Ethyl Acetate Soluble Fraction Research Articles

In-vitro and molecular docking studies of plant secondary metabolites isolated from Hypericum oblongifolium as antibacterial agents and lipoxygenase (5-LOX) inhibitors

Natural Products are bases for modern pharmaceuticals as they are precursors for many synthetic drugs. People also use natural products directly in the form of herbal medicines due to their less harmful effects. This research project aimed to investigate the new pharmacological potential of natural products found in Hypericum oblongifolium. Five compounds were identified from chloroform and ethyl acetate soluble fractions of H. oblongifolium. These compounds were identified as 21-β-hydroxyursolic acid (1), β-sitosterol (2), 7,4′-dihydroxy-5,3′-dimethoxyisoflavone (3), α-D-glucopyranosyl-6′-O-hexadecanoate (4), Quercetin-3′-O-ß-D-glucopyranoside (5). When these compounds were tested for antibacterial potential compound 5 showed maximum inhibition against gram-positive Bacillus subtilis and was highly active against Escherichia coli gram-negative bacterium. Minimum inhibition was shown by compounds 2 and 3 against Shigella flexneri among gram-negative bacteria. Compound 5 also exhibited the least binding energy in the molecular modeling of these compounds against Escherichia coli proteins GlcN-6-P synthase (PDB ID: 1MOQ). Compound 5 also showed good inhibition of lipoxygenase (5-LOX) enzyme with an IC50 value of 30.1 ± 0.20 as it exhibited maximum hydrogen bondings and minimum energy when docked with LOX-5 (PDB ID 3V99) using a patch dock.

Molecular level interaction, HOMO-LUMO, MEP, UV–Vis, Hirshfeld, topological analysis, and in-vitro of isoflavones from Eremostachys Vicaryi Benth. Ex Hook. f.

Eremostachys vicaryi Benth. Ex Hook. f. (Labiatae), a Pakistani (Baluchistan) herbal medicinal plant commonly known as “Stagh,” which has a sharp taste and smell. It acts as an antiseptic and antimicrobial agent and is used for fish poisoning, treating skin diseases, and holding a cooling effect. Following the assessment of the plant extract's cytotoxic and phytotoxic assays using phytochemical screening, the antioxidant and anti-inflammatory properties of the pure ingredients that were separated from the ethyl acetate soluble fractions have been evaluated. The bioassay-guided fractionation and purification of E. vicaryi B. crude methanol extract yielded two isoflavones (1–2) that showed promising antioxidative activities with DPPH and cytochrome-c-reduction assays. Compounds 1 and 2 have shown considerable anti-inflammatory efficacy, 1 being a good inhibitor of the superoxide within the living system. Initial assumptions on the one-electron transfer process indicated that Compound 1 has a smaller ionization potential value compared to Compound 2 and so has greater antioxidant potential than its predecessor, isoflavone. Moreover, the anti-inflammatory and antioxidant activity of isoflavones was also probed by first-principles calculations. Based on the findings, it has been concluded that the crude extract of E. vicaryi B. and its organic components exhibit noteworthy phytotoxic and cytotoxic properties. The isolated pure isoflavones 1 and 2 showed significant antioxidant and anti-inflammatory activities. Solute-solvent interactions were studied and reported for both compounds. An analysis was conducted on the UV–Vis electronic transition for different solvents. Using a Multiwave function analyzer the topological findings such as ELF, LOL, and RDG were done to comprehend the surface electron density, delocalization, localized orbital, and weak interaction in the compounds. The antioxidant activity is evaluated by radical methods using DPPH, and cytochrome-c-reduction assays using ascorbic acid as a standard. Through cell-based respiratory burst testing, the anti-inflammatory activity of standard indomethacin was examined. The ground state geometries of isoflavones and reference compounds were optimized by Density Functional Theory- (DFT) by computing various electronic properties. In-vitro bioassay results were validated by first-principles calculations.

