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A bispecific antibody to factors IXa and X restores factor VIII hemostatic activity in a hemophilia A model

Nature Medicine volume 18, pages 1570–1574 (2012)Cite this article

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Abstract

Hemophilia A is a bleeding disorder resulting from coagulation factor VIII (FVIII) deficiency. Exogenously provided FVIII effectively reduces bleeding complications in patients with severe hemophilia A. In approximately 30% of such patients, however, the 'foreignness' of the FVIII molecule causes them to develop inhibitory antibodies against FVIII (inhibitors), precluding FVIII treatment in this set of patients1,2,3. Moreover, the poor pharmacokinetics of FVIII, attributed to low subcutaneous bioavailability and a short half-life of 0.5 d, necessitates frequent intravenous injections3,4,5. To overcome these drawbacks, we generated a humanized bispecific antibody to factor IXa (FIXa) and factor X (FX), termed hBS23, that places these two factors into spatially appropriate positions and mimics the cofactor function of FVIII. hBS23 exerted coagulation activity in FVIII-deficient plasma, even in the presence of inhibitors, and showed in vivo hemostatic activity in a nonhuman primate model of acquired hemophilia A. Notably, hBS23 had high subcutaneous bioavailability and a 2-week half-life and would not be expected to elicit the development of FVIII-specific inhibitory antibodies, as its molecular structure, and hence antigenicity, differs from that of FVIII. A long-acting, subcutaneously injectable agent that is unaffected by the presence of inhibitors could markedly reduce the burden of care for the treatment of hemophilia A.

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Figure 1: Schematic illustrations of the action of FVIIIa or of a bispecific antibody as a cofactor promoting the interaction between FIXa and FX.
Figure 2: In vitro FVIII-mimetic cofactor activity of hBS23.
Figure 3: In vivo hemostatic activity and time profiles of plasma concentrations of hBS23.

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Acknowledgements

We thank T. Matsuura, T. Houjo, K. Kanisawa, R. Takemoto, T. Koike and M. Hiranuma for carrying out the in vivo experiments and M. Fujii, Y. Nakata, H. Ishida and F. Isomura for antibody generation and preparation. We also thank S. Ohtsu for carrying out in vitro experiments.

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Authors and Affiliations

  1. Fuji-Gotemba Research Laboratories, Chugai Pharmaceutical, Gotemba, Shizuoka, Japan.,

    Takehisa Kitazawa, Tomoyuki Igawa, Zenjiro Sampei, Atsushi Muto, Tetsuo Kojima, Tetsuhiro Soeda, Kazutaka Yoshihashi, Yukiko Okuyama-Nishida, Hiroyuki Saito, Hiroyuki Tsunoda, Tsukasa Suzuki, Hideki Adachi, Taro Miyazaki, Shinya Ishii, Mika Kamata-Sakurai, Takeo Iida, Aya Harada, Keiko Esaki, Miho Funaki, Chifumi Moriyama, Eriko Tanaka, Yasufumi Kikuchi, Tetsuya Wakabayashi, Manabu Wada, Masaaki Goto, Takeshi Toyoda, Atsunori Ueyama, Sachiyo Suzuki, Kenta Haraya, Tatsuhiko Tachibana, Yoshiki Kawabe & Kunihiro Hattori

  2. Department of Pediatrics, Nara Medical University, Kashihara, Nara, Japan.,

    Midori Shima

  3. Nara Medical University, Kashihara, Nara, Japan.,

    Akira Yoshioka

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  1. Takehisa Kitazawa
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Contributions

T. Kitazawa and T. Igawa led the pharmacological studies and the optimization of the bispecific antibody, respectively, in the program and wrote the manuscript. Z.S. designed the lead chimeric bispecific antibody and hBS23. T. Kojima led the lead identification. H.T. provided ideas on bispecific antibody engineering. T. Suzuki, H.A., T.M., S.I., M.K.-S. and T. Iida generated FVIII-, FIXa- and FX-specific antibodies. T. Soeda, Y.O.-N., A.H., M.F., C.M., E.T., T. Toyoda and A.U. performed the in vitro experiments. K.E. and S.S. performed the affinity analyses. Y. Kikuchi, T.W., M.W. and M.G. purified the bispecific antibody and the coagulation factor. A.M. and K.Y. performed the in vivo pharmacological study. K. Haraya and T. Tachibana performed the pharmacokinetic study. H.S. and Y. Kawabe provided direction and guidance for the various functional areas. M.S. and A.Y. provided advice on the program from the viewpoints of their medical expertise in hemophilia. K. Hattori provided the hypothesis and directed and organized the program.

Corresponding author

Correspondence to Takehisa Kitazawa.

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Competing interests

Declaration: T. Kitazawa, T. Igawa, Z.S., A.M., T. Kojima., T. Soeda, K.Y., Y.O.-N., H.S., T. Suzuki, T.M., C.M. and K. Hattori are employees of Chugai Pharmaceutical and inventors on the patents, patent applications or both relating to the bispecific antibodies to FIXa and FX, of which all rights have been assigned to the company. H.T., H.A., S.I., M.K.-S., T. Iida, A.H., K.E., M.F., E.T., Y. Kikuchi, T.W., M.W., M.G., T. Toyoda, A.U., S.S., K. Haraya, T. Tachibana and Y. Kawabe are employees of Chugai Pharmaceutical. M.S. receives consulting honoraria and research support from Chugai Pharmaceutical. A.Y. previously received research support from Chugai Pharmaceutical.

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Kitazawa, T., Igawa, T., Sampei, Z. et al. A bispecific antibody to factors IXa and X restores factor VIII hemostatic activity in a hemophilia A model. Nat Med 18, 1570–1574 (2012). https://doi.org/10.1038/nm.2942

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