Discovery and Validation of Lmj_04_BRCT Domain, a Novel Therapeutic Target: Identification of Candidate Drugs for Leishmaniasis

Abstract

Since many of the currently available antileishmanial treatments exhibit toxicity, low effectiveness, and resistance, search and validation of new therapeutic targets allowing the development of innovative drugs have become a worldwide priority. This work presents a structure-based drug discovery strategy to validate the Lmj_04_BRCT domain as a novel therapeutic target in Leishmania spp. The structure of this domain was explored using homology modeling, virtual screening, and molecular dynamics studies. Candidate compounds were validated in vitro using promastigotes of Leishmania major, L. amazonensis, and L. infantum, as well as primary mouse macrophages infected with L. major. The novel inhibitor CPE2 emerged as the most active of a group of compounds against Leishmania, being able to significantly reduce the viability of promastigotes. CPE2 was also active against the intracellular forms of the parasites and significantly reduced parasite burden in murine macrophages without exhibiting toxicity in host cells. Furthermore, L. major promastigotes treated with CPE2 showed significant lower expression levels of several genes (α-tubulin, Cyclin CYCA, and Yip1) related to proliferation and treatment resistance. Our in silico and in vitro studies suggest that the Lmj_04_BRCT domain and its here disclosed inhibitors are new potential therapeutic options against leishmaniasis.

Repurposing the Antibacterial Agents Peptide 19-4LF and Peptide 19-2.5 for Treatment of Cutaneous Leishmaniasis

“…We also showed that the treatment of Leishmania-infected macrophages with 19-2.5 and 19-4LF lowered the expression levels of several Leishmania genes implicated in proliferation (Cyclin 1 and Cyclin 6), pathogen virulence (gp63) and treatment resistance (yip1) In line with our results, these genes were similarly downregulated by CPE2, a recently reported leishmanicidal compound [47]. Importantly, our results demonstrated that 19-2.5 and 19-4LF retained their anti-parasitic potential in vivo since they dramatically reduced the parasite burden in the skin lesion as well as in the spleen of infected mice.…”

“…Inhibitors of cyclin-dependent kinases (CDKs) have been reported as suitable candidates for the treatment of leishmaniasis [62][63][64][65]. Recently discovered compounds with leishmanicidal activity have demonstrated the ability to inhibit the expression of such cyclins in Leishmania major [47]. On the other hand, terbinafine resistance locus protein (yip 1) codifying gene, has been detected to be overexpressed in drug-resistant Leishmania strains [42,66,67] Since the peptide 19-2.5 showed activity by inhibiting yip1 gene levels, it might be also considered as a potential candidate against resistant strains.…”

The BRCT Domain from the Homologue of the Oncogene PES1 in Leishmania major (LmjPES) Promotes Malignancy and Drug Resistance in Mammalian Cells

“…Thus, several data confirmed that BRCT domain from PES1 protein exhibited a functional importance in cancer development. In Leishmania , LmjPES harbors the firstly identified BRCT domain in these parasites, which has demonstrated to be a promising therapeutic target [ 39 ].…”

Challenges and Tools for In Vitro Leishmania Exploratory Screening in the Drug Development Process: An Updated Review

“…Other compounds were found to be inactive. This approach was also used by Peña-Guerrero et al, who validated the Lmj_04_BRCT domain as a novel therapeutic target in Leishmania by a structure-based drug discovery strategy, and experimentally validated in vitro a novel inhibitor (CPE2) against L. major, L. amazonensis, and L. infantum (promastigotes and intracellular amastigotes) among seven virtually selected compounds [149].…”