Identification of Major Bioactive Anti-inflammatory Compounds of Derris scandens Stem Using RAW 264.7 Cells and HPLC-UV Analysis.

Derris scandens (DS) is widely recognized for its therapeutic properties, specifically its analgesic effects, which significantly alleviate muscle pain. The chemical constituents of DS stem include various isoflavone derivatives. However, there is currently a lack of specified anti-inflammatory chemical markers and analytical methods for quality control. The present study aimed to evaluate the anti-inflammatory activity of DS and its constituents using the RAW 264.7 cell model. The expression of inflammatory genes such as inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), interleukin-6 (IL-6), and 5-lipoxygenase (5-LOX) was examined using quantitative RT-PCR. An high-performance liquid chromatography with a UV detection method was developed to quantitatively analyze genistein-7-O-[α-rhamnopyranosyl-(1 → 6)]-ß-glucopyranoside, genistein, derrisisoflavone A, lupalbigenin, and 6,8-diprenylgenistein in DS stem. The developed HPLC-UV method demonstrated high sensitivity with limits of detection and quantification ranging from 0.01 to 0.06 µg/mL and 0.03 to 0.18 µg/mL, respectively. The accuracy of the method ranged from 93.3 to 109.6%. Furthermore, the repeatability and reproducibility of the method were suitable, as indicated by the relative standard deviations of ≤ 3.02% and ≤ 6.22%, respectively. The DS extract notably inhibited NO production, exhibiting effects comparable to those of 500 µM diclofenac, and substantially suppressed the expression of iNOS, COX-2, IL-6, and 5-LOX of lipopolysaccharide (LPS)-induced genes. As to the pure isoflavone derivatives, the order of NO production inhibition was found to be genistein > lupalbigenin > derrisisoflavone A > 6,8-diprenylgenistein > genistein-7-O-[α-rhamnopyranosyl-(1 → 6)]-ß-glucopyranoside. Genistein, derrisisoflavone A, and 6,8-diprenylgenistein significantly suppressed the upregulation of all LPS-induced genes. Consequently, these compounds are recommended as anti-inflammatory markers for the quantitative chemical analysis of DS.

Study on Chemical Components from Derris taiwaniana and Their Lung Epithelial Cell Protect Effects.

The ethanol extract of roots of Derris taiwaniana gave two undescribed compounds, 3,3'-dimethoxy-5'-hydroxystilbene-4-O-ß-apiofuranosyl-(1→6)-ß-D-glucopyranoside (1) and 4',5-dihydroxy-3'-methoxyisoflavone-7-O-ß-apiofuranosyl-(1→6)-ß-D-glucopyranoside (2), along with thirty known components. Among them, compounds 14, 16-17, 23, 26-32 were isolated from this genus for the first time. Their structures were established based on physico-chemical properties and spectroscopic data, the lung epithelial cell protective effects were evaluated using NNK-induced MLE-12 cells. Among them, 2α,3α-epoxy-5,7,3',4'-tetrahydroxyflavan-(4ß-8-catechin) (30) showed the best significant protective effect, speculated to be the key component of D. taiwaniana that plays a protective role in lung epithelial cells.

A new anti-austerity agent, 4'-O-methylgrynullarin from Derris scandens induces PANC-1 human pancreatic cancer cell death under nutrition starvation via inhibition of Akt/mTOR pathway.

An ethanolic extract of Derris scandens flowers showed potent preferential cytotoxicity against PANC-1 human pancreatic cancer cells under nutrient-deprived condition, with a PC50 value of 0.7 µg/mL. Phytochemical investigation of this active extract led to the isolation of four prenylated isoflavones (1-4) including a new compound named 4'-O-methylgrynullarin (1). The structure elucidation of the new compound was achieved by HRFABMS and NMR spectroscopic analysis. The isolated compounds exhibited potent anti-austerity activity against four different human pancreatic cancer cell lines under nutrient-deprived conditions. The new compound 4'-O-methylgrynullarin (1) was also found to inhibit PANC-1 cell migration and colony formation under nutrient-rich condition. Mechanistically, compound 1 inhibited key survival proteins in the Akt/mTOR signaling pathway. Therefore, 4'-O-methylgrynullarin (1) can be considered as a potential lead compound for the anticancer drug development based on the anti-austerity strategy.

