Phytochemical Analysis and Profiling of Antioxidants and Anticancer Compounds from Tephrosia purpurea (L.) subsp. apollinea Family Fabaceae.

Tephorosia purpurea subsp. apollinea was extracted with methanol and n-hexane to obtain sub-fractions. The chemical compounds identified with GC-MS and HPLC in T. purpurea subsp. apollinea extracts showed antioxidant and anticancer properties. The antioxidant and anticancer activities were investigated using DDPH and ABTS assays, and MTT assay, respectively. Stigmasta-5,24(28)-dien-3-ol, (3 ß,24Z)-, 9,12,15-octadecatrienoic acid methyl ester, phytol, chlorogenic acid, and quercetin were the major chemical compounds detected in T. purpurea subsp. apollinea. These compounds possessed antioxidant and anticancer properties. The methanol extract showed antioxidant properties with DDPH and ABTS radical scavenging of 84% and 94%, respectively, relative to ascorbic acid and trolox. The anticancer effects of T. purpurea subsp. apollinea against the cancer cell lines MCF7 (IC50 = 102.8 ± 0.6 µg/mL), MG63 (IC50 = 118.3 ± 2.5 µg/mL), T47D (IC50 = 114.7 ± 1.0 µg/mL), HeLa (IC50 = 196.3 ± 2.3 µg/mL), and PC3 (IC50 = 117.7 ± 1.1 µg/mL) were greater than its anticancer effects against U379 (IC50 = 248.4 ± 7.5 µg/mL). However, it had no adverse effects on the normal cells (WI38) (IC50 = 242.9 ± 1.8 µg/mL). Therefore, the major active constituents presented in T. purpurea subsp. apollinea can be isolated and studied for their potential antioxidant and anticancer effects against breast, cervical, and prostate cancers and osteosarcoma.

Variation in Rotenone and Deguelin Contents among Strains across Four Tephrosia Species and Their Activities against Aphids and Whiteflies.

Botanical pesticides have received increasing attention for sustainable control of insect pests. Plants from the genus Tephrosia are known to produce rotenone and deguelin. Rotenone is known to possess insecticidal activities against a wide range of pests, but deguelin's activities remain largely inconclusive. On the other hand, the biosynthesis of rotenone and deguelin may vary in Tephrosia species. This study analyzed the rotenone and deguelin contents in 13 strains across 4 Tephrosia species over 4 growing seasons using HPLC. Our study shows that the species and even the strains within a species vary substantially in the biosynthesis of rotenone and deguelin, and their contents can be affected by the growing season. After identification of the LC50 values of chemical rotenone and deguelin against Aphis gossypii (Glover) and Bemisia tabaci (Gennadius), leaf extracts derived from the 13 strains were used to test their insecticidal activities against the 2 pests. The results showed that the extracts derived from 2 strains of T. vogelii had the highest insecticidal activity, resulting in 100% mortality of A. gossypii and greater than 90% mortality of B. tabaci. The higher mortalities were closely associated with the higher contents of rotenone and deguelin in the two strains, indicating that deguelin also possesses insecticidal activities. This is the first documentation of leaf extracts derived from 13 Tephrosia strains against 2 important pests of A. gossypii and B. tabaci. The strain variation and seasonal influence on the rotenone and deguelin contents call for careful attention in selecting appropriate strains and seasons to produce leaf extracts for the control of insect pests.

A new ß-hydroxydihydrochalcone from Tephrosia uniflora, and the revision of three ß-hydroxydihydrochalcones to flavanones.

The CH2Cl2/MeOH (1:1) extract of the stems of Tephrosia uniflora yielded the new ß-hydroxydihydrochalcone (S)-elatadihydrochalcone-2'-methyl ether (1) along with the three known compounds elongatin (2), (S)-elatadihydrochalcone (3), and tephrosin (4). The structures were elucidated by NMR spectroscopic and mass spectrometric data analyses. Elongatin (2) showed moderate antibacterial activity (EC50 of 25.3 µM and EC90 of 32.8 µM) against the Gram-positive bacterium Bacilus subtilis, and comparable toxicity against the MCF-7 human breast cancer cell line (EC50 of 41.3 µM). Based on the comparison of literature and predicted NMR data with that obtained experimentally, we propose the revision of the structures of three ß-hydroxydihydrochalcones to flavanones.

