An Inhibitory Antibody against Dipeptidyl Peptidase IV Improves Glucose Tolerance in Vivo

Tang, Jie; Majeti, Jiangwen Z.; Sudom, Athena; Xiong, Ye; Lü, Mei; Liu, Qiang; Higbee, Jared; Zhang, Yi; Wang, Yan; Wang, Wei; Cao, Ping; Xia, Zhengyuan; Johnstone, Sheree; Min, Xiaoshan; Yang, Xiaoping; Shao, Hui; Yu, Timothy; Sharkov, Nik; Walker, Nigel; Tu, Hua; Shen, Wenyan David; Wang, Zhong Lin

doi:10.1074/jbc.m112.396317

Cited by 7 publications

(12 citation statements)

References 42 publications

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“…DPP8 has D134 and Y135 in the same positions, which are disordered and not visible either in the unliganded crystal structure or in the peptide-complex structure. This segment offers itself as an epitope for antibodies with specific inhibitory properties in a similar approach as for DPP4 (47).…”

Section: Discussionmentioning

confidence: 99%

Structures and mechanism of dipeptidyl peptidases 8 and 9, important players in cellular homeostasis and cancer

Ross

Krapp²,

Augustin³

et al. 2018

Proc. Natl. Acad. Sci. U.S.A.

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Dipeptidyl peptidases 8 and 9 are intracellular N-terminal dipeptidyl peptidases (preferentially postproline) associated with pathophysiological roles in immune response and cancer biology. While the DPP family member DPP4 is extensively characterized in molecular terms as a validated therapeutic target of type II diabetes, experimental 3D structures and ligand-/substrate-binding modes of DPP8 and DPP9 have not been reported. In this study we describe crystal and molecular structures of human DPP8 (2.5 Å) and DPP9 (3.0 Å) unliganded and complexed with a noncanonical substrate and a small molecule inhibitor, respectively. Similar to DPP4, DPP8 and DPP9 molecules consist of one β-propeller and α/β hydrolase domain, forming a functional homodimer. However, they differ extensively in the ligand binding site structure. In intriguing contrast to DPP4, where liganded and unliganded forms are closely similar, ligand binding to DPP8/9 induces an extensive rearrangement at the active site through a disorder-order transition of a 26-residue loop segment, which partially folds into an α-helix (R-helix), including R160/133, a key residue for substrate binding. As vestiges of this helix are also seen in one of the copies of the unliganded form, conformational selection may contributes to ligand binding. Molecular dynamics simulations support increased flexibility of the R-helix in the unliganded state. Consistently, enzyme kinetics assays reveal a cooperative allosteric mechanism. DPP8 and DPP9 are closely similar and display few opportunities for targeted ligand design. However, extensive differences from DPP4 provide multiple cues for specific inhibitor design and development of the DPP family members as therapeutic targets or antitargets.

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Section: Discussionmentioning

confidence: 99%

Structures and mechanism of dipeptidyl peptidases 8 and 9, important players in cellular homeostasis and cancer

Ross

Krapp²,

Augustin³

et al. 2018

Proc. Natl. Acad. Sci. U.S.A.

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“…Preparation of IgG and Fab. mAb variants were expressed and purified as previously described 11 . Briefly, mAbs were expressed in 293 6E cells and captured on MabSelect SuRe columns, which had been equilibrated with HEPES buffered saline solution HBS (25 mM HEPES, 150 mM NaCl, pH 7.6).…”

Section: Methodsmentioning

confidence: 99%

“…Protein reagents and crystallography. Rat DPP-IV enzyme and Fab mutant proteins were expressed and purified as previously described 11 . The purified DPP-IV/Fab 11A19-sitagliptin hybrid complex was concentrated to 6.6 mg/ml in buffer containing 20 mM Tris 7.9 and 200 mM NaCl.…”

Section: Conjugation Of Dpp-iv Inhibitors To Engineered N30c 11a19 Famentioning

confidence: 99%

Structure-guided discovery of dual recognition chemibodies

Cheng¹,

Doherty²,

Johnstone³

et al. 2019

Acta Cryst Sect A

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Small molecules and antibodies each have advantages and limitations as therapeutics.Here, we present for the first time to our knowledge, the structure-guided design of "chemibodies" as small molecule-antibody hybrids that offer dual recognition of a single target by both a small molecule and an antibody, using DPP-IV enzyme as a proof of concept study. Biochemical characterization demonstrates that the chemibodies present superior DPP-IV inhibition compared to either small molecule or antibody component alone. We validated our design by successfully solving a co-crystal structure of a chemibody in complex with DPP-IV, confirming specific binding of the small molecule portion at the interior catalytic site and the Fab portion at the protein surface. The discovery of chemibodies presents considerable potential for novel therapeutics that harness the power of both small molecule and antibody modalities to achieve superior specificity, potency, and pharmacokinetic properties. Published: xx xx xxxx OPEN www.nature.com/scientificreports/ 2 ScIenTIfIc REPORTS | (2018) 8:7570 |

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“…We previously reported discovery of a panel of mouse monoclonal antibodies (mAbs) against rat DPP-IV that could complement known competitive small molecule inhibitors 11 . These inhibitory antibodies to rat DPP-IV showed notable in vivo efficacy in hyperglycemic Zucker fatty rats in terms of improving glucose tolerance.…”

Section: Introductionmentioning

confidence: 99%

“…To understand the molecular mechanism of the partial inhibition of the 11A19 antibody, we previously solved co-crystal structures of 11A19 Fab with DPP-IV catalytic domain (PDB code: 4FFV), which showed that the Fab does not access the catalytic site, but instead partially blocks the “side” opening, which is believed to be one of the entries to the catalytic site 11 . Interestingly, the structural data also revealed a possibility for small molecules to access the active site in the presence of the blocking antibody, 11A19.…”

Section: Introductionmentioning

confidence: 99%

Structure-guided Discovery of Dual-recognition Chemibodies

Cheng

Doherty

Johnstone

et al. 2018

Sci Rep

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Small molecules and antibodies each have advantages and limitations as therapeutics. Here, we present for the first time to our knowledge, the structure-guided design of “chemibodies” as small molecule-antibody hybrids that offer dual recognition of a single target by both a small molecule and an antibody, using DPP-IV enzyme as a proof of concept study. Biochemical characterization demonstrates that the chemibodies present superior DPP-IV inhibition compared to either small molecule or antibody component alone. We validated our design by successfully solving a co-crystal structure of a chemibody in complex with DPP-IV, confirming specific binding of the small molecule portion at the interior catalytic site and the Fab portion at the protein surface. The discovery of chemibodies presents considerable potential for novel therapeutics that harness the power of both small molecule and antibody modalities to achieve superior specificity, potency, and pharmacokinetic properties.

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An Inhibitory Antibody against Dipeptidyl Peptidase IV Improves Glucose Tolerance in Vivo

Cited by 7 publications

References 42 publications

Structures and mechanism of dipeptidyl peptidases 8 and 9, important players in cellular homeostasis and cancer

Structures and mechanism of dipeptidyl peptidases 8 and 9, important players in cellular homeostasis and cancer

Structure-guided discovery of dual recognition chemibodies

Structure-guided Discovery of Dual-recognition Chemibodies

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