Therapeutic Potential of Genus Pongamia and Derris: Phytochemical and Bioactivity.

Genus Pongamia and Derris belong to the Leguminosae family and are reported synonymously in literature. Although many compounds have been isolated from different plant parts but seed oil is known to produce non-edible medicinally important furanoflavonoids. The seed oil, commonly known as Karanj oil in Ayurvedic and Siddha traditional systems of medicine, is reported for the treatment of various skin infections and psoriasis. Several phytopharmacological investigations have proved the medicinal potential of furanoflavonoids in the skin and other disorders. Not only furanoflavonoids but several other important phenolic constituents such as chalcones, dibenzoylmethanes, aurones, isoflavones, flavanone dihydroflavonol, flavans, pterocarpans, rotenoids, coumarins, coumestans, stilbenoids and peltygynoids and their glycosides have been reported for different biological activities including antihyperglycemic, anti-inflammatory, anticancer, insecticidal, anti-alzheimer's, gastro protective, antifungal, antibacterial, etc. In the present review, the phytochemistry and pharmacological activities of the genera Pongamia and Derris have been summarized.

Anticariogenic Activities of Derris reticulata Ethanolic Stem Extract Against Streptococcus mutans.

BACKGROUND AND OBJECTIVE: Streptococcus mutans is a dominant causative pathogen of dental caries, which is a major oral health problem affecting million people worldwide. Derris reticulata is a medicinal plant possessing antimicrobial activity against several Gram-positive pathogenic bacteria. None the less, its effects on growth and cariogenic properties of S. mutans has not been clearly established. This study aimed to investigate the antibacterial and anti cariogenic activities of the D. reticulata ethanolic stem extract. MATERIALS AND METHODS: The TLC analysis was performed to authenticate the D. reticulata sample. Minimum inhibition concentration and minimum bactericidal concentration were determined by using broth dilution and drop plate methods, respectively. Sucrose dependent and sucrose independent-adherences, biofilm formation and glycolytic pH drop assays were performed to evaluate the anticariogenic activity. RESULTS: The ethanolic stem extract of D. reticulata possessed the antibacterial activity against S. mutans with the MIC and MBC of 0.875±0.250 and 1.750±0.500 mg mL-1, respectively. The extract at the lower concentrations of sub-MIC also had significant inhibitory actions against the cariogenic properties of S. mutans, including surface adherence, biofilm formation and glycolytic acid production. CONCLUSION: The D. reticulata stem extract had a substantial anticariogenic activities and thus potentially be developed as an oral health care product for dental caries prevention in the near future.

Anti-Angiogenic Activity of Rotenoids from the Stems of Derris trifoliata.

The plants in the genus Derris have proven to be a rich source of rotenoids, of which cytotoxic effect against cancer cells seem to be pronounced. However, their effect on angiogenesis playing a crucial role in both cancer growth and metastasis has been seldom investigated. This study aimed at investigating the effect of the eight rotenoids (1: -8: ) isolated from Derris trifoliata stems on three cancer cells and angiogenesis. Among them, 12a-hydroxyrotenone (2: ) exhibited potent inhibition on both cell growth and migration of HCT116 colon cancer cells. Further, anti-angiogenic assay in an ex vivo model was carried out to determine the effect of the isolated rotenoids on angiogenesis. Results revealed that 12a-hydroxyrotenone (2: ) displayed the most potent suppression of microvessel sprouting. The in vitro assay on human umbilical vein endothelial cells was performed to determine whether compound 2: elicits anti-angiogenic effect and its effect was found to occur via suppression of endothelial cells proliferation and tube formation, but not endothelial cells migration. This study provides the first evidence that compound 2: could potently inhibit HCT116 cancer migration and anti-angiogenic activity, demonstrating that 2: might be a potential agent or a lead compound for cancer therapy.

Stereocontrolled Semisyntheses of Elliptone and 12aß-Hydroxyelliptone.