A chalcone (Pongamol) and phytoconstituents of Tephrosia purpurea.

Tephrosia purpurea stem extract fractions shows more amount of Cyclohexane, 1-ethenyl-1-methyl-2,4-bis(1-methylethenyl)-, (1α,2ß,4ß)- (62.32%). Secondary metabolites contents were higher in stem than root of T. purpurea. It is the first time a chalcone (Pongamol) was crystallised from the column chromatography fraction of T. purpurea stem extract identified by using FT-IR and XRD. Quantitative analysis of T. purpurea stem and root reveals that total saponins, tannins and phenols (mg/g plant powder) contents were higher in the former than the latter part of the plant, respectively.

Deciphering the multi-scale mechanisms of Tephrosia purpurea against polycystic ovarian syndrome (PCOS) and its major psychiatric comorbidities: Studies from network pharmacological perspective.

Tephrosia purpurea (T. purpurea), a plant belonging to Fabaceae (pea) family, is a well-known Ayurvedic herb and commonly known as Sarapunkha in traditional Indian medicinal system. Described as "Sarwa wranvishapaka", i.e. having a capability to heal all types of wounds, it is particularly recognized for its usage in splenomegaly. Towards exploring the comprehensive effects of T. purpurea against polycystic ovarian syndrome (PCOS) and three comorbid neuropsychiatric diseases (anxiety, depression, and bipolar disorder), its constituent phytochemicals (PCs) were extensively reviewed and their network pharmacology evaluation was carried out in this study. The complex regulatory potential of its 76 PCs against PCOS is enquired by developing and analyzing high confidence tripartite networks of protein targets of each phytochemical at both pathway and disease association scales. We also developed a high-confidence human Protein-Protein Interaction (PPI) sub-network specific to PCOS, explored its modular architecture, and probed 30 drug-like phytochemicals (DPCs) having multi-module regulatory potential. The phytochemicals showing good binding affinity towards their protein targets were also evaluated for similarity against currently available approved drugs present in DrugBank. Multi-targeting and synergistic capacities of 12 DPCs against 10 protein targets were identified and evaluated using molecular docking and interaction analyses. Eight DPCs as a potential source of PCOS and its comorbidity regulators are reported in T. purpurea. The results of network-pharmacology study highlight the therapeutic relevance of T. purpurea as PCOS-regulator and demonstrate the effectiveness of the approach in revealing action-mechanism of Ayurvedic herbs from holistic perspective.

Isoflavones from the seedpods of Tephrosia vogelii and pyrazoisopongaflavone with anti-inflammatory effects.

Phytochemical investigation of Tephrosia vogelii seedpods led to the isolation of twelve compounds: vogelisoflavone A (1), vogelisoflavone B (2), isopongaflavone (3), onogenin, luteolin, 4',7-dihydroxy-3'-methoxyflavanone, trans-p-hydroxycinnamic acid, tephrosin, 2-methoxygliricidol, dehydrorotenone, 6a,12a-dehydro-α-toxicarol and pinoresinol. Compounds 1 and 2 are reported as new natural products. Isopongaflavone (3) was structurally modified using hydrazine to pyrazoisopongaflavone (4). These compounds were characterized based on their NMR and HRESIMS data. Further, four compounds (1-4) were evaluated for their anti-inflammatory effects in lipopolysaccharide (LPS)-stimulated peripheral blood mononuclear cells (PBMCs). Treatment of the LPS-stimulated PBMCs with the compounds at a concentration of 100 µM suppressed the secretion of interleukin IL-1ß interferon-gamma (IFN-γ), granulocyte macrophage-colony stimulating factor (GM-CSF) and tumour necrosis factor-alpha (TNF-α).

Prenylated Flavonoids from the Roots of Tephrosia rhodesica.

Five new compounds-rhodimer (1), rhodiflavan A (2), rhodiflavan B (3), rhodiflavan C (4), and rhodacarpin (5)-along with 16 known secondary metabolites, were isolated from the CH2Cl2-CH3OH (1:1) extract of the roots of Tephrosia rhodesica. They were identified by NMR spectroscopic, mass spectrometric, X-ray crystallographic, and ECD spectroscopic analyses. The crude extract and the isolated compounds 2-5, 9, 15, and 21 showed activity (100% at 10 µg and IC50 = 5-15 µM) against the chloroquine-sensitive (3D7) strain of Plasmodium falciparum.