Molecular networking-assisted isolation of chlorophenolic glycosides from the rhizomes of Curculigo orchioides and their inhibitory effect on α-glucosidase

Molecular networking analysis and in silico tools, such as Network Annotation Propagation (NAP) and MolNetEnhancer, were applied to explore bioactive constituents present in the ethyl acetate-soluble fraction of the rhizomes of Curculigo orchioides. Among the molecular networks, the most abundant cluster was classified as a phenolic glycoside using the ClassyFire module of MolNetEnhancer. Further, the major node in this cluster was accurately predicted as curculigine A using the in silico fragment analysis tool, NAP. Six undescribed chlorophenolic glycosides (1–6) and 11 known phenolic glycosides were isolated, using molecular networking-assisted isolation methods, and their structures were elucidated using 1D, 2D-NMR and HRESIMS. In particular, the structures of the isolated chlorophenolic glycosides, which have non-protonated aromatic rings, were determined using various NMR experiments, such as 1D-selective NOE, ROESY, and LR-HMBC, and acid hydrolysis. All isolated compounds were examined to determine their inhibitory effects on α-glucosidase and compounds 3, 8, 10, 11, 13, 14, and 16 revealed the IC50 values ranging from 19.6 to 35.5 μM. Their structure-activity relationships were also evaluated based on the analysis of their inhibitory effects and performance of molecular docking simulations.

Mechanism of antibacterial action of n-hexane and ethyl acetate soluble fractions of Psidium guajava leaves against Salmonella enterica serovar Typhi

Mechanism of antibacterial action of n-hexane and ethyl acetate soluble fractions of Psidium guajava leaves against Salmonella enterica serovar Typhi

Isolation and Characterization of Chemical Compounds from the Stem Bark Fractions of Citrus Grandis and Evaluation of Their Antimicrobial, Cytotoxic, and Antioxidant Activities

Citrus grandis belongs to the Rutaceae family and is a naturally occurring citrus plant. The fractionated crude extracts of C. grandis have been studied for the investigation of its phytochemicals and allowed for the evaluation of biological activities, especially antimicrobial, cytotoxic, and antioxidant activities. At first, we assessed the antibacterial and antifungal properties using the disc diffusion method and conducted cytotoxicity screening using the brine shrimp lethality bioassay. Among the raw extracts, we isolated two compounds named CG-F3-B1 and CG-F3-B2 through column chromatographic separation of the chloroform extract. By comparing their proton NMR spectra with published spectra, we identified the chemical structures of these compounds as Zeylenol and Limonin, respectively. Regarding antibacterial testing, we observed that the methanol soluble fractions and petroleum ether fractions exhibited mild to moderate activity against bacteria. Additionally, the ethyl acetate soluble fraction showed moderate antibacterial activity. In addition, the crude extract displayed moderate antifungal activity against specific fungi. Furthermore, the cytotoxic assay revealed a relevant cytotoxic effect of a crude extract with an LC50 value of 37 μg/ml. The antioxidant activity was determined using the DPPH assay, and the methanol soluble extract exhibited modest antioxidant activity with an IC50 value of 89 μg/ml. Our findings demonstrated that the stem bark fractions and isolated compounds from C. grandis exhibited various promising biological activities, including antibacterial, antifungal, cytotoxic, and antioxidant properties.
 Bioresearch Commu. 9(2): 1320-1329, 2023 (July)

Phytochemical and Biological Investigation of an Indigenous Plant of Bangladesh, Gynura procumbens (Lour.) Merr.: Drug Discovery from Nature.

Gynura procumbens (Lour.) Merr. (Family: Asteraceae) is a tropical Asian medicinal plant found in Thailand, China, Malaysia, Indonesia, and Vietnam. It has long been utilized to treat a variety of health concerns in numerous countries around the world, such as renal discomfort, constipation, diabetes mellitus, rheumatism, and hypertension. The chemical investigation resulted in the isolation and characterization of six compounds from the methanol (MeOH) extract of the leaves of Gynura procumbens, which were identified as phytol (1), lupeol (2), stigmasterol (3), friedelanol acetate (4), β-amyrin (5), and a mixture of stigmasterol and β-sitosterol (6). In-depth investigations of the high-resolution 1H NMR and 13C NMR spectroscopic data from the isolated compounds, along with comparisons to previously published data, were used to clarify their structures. Among these, the occurrence of Compounds 1 and 4 in this plant are reported for the first time. The crude methanolic extract (CME) and its different partitionates, i.e., petroleum ether (PESF), chloroform (CSF), ethyl acetate (EASF), and aqueous (AQSF) soluble fractions, were subjected to antioxidant, cytotoxic, thrombolytic, and anti-diabetic activities. In a DPPH free radical scavenging assay, EASF showed the maximum activity, with an IC50 value of 10.78 µg/mL. On the other hand, CSF displayed the highest cytotoxic effect with an LC50 value of 1.94 µg/mL compared to 0.464 µg/mL for vincristine sulphate. In a thrombolytic assay, the crude methanolic extract exhibited the highest activity (63.77%) compared to standard streptokinase (70.78%). During the assay for anti-diabetic activity, the PESF showed 70.37% of glucose-lowering activity, where standard glibenclamide showed 63.24% of glucose-reducing activity.

