High-resolution inhibition profiling and ligand fishing for screening of nucleoside hydrolase ligands in Moringa oleifera Lamarck

Rachel A de Faria; Pamella C O Oliveira; Mariana D P de Carvalho; Bruno S Peixoto; Vanessa G P Severino; Luzineide W Tinoco; Silvana V Rodrigues; Marcela C de Moraes

doi:10.1016/j.jpba.2022.114614

High-resolution inhibition profiling and ligand fishing for screening of nucleoside hydrolase ligands in Moringa oleifera Lamarck

J Pharm Biomed Anal. 2022 Mar 20:211:114614. doi: 10.1016/j.jpba.2022.114614. Epub 2022 Jan 29.

Authors

Rachel A de Faria¹, Pamella C O Oliveira¹, Mariana D P de Carvalho¹, Bruno S Peixoto¹, Vanessa G P Severino², Luzineide W Tinoco³, Silvana V Rodrigues⁴, Marcela C de Moraes⁵

Affiliations

¹ BioCrom, Organic Chemistry Department, Institute of Chemistry, Fluminense Federal University, Niterói, RJ, Brazil.
² Institute of Chemistry, Federal University of Goiás, Campus Samambaia, Esperança avenue, 74690900 Goiânia, Brazil.
³ Laboratory for Analysis and Development of Enzyme Inhibitors, Natural Products Research Institute, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ, Brazil.
⁴ Analytical Chemistry Department, Institute of Chemistry, Fluminense Federal University, Niterói, RJ, Brazil.
⁵ BioCrom, Organic Chemistry Department, Institute of Chemistry, Fluminense Federal University, Niterói, RJ, Brazil. Electronic address: mcmoraes@id.uff.br.

PMID: 35123329
DOI: 10.1016/j.jpba.2022.114614

Abstract

In Leishmania donovani, the causative protozoan of visceral leishmaniasis, nucleoside hydrolase enzyme (NH) is fundamental for the biosynthesis of its DNA and RNA. Therefore, LdNH is considered a potential target for the development of new leishmaniasis chemotherapy. Moringa oleifera Lamarck is a medicinal plant native to northeastern India with numerous pharmacological properties, including antileishmanial activity. Thus, this study aimed to explore the inhibitory activity of different extracts from M. oleifera leaves and flowers on LdNH. Using LdNH covalently immobilized on magnetic particles (LdNH-MPs), a novel activity assay was developed based on the direct quantification of the formed product by HPLC-DAD. This study screened 12 extracts from leaves and flowers of M. oleifera using different extraction methods. The hydroethanolic (70% ethanol) extract from flowers, obtained by infusion (FIEH) or ultrasound-assisted extraction (FUEH), exhibited respectively IC₅₀ values of 26.2 ± 4.63 µg/mL and 4.96 ± 0.52 µg/mL. The most promising extract (FUEH) was investigated by high-resolution LdNH inhibition profiling, which showed different regions of inhibition in the biochromatogram. A ligand fishing assay was attempted to pinpoint the bioactive compounds. Experimental conditions employed in the elution step of the ligand fishing assay did not result in ligands isolation. However, the analyses of the crude extract solution and the supernatants after the incubation with the active and inactive LdNH-MPs indicated missing peaks referring to compounds selectively retained in the active LdNH-MPs incubation. The missing peaks eluted in the same region that exhibits inhibition in the high-resolution LdNH inhibition profiling. The ligands were identified by UHPLC-MS/MS as palatinose, adenosine, 3-p-coumaroylquinic acid, 4-p-coumaroylquinic acid, hyperoside, quercetin-3-O-malonyl glycoside, and kaempferol-3-O-galactoside.

Keywords: High-resolution inhibition profiling; Leishmaniasis; Ligand fishing; Moringa; Nucleoside hydrolase; Screening assays.

MeSH terms

Ligands
Moringa oleifera*
N-Glycosyl Hydrolases
Plant Extracts / analysis
Plant Leaves / chemistry
Tandem Mass Spectrometry

Substances

Ligands
Plant Extracts
N-Glycosyl Hydrolases