Influence of Geographical and Seasonal Variations on Carbazole Alkaloids Distribution in Murraya koenigii: Deciding Factor of Its In Vitro and In Vivo Efficacies against Cancer Cells

Abstract

Murraya koenigii is a well-known Indian medicinal herb, and a carbazole alkaloid (mahanine) from this plant causes apoptosis in cancer cells. Here, we investigated how seasonal and geographical variations influence carbazole alkaloids composition and medicinal property of this plant against cancer cells in vitro and in vivo. Leaflets were collected from various places in different seasons for three years. A mahanine-enriched fraction (MEF) was prepared in two steps using ethanol and water. The best plant was selected based on the highest percent of mahanine. MEF prepared from leaflets of nine different locations showed a different concentration of identified markers (mahanine, mahanimbine, and koenimbine) which exhibited differential reduced metabolic activity against ovarian cancer, mahanine being the best. Our systematic study revealed that mahanine content was highest during September–December. Interestingly, MEF from southern part (tropical zone) exhibited 43 ± 2.5% mahanine compared to 2.7 ± 1.3% in northeastern part (subtropical zone) with five folds higher activity against PA1. Moreover, MEF reduced metabolic activity of sixteen cancer cell lines from nine different origins and significantly reduced tumor mass in lung and ovarian cancer xenograft models. Taken together, this is the first report demonstrating the marker’s content in these leaflets is highly dependent on location/season. A positive correlation between biological activity and mahanine concentration was established in MEF. Such a comprehensive study suggests that the selection of location and suitable season for collection of any plant materials with biologically active stable markers in sufficient quantity play a decisive role in determining the fate of their medicinal property.

Preclinical Development of Mahanine-Enriched Fraction from Indian Spice Murraya koenigii for the Management of Cancer: Efficacy, Temperature/pH stability, Pharmacokinetics, Acute and Chronic Toxicity (14-180 Days) Studies

“…A high content of mahanine in MEF M2 was further supported by in vitro activity against both ovarian and breast cancer cells. Earlier, we have reported the anticancer activity of MEF both in ovarian and lung cancer xenograft models [14]. Here, we showed the excellent in vivo efficacy of MEF M2 in the ovarian and breast cancer syngeneic models at a dose of 300 mg/kg BW.…”

“…Accordingly, water infusion before maceration with ethanol followed by the fractionation of the obtained extract with ethyl acetate gave the best results. Although Method 1 and our previously reported method [14] showed the highest yield (7.8 and 8.0 respectively), we have selected Method 2 because of its high percent and maximum amount of mahanine. A high content of mahanine in MEF M2 was further supported by in vitro activity against both ovarian and breast cancer cells.…”

“…We have earlier optimized a place and season for the collection of leaflets from the Murraya Koenigii plant having the highest amount of mahanine [14]. Accordingly, we procured the leaflets from Tamil Nadu in India (13.0675°N, 80.1951°E) between September and December.…”

“…In general, the qualities of five different MEFs were determined mainly based on the presence of the highest amount of mahanine, the most biologically active carbazole alkaloid by HPLC (Figure 1). Additionally, the HPLC fingerprints were compared for the presence of two more carbazole alkaloids, namely, mahanimbine and koenimbine which were characterized earlier [14]. The mahanine peak in MEF M2 was also confirmed by mass spectrometry.…”

“…In our previous study, we have systematically optimized the geographical location and season for the collection of this valuable plant with the highest mahanine content and prepared a fraction rich in mahanine with two more defined markers like mahanimbine and koenimbine [14]. Additionally, this fraction exhibited antiproliferative activity in both in vitro in sixteen different cancer cell lines of nine types of cancer and in vivo in ovarian and lung cancer xenograft mice models [14]. However, the efficacy of the fraction rich in mahanine was not tested in syngeneic ovarian and breast cancer models.…”