Chemical and Biological Investigations of the Leaf Extracts of Andrographis paniculata (Acanthaceae)

Introduction: Andrographis paniculata is a plant employed in traditional medicine among the natives of Owerri, Eastern Nigeria, in the management of several ailments. In this study the leaf was investigated for its chemical constituents and to validate its antimicrobial activity in ethnomedicine. Methods: The fresh leaf of the plant was steam-distilled to obtain the volatile oil which was subjected to FT-IR and GC-MS analysis. The powdered air-dried leaf was also extracted with methanol to obtain the crude extract which was subjected to phytochemical analysis. The crude methanolic extract was also partitioned into hexane, chloroform, ethylacetate and 50% methanol in chloroform to obtain the corresponding fractions. The crude extract and fractions were tested for activity against Staphylococcus aureus, Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Bacillus subtilis, Candida albicans and Aspergillus niger. The biologically active chloroform fraction was further purified using column chromatography and the fractions subjected to GC-MS analysis. Results: The steam distillation gave an oil and the GC-MS analysis indicated the presence of butylated hydroxytoluene, hexadecanoic acid, methyl ester, dibutyl phthalate, 8,11-octadecadienoic acid, methyl ester, 11-octadecenoic acid, methyl ester and bis (2-ethylhexyl) phthalate. Phytochemical analysis of the crude extract showed the presence of tannins, alkaloids, sterols/steroids, flavonoids, volatile oils, terpenoids, saponins and phenols. Among the extractives, the chloroform-soluble fraction showed the highest activity against S. aureus and B. subtilis while none of the extracts was active against C. albicans and A. niger. It was also found that the hexane-soluble fraction was inactive against all the test organisms while the ethylacetate-soluble fraction showed the broadest spectrum of anti-bacterial activity. Column chromatographic separation of the biologically active chloroform-soluble fraction gave fractions which on GC-MS analysis and by comparison with the standard library computer MS data led to the identification of some chemical constituents, including aromatic compounds, sterols, alicyclics and long-chain hydrocarbons and carbonyl compounds such as carboxylic acids, esters and lactones. Conclusion: The antimicrobial activity exhibited by the leaf extracts support the traditional medicinal use of the leaf to manage infections. Also, the presence of the identified phyto-constituents may be responsible for the ethno-medicinal properties of the plant.

Identification of pyrrolizidine alkaloids and flavonoid glycosides through HR-LCMS/MS analysis, biological screening, DFT and molecular docking studies on Heliotropium dasycarpum Ledeb.

The current study was carried out to reveal the chemical profile and biological screening of Heliotropium dasycarpum Ledeb, as well as the feasibility of its industrial application. Therefore, the methanolic extract (HdM) of H. dasycarpum was divided into n-hexane (HdH), ethyl acetate (HdE) and water (HdW) soluble fractions. All the fractions were inactive against acetylcholinesterase (AChE) enzyme, 3T3 and HeLa cell lines, but showed immunomodulatory effect up to 37 %. HdE fraction further displayed significant anti-urease activity with IC50 value of 74.072 ± 0.002, while HdM was found moderate inhibitor (63 %). Thus HdE was subjected to HR-LCMS/MS analysis in positive and negative ionization modes, and a qualitative analytical method with a data mining strategy was utilized. The secondary metabolites were identified by dereplication strategy using molecular networking as a bioinformatics tool, which disclosed the presence of 08 known alkaloids of heliotrine-type (1, 3, 5–10), and 05 (12–16) known flavonoids and flavonoid glycosides along with 03 new (2, 4 and 11) putative pyrrolizidine analogues. DFT simulations of identified compounds were performed using multiple quantum chemical and geometrical descriptors in order to determine their quantitative structure activity relationship. These secondary metabolites were docked against the enzyme urease which substantiated the inhibitory action of pyrrolizidine alkaloids and flavonoid glycosides. Furthermore, ADME analysis provided the base for the use of these compounds for further studies as drug leads and to unveil industrial application of H. dasycarpum in formulating products against gastritis, peptic ulcer and gastric cancer.

