SFN0011: a Novel Potent Inhibitor as a Next-generation Anti-angiogenic Molecule
Hamid Latifi Navid1 , Zahra-Soheila Soheili1 *, Mehdi Sadeghi2 , Shahram Samiei3 , Ehsan Ranaei Pirmardan4 , Sepideh Taghizadeh1 , Seyed Shahriar Arab5 , Saman Tajbakhsh6 , Fahimeh Zakeri1
- Department of Molecular Medicine, National Institute of Genetic Engineering and Biotechnology, Tehran, Iran
- Department of Medical Genetics, National Institute for Genetic Engineering and Biotechnology, Tehran, Iran
- Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine, Tehran, Iran
- Molecular Biomarkers Nano-Imaging Laboratory, Brigham and Women's Hospital, Boston, MA, USA
- Department of Biophysics, School of Biological Sciences, Tarbiat Modares University, Tehran
- Bahar Medical Lab Supervisor
Abstract: Current treatment for age-related macular degeneration (AMD) includes intravitreal injection of anti-vascular endothelial growth factors (anti-VEGFs). However, clinical experiences demonstrate that the efficacy of such therapies is restricted due to overlapping and compensatory alternative angiogenic pathways which culminate to escape mechanisms. Here, we introduce a new anti-angiogenic molecule (SFN0011) based on sFLT01 chimeric protein that neutralizes VEGFA, placental growth factor (PLGF) and a third party angiogenic factor simultaneously and reduces the initiation of associated signaling pathways.
Methods: We investigated sFLT01 molecule structural components via bioinformatic tools and got accessed to its amino acid and nucleotide sequences. We augmented the nucleotide sequence of sFLT01ʹs by another genetic syntax, against a nominated antigenic factor. Therefore, we analyzed the secondary and tertiary structures of the cognate tri-specific molecule with modeller and nanome. The best models were applied in docking analysis with cluspro. The cloning process of the construct in the AAV2 vector was performed and the result was confirmed by conventional PCR and restriction enzyme digestion. RNA extraction and culture condition media collection were performed following the transfection of HEK293T cell line by AAV2-SFN0011. Expression of the gene of interest and its protein output was evaluated by real-time PCR and western blotting respectively. To confirm the functional anti-angiogenic potency of the protein, tube formation assay, phospho-receptor assay, and ligand-receptor interaction ELISA were performed.
Results: We designed, constructed, and produced SFN0011 targeting VEGFA, PLGF, and a third party angiogenic factor that could efficiently inhibit tube formation, receptor phosphorylation and ligand-receptor interaction in vitro.
Conclusion: We propose that targeting various angiogenic pathways by SFN0011 may be a fundamental approach in development of the next generation antiangiogenic therapeutic drugs for retinal neovascular diseases.
