IR-775 - Hyptis loaded bioactive nanoparticles for enhanced phyto-photothermal therapy of breast cancer cells.

Photo-responsive therapy is an emerging treatment modality due to its bioimaging and therapeutic properties. Phototherapy induces localized hyperthermia and selectively eradicates cancer cells. The current study showed that multifunctional biodegradable liposome nanosystem (HIL NPs) containing Hyptis suaveolens bioactive molecules and IR-775, a NIR dye showed efficient bioavailability to cancer ells and allowed tumor ablation upon NIR laser irradiation. The resulting entities present in the nanosystem, i.e., bioactive molecules of Hyptis, serve as an anticancer agent, and IR-775 helps in the photothermal ablation of highly metastatic breast cancer cells. Hyptis suaveolens is a weed that grows rampantly, impeding the growth of neighboring plants; nonetheless, its bioactive compounds have demonstrated therapeutic benefits. The obtained HIL NPs, photothermally active liposome nanosystem showed a high fluorescence absorption peak in the NIR range and delivered a photothermal conversion efficiency of 55.20 % upon NIR laser irradiation. TEM and particle size analyzer revealed that HIL NPs have a size of 141 ± 30 nm with a spherical shape. The results of in-ovo (zebrafish) experiments have shown efficient bioimaging capabilities with minimal concentrations of HIL NPs compared to respective controls. Furthermore, in-vitro studies of HIL NPs against triple-negative breast cancer (4T1) indicated effective anticancer activity by a combined cytotoxic effect and hyperthermia. Tumor ablation was facilitated by reactive oxygen species production and hyperthermia, leading to DNA damage and apoptosis due to overexpression of É£-H2AX, Cathepsin B, and p53, which halted cancer cell proliferation. Therefore, HIL NPs demonstrated effective anticancer effects induced by combined phyto-photothermal therapy when evaluated against an in-vitro breast cancer model.

Impact of aqueous extracts of Cassia occidentalis, Eucalyptus camaldulensis and Hyptis suaveolens on the entomofauna and the seed yield of Gossypium hirsutum at Boklé (Garoua, Cameroon).

There is a frightening decline in the population pollinators around the world due to the over usage of synthetic pesticides, leading to the directly reduce of plant production. Plant extracts with insecticidal properties could be eco-friendly alternatives to synthetic pesticides in maintaining the pollinator population and the diversity of the ecosystem. The impact of aqueous extracts of Cassia occidentalis L., Eucalyptus camaldulensis Dehnh. and Hyptis suaveolens L. was investigated on the entomofauna and the seed yield of Gossypium hirsutum L. cotton. The study was carried out in RCBD, four times replicated: 3 extracts x 1 standard synthetic insecticide (TEMA) x 1 control x 4 groups of flowers (group 1: flowers free to insect visits, group 2: flowers protected from insects using gauze bags, group 3: protected flowers and opened exclusively to Amegilla sp. and group 4: protected flowers opened from time to time without any visit of insect). Gossypium hirsutum was found to be visited by the insects belonging to five orders, 10 families and 18 species. Amegilla sp.1 and Apis mellifera were the major pollinators during the rainy and dry seasons, respectively. The number and quality of seeds visited exclusively by Amegilla sp.1 were significantly improved by H. suaveolens extract. During the dry season, E. camaldulensis and H. suaveolens extracts as well as the standard insecticide improved the number of seeds and the percentage of normal seeds harvested from the flowers allowed to be visited by insects; that was probably due to their insecticidal effects which protected plants from pest damage. Therefore, aqueous extracts of E. camaldulensis and H. suaveolens are good candidates for incorporation in integrated pest management programs to minimize the risk of synthetic pesticides to pollinators, hence to increase the yield and the quality of seeds.

Assessment of larvicidal and genotoxic potentials of extracts of Hyptis suaveolens against Culex quinquefasciatus based on enzyme profile and RAPD-PCR assay.