Comparison of Antimicrobial and Membrane Stabilizing Properties of Leaf and Stem Extracts of Derris scandens (Roxb.) Benth. (Family: Fabaceae)

Derris scandens (Roxb.) Benth. (Family: Fabaceae), a recognized medicinal plant colloquially known as ‘Kalilata’ in Bangladesh, has been traditionally used as analgesic in Thailand. This study was intended to evaluate and compare the antimicrobial and membrane stabilizing potentials among the methanol extracts of D. scandens leaf and stem, and their fractionates. The stem and leaves of the plant were extracted by methanol separately and fractioned into aqueous, dicholoromethane (DCM), ethyl acetate and n-hexane soluble fractions. They were tested for antimicrobial activity by using the agar-well diffusion method where ciprofloxacin and fluconazole were used as references. The highest antimicrobial activity was observed against a Gram-negative bacteria (Escherichia coli) by the DCM soluble fraction of the leaf and stem extracts with 20 mm and 15 mm zones of inhibition, respectively. The n-hexane soluble fraction of methanol extract of stem revealed weak to moderate antimicrobial activity against Gram-positive bacteria. The ethyl acetate soluble fractionates of methanol extracts of leaf and stem showed similar antifungal property against Saccharomyces cerevisiae with a zone of inhibition of 12 mm. The membrane stabilizing potential of the extractives was estimated based on the prevention of hemolysis of RBC prompted by hypotonic solution as well as heat. The n-hexane and DCM soluble fractions of both extracts showed around 75% inhibition of hypotonic solution- and heatinduced hemolysis. All in all, the extracts and several fractionates showed moderate antimicrobial and membrane stabilizing properties and further investigations are warranted to find out active phytoconstituents responsible for these properties. Bangladesh Pharmaceutical Journal 26(1): 67-72, 2023 (January)

Evaluation of anti-inflammatory potential of extract, fractions and major compounds of Ponciri Fructus in LPS-induced RAW 264.7 cells

Dried immature fruit of Poncirus trifoliata (L.) Raf., known as Ponciri Fructus, is a well-known traditional Korean medicine for treatment of allergy and gastrointestinal disorders. In previous study, we reported the anti-obesity effects of the extracts, fractions and major compounds of Ponciri Fructus on 3 T3-L1 preadipocytes. In continuation, in this research, we evaluated their anti-inflammatory potential using lipopolysaccharide (LPS) induced RAW 264.7 cells. Results showed that the ethanol extract of fruits and hexane and ethyl acetate soluble fractions of the extract inhibited nitrous oxide (NO) production in LPS stimulated macrophage in all doses. Whereas, butanol fraction of extract showed significant inhibition in production of NO only at higher dose of 80 µg/mL. Some of the isolated compounds such as poncirin, naringin, imperatorin and phellopterin, significantly inhibited NO production in LPS-stimulated macrophages. Poncirin and imperatorin showed anti-inflammatory potential by inhibiting cyclooxygenase (COX)-2 protein as evidenced by downregulation of that protein in western blot assay. Imperatorin also inhibited inducible nitric oxide synthase (iNOS) responsible for production of NO in macrophages. In conclusion, Ponciri Fructus has strong anti-inflammatory potential and possesses many active flavonoids and coumarins contributing to this activity.

Antioxidant Activity Study of Zingiber Zerumbet Rhizome Extract and Fraction

Zingiber zerumbet is a member of the Zingiberaceae family with known pharmacological potentials. This study aims to characterize the phytochemical elements and evaluate the antioxidant potential of the methanol extract, soluble ethyl acetate, and n-hexane-soluble fractions of Zingiber zerumbet rhizome. Identify secondary metabolites for the alkaloids, saponins, flavonoids, polyphenols, tannins, and steroids/triterpenoids using the chemical reaction method in test tubes. Antioxidant activity was determined based on DPPH absorbance measurements using UV-Vis Spectrophotometry. The methanolic extract contained alkaloids, flavonoids, polyphenols/tannins, steroids, and triterpenoids. The ethyl acetate soluble fraction contained flavonoids and polyphenols/tannins, and the n-hexane soluble fraction contained alkaloids and flavonoids. The free radical scavenging activities of ascorbic acid, methanol extract, ethyl acetate, and the n-hexane soluble fraction were 7.79; 100,956; 69,476, and 77,780 ppm, respectively. The findings indicated that the methanolic extract, ethyl acetate, and n-hexane soluble fraction were suitable sources of natural antioxidants.