Operationally simple, stereocontrolled semisyntheses of the anticancer rotenoids elliptone and 12aß-hydroxyelliptone, isolated from Derris elliptica and Derris trifoliata, respectively, are described. Inspired by the work of Singhal, elliptone was prepared from rotenone via a dihydroxylation-oxidative cleavage, chemoselective Baeyer-Villiger oxidation, and acid-catalyzed elimination sequence. Elaboration of elliptone to 12aß-hydroxyelliptone was achieved via a diastereoselective chromium-mediated Étard-like hydroxylation. The semisynthesis of elliptone constitutes an improvement over previous methods in terms of safety, scalability, and yield, while the first synthesis of 12aß-hydroxyelliptone is also described.

5,7,4'-Trihydroxy-6,8-diprenylisoflavone and lupalbigenin, active components of Derris scandens, induce cell death on breast cancer cell lines.

BACKGROUND: Natural products are a potential source for cancer chemotherapeutic development. This current study was performed to investigate the anti-tumor potential of 5,7,4'-trihydroxy-6,8-diprenylisoflavone (TD) and lupalbigenin (LB), plant flavonoids found in Derris scandens Benth (family: Leguminosae), in cancer and normal cell lines. METHODS: The human breast cancer cell lines MCF-7, MDA-MB-231 and MDA-MB-468, the human colon cancer cell line SW-620, and the mouse fibroblast cell line L-929 were used to test their anti-cancer activity. Apoptotic cell levels were measured by staining with annexin-V and propidium iodide and Western blot analysis was performed to confirm the apoptotic mechanism. RESULTS: The results revealed that TD and LB showed specific cytotoxicity against MDA-MB-231 and MCF-7 cells. To elucidate mode of cell death via cytotoxic activities, breast cancer cell lines were treated. TD and LB induced MDA-MB-231 and MCF-7 cells to apoptosis, with the highest number of apoptotic cells at 24 and 72h, respectively. Furthermore, TD and LB inhibited cell cycle progression via up-regulation of p21. Both compounds stimulated apoptosis through down-regulation of bcl-2, up-regulation of bax and releasing of cytochrome C proteins. CONCLUSIONS: TD and LB have significant anti-cancer effects against human breast cancer cells via cell cycle arrest and the induction of apoptosis through mitochondria signaling pathways, and may be potential anti-cancer agents for the treatment of breast cancer.

[Studies on flavonoids from Derris eriocarpa].

Derris eriocarpa, a traditional Chinese medicine belonging to the family of Leguminosae, is widely distributed mainly over Yunnan, Guangxi and Guizhou of China. Modern pharmacological researches on this herb showed that it had extensive bioactivities, such as promoting urination, removing dampness and cough and reducing inspissated mucus and other biological activities. The extensive studies on the chemical constituents of this plant have resulted in the isolation of triterpenoids, steroids, fatty acid and others, but the flavone compounds haven't reported before. In our further research on the ethyl acetate of this plant, nine flavone compounds were obtained by column chromatography on silica gel, Sephadex LH-20, semi-prep HPLC, polyamide column chromatography and recrystallization for separation and purification. The structures were determined on the basis of extensive spectroscopic analysis, including MS, NMR experiments and comparison with spectroscopic data in the literature, respectively, as diosmetin (1), 3, 3'-di-O-methylquercetin (2), afromosin (3), 6, 3'-dihydroxy-7, 4'-dimethoxyisoflavone (4), odoratin (5), 7, 3'-dihydroxy-8, 4'-dimethoxyisoflavone (6), 6, 4'-dihydroxy-7, 3'-dimethoxyisoflavone (7), 5, 7, 4'-trihydroxy-3, 3', 5'-trimethoxyflavone (8), and alpinumisoflavone (9). All these compounds were isolated from Derris eriocarpa How for the first time. And the in vitro assays showed that compound 2 possessed moderate inhibitory activity against human cancer cells K562 and HEL.

Isolation and characterization of a new bioactive isoflavone from Derris eriocarpa.

Derris eriocarpa How is an important medicinal plant, which is used as Zhuang ethnomedicine and Dai ethnomedicine to treat various diseases. One new compound, 3',4'-di-O-methylene-5-hydroxy-7-methoxy-6-isopentenyl isoflavone (1) and a known synthetic but new naturally occurring compound trans-3,4,5-trimethoxy-4'-isopentenyloxyl-stilbene (2), together with five known compounds, 5,7-dimethoxy-6-(3-methyl-2-butenyl)-4'-hydroxyl isoflavones (3), robustone (4), trans-3,4,5,4'-tetramethoxy-stilbene (5), robustic acid (6), and robustin (7) were isolated from the stem of D. eriocarpa. Spectroscopic analysis revealed the chemical structures of compounds 1-7.. Compounds 1 and 3 exhibited significant scavenging activities against 1,1-diphenyl-2-picrylhydrazyl radical and superoxide anions. Compounds 1-3 exhibited potent antiproliferative activity on Hela cells.

