Engineered factor Xa variants retain procoagulant activity independent of direct factor Xa inhibitors

Daniël Verhoef, Koen M Visscher, C Ruben Vosmeer, Ka Lei Cheung, Pieter H Reitsma, Daan P Geerke, Mettine H A Bos

Research output: Scientific - peer-reviewArticle

Abstract

The absence of an adequate reversal strategy to prevent and stop potential life-threatening bleeding complications is a major drawback to the clinical use of the direct oral inhibitors of blood coagulation factor Xa. Here we show that specific modifications of the substrate-binding aromatic S4 subpocket within the factor Xa active site disrupt high-affinity engagement of the direct factor Xa inhibitors. These modifications either entail amino-acid substitution of S4 subsite residues Tyr99 and/or Phe174 (chymotrypsinogen numbering), or extension of the 99-loop that borders the S4 subsite. The latter modifications led to the engineering of a factor Xa variant that is able to support coagulation in human plasma spiked with (supra-)physiological concentrations of direct factor Xa inhibitors. As such, this factor Xa variant has the potential to be employed to bypass the direct factor Xa inhibitor-mediated anticoagulation in patients that require restoration of blood coagulation.A major drawback in the clinical use of the oral anticoagulants that directly inhibit factor Xa in order to prevent blood clot formation is the potential for life threatening bleeding events. Here the authors describe factor Xa variants that are refractory to inhibition by these anticoagulants and could serve as rescue agents in treated patients.

Original languageEnglish
Pages (from-to)528
JournalNature Communications
Volume8
Issue number1
DOIs
StatePublished - 13 Sep 2017

Cite this

Verhoef, Daniël; Visscher, Koen M; Vosmeer, C Ruben; Cheung, Ka Lei; Reitsma, Pieter H; Geerke, Daan P; Bos, Mettine H A / Engineered factor Xa variants retain procoagulant activity independent of direct factor Xa inhibitors.

In: Nature Communications, Vol. 8, No. 1, 13.09.2017, p. 528.

Research output: Scientific - peer-reviewArticle

@article{7c3a9ffce611476e8fefadd0e06515a4,
title = "Engineered factor Xa variants retain procoagulant activity independent of direct factor Xa inhibitors",
abstract = "The absence of an adequate reversal strategy to prevent and stop potential life-threatening bleeding complications is a major drawback to the clinical use of the direct oral inhibitors of blood coagulation factor Xa. Here we show that specific modifications of the substrate-binding aromatic S4 subpocket within the factor Xa active site disrupt high-affinity engagement of the direct factor Xa inhibitors. These modifications either entail amino-acid substitution of S4 subsite residues Tyr99 and/or Phe174 (chymotrypsinogen numbering), or extension of the 99-loop that borders the S4 subsite. The latter modifications led to the engineering of a factor Xa variant that is able to support coagulation in human plasma spiked with (supra-)physiological concentrations of direct factor Xa inhibitors. As such, this factor Xa variant has the potential to be employed to bypass the direct factor Xa inhibitor-mediated anticoagulation in patients that require restoration of blood coagulation.A major drawback in the clinical use of the oral anticoagulants that directly inhibit factor Xa in order to prevent blood clot formation is the potential for life threatening bleeding events. Here the authors describe factor Xa variants that are refractory to inhibition by these anticoagulants and could serve as rescue agents in treated patients.",
keywords = "Journal Article",
author = "Daniël Verhoef and Visscher, {Koen M} and Vosmeer, {C Ruben} and Cheung, {Ka Lei} and Reitsma, {Pieter H} and Geerke, {Daan P} and Bos, {Mettine H A}",
year = "2017",
month = "9",
doi = "10.1038/s41467-017-00647-9",
volume = "8",
pages = "528",
journal = "Nature Communications",
issn = "2041-1723",
publisher = "Nature Publishing Group",
number = "1",

}

Engineered factor Xa variants retain procoagulant activity independent of direct factor Xa inhibitors. / Verhoef, Daniël; Visscher, Koen M; Vosmeer, C Ruben; Cheung, Ka Lei; Reitsma, Pieter H; Geerke, Daan P; Bos, Mettine H A.

In: Nature Communications, Vol. 8, No. 1, 13.09.2017, p. 528.

Research output: Scientific - peer-reviewArticle

TY - JOUR

T1 - Engineered factor Xa variants retain procoagulant activity independent of direct factor Xa inhibitors

AU - Verhoef,Daniël

AU - Visscher,Koen M

AU - Vosmeer,C Ruben

AU - Cheung,Ka Lei

AU - Reitsma,Pieter H

AU - Geerke,Daan P

AU - Bos,Mettine H A

PY - 2017/9/13

Y1 - 2017/9/13

N2 - The absence of an adequate reversal strategy to prevent and stop potential life-threatening bleeding complications is a major drawback to the clinical use of the direct oral inhibitors of blood coagulation factor Xa. Here we show that specific modifications of the substrate-binding aromatic S4 subpocket within the factor Xa active site disrupt high-affinity engagement of the direct factor Xa inhibitors. These modifications either entail amino-acid substitution of S4 subsite residues Tyr99 and/or Phe174 (chymotrypsinogen numbering), or extension of the 99-loop that borders the S4 subsite. The latter modifications led to the engineering of a factor Xa variant that is able to support coagulation in human plasma spiked with (supra-)physiological concentrations of direct factor Xa inhibitors. As such, this factor Xa variant has the potential to be employed to bypass the direct factor Xa inhibitor-mediated anticoagulation in patients that require restoration of blood coagulation.A major drawback in the clinical use of the oral anticoagulants that directly inhibit factor Xa in order to prevent blood clot formation is the potential for life threatening bleeding events. Here the authors describe factor Xa variants that are refractory to inhibition by these anticoagulants and could serve as rescue agents in treated patients.

AB - The absence of an adequate reversal strategy to prevent and stop potential life-threatening bleeding complications is a major drawback to the clinical use of the direct oral inhibitors of blood coagulation factor Xa. Here we show that specific modifications of the substrate-binding aromatic S4 subpocket within the factor Xa active site disrupt high-affinity engagement of the direct factor Xa inhibitors. These modifications either entail amino-acid substitution of S4 subsite residues Tyr99 and/or Phe174 (chymotrypsinogen numbering), or extension of the 99-loop that borders the S4 subsite. The latter modifications led to the engineering of a factor Xa variant that is able to support coagulation in human plasma spiked with (supra-)physiological concentrations of direct factor Xa inhibitors. As such, this factor Xa variant has the potential to be employed to bypass the direct factor Xa inhibitor-mediated anticoagulation in patients that require restoration of blood coagulation.A major drawback in the clinical use of the oral anticoagulants that directly inhibit factor Xa in order to prevent blood clot formation is the potential for life threatening bleeding events. Here the authors describe factor Xa variants that are refractory to inhibition by these anticoagulants and could serve as rescue agents in treated patients.

KW - Journal Article

U2 - 10.1038/s41467-017-00647-9

DO - 10.1038/s41467-017-00647-9

M3 - Article

VL - 8

SP - 528

JO - Nature Communications

T2 - Nature Communications

JF - Nature Communications

SN - 2041-1723

IS - 1

ER -