Antioxidant, thrombolytic, cytotoxic and antibacterial activities of leaves of Vitex peduncularis

Vitex peduncularis (Verbenaceae), a perennial tree, is locally utilized for possessing multifaceted pharmacological properties including analgesic, antimalarial, anti-angina, antidiabetic, antioxidant, anti-inflammatory and hepatoprotective. Hence, this study aims to evaluate their antioxidant, thrombolytic, cytotoxic and antibacterial activities from the methanolic crude extract and its various solvent fractions. Data from antioxidant and thrombolytic assay displayed that Kupchan partitioned chloroform soluble fraction revealed the maximum phenolic content (73.39 mg of GAE/g of extract), DPPH free radical scavenging activity (26.69 μg/ml) and thrombolytic activity (37.35%). In brine shrimp lethality evaluation, the LC50 values indicate that the most cytotoxic compound was found to be from ethyl acetate soluble fraction (LC50 6.73 μg/ml). Finally, antibacterial activity of different fractions showed that both chloroform and methanolic fraction possessed mild activity against the tested bacteria. The potential biological effects of the crude fractions have been demonstrated demanding future studies of the isolated compounds to assess these activities. Bangladesh J. Sci. Ind. Res. 57(4), 239-246, 2022

Antimicrobial Activity of Smilax china L. Root Extracts against the Acne-Causing Bacterium, Cutibacterium acnes, and Its Active Compounds

The root of Smilax china L. is used in traditional Korean medicine. We found that the Smilax china L. root extract has strong antimicrobial activity against two Cutibacterium acnes strains (KCTC 3314 and KCTC 3320). The aim of this study was to identify the beneficial properties of Smilax china L. extracts for their potential use as active ingredients in cosmetics for the treatment of human skin acne. The high-performance liquid chromatography (HPLC) and liquid chromatography-hybrid quadrupole time-of-flight mass spectrometry (LC/QTOF/MS) methods were used to obtain the profile of secondary metabolites from the ethyl acetate-soluble fraction of the crude extract. Agar diffusion and resazurin-based broth microdilution assays were used to evaluate antimicrobial activity and minimum inhibitory concentrations (MIC), respectively. Among the 24 metabolites, quercetin, resveratrol, and oxyresveratrol were the most potent compounds against Cutibacterium acnes. Minimum inhibitory concentrations of quercetin, resveratrol, and oxyresveratrol were 31.25, 125, and 250 μg/mL, respectively.

Anti-Inflammatory Activities of Constituents from Cinnamomum insularimontanum Hayata Leaves and Their Mechanisms.

Cinnamomum insularimontanum is an endemic species of Taiwan. Although most Cinnamomum plants have significant biological activity, the bioactivity investment of C. insularimontanum is rare. Since inflammation plays an important role in many diseases, anti-inflammatory compounds can be developed into healthcare products. Therefore, we first conducted a study on the anti-inflammatory activity of C. insularimontanum leaves. First, we examined the antiinflammation activity of essential oil from C. insularimontanum leaves, and it revealed potent anti-inflammatory activity. A total of 23 volatile compounds were identified in C. insularimontanum leaves' essential oil by using GC/MS analysis. Among them were 1,8-cineole (35.94%), α-eudesmol (6.17%), pinene (7.55%), sabinene (5.06%), and isobornyl acetate (4.81%). According to previous studies, 1,8-cineole might be an anti-inflammation principal compound of C. insularimontanum leaves. Next, the ethanolic extracts of C. insularimontanum leaves also exhibited good anti-inflammatory activity. Two bioactive compounds, isoburmanol (F1) and burmanol (F2), were isolated from the ethyl acetate soluble fraction by using the bioactivity-guided separation protocol and spectroscopic analysis. F1 was obtained from C. insularimontanum for the first time, and F2 was isolated for the first time from natural resources. Both F1 and F2 could inhibit the production of nitric oxide (NO), and the IC50 values were 14.0 μM and 43.8 μM, RAW 264.7 cells after induction of lipopolysaccharide. Furthermore, F1 and F2 also revealed significant inhabitation effects on iNOS and COX-2 protein expression. The anti-inflammation activity of F1 and F2 was different from the common pathway of inhibiting NF-κB. Both of them could inhibit the production of NO and PGE2 by directly inhibiting the AP-1 (c-Jun) protein and then inhibiting the downstream iNOS and COX-2. Although both F1 and F2 possessed significant anti-inflammatory activity, the activity of F1 was better than F2. Through molecular docking simulation analysis, the results show that F1 and F2 interact with AP-1, inhibit the binding of AP-1 to DNA, and cause AP-1 to fail to transcribe the related factors of inflammation. The binding ability of AP-1 and F1 was stronger than F2, and that is the reason why F1 exhibited better activities in both downstream proteins and inflammatory cytokines. Based on the results obtained in this study, the essential oil and F1 and F2 isolated from C. insularimontanum leaves have good anti-inflammatory activities, and it is expected to be used as a reference for the development of medical care products in the future.