Well-aging: A new strategy for skin homeostasis under multi-stressed conditions.

BACKGROUND: Several studies evidenced significant increase of cortisol is the consequence of UV or emotional stress and leads to various deleterious effects in the skin. AIM: The well-aging, a new concept of lifestyle, procures an alternative to the anti-aging strategy. We demonstrated that Tephrosia purpurea extract is able to stimulate well-being hormones while reducing cortisol release. Furthermore, we hypothesized that the extract could positively influence the global skin homeostasis. METHOD: We evaluated the impact of the extract on cortisol, ß-endorphin, and dopamine, released by normal human epidermal keratinocytes (NHEKs). A gene expression study was realized on NHEKs and NHDFs. The protein over-expression of HMOX1 and NQO1 was evidenced at cellular and tissue level. Finally, we conducted a clinical study on 21 women living in a polluted environment in order to observe the impact of the active on global skin improvement. RESULTS: The extract is able to reduce significantly the cortisol release while inducing the production of ß-endorphin and dopamine. The gene expression study revealed that Tephrosia purpurea extract up-regulated the genes involved in antioxidant response and skin renewal. Moreover, the induction of HMOX and NQO1 expression was confirmed on NHDFs, NHEKs and in RHE. We clinically demonstrated that the extract improved significantly the skin by reducing dark circles, represented by an improvement of L*, a*, and ITA parameters. CONCLUSION: Tephrosia purpurea extract has beneficial effects on skin homeostasis through control of the well-being state and antioxidant defenses leading to an improvement of dark circles, a clinical features particularly impacted by emotional and environmental stress.

Tephrosia apollinea seed: a new rich source of essential polyunsaturated fatty acids, tocopherols, sterols, and squalene.

Tephrosia apollinea is a legume species, native to southwest Asia and northeast Africa, rich in bioactive flavonoids (hydrophilic compounds). T. apollinea seeds were not considered previously as a potential source of lipophilic compounds such as: essential fatty acids, tocopherols, sterols, and squalene, hence, the present study were performed. The oil yield in T. apollinea seeds amounted to 11.8% dw. The T. apollinea seed oil was predominated by the polyunsaturated fatty acids - linoleic (26.8%) and α-linolenic (22.7%). High levels were recorded also for oleic (27.6%) and palmitic (14.9%) acids. Four tocopherols and one tocotrienol, with the domination of γ-tocopherol (98%) were identified in T. apollinea seed oil. The ß-sitosterol (59%), Δ5-stigmasterol (21%) and campesterol (9%) were detected as main sterols in T. apollinea seed oil. The total content of tocochromanols, sterols, carotenoids and squalene in the T. apollinea seed oil was 256.7, 338.1, 12.5 and 1103.8 mg/100 g oil, respectively. T. apollinea seeds oil, due to the high concentration of lipophilic bioactive compounds can find a potential application in the food, cosmetic and pharmaceutical industry.

Therapeutic potential of ethyl acetate fraction of Tephrosia purpurea Linn. leaves in a rat model of gout.

OBJECTIVE: The present study is to determine the potential treatment effects of ethyl acetate fraction of Tephrosia purpurea Linn. leaves (EATP) against gout. METHODS: Gout in experimental rats was induced with potassium oxonate at the dose of 250 mg/kg (intraperitoneal injection) for 7 consecutive days; EATP was administered 1 h after administration of the potassium oxonate on each day of experiment. Potassium oxonate was discontinued on the 8th day; thereafter allopurinol (10 mg/kg, p.o.) and EATP (200 and 400 mg/kg, p.o.) were continued until day 14. The uric acid level was measured from serum and urine during the experiment. Other biochemical parameters were assessed, including blood and urine creatinine, erythrocyte sedimentation rate, and total protein. Blood urea nitrogen, serum aspartate aminotransferase serum alanine aminotransferase and alkaline phosphatase were also measured. The blood was analyzed for levels of malondialdehyde and the antioxidant enzymes such as superoxide dismutase, catalase and glutathione peroxidase. Histopathological and radiological changes in the ankle of rats were observed after completion of the experiment. RESULTS: EATP was able to decrease serum uric acid and creatinine level; it also reduced inflammation, oxidative stress and lysosomal enzyme level, which has a role in acute inflammation. EATP increased uric acid excretion through urine due to its uricosuric effect. CONCLUSION: EATP lowered the serum uric acid level and increased the urine uric acid level through excretion, which is useful in the treatment of gout. Hence the EATP was found to be helpful in the treatment of gout.

