The good and the bad: Alkaloid screening and brineshrimp bioassays of aqueous extracts of 31 medicinal plants of eastern Nicaragua

Context: Presence/absence tests for alkaloids of 31 medicinal vascular plant species from 31 genera and 26 families of eastern Nicaragua provided a baseline for bioactivity tests.

Objective: To determine the bioactivity and cytoxicity of aqueous extracts of widely used medicinal species in eastern Nicaragua.

Materials and methods: Ethnomedicinal applications were obtained from interviews of traditional healers. We used Dragendorff’s reagent to test alkaloids and brine shrimp for cytotoxicity of aqueous extracts.

Results: Twenty-nine of the 31 species tested positive for alkaloids. The median lethal concentration that kills 50% of the larvae within 24?h of contact with the extract (LC₅₀ was less than 1000 µg/mL for 4 (13%) species (the usual cytotoxic category), 1001–5000 µg/mL for 23 (74%) species, and between 5001–7500 µg/mL for the remaining 4 (13%) species.

Discussion and Conclusion: Twenty-five of the ethnomedicines contain alkaloids but are not cytotoxic. In contrast to first suppositions, we suggest that this is a good and desirable, and perhaps expected, outcome. Medicinal plants that are cytotoxic may obviously control or kill bacteria or other pathogens, but may also negatively affect the patient; some high alkaloid levels have been associated with carcinogens. Thus, perhaps the majority of effective medicinals should be expected to be noncytotoxic. We suggest that this is a new paradigm for consideration of the overall value and effectiveness of medicinals. Of course, medicinals also can be effective in numerous ways (e.g., organ stimulation or other physiological functions) other than simply as antimicrobials or antipathogens.     

Median Lethal Concentrations of Amazonian Plant Extracts in the Brine Shrimp Assay

In the current study, median lethal concentrations (LC₅₀) were obtained in the brine shrimp assay (BSA) for 31 methanol and water plant extracts representing 21 Amazonian plant species, most of which are used in traditional medicine. The following species presented extracts with LC₅₀ values less than 50 µg/ml: Croton cajucara Benth., Micrandropsis scleroxylon W.A. Rodrigues, Piper aduncum L., P. hostmannianum (Miq.) C. DC., P. tuberculatum Jacq., Protium aracouchini (Aubl.) Marchand, and Tapura amazonica Poepp. The methanol extract of T. amazonica Poepp., a species known to be toxic according to ethnobotanic reports, was the most active (LC₅₀ 1.2 µg/ml). The data presented are considered useful as general toxicity parameters, and their relevance in the context of discovery of new antitumor agents and potential sources of (new) bioactive compounds in the Amazonian flora is discussed.     

Two New Triterpene Glycosides from Dendrocalamus strictus

Abstract

The methanol extract of Gracilaria changii. B.M. Xia &; I.A. Abbott (Gracilariaciae), which exhibited antimicrobial activity, was tested for in vivo. brine shrimp lethality and in vitro.anticancer cell line activity. Both toxicity tests [brine shrimp test with LC₅₀ value 4.28 mg/mL (12 h), 3.13 mg/mL (24 h) and anticancer cell line activity with IC₅₀ value 44.10 μ g/mL] exhibited no significant toxicity result. These alga extracts, with high LC₅₀ and IC₅₀ values, are potential sources for novel antimicrobial compounds.     

Cytotoxic Activities of Salvia. of the Labiatae Family

ABSTRACT

Eight crude extracts of five Salvia. species were evaluated for cytotoxic activities against brine shrimps and four human cancer cell lines [human colon adenocarcinoma (HCA), HepG2, MCF-7, and human pancreatic carcinoma (HPC) along with a normal mouse cell line (areolar cells)] as a control. In the brine shrimp lethality test, all samples, except S. fruticosa. L. (Sifnos collection) and S. verbenaca. L. (Zante collection), were found to be highly active with ED₅₀ values less than 300?μg/ml. In the case of human cancer cell lines, S. fruticosa., collected from Kalymnos and Crete, were active against HCA cells with LC₅₀?=?60.4 and 40.1?μg/ml respectively. Interestingly, only one sample, S. fruticosa. collected from Kalymnos, was active against HepG2 cells with LC₅₀?=?68.1?μg/ml. In the case of MCF-7 cells, S. fruticosa. collected from three different locations (Kalymnos, Rhodos, and Crete) showed similar activity with LC₅₀?=?43.1, 41.1, and 42.3?µg/ml, respectively. All S. fruticosa. samples were found to be cytotoxic toward a normal mouse cell line when tested at 0.1?mg/ml. All the other samples had LC₅₀ values greater than 75?µg/ml,and were considered to be inactive.     

