Efficient extraction of cucurbitacins from Diplocyclos palmatus (L.) C. Jeffrey: Optimization using response surface methodology, extraction methods and study of some important bioactivities

Abstract

Diplocyclos palmatus (L.) C. Jeffrey is an important medicinal plant used in several reproductive medicines. It serves as a wide source of tetracyclic triterpens called cucurbitacins. Response surface methodology (RSM) with Box-Behnken design (BBD) was studied to optimize the production of cucurbitacins. RSM put forth the ideal conditions such as 1:30 SS ratio (g/mL), 80 rpm (mixing extraction speed), 150 µm mean particle size, 30 min extraction time and 50 °C using chloroform in continuous shaking extraction (CSE) and showed the highest cucurbitacin I (CUI) content (2.345 ± 0.1686 mg/g DW). Similarly, the highest yield of cucurbitacin B (CUB) (1.584 ± 0.15 mg/g DW) was recorded at ideal conditions (1:40 g/mL SS ratio and 60 min time and others similar to CUI). Among the tested extraction methods, the highest CUI, CUB, and CUI + B yield (1.437 ± 0.03, 0.782 ± 0.10, 2.17 ± 0.35 mg/g DW, respectively) as well as promising DPPH radical scavenging activity (25.06 ± 0.1 µgAAE/g DW) were recorded from the SBAE (steam bath assisted extraction). In addition, MAE and UAE revealed the highest inhibition of α-amylase (68.68%) and α-glucosidase (56.27%) enzymes, respectively. Fruit extracts showed potent anticancer activity against breast (MCF-7) and colon (HT-29) cancer cell lines (LC 50 -44.27 and 46.88 µg/mL, respectively). Our study proved that SS ratio, particle size and temperature were the most positively influencing variables and served to be the most efficient for the highest recovery of CUI and CUB. Based on the present study, the fruits of D. palmatus were revealed as a potent antioxidant, anti-diabetic and anticancer bio-resource that could be explored further to develop novel drug to manage diabetes, cancer and oxidative stress related disorders.The plants from the family Cucurbitaceae are commonly called as melons, squashes, and gourds which are traditionally used in the human diet. It comprises 122 genera and 940 species of which 31 genera and 94 species are found in India 1 . Diplocyclos palmatus (L.) C. Jeffrey is a slender-stemmed tendril climber commonly called as Shivlingi. Traditionally, this plant has been used in the folk medicine and possesses several activities such as gynaecological, anti-asthmatic, anti-convulsant, anti-venom, anti-inflammatory, androgenic and antioxidant 2-4 . The family Cucurbitaceae has been recognized as a rich source of bitter compounds, called cucurbitacins 5 . The cucurbitacins are highly unsaturated triterpenes containing many keto-, hydroxy-and acetoxy-groups. They are the major active components in traditional medicine, herbal remedies and Pharmacopoeia to treat various health issues such as hepatoprotective, anti-fertility, diabroticites cardiovascular and anti-inflammatory activities 6 . In a recent study, cucurbitacin I (CUI) has been identified as a strong inhibitor of the JAK2/STAT3 signaling pathway (a common oncogenic signaling pathway), which is constitutively activated in many types of cancer; hence, it is considered as a milestone in cancer therapy 7 . Similarly...

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