# Uncaria nervosa Elmer, a new herbal source for betulinic acid and ursolic acid: Metabolites profiling, isolation, and in vitro cytotoxicity studies against T47D breast cancer

## TL;DR

This study identifies betulinic acid and ursolic acid in the Indonesian plant Uncaria nervosa Elmer, which show potential as breast cancer treatments.

## Contribution

The first isolation and cytotoxicity evaluation of betulinic acid and ursolic acid from Uncaria nervosa Elmer for breast cancer treatment.

## Key findings

- Betulinic acid and ursolic acid were isolated from Uncaria nervosa Elmer leaves.
- Ursolic acid showed moderate cytotoxic activity against T47D breast cancer cells with an IC50 of 14.70±4.50 μg/ml.
- Nine compounds, including alkaloids and terpenoids, were identified in the ethanol extract of the plant.

## Abstract

Uncaria nervosa Elmer is an Indonesian herbal plant that is traditionally used for breast cancer. The results of phytochemical screening contained alkaloids, flavonoids, and terpenoids in the ethanol extract of this plant. Based on literature searches, reports regarding the bioactive compounds responsible for breast cancer have not been found. Further research is needed to understand the potential of
Uncaria nervosa Elmer as a breast cancer treatment and to identify the specific compounds responsible for its effects

This study aims to determine the metabolite profiling of ethanol extract, the isolation, characterization of bioactive compounds, and their bioactivity in T47D breast cancer cells. The research began by extracting the leaves by maceration using 70% ethanol, and then solid phase extraction was carried out using the solid phase extraction (SPE) method. In this study, the sorbent used was polyamide. The extract was analyzed using a tandem analysis technique based on LCMS using the MZmine and SIRIUS platforms. Isolation was carried out using column chromatography, and preparative recycling HPLC. Bioactive compounds were characterized using UV, HPLC, NMR, and 2D NMR, as well as bioactivity tests using the MTT method.

The results show that the extract contained N-[(1,3-dimethyl-2,6-dioxo-7-prop-2-ynylpurin-8-yl) amino] formamide, N-(3-phenylbutyl)hexan-2-amine, 1,1-Dichloro-1-nitrosopropane, ceratodictyol, betulinic acid, ursolic acid, 7-methyl-N-[6-[(7-methyl-6-oxooctanoyl) amino] hexyl]-6-oxononanamide, Nervisterol and 3,5,10-tris (acetyloxy)-2-hydroxy-4,14,16,16-tetramethyl-8-methylidene-13-oxo-15oxatetracyclo [9.4.1.0
1,
14.0
4,
9] hexadecan-7-yl 3-phenylprop-2-enoate. The ethanol extract of
Uncaria nervosa Elmer leaves contains nine compounds consisting of alkaloids, terpenoids, and fatty acid. The bioactive compounds that were successfully isolated were betulinic acid, and ursolic acid, with IC
50 values of ˃100 and 14,70±4,50 μg/ml, respectively. These compounds were reported in this plant for the first time.

Betulinic acid, and ursolic acid have been successfully isolated from leaves
Uncaria nervosa Elmer, and ursolic acid have moderate cytotoxic activity on T47D breast cancer cells.