Evaluation of the Ethnomedical Claims of Murraya koenigii.

Abstract

Based on sethnomedicine, Murraya koenigii. (L.) Spreng. is used as a stimulant, antidysentery, and for the management of diabetes mellitus. Twelve carbazole alkaloids were isolated from the stem, seed, and leaf of the plant growing in Nigeria and Sri Lanka. The methanol extracts were devoid of hypoglycemic activity, and some isolates decreased insulin secretion when they were subjected to both in vivo. and in vitro. (insulin secretion from INS-1 cells) antidiabetic tests. The cytotoxicity of the leaf and stem methanol extracts determined by the brine shrimp lethality bioassay were LC₅₀ 61.5 and 14.5?µg/ml, respectively. These extracts caused CNS depression in albino mice at the dose levels of 25–400?mg/kg. Also, they had an IC₅₀ of 34.0 and 35.0?µg/ml at 24?h, respectively, against trichomonas. These results confirmed the use of the plant as an antidysentery caused by trichomonas but refute the antidiabetic and stimulant ethnomedical claims for the plant. The differences observed in their alkaloidal composition suggested probable influence of geographical location on the elaboration of carbazole alkaloids in the plant and differences in the localization of carbazole alkaloids in the plant parts.     

Bioactivity of Extractives from Stachytarpheta urticaefolia

The n-hexane and methanol extracts of Stachytarpheta urticaefolia leaves exhibited significant antinociceptive effects (p < 0.01) in acetic acid-induced writhing model at a dose of 250 mg/kg b.w. Two compounds isolated from the plant, ipolamiide (1) and α-spinasterol (2), were also found to be potent inhibitors of acetic acid-induced writhing on mice at 50 mg/kg b.w. (p < 0.01). Abortifacient activity was demonstrated within 20 h of administration of a methanolic leaf extract at 200 mg/kg b.w. The methanolic leaf extract and compounds 1 and 2 were also tested for in vitro antibacterial activities against 18 bacterial strains. The extract showed mean zones of inhibition ranging from 17 to 30 mm at 500 µg/disk, but the compounds demonstrated only mild or no in vitro antibacterial activity even at 400 µg/disk. Finally, in a cytotoxicity study, the n-hexane and methanol extracts of leaves and root bark of the plant and the purified compounds (1, 2) exhibited LC₅₀ values of 5.42, 4.42, 17.15, 5.38, 3.93, and 13.65 µg/ml, respectively, against the brine shrimp nauplii, whereas the positive control, vincristine sulfate, showed an LC₅₀ of 0.583 µg/ml.     

Maculosin,a non-toxic antioxidant compound isolated from Streptomyces sp. KTM18

ContextStreptomyces species are prolific sources of bioactive secondary metabolites known especially for their antimicrobial and anticancer activities.ObjectiveThis study sought to isolate and characterize antioxidant molecules biosynthesized by Streptomyces sp. KTM18. The antioxidant potential of an isolated compound and its toxicity were accessed.Materials and methodsThe compound was purified using bioassay-guided chromatography techniques. Nuclear magnetic resonance (NMR) experiments were carried out for structure elucidation. The antioxidant potential of the isolated compound was determined using DPPH free radical scavenging assay. The toxicity of the isolated compound was measured using a brine shrimp lethality (BSL) assay.ResultsEthyl acetate extract of Streptomyces sp. KTM18 showed more than 90% inhibition of DPPH free radical at 50 µg/mL of the test concentration. These data were the strongest among 13 Streptomyces isolates (KTM12–KTM24). The active molecule was isolated and characterized as maculosin (molecular formula, C₁₄H₁₆N₂O₃ as determined by the [M + H]⁺ peak at 261.1259). The DPPH free radical scavenging activity of pure maculosin was higher (IC₅₀, 2.16 ± 0.05 µg/mL) than that of commercial butylated hydroxyanisole (BHA) (IC₅₀, 4.8 ± 0.05 µg/mL). No toxicity was observed for maculosin (LD₅₀, <128 µg/mL) in brine shrimp lethality assay (BSLA) up to the compound’s antioxidant activity (IC₅₀) concentration range. The commercial standard, berberine chloride, showed toxicity in BSLA with an LD₅₀ value of 8.63 ± 0.15 µg/mL.ConclusionsMaculosin may be a leading drug candidate in various cosmetic and therapeutic applications owing to its strong antioxidant and non-toxic properties.     