Vector control strategies have focused on the development of effective and ecofriendly alternatives. In the present study, investigation of larvicidal and genotoxic effects of leaves of Hyptis suaveolens from four different extraction solvents (aqueous, hexane, methanol and acetone) on fourth instar larvae of Culex quinquefasciatus was carried out. Extraction was done using soxhlet apparatus and the characteristics functional group of active constituents were identified using Fourier Transform Infrared spectrophotometer. Larvicidal activities were screened using three different concentrations (50, 150 and 250 mg/mL) following WHO standard protocol and mortality was recorded after 24, 48 and 72 hr. Hexane extract showed the highest mortality (27.92, 38.75, 90.42 %; LC50: 272.5, 191.3, 114.8 mg/mL), followed by aqueous extract (20.83, 34.58, 59.58 %; LC50: 496.6, 392.9, 208.1 mg/mL) and acetone extract (20.83, 32.08, 59.58 %; LC50: 1111.2, 393.6, 266.1 mg/mL) and methanol extract (17.92, 29.17, 52.92 %; LC50: 466.0, 400.1, 272.3 mg/mL). Enzyme profile such as superoxide dismutase (SOD), glutathione-S-transferase (GST), catalase (CAT), glutathione (GSH) and alkaline phosphatase (ALP) were significantly altered in the larvae exposed to the four extracts. Phytochemical screening of all solvents extract revealed the presence of saponins, flavonoids, terpenoids and alkaloids as common constituents. Random Amplified Polymorphic DNA-Polymerase Chain Reaction (RAPD-PCR) profile implied hexane and aqueous extracts altered the DNA of larvae. Furthermore, FTIR Spectroscopic analysis revealed phenols, alcohols, aliphatic primary amines and saponins as the major groups in the extracts. Conclusively, this study established the lethal potential of extracts of H. suaveolens as alternative plant-based and eco-friendly larvicide against Cx. quinquefasciatus.

Karyomorphological effects of two new oil formulations on Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae).

The cotton bollworm Helicoverpa armigera is a serious pest of many economically important crops. Since this pest has become resistant to the conventional synthetic insecticides, newer compounds and formulations are being developed against this insect pest. Many natural compounds isolated from the plants were tested against this pest. Among them Hyptis suaveolens and Melochia corchorifolia showed insecticidal properties against H. arnigera. Based on bioefficacy studies, caryophyllene and ß-sitosterol were isolated from H. suaveolens and M. corchorifolia respectively. The isolated natural compounds were further developed as formulations in various combinations with neem (Azadirachta indica) and karanj (Pongamia pinnata) oils. The present study indicated that the formulations influenced the karyomorphology of H. armigera.

Hyptis suaveolens (L.) Poit protects colon from TNBS-induced inflammation via immunomodulatory, antioxidant and anti-proliferative mechanisms.

ETHNOPHARMACOLOGICAL RELEVANCE: In folk medicine Hyptis suaveolens (Lamiaceae) has been reported to relieve respiratory and gastrointestinal infections, indigestion, cold, pain, fever, cramps, skin diseases, gastric ulcer and inflammatory disorders. This study investigated the effects and the mechanisms of action of Hyptis suaveolens (L.) Poit (Lamiaceae) ethanol extract (Hs-EtOH) and hexane phase (Hs-HexF) against intestinal inflammation. MATERIAL AND METHODS: Acute and relapse TNBS-induced ulcerative colitis protocols were used to evaluate intestinal anti-inflammatory activity. Damage evaluations, biochemical, histological and immunostaining parameters were determined. RESULTS: Both extracts decreased macroscopic colonic inflammation and the area of lesion induced by TNBS. Nevertheless, only Hs-HexF was able to reduce colonic wall thickness, edema and diffuse inflammatory cell infiltration and to prevent GSH depletion in the acute model of ulcerative colitis. In the chronic phase with relapse of colonic ulceration, yet again only Hs-HexF significantly attenuated inflammatory parameters and presented a decrease in nitrite/nitrate, MDA, MPO, IL-1-ß and TNF-α and increased levels of SOD, CAT, GSH and IL-10. Hs-HexF also significantly reduced positive cells immunostained for PCNA. CONCLUSION: The data indicate intestinal anti-inflammatory activity for H. suaveolens, due to the participation of the antioxidant system, decreased neutrophil infiltration and cytokine modulation, as well as, owing to regulation of cell proliferation.

Chemical Composition, Antioxidant and Cytotoxic Activities of Hyptis suaveolens (L.) Poit. Essential Oil on Prostate and Cervical Cancers Cells.

