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Cyclic peptide analogs of 558–565 epitope of A2 subunit of Factor VIII prolong aPTT. Toward a novel synthesis of anticoagulants

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Abstract

Novel anticoagulant therapies target specific clotting factors in blood coagulation cascade. Inhibition of the blood coagulation through Factor VIII–Factor IX interaction represents an attractive approach for the treatment and prevention of diseases caused by thrombosis. Our research efforts are continued by the synthesis and biological evaluation of cyclic, head to tail peptides, analogs of the 558–565 sequence of the A2 subunit of FVIII, aiming at the efficient inhibition of Factor VIIIa–Factor IXa interaction. The analogs were synthesized on solid phase using the acid labile 2-chlorotrityl chloride resin, while their anticoagulant activities were examined in vitro by monitoring activated partial thromboplastin time and the inhibition of Factor VIII activity. The results reveal that these peptides provide bases for the development of new anticoagulant agents.

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Abbreviations

AcOH:

Acetic acid

aPTT:

Activated partial thromboplastin time

Boc:

tert-Butoxycarbonyl

Bzl:

Benzyl group

CLTR-Cl:

2-Chlorotrityl chloride resin

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

PPP:

Platelet poor plasma

PyBOP:

Benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate

PT:

Prothrombin time

rFVIII:

Recombinant factor VIII

But :

tert-Butyl group

RP-HPLC:

Reversed-phase high-performance liquid chromatography

TBTU:

2-(1H-Benzotriazole-1-yl)-1,1,3,3-tetramethylammonium tetrafluoroborate

TES:

Triethylsilane

TFA:

Trifluoroacetic acid

TFE:

2,2,2-Trifluoroethanol

TLC:

Thin layer chromatography

Tol:

Toluene

Trt:

Trityl, triphenylmethyl group

vWF:

Von Willebrand factor

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Acknowledgments

Anticoagulant assays were performed under the supervision of Dr. Emmanouil A., University Hospital of Rio 26504, Patras, Greece. This Research Project is co-financed: 80 % by European Union—European Social Fund and 20 % by General Secretary of Research & Technology (PENED 03ED569).

Conflict of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Department of Chemistry, University of Patras, Patras, Greece

    C. Anastasopoulos, Y. Sarigiannis & G. Stavropoulos

Authors
  1. C. Anastasopoulos
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  2. Y. Sarigiannis
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  3. G. Stavropoulos
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Corresponding author

Correspondence to G. Stavropoulos.

Additional information

Abbreviations of common amino acids are in accordance with the recommendations of IUPAC-IUB Joint Commission on Biochemical Nomenclature: Arch Biochem Biophys 206 (1988) v–xxii, J Biol Chem 264 (1989) 668–673, J Peptide Sci 12 (2006) 1–8, Amino Acids, Pept. Proteins, (2012) 37, xi–xviii.

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Anastasopoulos, C., Sarigiannis, Y. & Stavropoulos, G. Cyclic peptide analogs of 558–565 epitope of A2 subunit of Factor VIII prolong aPTT. Toward a novel synthesis of anticoagulants. Amino Acids 46, 1087–1096 (2014). https://doi.org/10.1007/s00726-014-1673-7

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  • DOI: https://doi.org/10.1007/s00726-014-1673-7

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