Investigation of nepetolide as a novel lead compound: Antioxidant,antimicrobial, cytotoxic,anticancer, anti-inflammatory,analgesic activities and molecular docking evaluation

In the present study, we describe various pharmacological effects and computational analysis of nepetolide, a tricyclic clerodane-type diterpene, isolated from Nepeta suavis. Nepetolide concentration-dependently (1.0–1000?µg/mL) exhibited 1,1-diphenyl,2-picrylhydrazyl free radical scavenging activity with maximum effect of 87.01?±?1.85%, indicating its antioxidant potential, as shown by standard drug, ascorbic acid. It was moderately active against bacterial strain of Staphylococcus aureus. In brine shrimp’s lethality model, nepetolide potently showed cytotoxic effect, with LC₅₀ value of 8.7?µg/mL. When evaluated for antitumor activity in potato disc tumor assay, nepetolide exerted tumor inhibitory effect of 56.5?±?1.5% at maximum tested concentration of 1000?µg/mL. Nepetolide at 20?mg/kg reduced carrageenan-induced inflammation (P?<?.001 vs. saline group) in rat paw. Nepetolide dose-dependently (100–500?mg/kg) decreased acetic acid evoked writhes, as exhibited by diclofenac sodium. In-silico investigation of nepetolide was carried out against cyclooxygenase-2, epidermal growth factor receptor and lipoxygenase-2 targets. Virtual screening through Patchdock online docking server identified primarily hydrophobic interactions between ligand nepetolide and receptors proteins. Enhanced hydrogen bonding was predicted with Autodock showing 6–8 hydrogen bonds per target. These results indicate that nepetolide exhibits antioxidant, antibacterial, cytotoxic, anticancer, anti-inflammatory and analgesic activities and should be considered as a lead compound for developing drugs for the remedy of oxidative stress-induced disorders, microbial infections, cancers, inflammations and pain.     

Toxicity study of Vernonia cinerea

The methanol extract of Vernonia cinerea Less (Asteraceae), which exhibited antimicrobial activity, was tested for toxicity. In an acute toxicity study using mice, the median lethal dose (LD₅₀) of the extract was greater than 2000?mg/kg, and we found no pathological changes in macroscopic examination by necropsy of mice treated with extract. As well as the oral acute toxicity study, the brine shrimp lethality test was also done. Brine shrimp test LC₅₀ values were 3.87?mg/mL (6?h) and 2.72?mg/mL (24?h), exhibiting no significant toxicity result. In conclusion, the methanol extract of V. cinerea did not produce toxic effects in mice and brine shrimp.     

Analysis of Peganum harmala,Melia azedarach and Morus alba extracts against six lethal human cancer cells and oxidative stress along with chemical characterization through advance Fourier Transform and Nuclear Magnetic Resonance spectroscopic methods towards green chemotherapeutic agents

Traditional medicines implicate consumption of plant crude extracts, which may consist of extensive phytochemical diversity. Overall, the most biologically active extract of Peganum harmala (seeds) exhibited significant cytotoxic activity on Artemia salina with LC₅₀ value of 61.547 µg/mL, while P. harmala (roots) [LC₅₀ = 124.229 µg/mL] and M. azedarach (fruits) [LC₅₀ = 147.813 µg/mL] showed moderate cytotoxic potential. P. harmala (seeds) extract also showed the maximum antitumor potential with 52.278 µg/mL LC₅₀. Branches of P. harmala and Morus alba were not active in both bioassays. These outcomes were further reinforced by the levels of phenolics and flavonoids checked against gallic acid and quercetin equivalents, respectively, by standard curves. Current study aims to isolate, structurally characterize and analyze the bioactive compound from plant extracts by using chromatographic and spectrophotometric techniques. Bioactivity guided isolation of extracts led to the isolation of PH-HM-16 from ethyl acetate fraction P. harmala seeds. Chemical structure of PH-HM-16 was elucidated by ESI-MS, ¹H NMR, ¹³C NMR, HSQC and IR spectrum. The results demonstrated significant positive anticancer activities against six human cancer cell lines assessed through MTT cancer cell growth inhibition assay. PH-HM-16 was most effective against prostate cancer cell lines [IC₅₀ = 17.63 µg/mL] followed by breast cancer cell line MCF7 [IC₅₀ value of 41.81 µg/mL]. IC₅₀ value of PH-HM-16 against human myeloid leukemia cell line HL-60 and human colorectal tumor cells HCT-116 was observed as 68.77 µg/mL and 71.54 µg/mL respectively. The IC 50 value of PH-HM-16 compound was not significant against human gastric cancer SGC-7901 (111.89 µg/mL) and human lung adenocarcinoma epithelial cell line A549 (176.04 µg/mL). Isolated bioactive metabolite PH-HM-16 possesses significant antitumor potential so this could be the first step to develop an effective anticancer agent. Hence, this compound represents a promising potential to be chemically standardized or developed into pharmaceuticals for the chemoprevention and/or the treatment of certain types of cancer, especially as adjuvant phytotherapeutics in conventional chemotherapy.     