BACKGROUND AND OBJECTIVE: Hyptis suaveolens is an aromatic plant used in traditional medicine in Burkina Faso for management of various diseases including wounds and inflammatory diseases. Thus, the objective of this work was to characterize the chemical composition, antioxidant and cytotoxic activity of Essential Oil (EO) of H. suaveolens from Burkina Faso on cultured cancer cells. MATERIALS AND METHODS: The chemical composition of EO was determined by GC/FID and GC/MS analysis and the antioxidant activity was evaluated through inhibition of DPPH radicals and ABTS +• radical cations. The cytotoxic activity in prostate cancer cells (LNCaP) and cervical cancer cells (HeLa) of EO was evaluated by MTT assay and effect on cells cycle by flow cytometry analysis. RESULTS: A total of 58 compounds were identified in the EO of H. suaveolens of which the major compounds identified are Sabinene 14.03%, ß-Pinene 5.92%, Limonene 4.40%, Eucalyptol 12.78%, Trans-Oxide of Linalol 5.43%, ß-Caryophyllene 11.27%, Germacrene-D 3.04% and Bicyclogermacrene 8.08%. The EO of H. suaveolens showed antioxidant activity and concentration dependent antiproliferative activities with G0/G1 arrest on LNCaP and HeLa cells. CONCLUSIONS: This work help to justify some uses of H. suaveolens in traditional medicine in Burkina Faso and also, presents a promising new application for the essential oil of H. suaveolens in prostate and cervical cancer research.

In vitro Antibiotic and Modulatory Activity of Mesosphaerum suaveolens (L.) Kuntze against Candida strains.

The emergence of fungal resistance to commercial drugs has been a major problem for the WHO. In this context, research with natural products is promising in the discovery of new active substances. Thus, this work evaluated the antifungal effect of a medicinal plant (i.e., Mesosphaerum suaveolens) against strains of the genus Candida, tested the combined effect with the drug fluconazole, and, finally, determined the phenolic constituents present in the species. Initially, aqueous extracts of leaves (AELMs) and aerial parts (AEAPMs) of the species were prepared. For microbiological assays, the minimum fungicidal concentration was determined by broth microdilution, and the combined effect of fluconazole extracts were verified by sub-inhibitory microdilution concentrations (CFM/8) followed by spectrophotometric readings which were used to determine the IC50. HPLC detected the presence of flavonoids and phenolic acids, detecting eight compounds present in the samples of which caffeic acid and quercetin were major components. The AELMs modulated fluconazole activity since it decreased fluconazole's IC50 from 7.8 µg/mL to an IC50 of 4.7 µg/mL (CA LM 77) and from 28.8 µg/mL to 18.26 µg/mL (CA INCQS 40006) for the C. albicans strains. The AEAPMs were able to potentiate the effect of fluconazole more effectively than the AELMs. Such an effect was significant for the 16 µg/mL concentration for CA LM 77 and 32 µg/mL for CA INCQS 40006. The AEAPMs as well as the AELMs presented clinically relevant activities for C. tropicalis strains. For the C. tropicalis LM 23 strain, the AEPMs obtained an IC50 of 25 µg/mL and the AELMs an IC50 of 359.9 µg/mL.

Chemical investigation of Hyptis suaveolens seed, a potential antihyperuricemic nutraceutical, with assistance of HPLC-SPE-NMR.

The seed of Hyptis suaveolens, commonly known as wild flour ball (san fen yuan) in Taiwan, serves as a main refreshing drink substance in several regions. This study investigated firstly its secondary metabolites, leading to the isolation of five major caffeoylquinic acid derivatives (1-5) from the ethanol extract. In addition, ten minors, including three caffeoylquinic acid derivatives (12-14), were characterized via assistance of HPLC-SPE-NMR. Of these isolates, sodium 4,5-dicaffeoylquinate (2) and methyl 3,5-dicaffeoylquinate (4) showed moderate inhibitory activity against xanthine oxidase with the respective IC50 values of 69.4 µM and 92.1 µM (c.f. allopurinol IC50 28.4 µM). Quantitative HPLC analysis of the EtOH extract indicates the content of sodium 3,5-dicaffeoylquinate (1) and sodium 4,5-dicaffeoylquinate (2) to be 0.1% and 0.08% (w/w, dry seed), respectively. This study not only discloses the bioactive constituents, but also demonstrates the potential of H. suaveolens seed as an antihyperuricemic nutraceutical.

Bush mint (Hyptis suaveolens) and spreading hogweed (Boerhavia diffusa) medicinal plant extracts differentially affect activities of CYP1A2, CYP2D6 and CYP3A4 enzymes.

