Biological Activities of Withanolides from Datura innoxia

Aims: This study aimed to explore the therapeutic potential of Datura innoxia through the chemoinformatic and antibacterial evaluation of withanolides extracted from it. Study Design: The pharmacokinetic and pharmacodynamic properties and drug-likeness of the withanolides—withametelinol A, withametelinol B, witharifeen, withametelin, dinoxin B, and daturalicin—of D. innoxia were analyzed using the SwissADME program. Schrodinger software was used to target and evaluate their antibacterial potentialities through docking studies. The penicillin-binding protein, DNA gyrase, efflux pump protein, and quorum sensing regulators of S. aureus and E. coli were selected as target proteins for assessing protein–ligand interactions. All observations were comparatively analyzed with the properties of withanolide A and withaferin A, the best-known withanolides. Most active dinoxin B withanolide (12500–100000 μg/ml) extracted from leaves of Datura innoxia; was subjected to antibacterial assay against methicillin-resistant S. aureus (MRSA) and multi-drug resistant(MDR) E. coli isolated from the urine samples of urinary tract infected patients. Results: In-silico studies revealed the therapeutical properties of various withanolides present in D. innoxia. In particular, the drug-likeness and antibacterial properties of withametelin and dinoxin B were significantly and remarkably high due to their binding affinity toward cell membrane proteins. Docking studies have shown that the efflux pump protein of E. coli and penicillin-binding proteins of S. aureus to be the ligand -interaction targets. A significant antibacterial assay revealed that the MRSA isolates were susceptible to dinoxin B, with a zone of inhibition of 21±0.5 mm to 24±0.5 mm, and the bacteria were susceptible at a concentration rate of ≤ 12.5 mg/ml. Conclusion: It is crucial to bring awareness of the therapeutical importance of D. innoxia and to preserve this vital plant from being largely destroyed. As computational studies promote the effective selection of drug molecules, this research also helps to select the best compound for further clinical analysis.

Anti-Inflammatory Potential of Daturaolone from Datura innoxia Mill.: In Silico, In Vitro and In Vivo Studies

Exploration of leads with therapeutic potential in inflammatory disorders is worth pursuing. In line with this, the isolated natural compound daturaolone from Datura innoxia Mill. was evaluated for its anti-inflammatory potential using in silico, in vitro and in vivo models. Daturaolone follows Lipinski’s drug-likeliness rule with a score of 0.33. Absorption, distribution, metabolism, excretion and toxicity prediction show strong plasma protein binding; gastrointestinal absorption (Caco-2 cells permeability = 34.6 nm/s); no blood–brain barrier penetration; CYP1A2, CYP2C19 and CYP3A4 metabolism; a major metabolic reaction, being aliphatic hydroxylation; no hERG inhibition; and non-carcinogenicity. Predicted molecular targets were mainly inflammatory mediators. Molecular docking depicted H-bonding interaction with nuclear factor kappa beta subunit (NF-κB), cyclooxygenase-2, 5-lipoxygenase, phospholipase A2, serotonin transporter, dopamine receptor D1 and 5-hydroxy tryptamine. Its cytotoxicity (IC50) value in normal lymphocytes was >20 µg/mL as compared to cancer cells (Huh7.5; 17.32 ± 1.43 µg/mL). Daturaolone significantly inhibited NF-κB and nitric oxide production with IC50 values of 1.2 ± 0.8 and 4.51 ± 0.92 µg/mL, respectively. It significantly reduced inflammatory paw edema (81.73 ± 3.16%), heat-induced pain (89.47 ± 9.01% antinociception) and stress-induced depression (68 ± 9.22 s immobility time in tail suspension test). This work suggests a possible anti-inflammatory role of daturaolone; however, detailed mechanistic studies are still necessary to corroborate and extrapolate the findings.

PALYNOMORPHOLOGICAL STUDY OF WEEDY MELLIFEROUS (BEE VISITED) PLANTS USING LIGHT MICROSCOPIC TECHNIQUES FROM SOUTHERN KHYBER PAKHTUNKHWA, PAKISTAN

Pollen morphology of 10 different weedy bee foraged plants belong to 10 various families from Southern Khyber Pakhtunkhwa were collected, identified and studied using light microscopy (LM). The plants were Asphodelus tenuifolius, Euphorbia helioscopia, Parthenium hysterophorus, Rhazya stricta, Datura innoxia, Eruca sativa, Convolvulus arvensis, Anagallis arvensis, Galium aparine, and Anethum graveolens. Slides for Light microscopic studies were prepared with the help of acetic acid, glycerin jelly and anthers of flowers. Pollen grain recorded ranged from monocolpate to hexacolporate and from psilate to echinate which were important systematic significance. Pollen size, shape, P/E ratio, exine thickness, number of colpi, number of pores, equatorial diameter, polar diameter, colpus width, colpus length, spines number, length and width of spines were examined with the help of light microscopy and all these values were analyzed statistically using software SPSS. This research provide a data to the optimal utilization of bee foraged weed plants by honeybees and identification of bee flora for the beekeeping business and honey production. Results revealed that weedy melliferous flora of study area is very helpful for botanical origin, geographical origin of bee species and adulterations found in honey.