Analysis on safety assessment of Tephrosia vogelii Hook to Apis cerana cerana.

Tephrosia vogelii Hook was excellent insecticidal plant, it was introduced into China and planted over a large area in Guangdong province. The main active components of T. vogelii was rotenone and it widely found in leaves and pods of T. vogelii. This paper study of the safety assessment of T. vogelii flowers to worker bees. In this paper, the content of rotenone in T. vogelii petal, nectar, pollen, pistil, and stamen samples were investigated by HPLC, and tested the toxicity of T. vogelii flowers for Apis cerana cerana during 24 h. The dissipation and dynamic of rotenone in A. c. cerana different biological compartments were investigated under indoor conditions during 24 h. The results showed, The LT50 of T. vogelii flowers to worker bees were collected from the eastern, western, southern, northern and top were 13.95, 24.17, 12.55, 26.48, and 18.84 h, the haemolymph of worker bees have the highest content of rotenone, the least accumulation of rotenone in workers bee's thorax, and the rate of dissipation was slowly during the whole study. In conclusion, the results showed the T. vogelii create security risks to worker bees under some ecosystems.

A Validated HPTLC Densitometric Method for Determination of Lupeol, ß-Sitosterol and Rotenone in Tephrosia purpurea: A Seasonal Study.

Tephrosia purpurea (L.) Pers., commonly known as "sarpunkha" and "wild indigo", is being used in traditional systems of medicine to treat liver disorders, spleen and kidney. In the present study, a validated High Performance Thin Layer Chromatography (HPTLC) method was established for the estimation of lupeol, ß-sitosterol and rotenone in various extracts of T. purpurea with the aim to see the effect of seasons on the quantity of aforesaid phytoconstituents. The plant material was collected in summer (April), rainy (August) and winter (December) during 2013-2014 from Lucknow, India. The method was validated in terms of precision, repeatability, specificity, sensitivity linearity and robustness. The method permits reliable quantification and showed good resolution on silica gel with toluene-ethyl acetate-formic acid (9:1:1 v/v/v) as mobile phase, and characteristic bands of ß-sitosterol, rotenone and lupeol were observed at Rf 0.38, 0.45 and 0.52, respectively. The content of aforesaid phytoconstituents varies from season to season and extract to extract. Our finding indicated that winter season (December) may not be appropriate for collection of T. purpurea for the preparation of therapeutic formulations because of the high content of rotenone, a known insecticide that is responsible for Parkinson's disease and associated with heart failure, fatty liver and liver necrosis.

Regulating role of ethyl acetate fraction of Tephrosia tinctoria pers. in carbohydrate metabolism and oxidative stress in diabetic rats.

Traditionally, Tephrosia tinctoria (TT) is used as antimicrobial, larvicidal, antidiabetic and antioxidant agents. In the present study, the diabetes was induced in male albino - rats using alloxan (140 mg/kg b.w.) and treated with ethyl acetate extract (100 and 200 mg/kg b.w.) of T. tinctoria (TTEA) orally for 28 days. The effects of TTEA on biochemical paramets were studied. The results of this comprehensive study revealed that the TTEA significantly restored the altered parameters in alloxan-induced diabetic rats. The diabetes-induced elevated levels of glycogenolysis and gluconeogenesis metabolism were significantly controlled by TTEA. In the same, the decreased glycogenesis, glycolysis and TCA cycle metabolisms were significantly increased by TTEA. The enzymatic and non- enzymatic antioxidant levels were significantly elevated in TTEA treated diabetic rats and also the total non-enzymatic antioxidants capacity of serum and liver homogenate were calculated though DPPH inhibition. To conclude, this study demonstrates the anti-diabetic effect of TTEA possibly mediated through regulating carbohydrate metabolism and antioxidant status.