ETHNO-PHARMACOLOGICAL RELEVANCE: Hyptis suaveolens (L) Poit and Boerhavia diffusa Linn are medicinal herbal plants commonly found in the tropics and sub-tropics. They are used to treat various conditions among them boils, dyslipidaemia, eczema, malaria, jaundice and gonorrhoea. Thus, the herbal medicinal extracts are now found as part of some commercial herbal formulations. There has not been adequate characterization of these medicinal herbs on their effects on drug metabolising enzymes. AIM OF THE STUDY: To investigate the effects of extracts of Hyptis suaveolens (HS) and Boerhavia diffusa (BD) on activity of drug metabolising enzymes, CYP1A2, CYP2D6 and CYP3A4, as well predict their potential for herb-drug interaction. A secondary aim was to identify constituent compounds such as polyphenolics, in the crude extract preparations of Hyptis suaveolens and Boerhavia diffusa and measure them for activity. MATERIALS AND METHODS: CYP450 inhibition assays using recombinant CYP450 (rCYP) and fluorescence screening employing individual isozymes (CYP1A2, CYP2D6 and CYP3A4) were used to determine reversible- and time-dependent inhibition (TDI) profiles of extracts of Hyptis suaveolens and Boerhavia diffusa. Inhibition kinetic parameters, Ki and Kinact were also estimated. UPLC-MS employing a Synapt G2 (ESI negative) coupled to a PDA detector was used to identify polyphenolic compounds in crude extracts of Hyptis suaveolens and Boerhavia diffusa. RESULTS: The inhibitory potency of Hyptis suaveolens and Boerhavia diffusa extracts varied among the different enzymes, with CYP1A2 (3.68 ± 0.10µg/mL) being the least inhibited by HS compared to CYP2D6 (1.39 ± 0.01µg/mL) and CYP3A4 (2.36 ± 0.57µg/mL). BD was most potent on CYP3A4 (7.36 ± 0.94µg/mL) compared to both CYP2D6 (17.79 ± 1.02µg/mL) and CYP1A2 (9.48 ± 0.78µg/mL). Extracts of Hyptis suaveolens and Boerhavia diffusa exhibited TDIs on all CYPs. The most prominent phenolic candidates identified in both medicinal herbs using UPLC-MS analysis included caffeic acid, rutin, quercetin, citric acid, ferulic acid and gluconic acid. These phenolic compounds are thought to potentially give HS and BD their therapeutic effects and inhibitory characteristics affecting CYP450 activities. In vivo predictions showed the potential for HS and BD extracts to cause significant interactions if co-administered with other medications. CONCLUSIONS: The study reveals that crude aqueous extracts of HS and BD potentially inhibit drug metabolising isozymes CYP1A2, CYP2D6 and CYP3A4 in a reversible and time-dependent manner. Thus care should be taken when these extracts are co-administered with drugs that are substrates of CYP1A2, CYP2D6 and CYP3A4.

Evaluation of phytosynthesised silver nanoparticles from leaf extracts of Leucas aspera and Hyptis suaveolens and their larvicidal activity against malaria, dengue and filariasis vectors.

The present study deals with the green synthesis of silver nanoparticle from the aqueous leaf extracts of Leucas aspera and Hyptis suaveolens as reducing agent and to investigate the larvicidal activity of synthesized silver nanoparticles. The synthesized silver nanoparticles were characterized by Ultraviolet and visible absorption spectroscopy (UV), Fourier transform-infrared spectroscopy (FT-IR), X-ray spectroscopy (XRD), Field emission scanning electron microscope (FESEM) and High-resonance transmission electron microscopy (HRTEM) analysis. The nanoparticles are spherical, hexagonal, triangular and polyhedral in shape and the size of the Silver nanoparticles (AgNPs) of L. aspera was found to be in the range of 7-22 nm and AgNPs of H. suaveolens was 5-25 nm. Larvicidal bioassay with synthesized AgNPs synthesized from L. aspera and H. suaveolens extract, showed 100% mortality at 10 mg/L against Aedes aegypti, Anopheles stephensi and Culex quinquefasciatus with LC50 of 4.02, 4.69, 5.06 mg/L and LC90 of 11.22, 12.09, 12.74 mg/L and LC50 of 4.63, 4.04, 3.52 mg/L and LC90 of 12.07, 10.99, 09.61 respectively. These results suggest that the synthesized AgNPs of L. aspera and H. suaveolens have the potential to be used as an ideal eco-friendly agent for the control of the mosquito larvae.

Molecular dimensions and structural features of neutral polysaccharides from the seed mucilage of Hyptis suaveolens L.