Phytochemical profiling, in vitro biological activity, docking studies, and cytotoxicity assessments of Rondeletia odorata Jacquin: An unexplored plant of the coffee family

Rondeletia odorata Jacquin is a flowering plant that belongs to the coffee family. As a rich source of polyphenols with significant antioxidant potential, R. odorata may have health benefits. Therefore, in the current work, ethanolic extract of aerial parts and its n-hexane, ethyl acetate, and n-butanol soluble fractions were analyzed for their antioxidant potential and various enzyme inhibition properties. The total phenolic and flavonoid contents of the crude ethanol extract (ROE) and its n-hexane (ROH), ethyl acetate (ROEA), and n-butanol (ROB) fractions were determined spectrophotometrically, while metabolic profiling was established through UHPLC-MS analysis, which revealed the presence of 58 phytochemicals. Total phenolic and flavonoid contents of ROE extract were measured as 51.92mg GA.Eq./g of dry extract and 52.35mg Qu.Eq./g of the dry extract, respectively. In the DPPH radical scavenging activity assay, ROE and ROEA showed the highest potential with values of 62.13 ± 0.62 and 76.31% ± 1.86%, respectively, comparable to quercetin (80.89% ± 0.54%). Similarly, in the FRAP assay, the same pattern of the activity was observed with ROE and ROEA, which displayed absorbance values of 1.32 ± 0.01 and 0.80 ± 0.02 at 700nm, respectively, which are comparable (1.76 ± 0.02) with the reference compound quercetin, whereas the ROH showed maximum metal-chelating capacity (62.61% ± 1.01%) among all extracts and fractions. Antibacterial activity assay indicated that the ROEA fraction was the most active against Serratia marcescens, Stenotrophomonas maltophilia, Bacillus subtilis, Klebsiella pneumonia, and Staphylococcus aureus, while the rest of the fractions showed good to moderate activity. Enzyme inhibition assays showed that ROEA fraction exhibited the highest activity with IC50 values of 2.78 ± 0.42 and 3.95 ± 0.13mg/mL against urease and carbonic anhydrase (CA), respectively. Furthermore, the docking studies of some of the major compounds identified in the extract revealed a strong correlation with their inhibitory activity. All extracts and fractions were also tested for their thrombolytic activity, and the ROB fraction showed a notable potential. Antiviral assay led to remarkable outcomes. Thus, it can be inferred that aerial parts of R. odorata are potential sources of bioactive components with several significant pharmacological activities.

Lanostane Triterpenoids and Ergostane Steroids from Ganoderma luteomarginatum and Their Cytotoxicity.

Macrofungus Ganoderma luteomarginatum is one of the main species of Ganoderma fungi distributed in Hainan province of China, the fruiting bodies of which have been widely used in folk as a healthy food to prevent tumors. To explore the potential cytotoxic constituents from G. luteomarginatum, the phytochemical investigation on the ethyl acetate soluble fraction of 95% ethanolic extract from the fruiting bodies of this fungus led to the isolation of twenty-six lanostane triterpenoids (1–26), including three undescribed ones (1–3), together with eight ergostane steroids (27–34). The structures of three new lanostane triterpenoids were elucidated as lanosta-7,9(11)-dien-3β-acetyloxy-24,25-diol (1), lanosta-7,9(11)-dien-3-oxo-24,26-diol-25-methoxy (2), and lanosta-8,20(22)-dien-3,11,23-trioxo-7β,15β-diol-26-oic acid methyl ester (3) by the analysis of 1D, 2D NMR, and HRESIMS spectroscopic data. All isolates were assayed for their cytotoxic activities using three human cancer cell lines (K562, BEL-7402, and SGC-7901) and seven lanostane triterpenoids (1, 2, 7, 13, 18, 22, and 24), and one ergostane steroid (34) showed definite cytotoxicity with IC50 values that ranged from 6.64 to 47.63 μg/mL. Among these cytotoxic lanostane triterpenoids, compounds 2 and 13 showed general cytotoxicity against three human cancer cell lines, while compounds 1 and 18 exhibited significant selective cytotoxicity against K562 cells with IC50 values of 8.59 and 8.82 μg/mL, respectively. Furthermore, the preliminary structure–cytotoxicity relationships was proposed.

