Estudo comparativo da farmacocinética pré-clínica do LASSBio-1920 livre e encapsulado: perspectivas como agente anticâncer

Abstract

Cancer refers to multiple diseases characterized by the uncontrolled proliferation of cells capable of invading adjacent tissues and causing metastases the leading cause of mortality among cancer patients. According to the National Cancer Institute (INCA), Brazil is expected to register approximately 704,000 new cancer cases in 2025, with the highest incidence of breast, prostate, colon, and rectal cancers. Most deaths occur in low- and middle-income countries, where access to early diagnosis and treatment is still limited. Major risk factors include smoking, excessive alcohol consumption, a sedentary lifestyle, poor diet, and viral infections such as HPV. To reduce mortality, it is essential to develop new therapies that can undergo preclinical (in vitro and in vivo) and clinical studies before being used in humans. LASSBio-1920, a derivative of combretastatin A-4 (CA4), has shown promise in combating tumor cells in vitro. It exhibits potent cytotoxic activity in vitro, with selectivity in tumor cell lines and IC₅₀ values in the nanomolar range. Although its mechanism of action has been elucidated, the molecule has low water solubility. Therefore, the project aimed to develop a nanoformulation of LASSBio-1920 for future in vivo testing. This study aimed to characterize the pharmacokinetic profile of the free molecule and the nanoformulation of LASSbio-1920 through in vitro and in vivo assays. The nanoformulations and quality control of LASSBio-1920 were performed prior to each study. The cellular uptake study in HCT-116 cells was performed using an LC/MS apparatus and flow cytometry to determine intracellular quantification at different time points. The data demonstrated that the nanoformulation allowed maximum quantification of LASSBio-1920 after 2 hours of plating with the nanoformulation. Regarding the in vivo studies, the animals were divided into three groups. The first group received the nanoformulation at a dose equivalent to 2 mg/mg; the second and third groups received doses of 2 mg/kg and 10 mg/kg, respectively, using the free molecule. LASSbio-1920 was then quantified in the blood of these animals, and it was observed that the molecule was eliminated more rapidly when nanoformulated in liposomes compared to the free molecule, presenting lower AUC0-inf and T1/2, as well as a higher CL clearance than the free molecule, factors likely linked to the components of the liposomal nanoformulation. Regarding the free molecule, as administered at two different concentrations, the data possibly indicate linear elimination of the molecule, as the elimination half-life (T1/2) showed similar values at both application doses. Finally, a metabolic cage experiment was performed, applying 2mg/kg of LASSBio-1920 nanoformulation and then dosing this molecule in the urine at different times, showing a decline in concentration over time, suggesting renal and concentration-dependent elimination, as well as dosing in the feces, not showing relevant quantification after 24 hours of collection.