Molecular Modeling Drug Screening Platform (MMDSP): A model preclinical and molecular insights for the development of new antilipidemic drug targeting PCSK9

• Grant number: 23/15933-4
• Support Opportunities: Research Grants - Young Investigators Grants
• Start date: January 01, 2026
• End date: December 31, 2030
• Field of knowledge: Health Sciences - Pharmacy
• Principal Investigator: Glaucio Monteiro Ferreira
• Grantee: Glaucio Monteiro Ferreira
• Host Institution: Instituto de Química (IQ). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• City of the host institution: São Paulo
• Associated researchers: Alcindo Aparecido dos Santos ; Mario Hiroyuki Hirata

Abstract

The main cause of reduced adherence of treatment of hypercholesterolemia are related with a side effect of statins causing muscle pain. Consequently, the effective treatment efficacy by statins is compromised. Association of statin with ezetimibe, lowing amount of statin is another pharmacological alternative to treat this disease. Lately with description of another mechanism of hypercholesterolemia with gain of function of PCSK9, responsible to control LDLr recicling in the cells, can be an important mechanism to increase cholesterol in plasma. Also, the variants in Apo B gene can be responsible to hypercholesterolemia. Inhibiting the activity of the PCSK9 enzyme cause decrease, plasma cholesterol concentration. This process is effective using monoclonal antibody against this protein as shown in many research. However, the cost effective using this treatment is very high and not useful for large amount of patients and useful for Brazilian Health system (SUS). This research proposal aimed to try identifying potential drug candidates through Structure-Based Virtual Screening (SBVS), selecting compounds with low cytotoxicity and high potency for PCSK9 inhibition. Previous selected compounds using SBVS were evaluated the efficacy using immunochemical method. Additionally, a Thermal Shift Assay (TSA) was performed using SYPRO Orange® as the fluorophore to calculate the melting temperature (Tm) data. Previous results showed by virtual screening two highly active ligands, namely LBMAD10 and LBMAD13, which had PCSK9 potent inhibition with IC50 values of 32.5 nM and 31.7 nM, respectively. Molecular dynamics simulations demonstrated strong interactions between these ligands with PCSK9, confirming their effective potential as inhibitors. Our study shown specifically, LBMAD10 interaction with Asp367, while LBMAD13 interacted with Trp156 in the P'Helix region of the PCSK9, impacting a negatively charged well and low LDL receptor recognition, respectively. Principal Component Analysis (PCA) further supported the inhibitory effects of the compounds on PCSK9. To confirm the potential benefit of these compound we propose evaluated in vitro and preclinical study using OMICS tool to find out metabolic influence of this activity is safety. Studies of genotoxicity, hepatotoxicity, nephrotoxicity, neurotoxicity, cardiotoxicity, and alteration in lung and immune functions. In conclusion, this study will evaluate the efficacy and applicability of 10 compounds, based in previous results of most active compounds, LBMAD10 and LBMAD13. These results highlight the potential of these process to be useful and minimize time consuming and warrant further evaluation in preclinical tests. (AU)