Total Bioactive Contents, Metabolic Profiling, Docking Studies, Antioxidant and Enzyme Inhibition Activities of Convolvulus Arvensis L. and Multivariate Analysis to Unravel a Potential Herb as Natural Resource for Pharmaceutical Industry.

Convolvulus arvensis L. is an evergreen herb growing in various regions of Pakistan. Despite of several medicinal properties associated to this herb, it was not investigated scientifically for its bioactive compounds and detailed pharmaceutical properties. Therefore, its methanolic extract was divided into hexane (CA-H), chloroform (CA-C), ethyl acetate (CA-E) and butanol (CA-B) soluble fractions. CA-H and CA-C were found rich in phenolics (30.73±0.63 and 20.15±0.59 mg GAE/g of the extract, respectively), and the same fractions exhibited significant antioxidant activities (DPPH: 5.23±0.11 & 12.34±0.17 mg TE/g extract, respectively; ABTS: 36.82±0.04 & 56.74±0.61 mg TE/g extract, respectively). Also in CUPRAC activity assay, CA-H and CA-C exhibited highest activities as 87.30±0.46 and 56.74±0.61 mg TE/g extract, respectively, while CA-C was most active in FRAP activity assay with value of 40.21±2.19 mg TE/g extract. Total antioxidant capacity (1.23±0.033 mmol TE/g extract) was also found higher for CA-C, while CA-H activity was also comparable, however, CA-H showed higher metal chelating activity (22.74±0.001 mg EDTAE/g extract) than that of CA-C (17.55±0.22 mg EDTAE/g extract). These activities clearly revealed a direct relation between antioxidant potential and phenolic contents of CA-H and CA-C. In AChE and BChE inhibitory assay, CA-H and CA-E showed better inhibition (AChE: 8.24±0.77 & 4.46±0.007 mg GALAE/g extract; BChE: 5.40±0.02 & 1.92±0.24 mg GALAE/g extract) as compared to other fractions, whereas, against tyrosinase, CA-B was most active (37.35±0.53 mg KAE/g extract). CA-H and CA-C also showed higher inhibitory potential (0.98±0.08 & 0.58±0.01 mmol ACAE/g extract) against α-Amylase; while against α-Glucosidase, CA-E was the most active fraction. UHPLC/MS analysis of the methanolic extract of C. arvensis disclosed the presence of 62 compounds as sterols, triterpenes, flavonoids, fatty acids, alkaloids and coumarins. In Multivariate Analysis, the total phenolic contents were correlated strongly with all antioxidant assays except FRAP and DPPH. Regarding enzyme inhibitory properties, only AChE, BChE and α-amylase were correlated with the total phenolic contents in the extracts. Docking analyses confirmed these findings, as identified compounds had high binding free energy and inhibition constants with the enzymes studied. It was finally concluded that C. arvensis is a potential industrial crop, which can be a component of nutraceuticals and functional foods, if evaluated for its toxicity.

Chemico-Biological Profiling of Blumea lacera (Burm.f.) DC. (Family: Asteraceae) Provides New Insights as a Potential Source of Antioxidant, Cytotoxic, Antimicrobial, and Antidiarrheal Agents.

Blumea lacera (Burm.f.) DC., popular for its traditional use in different diseases, was employed in phytochemical and biological investigations. The chemical studies led to the isolation of acyclic diterpene-phytol (1) along with two fatty acids-linolenic acid (2) and oleic acid (3). All the structures were determined by 1H NMR spectroscopic analysis and first time reported from this plant. Different fractions of crude methanol extract were subjected to antioxidant, cytotoxicity, antimicrobial, and antidiarrheal assays. The molecular docking studies have been implemented using PyRx, UCSF Chimera, Discovery Studio, and online tools. In addition, The ADME/T analysis and PASS prediction were implemented by using PASS online tools. In the molecular docking study of antioxidant, cytotoxicity, antimicrobial, and antidiarrheal activity, the compounds showed strong binding affinity ranging from −4.5 to −6.2 kcal/mol. Again, all three isolated compounds met the preconditions of Lipinski's five rules for drug discovery. In DPPH free radical scavenging assay, the pet-ether and chloroform soluble fraction showed noteworthy antioxidant activity sowing promising IC50 values (10.76 μg/ml and 11.77 μg/ml, respectively), compared to the standard (6.05 μg/ml) with a total phenolic content range of 7.33–40.33 mg of GAE/gm. The pet-ether soluble fraction revealed substantial cytotoxicity showing an LC50 value of 1.03 μg/ml, compared to the standard (0.93 μg/ml). Besides, ethyl acetate soluble fraction showed moderate activity against both Gram-positive and Gram-negative bacteria, while both ethyl acetate and pet-ether soluble fraction showed excellent dose-dependent antidiarrheal activity.

