Amide-functionalized 1,2,4-Triazol-5-amines as Covalent Inhibitors of Blood Coagulation Factor XIIa and Thrombin

Abstract

To counteract thrombosis, new safe and efficient antithrombotics are required. We herein report the design, synthesis, and biological activity of a series of amide-functionalized acylated 1,2,4-triazol-5-amines as selective inhibitors of blood coagulation factor XIIa and thrombin. The introduction of an amide moiety into the main scaffold of 3-aryl aminotriazoles added certain three-dimensional properties to synthesized compounds and allowed them to reach binding sites in FXIIa and thrombin previously unaddressed by non-functionalized 1,2,4-triazol-5-amines. Among synthesized compounds, one quinoxaline-derived aminotriazole bearing N-butylamide moiety inhibited FXIIa with the IC50 value of 28 nM, whereas the N-phenylamide-derived aminotriazole inhibited thrombin with the IC50 value of 41 nM. Performed mass-shift experiments and molecular modeling studies proved the covalent mechanism of FXIIa and thrombin inhibition by synthesized compounds. In plasma coagulation tests, developed aminotriazoles showed anticoagulant properties mainly affecting the intrinsic blood coagulation pathway, activation of which is associated with thrombosis but is negligible for hemostasis.

Microwave-Assisted Synthesis, Structure, and Preliminary Biological Evaluation of Novel 6-Methoxy-5,6-dihydro-5-azapurines

“…We found that no efficient synthesis toward unsymmetrical aminotriazole-based N , N ′-disubstituted formamidines 20 is known. Recently, we reported practical synthetic methods for 1,2,4-triazol-5-amines − that seemed to be suitable starting materials for formamidines 20 . Thereby, we initiated the development of an efficient and universal approach allowing access to desired N , N ′-disubstituted formamidines 20.…”

“…HRMS was performed with a MicrOTOF-QII (Bruker) instrument. The HPLC method to determine the purity of compounds, as well as the yields in method development procedures, is as follows: equipment 1, pump L-7100, degasser L-7614, autosampler L-7200, UV detector L-7400, interface D-7000, data transfer D-line, data acquisition HSMS software (LaChrom, Merck Hitachi); equipment 2, pump LPG-3400SD, degasser DG-1210, autosampler ACC-3000T, UV detector VWD-3400RS, interface Dionex UltiMate 3000, data acquisition Chromeleon 7 (Thermo Fisher Scientific); LiChrospher 60 RPselect B (5 μm) column, LiChroCART 250–4 mm cartridge; flow rate 1.0 mL/min; injection volume 5.0 μL; detection at λ = 210 nm; solvents A (demineralized water with 0.05% (v/v) trifluoroacetic acid) and B (acetonitrile with 0.05% (v/v) trifluoroacetic acid); gradient elution (% A), 0–4 min 90%, 4–29 min gradient from 90 to 0%, 29–31 min 0%, 31–31.5 min gradient from 0 to 90%, and 31.5–40 min 90% . The purity of compounds 21l – q was analyzed without trifluoroacetic acid addition to solvents.…”

“…and B (acetonitrile with 0.05% (v/v) trifluoroacetic acid); gradient elution (% A), 0−4 min 90%, 4−29 min gradient from 90 to 0%, 29−31 min 0%, 31−31.5 min gradient from 0 to 90%, and 31.5−40 min 90%. 42 The purity of compounds 21l−q was analyzed without trifluoroacetic acid addition to solvents. The purity of all test compounds was greater than 95%.…”

New Triazole-Based Potent Inhibitors of Human Factor XIIa as Anticoagulants

“…Very few potent and selective small molecule inhibitors of FXIIa are in development (Figure ). − We previously reported inhibitor “ RA ” as a potent and selective inhibitor of FXIIa . In this study, we combined a small library of 32 triazole and triazole-like analogs of inhibitor RA to be evaluated for FXIIa inhibition using a chromogenic substrate hydrolysis assay under physiological conditions.…”

An automated analysis method enabling the screening of covalent thrombin and factor XIIa inhibitors via liquid chromatography‐mass spectrometry

“…2,3 As this formed thrombus blocks the blood supply to important organs completely or partially, thrombosis can trigger major cardiovascular disorders, which represent leading global causes of mortality. 1,4,5 Conventionally, anticoagulants are used to prevent thrombosis. 6 As anticoagulants also prevent blood coagulation when it is desired, an increased risk for internal bleeding is a major disadvantage of an anticoagulant application.…”