The seed mucilage of Hyptis suaveolens L. includes acid - and neutral heteropolysaccharides in a ratio of about 1:1. The anionic charged fraction responsible for swelling and viscous behaviour possesses an average molar mass of Mw=350kg/mol, Mn=255kg/mol. The neutral polysaccharide fraction shows an average molar mass of Mw=47kg/mol and Mn=28kg/mol and is composed of d-Galp-, d-Glcp- and d-Manp residues in a molar ratio of about 3:2:1. The structural features present galactoglucan (30%) and galactoglucomannan (70%) with a high level of terminal ß-linked d-Galp residues (18%). Structural details of galactoglucomannan are derived by combined enzymatic and chemical methods as well as NMR spectroscopy. Sequences of octa/nonasaccharide ß-d-Glcp-(1→4)[ß-d-Galp-(1→2)-α-d-Galp-(1→6)]-ß-d-Manp-(1→4)-ß-d-Glcp-(1→4)-ß-d-Glcp-(1→4)[ß-d-Galp-(1→2)-α-d-Galp-(1→6)]-ß-d-Manp and lower mass tetrasaccharide repeating units ß-d-Glcp-(1→4)[ß-d-Galp-(1→2)-α-d-Galp-(1→6)]-ß-d-Manp were found. The level of the prebiotic activity is related to the availability of ß-linked d-Galp residues in the side chains of the molecules.

Prebiotic potential of neutral oligo- and polysaccharides from seed mucilage of Hyptis suaveolens.

Prebiotics are selectively fermented by the gastrointestinal microflora, resulting in benefits to human health. The seed mucilage of Hyptis suaveolens contains neutral and acidic polysaccharides in a ratio of 1:1. The neutral polysaccharides consist of galactose, glucose and mannose whereas the acidic polysaccharides contain fucose, xylose and 4-O-methylglucuronic acid -residues. The growth of probiotics in the presence of total, acidic or neutral polysaccharides and oligosaccharides was tested using turbidity measurements. The majority (11 out of 14) of the tested probiotic strains significantly grew in the neutral fraction. Growth occurred with some time delay, but may be longer lasting than with other lower molecular prebiotics. The extent of growth increased with neutral polysaccharides from H. suaveolens corresponding to the externally available galactose units (20%). In conclusion, neutral poly- and oligosaccharides from H. suaveolens have a prebiotic potential characterized by a delayed but long lasting effect.

Genotoxicity assessment of two common curing weeds: Hyptis suaveolens (L.) Poir. and Leucas indica (L.) R. Br.

Hyptis suaveolens and Leucas indica, two common weeds were selected for the present study, to reveal their probable cytotoxic potential. The meristematic root tips of Allium cepa were used for testing the cytotoxic property of the aqueous leaf extracts containing both polar and non-polar compounds, and that containing polar compounds alone, at different concentrations (0.125, 0.25, 0.5, 1 and 2 %) and at different time durations, using distilled water as negative control. Mitotic squash preparations were made using a standard protocol. The mitotic index of the treated root tip cells was found to be decreasing and the abnormality percentage was found to be increasing with increase in extract concentration when compared with the control. Maximum cytotoxicity was observed in the extract containing both polar and non-polar compounds. Both the tested plants were found to be cytotoxic. The abnormalities noticed were of both clastogenic (nuclear lesions, nuclear fragmentation, etc.) and non-clastogenic (aberrant cell wall formation at cytokinesis, ball metaphase, etc.) types. Both plant extracts were found to significantly (P < 0.05) inhibit root growth of Allium cepa with an EC50 value of 1.92 % (R(2) = 0.594) for Hyptis and 1.58 % (R(2) = 0.757) for Leucas. The results were also analysed statistically by using analysis of variance followed by appropriate post hoc tests. These two weeds are aromatic plants comprising of essential oils that are volatile, natural complex compounds characterized by a strong odour and formed as secondary metabolites. In nature, essential oils play an important role in the protection of the plants as insecticides by reducing their appetite for such plants. When specifically targeted the concept of effectively exploiting these weeds for the formulation of herbal insecticides/pesticides may be possible in the near future.

Purification and biochemical characterization of 11S globulin from chan (Hyptis suaveolens L. Poit) seeds.

Chan (Hyptis suaveolens) is a Mesoamerican crop highly appreciated since the pre-Hispanic cultures. Its proteins are a good source of essential amino acids; however, there are no reports on the properties of its individual proteins. In this study, the 11S globulin (Hs11S) was purified and biochemically characterized. The molecular weight of native Hs11S was about 150-300 kDa with isoelectric points of 5.0-5.3, composed by four monomers of 53.5, 52, 51.1 and 49.5 kDa, each formed by one acidic subunit and one basic subunit linked by a disulfide bond. Dynamic light scattering, size exclusion chromatography and native PAGE show that Hs11S is assembled in different oligomeric forms. LC-MS/MS analysis confirmed its identity. Hs11S presents antigenic determinants in common with lupin 11S globulin. Carbohydrate moieties or phosphate groups linked to Hs11S were not detected. This information is very useful in order to exploit and utilize rationally chan 11S globulin in food systems.