GC-MS investigation and toxicological evaluation of alkaloids from Leptadenia pyrotechnica

GC-MS analysis of isolated alkaloids from aerial parts of Leptadenia pyrotechnica (Forsk.) Decne (Asclepiadaceae) was performed. Twenty-four alkaloids and six simple amines were detected for the first time in this plant. Almost all of the alkaloids belonged to pyridine, pyrrole, pyrazine, and indole types. The acute LC₅₀ of the total alkaloids and alcohol extracts estimated by means of brine shrimp toxicity test were 63.09 and 11.89?ppm, respectively. The antitumor activities of these extracts, using potato disc screen, showed good activity represented by ?33.6% and ?49.3%, respectively.     

Brine shrimp cytotoxicity and antimalarial activity of plants traditionally used in treatment of malaria in Msambweni district

Abstract

Context: In Kenya, most people use traditional medicine and medicinal plants to treat many diseases including malaria. To manage malaria, new knowledge and products are needed. Traditional herbal medicine has constituted a good basis for antimalarial lead discovery and drug development.

Objectives: To determine in vivo antimalarial activity and brine shrimp toxicity of five medicinal plants traditionally used to treat malaria in Msambweni district, Kenya.

Materials and methods: A 0.2?ml saline solution of 100?mg/kg aqueous crude extracts from five different plant parts were administered orally once a day and evaluated for their in vivo chemosuppressive effect using Plasmodium berghei berghei-infected Swiss mice for four consecutive days. Their safety was also determined using Brine shrimp lethality test: Grewia trichocarpa Hochst ex A. Rich (Tiliaceae) root, Dicrostachys cinerea (L) Wight et Am (Mimosaceae) root, Tamarindus indica L. (Caesalpiniaceae) stem bark, Azadirachta indica (L) Burn. (Meliaceae) root bark, and Acacia seyal Del. (Mimosaceae) root.

Results: Parasitaemia was as follows: A. indica, 3.1%; D. cinerea, 6.3%; T. indica, 25.1%; A. seyal, 27.8%; and G. trichocarpa, 35.8%. In terms of toxicity, A. indica root bark extract had an LC₅₀ of 285.8?µg/ml and was considered moderately toxic. T. indica stem bark extract and G. trichocarpa root extract had an LC₅₀ of 516.4 and 545.8?µg/ml, respectively, and were considered to be weakly toxic while A. seyal and D. cinerea root extracts had a LC₅₀ >1000?µg/ml and were, therefore, considered to be non-toxic.

Discussion and conclusion: All extracts had antimalarial activity that was not significant compared to chloroquine (p?≥?0.05). No extract was toxic to the arthropod invertebrate, Artemia salina L. (Artemiidae) larvae, justifying the continued use of the plant parts to treat malaria.     