Investigation on apposite chemical marker for quality control of Tephrosia purpurea (L.) Pers. by means of HPTLC-chemometric analysis.

Application of phytochemicals as the markers for quality assurance of the herbal medicinal material is one of the prominently used approach. In the present study, six major chemical compounds i.e. rutin, quercetin, lupeol, ß-sitosterol, rotenone, deguelin of therapeutically important plant Tephrosia purpurea were tested for their significance to be used as chemical markers in analytical methods. Plants were collected from different locations. Extraction procedures as well as HPTLC analytical methods have been optimized for each compound. All these methods have been validated in terms of linearity, intraday-interday precision, LOD, LOQ, accuracy and repeatability. Metabolites have been quantified and quantitative data has been subjected to chemometric analysis. Results revealed that Rf values of all the compounds are between 0.3 and 0.4. Rutin, lupeol and ß-sitosterol gave desirable response and other three compounds were found undetectable in HPTLC. Content of rutin, beta-sitosterol and lupeol ranged from 0.095 to 0.84, 0.043 to 0.125, 0.023 to 0.045 w/w % respectively. Among them, rutin content was highest in all samples. Quantitative measurements combined with chemometric analysis displayed chemodiversity in the samples. In addition, principal component analysis ranked the marker as in order of their significance rutin > ß-sitosterol > lupeol. Results indicate rutin to be most preferable chemical marker for the Tephrosia. Furthermore, application of all the three compounds combined as the multimarker system should be preferred for standardization of T. purpurea.

In vitro oxidative stress regulatory potential of Citrullus colocynthis and Tephrosia apollinea.

The present study investigates the potential role of medicinal plants Citrullus colocynthis and Tephrosia apollinea in ameliorating the oxidative stress developed during the generation of reactive oxygen species. Organic extracts of different organs (leaf, stem and root) of these medicinal plants obtained in n-hexane, chloroform, n-butanol and water were assayed for radical scavenging, total antioxidant capacity, anti-lipid peroxidation and reduced glutathione. The total phenolic content (TPC) of both selected medicinal plants was also evaluated. The results indicated that extracts of T. apollinea leaf, stem and root have higher TPC compared to those of C. colocynthis. Similarly, the results of the present study revealed higher bioactivity of C. colocynthis than that of T. apollinea in various antioxidant assays. Various plant parts of each plant were also compared.

Antiplasmodial prenylated flavanonols from Tephrosia subtriflora.

The CH2Cl2/MeOH (1:1) extract of the aerial parts of Tephrosia subtriflora afforded a new flavanonol, named subtriflavanonol (1), along with the known flavanone spinoflavanone B, and the known flavanonols MS-II (2) and mundulinol. The structures were elucidated by the use of NMR spectroscopy and mass spectrometry. The absolute configuration of the flavanonols was determined based on quantum chemical ECD calculations. In the antiplasmodial assay, compound 2 showed the highest activity against chloroquine-sensitive Plasmodium falciparum reference clones (D6 and 3D7), artemisinin-sensitive isolate (F32-TEM) as well as field isolate (KSM 009) with IC50 values 1.4-4.6 µM without significant cytotoxicity against Vero and HEp2 cell lines (IC50 > 100 µM). The new compound (1) showed weak antiplasmodial activity, IC50 12.5-24.2 µM, but also showed selective anticancer activity against HEp2 cell line (CC50 16.9 µM).

Brown Stink Bug Mortality by Seed Extracts of Tephrosia Vogelii Containing Deguelin and Tephrosin

ABSTRACT Extracts of the seeds of Tephrosia vogelii Hook. f. were studied in relation to its chemical composition and toxicity to the brown stink bug Euschistus heros (F.). The extracts were obtained in ethyl acetate and ethanol in the sequence according to the polar nature of the solvents. Extracts were sprayed in concentration of 1.0, 2.5, 5.0, 7.5 and 10% on third-instars nymphs and adults, and mortality was recorded. Presence two rotenoids in ethyl acetate was detected, with analyzed with gas chromatography-mass spectrometry (GC-MS). Crude fraction analyses confirmed the presence of these rotenoids (tephrosin - 2.71% in ethyl acetate and 3.66% in methanol; and deguelin - 10.46% in ethyl acetate and 1.22% in methanol) and three other rotenoids in small amounts. Eight days after applications, ethyl acetate caused more stink bugs mortality and on less time than ethanol extract, because great quantity of rotenoids, as polarity. Concentrations above to 1 and 2.5% of the ethyl acetate extracts caused mortality above 80% of the nymphs and adults of E. heros, respectively. Concentration were considered high, thus chemist analyzes demonstrated high rotenoids presence. In conclusion, seed T. vogelli extracts, rich in deguelin and tephrosin (3:1), cause mortality of E. heros, however, high concentration are necessary.