Lupalbigenin from Derris scandens Sensitizes Detachment-induced Cell Death in Human Lung Cancer Cells.

BACKGROUND/AIM: The ability of cancer cells to resist to anoikis has been shown to augment cancer cell metastasis in many cancers. In search for potential substances for anti-metastatic approaches, this study aimed to investigate anoikis-sensitizing activity of lupalbigenin, extracted from Derris scandens. MATERIALS AND METHODS: Human lung cancer cells were treated with non-cytotoxic concentrations of lupalbigenin in a detachment condition. Anoikis was evaluated at various time points using MTT viability assays. The effect of lupalbigenin on anchorage-independent growth was performed by soft-agar assay. The survival signaling proteins, as well as regulatory proteins of apoptosis and metastasis, were examined by western blot analysis. RESULTS: Lupalbigenin treatment significantly down-regulated survival proteins, including protein kinase B (pAKT/AKT) and extracellular signal-regulated kinase (pERK/ERK), as well as anti-apoptotic protein B-cell lymphoma 2 (BCL-2), resulting in the enhancement of the cellular response to anoikis and the decrease of growth and survival in an anchorage-independent condition. CONCLUSION: Lupalbigenin sensitizes detachment-induced cell death in human lung cancer cell through down-regulation of pro-survival proteins.

Antioxidant, α-glucosidase inhibitory activity and sub-chronic toxicity of Derris reticulata extract: its antidiabetic potential.

BACKGROUND: Antidiabetic activity of Derris reticulata extract on alloxan-induced diabetic rats has been reported. The extract was found to lower blood glucose and inhibit intestinal glucose absorption. The aim of this study was to further investigate mechanisms underlying the antihyperglycemic activity of D. reticulata extract in vitro. METHODS: The aqueous extract was obtained from D. reticulata stem. Phytochemical screening, total phenolic, and flavanoid contents were examined. ABTS and DPPH scavenging assays, and FRAP method were used to determine in vitro antioxidant activities. Measurement of cell viability on alloxan-induced cellular damage was performed in the insulin-secreting RINm5F cells by MTT assay. The effects of the extract on α-glucosidase activity and insulin release were studied. In addition, sub-chronic toxicity test in rats was also conducted. RESULTS: The results revealed that the extract, which consisted of terpenoids, saponins, tannins and flavonoids, possessed moderate radical scavenging activities. Pre-treatment of RINm5F cells with the extract was also found to exert moderate, but significant, in vitro protection against alloxan, an oxidative stress producing agent. Unlike glibenclamide, the extract did not stimulate insulin secretion. However, the extract was found to inhibit α-glucosidase activity similar to acarbose. It was found that in sub-chronic toxicity studies D. reticulata extract did not cause mortality or produce any remarkable haematological, biochemical and histopathological adverse effects in rats. CONCLUSIONS: The data suggest that the possible mechanisms underlying antihyperglycemic activity of D. reticulata extract are cytoprotective effect on pancreatic cells, presumably by its antioxidant activity, and inhibition of α-glucosidase. Sub-chronic toxicity study also provides scientific evidence to corroborate the safety of this plant as an alternative antidiabetic agent.

Cytotoxicity of compounds from the fruits of Derris indica against cholangiocarcinoma and HepG2 cell lines.

Two new compounds, derrivanone (1) and derrischalcone (2), were isolated from the crude hexane extract of the fruits of Derris indica. In addition, 14 known compounds were isolated from the fruits of this plant. Chalcones 2-4 showed strong cytotoxicity against cholangiocarcinoma cell line (M156) and human hepatoma HepG2 cells. In addition, flavanones 15 and 16 exhibited potent and high cytotoxic efficacy.

Consumer and farmer safety evaluation of application of botanical pesticides in black pepper crop protection.