Antibacterial,antifungal and cytotoxic activities of salviasperanol isolated from Amorphophallus campanulatus

Salviasperanol, a diterpenoid isolated from Amorphophallus campanulatus (Roxb.) Bl. (Araceae) was studied for in vitro antibacterial, antifungal, and cytotoxic activities. The disc diffusion technique was used to determine in vitro antibacterial and antifungal activities. Cytotoxicity was determined against brine shrimp nauplii. In addition, minimal inhibitory concentration (MIC) was determined using serial dilution technique to determine antibacterial potency. The MIC values against these bacteria ranged from 8 to 64 μg/mL. The compound showed significant antibacterial activity against four Gram-positive bacteria (Bacillus subtilis, Bacillus megaterium, Staphylococcus aureus, Streptococcus β-haemolyticus) and six Gram-negative bacteria (Escherichia coli, Shigella dysenteriae, Shigella sonnei, Shigella flexneri, Pseudomonus aeruginosa, Salmonella typhi). The compound showed weak antifungal activity against a number of fungi. In the cytotoxicity determination, the LC₅₀ of the compound against brine shrimp nauplii was 8.02 μg/mL.     

Evaluating the Teratogenicity of Ritodrine and Nifedipine using a Frog Embryo Teratogenesis assay (FETAX)

The Frog Embryo Teratogenesis Assay—Xenopus (FETAX) was used to assess the teratogenic potential of two tocolytics. Embryos of the South African clawed frog, Xenopus laevis, were exposed to ritodrine or nifedipine. Exposure media were changed and monitored at 24-hour intervals. The 96-hour LC₅₀ (Lethal concentration), the 96-hour EC₅₀ (Malformation), and the No Observable Adverse Effect Concentrations (NOAEC) and the Lowest Observable Adverse Effect Concentration (LOAEC) for mortality, malformation and length were determined for each drug. Nifedipine was determined to be the more toxic and teratogenic than ritodrine, with a LC₅₀ of 0.606?µg/L, an EC₅₀ of 0.006?µg/L, and a teratogenicity Index (TI) value (LC₅₀/EC₅₀) of 101. On the other hand, the LC₅₀ of ritodrine was 28.571?mg/L. In addition; the LC₅₀, EC₅₀ and TI values for nifedipine in the 5?mg/L ritodrine?+?nifedipine combination group were determined as 1.050?µg/L, 0.868?µg/L and 1.5 respectively. For ritodrine, the NOAEC and LOAEC values were determined as 2?mg/L and 4?mg/L, respectively. For the nifedipine and the ritodrine?+?nifedipine groups; while the LOAEC values of these groups were 0.0001?µg/L and 0.1?µg/L, respectively. NOAEC value couldn’t be determined. Our results demonstrated that nifedipine administration was associated with higher levels of teratogenic and toxic effects. However, the ritodrine?+?nifedipine combination form reduced the toxic and teratogenic effects of nifedipine on Xenopus embryos. Further studies should be conducted in order to investigate the optimal combination concentrations of these substances for the treatment of preterm labor.     

Antimycoplasmal activity of some plant species from northern Nigeria compared to the currently used therapeutic agent

Context:?Mycoplasma spp. are obligate parasites of humans and animals. But due to the special requirements needed to culture Mycoplasma in the laboratory, little or no research has been done to evaluate the efficacy of medicinal plants on the organism.

Objective:?To screen medicinal plants traditionally used to treat infections for possible antimycoplasmal and cytotoxic activities.

Materials and methods:?Acetone extracts of 21 Nigerian medicinal plants were analyzed for antimycoplasmal and cytotoxicity activities using the metabolic inhibition and colorimetric methods, respectively. The extract with the best antimycoplasmal activities was also analyzed for its phytochemical constituents using the desktop method.

Results:?Calotropis procera (Aiton) R.Br (Asclepiadaceae) extract had the best antimycoplasmal effect with a minimum inhibitory concentration (MIC) of 80 µg/mL and minimum mycoplasmacidal concentration (MMC) of 160 µg/mL. This extract contained saponins, tannins, cardiac glycosides, alkaloids, and flavonoids. The extract of Vernonia amygdalina Delile (Compositae) was the most cytotoxic with median lethal concentration (LC₅₀) of approximately 17 µg/mL, and that of Anacardium occidentale L. (Anacardiaceae) was the least cytotoxic with an LC₅₀ of approximately 1919 µg/mL.

Discussion:?Calotropis procera is a promising plant for an alternative antimycoplasmal agent because the crude acetone extract had a higher mycoplasmacidal activity than the conventional drug tylosin, which is currently used in treatment of the disease in Nigeria.

Conclusion:?The crude extract of Calotropis procera is worth investigating for the development of a potent agent against cattle Mycoplasma, which has long defied solution by conventional chemotherapy.     

