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Antibodies targeting the catalytic zinc complex of activated matrix metalloproteinases show therapeutic potential

Abstract

Endogenous tissue inhibitors of metalloproteinases (TIMPs) have key roles in regulating physiological and pathological cellular processes1,2,3. Imitating the inhibitory molecular mechanisms of TIMPs while increasing selectivity has been a challenging but desired approach for antibody-based therapy4. TIMPs use hybrid protein-protein interactions to form an energetic bond with the catalytic metal ion, as well as with enzyme surface residues5,6,7. We used an innovative immunization strategy that exploits aspects of molecular mimicry to produce inhibitory antibodies that show TIMP-like binding mechanisms toward the activated forms of gelatinases (matrix metalloproteinases 2 and 9). Specifically, we immunized mice with a synthetic molecule that mimics the conserved structure of the metalloenzyme catalytic zinc-histidine complex residing within the enzyme active site. This immunization procedure yielded selective function-blocking monoclonal antibodies directed against the catalytic zinc-protein complex and enzyme surface conformational epitopes of endogenous gelatinases. The therapeutic potential of these antibodies has been demonstrated with relevant mouse models of inflammatory bowel disease8. Here we propose a general experimental strategy for generating inhibitory antibodies that effectively target the in vivo activity of dysregulated metalloproteinases by mimicking the mechanism employed by TIMPs.

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Figure 1: Mimicry of the zinc-protein motif of MMPs.
Figure 2: Characterization of the antibodies SDS3 and SDS4.
Figure 3: Structural analyses of the inhibitory antibody SDS3.
Figure 4: Prophylactic and therapeutic treatment with SDS3 protects C57BL/6J mice against DSS-induced colitis.

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Acknowledgements

We would like to thank D. Tawfik and I. Shachar for critical reading of the manuscript, K. Vyacheslav for assistance in optical imaging data collection and analysis, and C. Power, O. Leger and O. Lightner for technical support in antibody production. This work was supported by the Israel Science Foundation, the Minerva Foundation, the Helmsley Foundation, the Kimmelman Center at the Weizmann Institute and funds from C. Berall and L. Berall, M. Steinberg and C. Adelson. I.S. is the Incumbent of the Maurizio Pontecorvo Professorial Chair.

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N.S.-P. designed, performed and analyzed the antibody biophysical analyses, animal experiments and wrote the manuscript. R.K., R.A.-Y. and A.S. designed, synthesized and analyzed the Zn-tripod antigen. R.M. performed the animal experiments. O.D. and H.R. collected X-ray diffraction data, O.D. supervised crystallization experiments and determined the crystal structure. M.E. performed the computational docking analysis. O.B. performed pathological analysis of tissue samples. T.S. assisted with figures and experimental design. T.D. assisted and provided technical support with antibody generation. I.S. designed the experiments, analyzed the data and wrote the manuscript.

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Correspondence to Irit Sagi.

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The authors declare no competing financial interests.

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Supplementary Figures 1–10, Supplementary Table 1 and Supplementary Methods (PDF 2463 kb)

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Sela-Passwell, N., Kikkeri, R., Dym, O. et al. Antibodies targeting the catalytic zinc complex of activated matrix metalloproteinases show therapeutic potential. Nat Med 18, 143–147 (2012). https://doi.org/10.1038/nm.2582

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