This study presents a consumer and farmer safety evaluation on the use of four botanical pesticides in pepper berry crop protection. The pesticides evaluated include preparations from clove, tuba root, sweet flag and pyrethrum. Their safety evaluation was based on their active ingredients being eugenol, rotenone, ß-asarone and pyrethrins, respectively. Botanical pesticides from Acorus calamus are of possible concern because of the genotoxic and carcinogenic ingredient ß-asarone although estimated margins of exposure (MOE) for consumers indicate a low priority for risk management. For the other three botanical pesticides the margin of safety (MOS) between established acute reference doses and/or acceptable daily intake values and intake estimates for the consumer, resulting from their use as a botanical pesticide are not of safety concern, with the exception for levels of rotenone upon use of tuba root extracts on stored berries. Used levels of clove and pyrethrum as botanical pesticides in pepper berry crop production is not of safety concern for consumers or farmers, whereas for use of tuba root and sweet flag some risk factors were defined requiring further evaluation and/or risk management. It seems prudent to look for alternatives for use of sweet flag extracts containing ß-asarone.

Prenylated flavonoids from the leaves of Derris malaccensis and their cytotoxicity.

A new prenylated isoflavone, pomiferin-4'-O-methyl ether, and a new prenylated chalcone, 2',4'-dihydroxy-4-methoxy-3'-(2-hydroxy-3-methylbut-3-enyl)chalcone, together with four known flavonoids, were isolated from the leaves of Derris malaccensis. All isolated compounds were evaluated for their cytotoxicity.

Ação inseticida do extrato de Derris amazonica Killip para Cerotoma arcuatus Olivier (Coleoptera: Chrysomelidae) / Insecticide action of the extract of Derris amazonica Killip for Cerotoma arcuatus Olivier (Coleoptera: Chrysomelidae)

A abundância e o potencial inseticida de Derris amazonica e a necessidade de controle de Cerotoma arcuatus Olivier (Coleoptera: Chrysomelidae) na cultura do feijão-caupi (Vigna unguiculata L. Walp) estimularam a realização desta pesquisa, que objetivou avaliar a ação inseticida do extrato de D. amazonica a adultos de C. arcuatus em condições de laboratório. Os bioensaios testaram as vias de intoxicação por ingestão de folhas contaminadas, contato com superfície contaminada e aplicação tópica, com delineamento experimental inteiramente casualizado, com quatro repetições. Os valores de mortalidade e consumo foliar dos insetos foram submetidos à análise de regressão, sendo utilizada a análise de Probit para determinação das CL50, da DL50 e dos TL50. O extrato de D. amazonica, contendo 3,7% de rotenona, foi tóxico para adultos de C. arcuatus via ingestão de folhas contaminadas (CL50=15,14 µL do extrato.mL-1 de água), superfície contaminada (CL50=0,45 µL do extrato.cm-2) e aplicação tópica (DL50=1,44 µL do extrato.g-1 do inseto). Mortalidades de adultos de C. arcuatus superiores a 80% e os menores tempos letais médios foram obtidos na concentração de 5% (v v-1) do extrato em todos os bioensaios. O consumo foliar de adultos de C. arcuatus foi inversamente proporcional a concentração do extrato quando expostos por via de ingestão foliar ou aplicação tópica, sendo inclusive observada inibição da alimentação dos indivíduos. O extrato de D. amazonica é tóxico para C. arcuatus e inibe a alimentação dos insetos a partir da concentração de 1% (v v-1).

The abundance and insecticidal potential of Derris amazonica in addition to need of controlling Cerotoma arcuatus for bean crop stimulated this research. The objective of this work was to evaluate insecticide action of the extract of D. amazonica to adults of C. arcuatus in laboratory conditions. The bioassays were carried out using three distend methodologies: leaf intake, contact in treated surface (filter paper) and topical application. A completed randomized experimental design was used with four replications. Mortality values and leaf consumption of the insects were subjected to regression analyses, being the Probit analyses used to determine of the i.e., LC50, LT50 and LD50. The extract of D. amazonica containing 3.7% of rotenone was toxic to adults C. arcuatus when exposed to treated leaves (LC50 = 15.14 µl.mL-1), treated surface (LC50 = 0.45 µl.cm-2) and subjected to topical exposure (LD50 = 1.44 µl.g-1). In all bioassays the adults mortality was higher than 80% with lower median lethal times obtained with 5% (v.v-1) concentrations of the extract. Leaf consumption by adults C. arcuatus was inversely proportional to the concentration of the extract when exposed by leaf intake or topical application, also being observed inhibition of feeding individuals. The extract of D. amazonica is toxic to C. arcuatus and inhibits the feeding of insects from the concentration of 1% (v v-1).

