Development of a novel anti-vegfr2, proline/alanine/serine fused adnectin with enhanced pharmacokinetic properties

Safieh Aghaabdollahian,1,* Reza ahangari cohan,2 Dariush norouzian,3 Fatemeh davami,4 Mohammad reza asadi karam,5 Behrouz vaziri,6

1. Department of Nanobiotechnology, Advanced Technology Group, Pasteur Institute of Iran, Tehran, Iran
2. Department of Nanobiotechnology, Advanced Technology Group, Pasteur Institute of Iran, Tehran, Iran
3. Department of Nanobiotechnology, Advanced Technology Group, Pasteur Institute of Iran, Tehran, Iran
4. Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran
5. Department of Molecular Biology, Pasteur Institute of Iran, Tehran, Iran
6. Department of Biotechnology, Pasteur Institute of Iran, Tehran, Iran

Abstract


Introduction

The key pathological role of vegf receptor 2 (vegfr2) signaling in the angiogenesis and metastasis of solid tumors has prompted the development of inhibitors with minimal bystander effects. recently, adnectin c –a protein scaffold derivated from 10th domain of domain of fibronectin type iii- has attracted attention for cancer treatment. to overcome the problematic properties of adnectin, such as its short half-life, a novel form of adnectin c has been designed by its fusion to a biodegredable polymeric peptide containing pro/ala/ser repetitive sequence with a length of 200 residues.

Methods

E. coli-expressed recombinant fused and unfused proteins were compared in terms of particle size, zeta potential, charge, intact mass, and freeze-thaw stability using standard methods. pharmacokinetic profile was assessed after single dose by intravenous injection into female balb/c mice.

Results

Dynamic light scattering analysis of pasylated adnectin c revealed an approximate 2-fold increase in particle size with a slight change in the net charge. additionally, fusion of the pas sequence improved its stability against the freeze-thaw conditions. pharmacokinetic studies in mice have shown an improvement in the pharmacokinetic profile of pas-fused proteins that could be explained by an increase in the hydrodynamic volume of the protein. a remarkable increase in the terminal half-life by a factor of 4.58 of the native protein was observed after pasylation.

Conclusion

The results suggest that pasylation could offer a superior half-life extension strategy for developing adnectin-derived drugs with improved patient compliance.

Keywords

Adnectin; angiogenesis; pasylation; pharmacokinetic; vegfr2