PB0735 - Novel synthetic Nilotinib analogues with improved cytostatic properties and potent antiplatelet activity

Monday, June 26, 2023

18:30 – 19:30 ET

Room: Exhibition

Co-Author(s)

LP

Louisa M. Pechlivani, MSc, PhD candidate

Chemist, PhD Candidate
Atherothrombosis Research Centre/Laboratory of Biochemistry, Department of Chemistry, University of Ioannina, Ioannina, Greece
Ioannina, Ioannina, Greece
NN

Nikoleta Ntemou

Chemist, PhD
Section of Organic Chemistry and Biochemistry, Department of Chemistry, University of Ioannina, Ioannina, Greece
Ioannina, Ioannina, Greece
PV

Pinelopi Voulgari

Chemist, PhD
Section of Organic Chemistry and Biochemistry, Department of Chemistry, University of Ioannina, Ioannina, Greece
Ioannina, Ioannina, Greece
KS

Konstantinos Skobridis

Professor of Organic Chemistry, PhD
Section of Organic Chemistry and Biochemistry, Department of Chemistry, University of Ioannina, Ioannina, Greece
Ioannina, Ioannina, Greece

Presenting Author(s)

Alexandros D. Tselepis, MD, PhD

Emeritus Professor of Biochemistry-Clinical Chemistry, MD, PhD
Atherothrombosis Research Centre/Laboratory of Biochemistry, Department of Chemistry, University of Ioannina, Ioannina, Greece
Ioannina, Ioannina, Greece

Background: Nilotinib is a Bcr-Abl tyrosine kinase inhibitor used to treat various malignancies. We synthesized new inhibitors and demonstrated their antiplatelet and cytostatic activities (Pechlivani L. et al. PB0755. ISTH 2022 Congress).

Aims: In this study we aimed to further evaluate their antiplatelet potency and their profile towards cancer cell functionality by studying the phosphorylation of the Src protein-tyrosine kinase, a proto-oncogene that plays a key role in cell growth and migration.

Methods: Three novel Bcr-Abl tyrosine kinase inhibitors were designed, as we previously described and they differentiate at their final phenyl ring, having a halogen atom, F or Cl at the 4th position (Figure). Light Transmittance Aggregometry and Flow Cytometry in the presence of Nilotinib and the inhibitors was performed in platelet-rich plasma (PRP) activated with Arachidonic acid (AA), in order to evaluate the platelet aggregation and the P-selectin and PAC-1 expression. After the 48h incubations with 10μΜ of the synthetic compounds, confluent hepatoma HepG2 cells, in culture, were lysed and the Src phosphorylation (Tyr416) was evaluated with ELISA.

Results: Compounds 1, 2 and 3 inhibited platelet aggregation induced by AA to a similar extent and the IC50 values were 1.50±1.08μΜ*, 0.94±0.66μΜ* and 1.73±0.73μΜ, respectively, whereas the IC50 value for Nilotinib was 3.94±2.09μΜ (*p < 0.05 compared with Nilotinib). They also exhibited strong inhibitory effect on P-selectin and PAC-1 expression (Table). Nilotinib and the compounds inhibited the phosphorylation of Src by 11.18%, 34.33%, 20.53% and 26.85%, respectively.

Conclusion(s): All compounds exhibited more potent antiplatelet activity, compared to Nilotinib, while the compound-1 demonstrated the strongest inhibitory effect towards the Src phosphorylation. These results indicate considerable potential to develop new inhibitors with potent antiplatelet activity and improved cytostatic properties to treat cancer and effectively prevent cancer-associated thrombosis.