Phytochemical and Pharmacological Profiling of Heritiera fomes Buch. Ham. Deciphered Thrombolytic, Antiarthritic, Anthelmintic, and Insecticidal Potentialities via In Vitro Approach.

Medicinal plants have been crucial in treating various chronic ailments since ancient times. The objective of this study was to evaluate in vitro pharmacological properties of petroleum ether, chloroform, and ethyl acetate soluble fractions of ethanolic extract (leaf, bark, and root) of Heritiera fomes Buch. Ham., including the phytochemical screening of the plant. Thrombolytic and antiarthritic properties were assessed through the clot lysis and protein denaturation experimental method, correspondingly. Anthelmintic and insecticidal activities were studied against Pheretima posthuma and Tribolium castaneum, respectively. The phytochemical analysis exhibited numerous active phytochemicals in different solvent fractions. In thrombolytic investigation, among all crude extracts, ethanolic leaf extract showed the highest 33.12 ± 7.52% clot lysis as compared to standard streptokinase (67.77 ± 9.78%). In antiarthritic assay, all the tested samples exhibited noteworthy protein denaturation in dose-dependent manner (100–500 μg/mL), whereas the utmost percentage inhibition was noticed for chloroform extract of roots (63.28 ± 5.96% at 500 μg/mL). All crude extracts exhibited a significant anthelmintic activity in different concentrations (25–75 mg/mL) and revealed paralysis and death of earthworms in comparison with albendazole; ethanolic extract of the bark was found to be more potent at the highest dose. For the insecticidal test, ethanolic extract of the leaf showed the utmost mortality rate (73%). The outcomes of the investigation confirmed the potential thrombolytic, antiarthritic, anthelmintic, and insecticidal activities of the different extracts of H. fomes, and hence, advanced studies on the isolation and identification of active phytocompounds are highly needed for new drug development.

Efficacy of Phytochemicals Derived from Roots of Rondeletia odorata as Antioxidant, Antiulcer, Diuretic, Skin Brightening and Hemolytic Agents-A Comprehensive Biochemical and In Silico Study.

Roots of Rondeletia odorata are a rich source of phytochemicals with high antioxidant potential and thus may possess health benefits. This study used the LC-MS technique to identify phytoconstituents in R. odorata roots extract/fractions. Results revealed that n-butanol fraction and ethanolic extract contained total phenolic and flavonoid contents with values of 155.64 ± 0.66 mgGAE/g DE and 194.94 ± 0.98 mgQE/g DE, respectively. Significant potential of antioxidants was observed by DPPH, CUPRAC and FRAP methods while the ABTS method showed moderate antioxidant potential. Maximum % inhibition for urease, tyrosinase and carbonic anhydrase was shown by ethanolic extract (73.39 ± 1.11%), n-butanol soluble fraction (80.26 ± 1.59%) and ethyl acetate soluble fraction (76.50 ± 0.67%) which were comparable with thiourea (standard) (98.07 ± 0.74%), kojic acid (standard) (98.59 ± 0.92%) and acetazolamide (standard) (95.51 ± 1.29%), respectively, while all other extract/fractions showed moderate inhibition activity against these three enzymes. Hemolytic activity was also observed to range from 18.80 ± 0.42 to 3.48 ± 0.69% using the standard (triton X-100) method. In total, 28 and 20 compounds were identified tentatively by LC-MS analysis of ethanolic extract and n-butanol soluble fraction, respectively. Furthermore, molecular docking was undertaken for major compounds identified by LC-MS for determining binding affinity between enzymes (urease, tyrosinase and carbonic anhydrase) and ligands. It was concluded that active phytochemicals were present in roots of R. odorata with potential for multiple pharmacological applications and as a latent source of pharmaceutically important compounds. This should be further explored to isolate important constituents that could be used in treating different diseases.