An improved brine shrimp larvae lethality microwell test method

This article described an improved brine shrimp larvae lethality microwell test method. A simply designed connecting vessel with alternative photoperiod was used to culture and collect high yield of active Artemia parthenogenetica nauplii for brine shrimp larvae lethality microwell test. Using this method, pure A. parthenogenetica nauplii suspension was easily cultured and harvested with high density about 100–150 larvae per milliliter and the natural mortality was reduced to near zero by elimination of unnecessary artificial disturbance. And its sensitivity was validated by determination of LC₅₀–24?h of different reference toxicants including five antitumor agents, two pesticides, three organic pollutants, and four heavy metals salts, most of which exhibited LC₅₀–24?h between 0.07 and 58.43?mg/L except for bleomycin and mitomycin C with LC₅₀–24?h over 300?mg/L.     

Identification and comparison of the volatile constituents of fresh and dried leaves of Spondias mombin found in North-central Nigeria: in vitro evaluation of their cytotoxic and antioxidant activities

Context: Various studies have shown that the leaf extracts of Spondias mombin Linn (Anacardiaceae) possess pharmacological properties such as antioxidant and antiviral effects. However, no biological activity from its essential oil has been reported in literature.

Objective: To analyse the chemical constituents, cytotoxic activity and antioxidant capability of the essential oils from fresh and dried leaves of S. mombin.

Materials and methods: Hydrodistillation using Clevenger-type apparatus was employed to obtain the essential oil. Oil analysis was performed using an HP 6890 Gas Chromatograph coupled with an HP 5973 Mass Selective Detector. The cytotoxicity bioassay was carried out using the brine shrimp lethality test (10,000–0.01?μg/mL). Additionally, the reactive oxygen species scavenging potential of the two S. mombin oils (1000–200?μg/mL) were investigated using a hydroxyl radical scavenging and ferric iron reducing system.

Results: Chemical analysis of essential oils from S. mombin revealed the presence of 41 compounds, with predominance of monoterpenoids, sesquiterpenoids and non-terpenoids derivatives. In both fractions, the principal component was β-caryophellene (27.9–30.9%), followed by γ-cadinene (9.7–12.3%). There was an increase in the oxygenated monoterpenoid contents and a concomitant decrease in the amounts of sesquiterpenoids hydrocarbons observed on drying the leaves. The oil obtained from the fresh leaves was more active than that obtained from dried leaves, with LC₅₀ values (from the brine shrimp lethality assay) of 0.01 and 4.78?μg/mL, respectively. The two oils (from fresh and dried leaves) at 1.0?mg/mL scavenged hydroxyl radical by 83% and 99.8%, respectively. Moreover, they reduced ferric ion significantly and compared favourably with vitamin C.

Conclusions: Essential oil derived from the leaves of S. mombin could hold promise for future application in the treatment of cancer-related diseases.     

Antioxidant and cytotoxic potential of a new thienyl derivative from Tagetes erecta roots

Context: The search for newer compounds against pathogenic species continues unabated due to drug resistance. Traditionally, Tagetes erecta Linn. (Compositae) has been used for the treatment of various parasitic and microbial diseases.

Objective: To evaluate the antioxidant activity of the ethanol extract of Tagetes erecta roots and its cytotoxicity against prostate and HeLa cancer cell lines followed by activity-guided isolation.

Materials and Methods: The antioxidant screening was carried out using diphenylpicrylhydrazyl (DPPH) radical scavenging assay with serial concentrations ranging from 2 to 100 µg/mL, and cytotoxicity was evaluated against prostate (PC-3) and HeLa cell lines using microculture tetrazolium test (MTT) assay with concentrations ranging from 500 to 1.89 µg/mL. Isolation of the ethanol extract was carried out using column chromatography whereby 21 isolates were obtained (T₁-T₂₁), and the most active isolate was subjected for characterization using ultraviolet (UV), infrared (IR), nuclear magnetic resonance (NMR), and mass spectroscopic techniques.

Results: The ethanol extract scavenged DPPH free radicals thereby exhibiting antioxidant activity with an IC₅₀ of 35.9 µg/mL. In addition, the extract conferred noticeable cytotoxicity against the HeLa (LD₅₀ of 164.28 µg/mL) and PC-3 cell lines (LD₅₀ of 407.3 µg/mL). Among all the isolates, T₃ showed antioxidant activity with IC₅₀ of 11.56 µg/mL and cytotoxicity with LD₅₀ of 12.5 µg/mL against HeLa and 30.25 µg/mL against PC-3 cell lines and was characterized as 2-ethynyl-5-(thiophen-2-yl) thiophene.

