*Article* **Syk Inhibitors: New Computational Insights into Their Intraerythrocytic Action in** *Plasmodium falciparum* **Malaria**

**Giuseppe Marchetti 1,\*, Alessandro Dessì 2,\*, Roberto Dallocchio 2, Ioannis Tsamesidis 1,3, Maria Carmina Pau 1, Francesco Michelangelo Turrini 4 and Antonella Pantaleo 1**


Received: 17 July 2020; Accepted: 18 September 2020; Published: 23 September 2020

**Abstract:** Resistance to antimalarial drugs has spread rapidly over the past few decades. The WHO recommends artemisinin-based combination therapies for the treatment of uncomplicated malaria, but unfortunately these approaches are losing their e fficacy in large areas of Southeast Asia. In 2016, artemisinin resistance was confirmed in 5 countries of the Greater Mekong subregion. We focused our study on Syk inhibitors as antimalarial drugs. The Syk protein is present in human erythrocytes, and the membrane of protein band 3 is its major target following activation by oxidant stress. Tyr phosphorylation of band 3 occurs during *P. falciparum* growth, leading to the release of microparticles containing hemicromes and structural weakening of the host cell membrane, simplifying merozoite reinfection. Syk inhibitors block these events by interacting with the Syk protein's catalytic site. We performed in vitro proteomics and in silico studies and compared the results. In vitro studies were based on treatment of the parasite's cellular cultures with di fferent concentrations of Syk inhibitors, while proteomics studies were focused on the Tyr phosphorylation of band 3 by Syk protein with the same concentrations of drugs. In silico studies were based on di fferent molecular modeling approaches in order to analyze and optimize the ligand–protein interactions and obtain the highest e fficacy in vitro. In the presence of Syk inhibitors, we observed a marked decrease of band 3 Tyr phosphorylation according to the increase of the drug's concentration. Our studies could be useful for the structural optimization of these compounds and for the design of novel Syk inhibitors in the future.

**Keywords:** band 3; red blood cells; antimalarial drugs; molecular docking; molecular dynamics