Experimental therapeutic assays of Tephrosia vogelii against Leishmania major infection in murine model: in vitro and in vivo.

BACKGROUND: Conventional targeted leishmanicidal chemotherapy has persistently remained prohibitive for most economically deprived communities due to costs, associated time to accessing health services and duration for successful treatment programme. Alternatives are bound to be incorporated in rational management of leishmaniasis by choice or default due to accessibility and cultural beliefs. Therefore, there is need to rigorously investigate and appraise the activity of medicinal compounds that may have anti-leishmanicidal activity especially in the context of products that are already being utilized by the populations for other ailments but have limited information on their therapeutic value and possible cytoxicity. Hence, the study examined both in vivo and in vitro response of L. major infection to Tephrosia vogelii extracts in BALB/c mice as the mouse model. METHODS: A comparative study design was applied for the in vivo and in vitro assays of the extract with Pentostam (GlaxoSmithKline, UK) and Amphotericin B [Fungizone™, X-Gen Pharmaceuticals (US)] as standard drugs. RESULTS: In BALB/c mice where the chemotherapeutic extract was administered intraperitoneally, there was significantly (p < 0.05) larger reduction in lesion size and optimal control of parasite burden than those treated orally. However, standard drugs showed better activity. Tephrosia vogelii had 50% inhibitory concentration (IC50) and IC90 of 12 and 68.5 µg/ml respectively, while the standard drugs had IC50 and IC90 of 5.5 and 18 µg/ml for Pentostam and 7.8 and 25.5 µg/ml for Amphotericin B in that order. In the amastigote assay, the infection rates decreased with increase in chemotherapeutic concentration. The multiplication indices for L. major amastigotes in macrophages treated with 200 µg/ml of the standard drugs and extract were significantly different (p < 0.05). 200 µg/ml of T. vogelii extract showed a multiplication index of 20.57, 5.65% for Amphotericin B and 9.56% for Pentostam. There was also significant difference (p < 0.05) in levels of Nitric oxide produced in the macrophages. CONCLUSIONS: The findings demonstrated that T. vogelii extract has anti-leishmanial activity and further assays should be done to ascertain the active compounds responsible for anti-leishmanial activity.

Four Prenylflavone Derivatives with Antiplasmodial Activities from the Stem of Tephrosia purpurea subsp. leptostachya.

Four new flavones with modified prenyl groups, namely (E)-5-hydroxytephrostachin (1), purleptone (2), (E)-5-hydroxyanhydrotephrostachin (3), and terpurlepflavone (4), along with seven known compounds (5-11), were isolated from the CH2Cl2/MeOH (1:1) extract of the stem of Tephrosia purpurea subsp. leptostachya, a widely used medicinal plant. Their structures were elucidated on the basis of NMR spectroscopic and mass spectrometric evidence. Some of the isolated compounds showed antiplasmodial activity against the chloroquine-sensitive D6 strains of Plasmodium falciparum, with (E)-5-hydroxytephrostachin (1) being the most active, IC50 1.7 ± 0.1 µM, with relatively low cytotoxicity, IC50 > 21 µM, against four cell-lines.

A kaempferol triglycoside from Tephrosia preussii Taub. (Fabaceae).

A phytochemical investigation of the MeOH extract of twigs and leaves of Tephrosia preussi was carried out to give a new kaempferol triglycoside, named tephrokaempferoside (1), together with five known compounds: tephrosin (2), betulinic acid (3), lupeol (4), ß-sitosterol (5) and 3-O-ß-d-glucopyranoside of ß-sitosterol (6). The structure of the new compound was characterised by analyses of NMR (1D and 2D) and MS data, and chemical conversion. Tephrokaempferoside (1) had weak antibacterial activity against Klebsiella pneumoniae with an MIC value of 150 µg/mL.