Discussion: The new thienyl compound (T₃) exhibited profound antioxidant activity and cytotoxicity at relatively lower concentrations than the extract.

Conclusion: The observations provide support for the ethnobotanical use of the plant.     

Evaluation of seeds ethanolic extracts of Triplaris gardneriana Wedd. using in vitro and in vivo toxicological methods

ABSTRACT

Triplaris gardneriana Wedd. is a tree used in folk medicine to treat venereal diseases and inflammation as well as a source of biological compounds with antioxidant capacity. In order to assess the safety of these bioactive compounds, the present study aimed to determine the toxicity of an ethanolic extract of T. gardneriana, (EETg). Toxicological tests included hemolytic activity, toxicity toward the brine shrimp Artemia, cytotoxicity against breast cancer cells (MCF7) and acute oral toxicity in rodents. In addition, toxicogenomics techniques were used to determine genome expression in MCF7 cells exposed to EETg. The results showed that the extract exhibits approximately 60% of hemolytic activity at the highest tested concentration (64 µg/ml) and toxicity against nauplii of Artemia sp. (LC₅₀ of 67.85 µg/ml). Further, EETg appears to be cytotoxic to MCF7 (cell viability reduced to 40% at 250 µg/ml after 24 hr). Genomic data demonstrated differential expression of 14 genes. Data analysis indicated possible altered pathways (e.g., xenobiotic metabolism), possible adverse health risks (e.g., hepatotoxicity), and drugs with similar gene expression profile (e.g., antimicrobials). The investigation provides important information on potentially adverse aspects of EETg, which need to be considered prior to the therapeutic utilization of this plant.

Abbreviations: EETg: ethanolic extract of T. gardneriana seeds; MCF7: michigan cancer foundation-7 which refers to a human breast cell line (adenocarcinoma); NGS: next-generation sequencing; edgeR: empirical analysis of digital gene expression data in R; Consensus: consensus path database; FDR: false discovery rate; NCBI: national center for biotechnology information; KEGG: kyoto encyclopedia of genes and genomes; Ingenuity: ingenuity pathway analysis software; CMAP: connectivity map; OECD: organization for economic co-operation and development; HL-60: human promyelocytic leukemia cells; PC3: prostate cancer cells.     

Bioactivity-guided isolation of cytotoxic constituents from three medicinal plants

Abstract

Context: The ethanol extracts and their fractions of three Indian medicinal plants, Ervatamia coronaria (Jacq.) Stapf, (Apocynaceae), Mimosa pudica L. (Mimosaceae) and Caesalpinia bonduc (L.) Roxb. (Caesalpiniaceae) were tested for their cytotoxic activity in the brine shrimp lethality (BSL) bioassay and in various cancer cell lines. The plants were selected based on their traditional use in the treatment of cancer/tumors.

Objectives: To investigate the in vitro cytotoxicity of Ervatamia coronaria, Mimosa pudica and Caesalpinia bonduc.

Materials and methods: Ethanolic extracts and their fractions of E. coronaria, M. pudica and C. bonduc were subjected to cytotoxicity studies using BSL bioassay method with concentrations of 10, 50, 100, 500 and 1000?µg/ml. The alkaloid fraction of E. coronaria with significant cytotoxicity in BSL bioassay was subjected to in vitro cytotoxicity studies with HT-29, A-549, HepG-2, MCF-7 and L-6 cell lines at concentrations of 12.5, 25, 50, 100 and 200?µg/ml and a DNA fragmentation study using the HT-29 cell line.

Results: The alkaloid fractions of E. coronaria and M. pudica showed significant cytotoxicity with LC₅₀ values of 65.83 and 85.10?µg/ml in the BSL bioassay, respectively. The purified alkaloid fraction of E. coronaria exhibited highest cytotoxicity in HT-29, A-549 and MCF-7 cell lines with IC₅₀ values of 32.5, 47.5 and 72.5?µg/ml, respectively, and induced DNA fragmentation in the HT-29 cell line at a concentration of 65?µg/ml.

Conclusion: The alkaloid fraction of E. coronaria exhibited significant cytotoxicity. Alkaloids such as ervatamine, apparicine and coronaridine that were earlier reported may be responsible for this activity.