INVESTIGATION NON-ALKALOIDAL BIOLOGICAL ACTIVE COMPOUNDS OF DATURA INNOXIA

The purpose of the investigation is to study non-alkaloidal biological active compounds of various organs, the antimicrobial activity of plant oil and ethanolic extracts of Datura innoxia Mill. from nightshades (Solanaceae). D. innoxia grows in the natural environmental conditions of the Republic of Azerbaijan. We have studied presence of several biological active compounds – triterpenoic acids, essential oils, fatty acids, sterols and amino acids in the content of various organs of plant. Oil was obtained by using Soxhlet apparatus and investigated by GC method. After removing the oil from the seeds, triterpenoic acids - oleanolic and ursolic acids were isolated and identified by TLC. Essential oils were obtained from plant leaves by hydrodistillation and studied by GC-MS method. 20 amino acids were identified in the aerial part of the plant by the HPLC method, with amino acid analyzer L-8800 (Hitachi, Ltd.), 12 of them are non-essential, 8 are essential. The antimicrobial activity of plant oils of seed and ethanolic extracts of some parts (roots, stems, leaves, flowers, fruits and seeds) were studied on Staphylococcus aureus, Esherichia coli, Pseudomonas aeruginosa, Bacillus anthracoides, Klebsiella pneumoniae and Candida albicans. The significant activity of some extracts (stems, leaves, fruits and seeds) was observed against S. aureus, E. coli, C. albicans, B. anthracoides and plant oil against C. albicans.

Histological and body weight effects of Datura innoxia seeds and leaves extracts in rats

The study aim to determine the chemical compounds in aqueous and methanolic extracts of Datura innoxia seeds and leaves and to evaluate their toxic effects on experimental rats. Seeds and leaves were collected from El-Obied, North Kordofan State, Sudan, in October, 2016. The aqueous and methanol extracts were carried out by using maceration method and soxhlet apparatus respectively. Sixty five male Albino Wistar rats, three months old and with an average body weight ranged 110-120 g, were randomly divided into thirteen Groups, consisting of five rats in each Group. Group 1 served as control and fed with normal rats’ food and water for thirty days. Groups 2, 6 and 10 administered aqueous seeds extract, Groups 4, 8 and 12 received methanol seeds extract, Groups 3, 7 and 11 received aqueous leaves extracts, Groups 5, 9 and 13 received methanol leaves extract, all the Groups received the same type of extract were administered 40, 60 and 80 mg/kg body weight respectively. The extracts administered to the rats intra gastrically using cathodal tube daily for thirty days. The effects of oral administration of leaves and seeds extracts to 60 healthy rats over 30 days were evaluated by histological studies and body weight changes. The analysis by gas chromatography-mass spectrometry (GC-MS) of aqueous and methanolic extracts revealed the presence of alkaloids (Scopolamine, atropine and Hyoscyamine), fatty acids, esters, amides, amino acids, ketones, coummarins, terpinoids, phenols, alcohols and hydrocarbons compounds. The histological results showed that administration of extracts caused pathologic changes in the organs studied. The treated Groups had lower (p ≤ 0.05) body weight gains than control Group. The study concluded that the toxicity of seeds and leaves (methanolic and aqueous) extracts are nearly have the same toxic effects on rats due to their same active ingredients (alkaloids) and the oral administration of the extracts was found to be safe up to 40 mg/kg.

In Vitro Cytotoxic Evaluation and Apoptotic Effects of Datura innoxia Grown in Saudi Arabia and Phytochemical Analysis

Datura innoxia is an important species of Solanaceae family with several purposes in folk medicine. This study intends to explore the cytotoxic effect of D. innoxia on various cancer cell proliferation. D. innoxia ethanolic extract’s effect on the progression of the cell cycle and the induction of apoptosis were investigated by flow cytometry. Further, real-time PCR was employed to confirm apoptosis initiation. In addition, active phytochemicals of D. innoxia was identified by gas chromatography–mass spectroscopy (GC-MS). The cell viability study revealed that the ethanolic extract of D. innoxia demonstrated potent cytotoxicity, with an IC50 value of 10 μg/mL against LoVo colon cancer cells. Cell cycle staining with propidium iodide revealed that D. innoxia treatment leads to cell accumulation in the sub-G1 phase. Using the Annexin V-FITC/PI assay, the ethanolic extract was found to cause a dose-dependent increase in early and late apoptosis when compared to control cells. Apoptosis as the mode of cell death was also confirmed by the increased expression of p53, bax and caspase-8, -9, and -3 along with downregulation of Bcl-2. GC-MS analysis displayed that 3,5-Dihydroxybenzoic acid (16.53%), heneicosyl formate (14.14%), 2,3-dimethyl-3-pentanol (12.89%), 2-hydroxy-4-methyl pentanoic acid (5.19%) were the main phytoconstituents. These findings conclude that D. innoxia causes cell death through apoptosis, suggesting more attention should be paid to further exploration of the active components from D. innoxia responsible for the observed activities.

Cytotoxic withanolides from Datura innoxia

Chemical investigation of the aerial parts (except fruits) of the medicinal, hallucinogen and toxic plant Datura innoxia Mill. [Solanaceae] led to the isolation of the new withanolide, dinnoxolide A (1), along with the known compounds 21,27-dihydroxy-1-oxowitha-2,5,24-trienolide (2), daturamalakin B (3) and withametelin (4). Their structures were established by analysis of their spectroscopic data, including 1D and 2D NMR experiments and MS. Compounds 2 and 3 were isolated as natural products for the first time and the name dinnoxolide B was given to compound 2. The four withanolides showed in vitro cytotoxic activity against U251 (glioblastoma) and SK-LU-1 (lung adenocarcinoma) human cancer cell lines, with IC50 values ranging from 1.2 to 19.6 µM, and also against the noncancerous monkey kidney cell line (COS-7), with IC50 values ranging from 5.0 to 19.7 µM. Compound 4 was two times more active than the reference compound, etoposide, against lung adenocarcinoma cells.