Pharmacological Profiling of Calotropis Procera and Rhazya Stricta: Unraveling the Antibacterial and Anti-Cancer Potential of Chemically Active Metabolites

Abstract

Background: The increasing prevalence of cancer and bacterial resistance necessitates more effective anti-cancer and anti-bacterial treatments. This study explores the potential of medicinal plants, specifically Calotropis procera (C. procera) and Rhazya stricta (R. stricta), in addressing this need, aiming to uncover new therapeutic interventions. Methods: Various extraction methods for the leaves of C. procera and R. stricta were employed to investigate the anti-bacterial and anti-cancer properties of these herbs. For anti-bacterial testing, extracts were prepared using water, chloroform, and ethyl acetate, and their activity against methicillin-resistant Staphylococcus aureus (S. aureus) (MRSA) and Escherichia coli (E. coli) was assessed. The anti-cancer potential was evaluated through MTT cytotoxicity assays on various cancer cell lines and further testing using high-content imaging (HCI)-Apoptosis Assay and the ApoTox-GloTM Triplex Assay. Liquid chromatography-mass spectrometry (LC-MS) was used to identify the secondary metabolites of C. procera, and computational predictions were performed to assess the activity of these metabolites. Results: The leaf extracts of both C. procera and R. stricta demonstrated antibacterial activity against S. aureus and E. coli. The C. procera ethyl acetate extract exhibited potent anti-cancer effects on several cancer cell lines. The research also revealed a dose-dependent induction of apoptosis and a decline in cell viability. Computational predictions suggested the identified metabolites were active as nuclear receptor ligands and enzyme inhibitors, with good oral bioavailability. Most metabolites were found to be immunologic and cytotoxic, except for proceragenin and calotropone, which were determined to be non-cardiotoxic. Conclusion: The study's findings demonstrate the remarkable cytotoxic and antibacterial effects of C. procera extracts prepared using ethyl acetate. These results pave the way for further in vitro studies to explore the full potential of these extracts and highlight the presence of chemically active metabolites in C. procera, which hold promise as lead molecules for the development of novel therapies targeting bacterial infections and cancer while minimizing potential side effects.

Keywords: Anti-bacterial; Anti-cancer; Calotropis procera, Rhazya stricta, Apocynaceae, In silico.

Figure 1

Antimicrobial activity of A) C. procera and B) R. stricta extracts using the agar well diffusion assay. Inhibition zones (mm) were determined for water (A), chloroform (C), and ethyl acetate (E) extracts of each plant/extract. Ampicillin served as a positive control, and the experiments were conducted with four replicates (n=4). The statistical significance of ^** is indicated by a p-value of 0.001, the significance of ^*** is indicated by a p-value of 0.0001, while the significance of ^**** is denoted by a p-value of less than 0.0001,

Figure 2

MTT Assay Results for C. procera samples Aganis Several Cancer Cell Lines.

Figure 3

HCl Apoptosis Assay of Ethyl Acetate extract of C. procera against A) KAIMRC2 and B) HCT8 cell lines.

Figure 4

HCl Apoptosis Assay of A) Ethyl Acetate extract of C. procera and Mitoxantrone against KAIMRC2 and B) HCT8 cancer cell lines.

Figure 5

Base Peak Chromatogram of C. procera Ethyl Acetate Extract Was Extracted and Tentatively Identified Secondary Metabolites Are (A) proceragenin, (B) calotoxin, (C) uscharin, (D) calotropone, (E) gofrusid, (F) calactin, and (G) coroglaucigenin. Means M/Z Implies Measured M/Z.

Figure 6

Swiss Target Prediction of Identified Metabolites of C. procera in Ethyl Acetate Extract.

Figure 6

Swiss Target Prediction of Identified Metabolites of C. procera in Ethyl Acetate Extract.

Figure 7

Radar Charts for Oral Bioavailability of the Identified Metabolites of C. procera in Ethyl Acetate Extract.