Amino Acids

, Volume 44, Issue 4, pp 1159–1165 | Cite as

A novel approach in potential anticoagulants from peptides epitope 558–565 of A2 subunit of factor VIII

  • C. Anastasopoulos
  • Y. Sarigiannis
  • G. Stavropoulos
Original Article

Abstract

Factor VIII, a human blood plasma protein, plays an important role during the intrinsic pathway of blood coagulation cascade after its activation by thrombin. The activated form of FVIII acts as cofactor to the serine protease Factor IXa, in the conversion of the zymogen Factor X to the active enzyme Factor Xa. The Ser558–Gln565 region of the A2 subunit of Factor VIII has been shown to be crucial for FVIIIa–FIXa interaction. Based on this, a series of linear peptides, analogs of the 558–565 loop of the A2 subunit of the heavy chain of Factor VIII were synthesized using the acid labile 2-chlorotrityl chloride resin and biologically evaluated in vitro by measuring the chronic delay of activated partial thromboplastin time and the inhibition of Factor VIII activity, as potential anticoagulants.

Keywords

Factor VIII Anticoagulant activity Activated partial thromboplastin time Synthetic peptides Blood coagulation cascade FVIII-FIX interaction 

Abbreviations

AcOH

Acetic acid

aPTT

Activated partial thromboplastin time

Boc

Tert-butoxycarbonyl

Bzl

Benzyl group

CLTR-Cl

2-chlorotrityl chloride

DCM

Dichloromethane

DIC

N,N′-diisopropylcarbodiimide

DIPEA

Diisopropylethylamine

DMF

N,N′-dimethylformamide

dPPP

Deficient platelet poor plasma

ESI-MS

Electrospray ionization mass spectrometry

Fmoc

9-Fluorenylmethyloxycarbonyl group

FVIII

Factor VIII

FIX

Factor IX

HOBt

1-hydroxybenzotriazole

i-PrOH

2-propanol

Me

Methyl group

MeCN

Acetonitrile

MeOH

Methanol

MBHA

4-methylbenzhydrylamine

PPP

Platelet poor plasma

PyBOP

(Benzotriazol-1-yloxy)-tris(pyrrolidino)phosphonium hexafluorophosphate

PT

Prothrombin time

rFVIII

Recombinant Factor VIII

tBu

Tert-butyl group

RP-HPLC

Reversed phase high performance liquid chromatography

TES

Triethylsilane

TFA

Trifluoroacetic acid

TFE

2,2,2-trifluoroethanol

TLC

Thin layer chromatography

Tol

Toluene

Trt

Trityl group

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Copyright information

© Springer-Verlag Wien 2013

Authors and Affiliations

  • C. Anastasopoulos
    • 1
  • Y. Sarigiannis
    • 1
  • G. Stavropoulos
    • 1
  1. 1.Department of ChemistryUniversity of PatrasPatrasGreece

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