HDX-MS study on garadacimab binding to activated FXII reveals potential binding interfaces through differential solvent exposure

Ow, Saw Yen; Kapp, Eugene A.; Tomasetig, Vesna; Zalewski, Anton; Simmonds, Jason; Panousis, Con; Wilson, Michael J.; Nash, Andrew D.; Pelzing, Matthias

doi:10.1080/19420862.2022.2163459

Cited by 5 publications

(5 citation statements)

References 42 publications

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“…Consistent with the previous HDX-MS data (Ow et al, 2023), garadacimab inhibits βFXIIa by binding to the βFXIIa loops neighbouring the S1 catalytic pocket entrance and preventing substrate binding through steric hindrance. The binding of the garadacimab Fab to βFXIIa buries a surface area of ∼1054 Å 2 , with ∼700 Å 2 from the heavy and ∼355 Å 2 from the light chain.…”

Section: Resultssupporting

confidence: 91%

“…Specifically, we demonstrate that the interfaces neighbouring the S1 catalytic pocket entrance (e.g. 60-loop, 99-loop, 180-loop and 220-loop) overlapped with regions shown to be blocked by the CDR-H3, CDR-H2 and CDR-L1 of garadacimab (Ow et al, 2023).…”

Section: Discussionmentioning

confidence: 89%

“…The interactions mapped using the cryo-EM structure of Garadacimab Fab-βFXIIa were aligned against binding interfaces identified in our recent report using HDX-MS (Ow et al, 2023). Highlighted HDX-MS interfaces on βFXIIa show very good agreement with contact regions observed in the cryo-EM structure.…”

Section: Discussionmentioning

confidence: 99%

“…This provided evidence that garadacimab may in fact obstruct entry of the βFXIIa S1 pocket (Ow et al, 2023). Through solvent accessibility data coupled with HDX rate, we could highlight putative residues forming the binding interface.…”

Section: Introductionmentioning

confidence: 94%

“…Simultaneously, a hydrogen-deuterium exchange mass spectrometry (HDX-MS) study to elucidate the potential binding interface of garadacimab on βFXIIa was initiated. This provided evidence that garadacimab may in fact obstruct entry of the βFXIIa S1 pocket (Ow et al, 2023). Through solvent accessibility data coupled with HDX rate, we could highlight putative residues forming the binding interface.…”

Section: Introductionmentioning

confidence: 99%

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Structural basis for inhibition of βFXIIa by garadacimab

Drulyte,

Ghai,

et al. 2023

Preprint

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FXIIa is the principal initiator of the plasma contact system and can activate both procoagulant and proinflammatory pathways. Its activity is important in the pathophysiology of Hereditary Angioedema (HAE). Here, we describe a high-resolution cryo-EM structure of the beta-chain from FXIIa (βFXIIa) complexed with the Fab fragment of garadacimab. Garadacimab binds to βFXIIa through an unusually long CDR-H3 that inserts into the S1 pocket in a non-canonical way. This atypical structural mechanism is likely the primary contributor in the inhibition of activated FXII proteolytic activity in HAE. Garadacimab Fab-βFXIIa structure also reveals critical determinants of high-affinity binding of garadacimab to activated FXII. Structural analysis with other bonafide FXIIa inhibitors, such as benzamidine and C1-INH, reveals a conserved mechanism of βFXIIa inhibition, a novel finding of this study. In summary, garadacimab Fab-βFXIIa structure provides crucial insights into its mechanism of action and delineates primary and auxiliary paratopes/epitopes. This work reaffirms the benefits of cryo-EM as a primary tool to study antigen-antibody complexes in near native state, particularly where other methods have fallen short.

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Section: Resultssupporting

confidence: 91%

Section: Discussionmentioning

confidence: 89%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 94%

Section: Introductionmentioning

confidence: 99%

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Structural basis for inhibition of βFXIIa by garadacimab

Drulyte,

Ghai,

et al. 2023

Preprint

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Pharmacokinetics, Pharmacodynamics, and Safety of Subcutaneous and Intravenous Garadacimab Following Single‐Dose Administration in Healthy Japanese and White Adults

Glassman,

Lawo,

Bica

et al. 2024

The Journal of Clinical Pharma

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Garadacimab, an activated factor XII (FXIIa) inhibitor monoclonal antibody, is being evaluated for the long‐term prophylaxis of hereditary angioedema. Here, we report the results from a two‐part, phase 1, open‐label, single ascending dose study assessing the pharmacokinetics (PK), pharmacodynamics, safety, and tolerability after subcutaneous (SC) and intravenous (IV) administration of garadacimab in healthy Japanese and White participants. Part 1 assessed garadacimab PK after SC administration of a 200 mg dose in weight‐matched White and Japanese participants, and 600 mg dose in Japanese participants. Part 2 assessed 3 and 10 mg/kg IV doses in Japanese participants. Follow‐up for safety was over 84 days post‐dose. Overall, 37 participants received garadacimab dosing and 36 completed the study, with one participant lost to follow‐up. Following SC administration, time to maximum plasma concentration (tmax) occurred at 7 days post‐dose, and garadacimab exposure, based on maximum plasma concentration (Cmax) and area under the plasma concentration–time curve (AUC), increased less than 3‐fold when tripling the dose. PK was comparable between Japanese and White participants, with geometric mean ratios for Cmax and AUC close to 100%. Following IV administration, tmax occurred at the end of infusion, and garadacimab exposure increased in a dose‐proportional manner. Inhibition of FXIIa‐mediated kallikrein activity versus baseline was observed in all participants receiving the SC and IV doses. No anti‐drug antibodies against garadacimab were reported. Consistent with pivotal phase 3 (VANGUARD) outcomes, no safety concerns and no difference in the safety profile of garadacimab were observed between healthy Japanese and White participants.

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Structural basis for the inhibition of βFXIIa by garadacimab

Drulyte,

Ghai,

et al. 2024

Structure

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HDX-MS study on garadacimab binding to activated FXII reveals potential binding interfaces through differential solvent exposure

Cited by 5 publications

References 42 publications

Structural basis for inhibition of βFXIIa by garadacimab

Structural basis for inhibition of βFXIIa by garadacimab

Pharmacokinetics, Pharmacodynamics, and Safety of Subcutaneous and Intravenous Garadacimab Following Single‐Dose Administration in Healthy Japanese and White Adults

Structural basis for the inhibition of βFXIIa by garadacimab

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