Cancer-relevant biochemical targets of cytotoxic Lonchocarpus flavonoids: a molecular docking analysis.

A molecular docking investigation has been carried out on cytotoxic prenylated flavonoids from Lonchocarpus haberi with cancer-relevant chemotherapeutic targets known to be inhibited by flavonoids. Two molecular docking programs, Molegro and ArgusDock, were used to compare the binding energies of Lonchocarpus flavonoids with other flavonoids, inhibitors, or known ligands, to aromatase (CYP 19), fatty acid synthase (FAS), xanthine oxidase (XO), cyclooxygenases (COX-1 and COX-2), lipoxygenase (LOX-3), ornithine decarboxylase (ODC), protein tyrosine kinase (PTK), phosphoinositide 3-kinase (PI3K), protein kinase C (PKC), topoisomerase II (ATP binding site), ATP binding cassette (ABC) transporter, and phospholipase A(2) (PLA). The Lonchocarpus flavonoids examined in this study exhibited docking energies comparable to or stronger than other flavonoids that had been previously shown to be effective inhibitors of these enzymes. Furthermore, prenylated flavonoids, such as the Lonchocarpus flavonoids and xanthohumol, generally showed greater binding energies than the non-prenylated flavonoids. We conclude, therefore, that the Lonchocarpus flavonoids possibly owe their cytotoxic activity by inhibition of one or more of these enzymes.

Purification and biochemical characterization of a serine proteinase inhibitor from Derris trifoliata Lour. seeds: insight into structural and antimalarial features.

A potent serine proteinase inhibitor was isolated and characterized from the seeds of the tropical legume liana, Derris trifoliata (DtTCI) by ammonium sulfate precipitation, ion exchange chromatography and gel filtration chromatography. SDS-PAGE as well as MALDI-TOF analysis showed that DtTCI is a single polypeptide chain with a molecular mass of approximately 20 kDa. DtTCI has three isoinhibitors (pI: 4.55, 5.34 and 5.72) and, inhibited both trypsin and chymotrypsin in a 1:1 molar ratio. Both Dixon plots and Lineweaver-Burk double reciprocal plots revealed a competitive inhibition of trypsin and chymotrypsin activity, with inhibition constants (K(i)) of 1.7x10(-10) and 1.25x10(-10) M, respectively. N-terminal sequence of DtTCI showed over 50% similarity with numerous Kunitz-type inhibitors of the Papilionoideae subfamily. High pH amplitude and broad temperature optima were noted for DtTCI, and time course experiments indicated a gradual loss in inhibitory potency on treatment with dithiothreitol (DTT). Circular Dichroism (CD) spectrum of native DtTCI revealed an unordered structure whereas exposure to thermal-pH extremes, DTT and guanidine hydrochloride (Gdn HCl) suggested that an abundance of beta-sheets along with intramolecular disulfide bonds provide conformational stability to the active site of DtTCI, and that severity of denaturants cause structural modifications promoting inhibitory inactivity. Antimalarial studies of DtTCI indicate it to be a potent antiparasitic agent.

Synthesis and evaluation of derrubone and select analogues.

Recently, we reported that the natural product derrubone exhibits Hsp90 inhibitory activity. Due to its unique architectural scaffold and proposed rapid assembly, the synthesis of this natural product was pursued with the aim of identifying structure--activity relationships. Synthesis of the natural product was accomplished in eight highly convergent steps, which led to a facile method for the construction of related compounds. Biological evaluation of derrubone and its analogues identified several compounds that exhibit low micromolar inhibitory activity against breast and colon cancer cell lines.

Cytotoxic and antiprotozoal activity of flavonoids from Lonchocarpus spp.

A number of flavonoids isolated from Lonchocarpus spp. were evaluated for their antiprotozoal and cytotoxic activity. Flavone 6 and chalcone 7 were found to be the most active against Leishmania parasites and against cell cultures of Leukemia P388DI and adenocarcinoma prostate PC-3.