Non-cytotoxic 1,2,3-triazole tethered fused heterocyclic ring derivatives display Tax protein inhibition and impair HTLV-1 infected cells

Abstract

Human T cell lymphotropic virus type 1 (HTLV-1) is a human retrovirus that infects approximately 10-20 million people worldwide and causes an aggressive neoplasia (adult T-cell leukemia/lymphoma -ATL). Therapeutic approaches for the treatment of ATL have variable effectiveness and poor prognosis, thus requiring strategies to identify novel compounds with activity on infected cells. In this sense, we initially screened a small series of 25 1,2,3-triazole derivatives to discover cell proliferation inhibitors and apoptosis inducers in HTLV-1-infected T-cell line (MT-2) for further assessment of their effect on viral tax activity through inducible-tax reporter cell line (Jurkat LTR-GFP).Eight promising compounds (02, 05, 06, 13, 15, 21, 22 and 25) with activity ≥ 70% were initially selected, based on a suitable cell-based assay using resazurin reduction method, and evaluated towards cell cycle, apoptosis and Tax/GFP expression analyses through flow cytometry. Compound 02 induced S phase cell cycle arrest and compounds 05, 06, 22 and 25 promoted apoptosis. Remarkably, compounds 22 and 25 were also able to reduce GFP expression in an inducible-tax reporter cell, which suggests an effect on Tax viral protein. More importantly, compounds 02, 22 and 25 were not cytotoxic in human hepatoma cell line (Huh-7). Therefore, the discovery of 3 active and non-cytotoxic compounds against HTLV-1-infected cells can potentially contribute, as an initial promising strategy, to the development process of new drugs against ATL.

New Perspectives about Drug Candidates Targeting HTLV-1 and Related Diseases

“…They could decrease the metabolic activity of the HTLV-1-infected cell line (MT-2), promoting the significant activation of effector caspase-3/7 compared to the control, and/or interfering in LTR transactivation and/or Tax expression (Table 1). The compounds can potentially contribute as a drug for ATLL patients [22].…”

Identificação e caracterização funcional de compostos sintéticos indutores do efeito imunomodulador em células estromais mesenquimais

“…Para moduladores de mecanismos biológicos, por exemplo, utiliza-se técnicas relacionados à colorimetria, absorbância, luminescência e fluorescência em ensaios direcionados a expressão gênica, interação receptorligante, interações de proteínas ou ensaios enzimáticos (ALVES et al, 2011;BARA et al, 2015;BREY et al, 2011;CRUZ et al, 2013;DOS SANTOS et al, 2020;ENTZEROTH;FLOTOW;CONDRON, 2009;HAJARE et al, 2013;HUANG et al, 2008;LO CICERO et al, 2016;WALMSLEY et al, 2015;YANG et al, 2016).…”

“…Ao passar por esta diluição, cada composto apresentou-se a 300 µM e DMSO 3% em uma solução com 66 µL. Adicionalmente, os controles foram também preparados, sendo o controle positivo TNF-ɑ a 600 ng/mL e negativo DMSO a 3%.Esta metodologia de preparo da biblioteca para triagem foi elaborada no artigo de dosSantos et al (2020). raspadas e posteriormente todo o conteúdo do poço foi homogeneizado por pipetagem manual.…”