjugate o human cancer cell"Anticancer Res.. 19. 4161-4166 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Futami J.: "Inhibition of cell growth by a fused protein of human ribonuclease 1 and human fibroblast growth factor"Protein Engineering. 12. 1013-1019 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Psarras K: "Human Pancreatic RNase1-human EGF fusion: an entirely human immunotoxin analogy with cytotoxic properties against squamous cell rowth"Protein ENgineering. 11. 1285-1292 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Kaneta Y: "Effect of gelonin immunoconjugate with monoclenal antibody MSN-1 to extometrial adenocarcinoma on antigen-producing-lumor cells in vivo"Jpn. J. Cancer Res.. 89. 583-588 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Suwa T.: "Magnetic resonance imaging of esophageal squamous cell carcinoma using magnotite particles coated with anti-EGF antibody"Int J. Cancer. 75. 626-634 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Shin C. et al.: "Vascular endothelial qrowth factor expression predicts outcome and lymph node metastasis in squamous cell carcinoma of the esophaqus"Clin. Cancer Res. (in press).

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Psarras K. et al: "Tarteting IL-2 receptors with an entirely human immunotoxin analog: human pancreatic Rnase-human IL-2 fuion"Cvtokine. (in press).

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Futami. J. et al: "Inhibition of cell qrowth by a fuxed of human ribonucleasel and human fibroblast qrowth factor"Protein Eng. 12. 1013-1019 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Suwa T. et al: "Epidermal qrowth factor receptor-dependent cytotoxic effect of anti-EGFR antibbody-ribonuclease conjugate on human cancer cells"Anticancer Res. 19. 4169-4166 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Koyanagi K. et al: "Clinical singnificance of telomerase acitivity in non-cancerous epitelial region of the esophaqeal cancer"Br. J. Surgery. 86. 674-679 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Psarras K. et al.: "Human pancreatic Rnase 1- human epidermal growth factor fusion: an enntirely human immunotoxin analog with cytotoxic properities against sguamous cell carcinoma"Protein Eng. 11. 1285-1292 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Kaneta Y. et al: "Effects of gelonin immunotoxin conjugate with monoclonal antibody MSN-1 to endometrial edenocarcinomas on antigen-producing tumors cells in vivo"Jpn. J. Cancer Res. 89. 583-588 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Suwa T. et al: "Magnetic resonance imaging of esophageal sguamous cell carcinoma using magnetite particles coated with anti-epidermal growth factor receptor antibody"Int. J. Cancer. 75. 626-634 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Ueda M.. Et al.: "Molecular tarteting for epidermal growth factor receptor expressed on cancer cells by human fusion protien"Breast Cancer. 4. 253-255 (1997)

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 2001-10-23  

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