Evaluation of glucose utilization capacity of bioactivity-guided fractions of Barleria prionitis Linn and Hyptis suaveolens (L.) Poit in isolated rat hemidiaphragm.

INTRODUCTION: Diabetes mellitus (DM) is a chronic disease characterized by high blood glucose levels due to absolute or relative circulating insulin levels. Plants represent a major potential source of drugs for treating diabetes. The study of plants having antidiabetic activity may give a new approach in the treatment of DM. AIM: This study was aimed to investigate the glucose utilization capacity of bioactivity-guided fractions of Barleria prionitis and Hyptis suaveolens. MATERIALS AND METHODS: The dried coarse powdered plant material was extracted in aqueous ethanol by cold maceration; further, ethanolic extracts were fractionated using solvents of varying polarity and were investigated in isolated rat hemidiaphragm using 0.1% and 0.2% concentrations of plant extracts. RESULTS: The tested fractions of both plants showed significant and dose-dependent increased glucose uptake capacity and was found to be maximum with petroleum ether and aqueous ethanolic extracts of both plants; 0.2% concentration of both plant extracts is superior in activity when compared to 0.1% of the test extracts. H. suaveolens has produced more glucose utilization capacity when compared to B. prionitis. CONCLUSION: The activity of H. suaveolens is comparable to standard insulin (P < 0.01). Both the plant materials have some extra pancreatic mechanism like glucose uptake by peripheral tissue.

Activity studies of sesquiterpene oxides and sulfides from the plant Hyptis suaveolens (Lamiaceae) and its repellency on Ixodes ricinus (Acari: Ixodidae).

Hyptis suaveolens (Lamiaceae), a plant traditionally used as a mosquito repellent, has been investigated for repellent properties against nymphs of the tick Ixodes ricinus. Essential oils and volatile compounds of fresh and dried leaves, from plants originating from Laos and Guinea-Bissau, were identified by GC-MS and tested in a tick repellency bioassay. All the essential oils were strongly repellent against the ticks, even though the main volatile constituents differed in their proportions of potentially tick repellent chemicals. (+)/(-)-sabinene were present in high amounts in all preparations, and dominated the emission from dry and fresh leaves together with 1,8-cineol and α-phellandrene. 1,8-Cineol and sabinene were major compounds in the essential oils from H. suaveolens from Laos. Main compounds in H. suaveolens from Guinea-Bissau were (-)-sabinene, limonene and terpinolene. Among the sesquiterpene hydrocarbons identified, α-humulene exhibited strong tick repellency (96.8 %). Structure activity studies of oxidation or sulfidation products of germacrene D, α-humulene and ß-caryophyllene, showed increased tick repellent activity: of mint sulfide (59.4 %), humulene-6,7-oxide (94.5 %) and caryophyllene-6,7-oxide (96.9 %). The substitution of oxygen with sulfur slightly lowered the repellency. The effects of the constituents in the oils can then be regarded as a trade off between the subsequently lower volatility of the sesquiterpene derivatives compared to the monoterpenes and may thus increase their potential usefulness as tick repellents.

Hedgehog inhibitors from Artocarpus communis and Hyptis suaveolens.

The hedgehog (Hh) signaling pathway plays crucial roles in cell maintenance and proliferation during embryonic development. Naturally occurring Hh inhibitors were isolated from Artocarpus communis and Hyptis suaveolens using our previously constructed cell-based assay system. Bioactivity guided fractionation led to the isolation of 15 compounds, including seven new compounds (4, 5, 6, 7, and 9-11). The isolated compounds showed cytotoxicity against a cancer cell line (PANC1) in which Hh signaling was abnormally activated. Several compounds (12-14; GLI1 transcriptional inhibition IC50=7.6, 4.7, and 4.0 µM, respectively) inhibited Hh related protein (BCL2) expression. Moreover, compounds 1, 12, and 13 disrupted GLI1 and DNA complex formation.

Hyptis suaveolens (L.) Poit (Lamiaceae), a medicinal plant protects the stomach against several gastric ulcer models.

ETHNOPHARMACOLOGICAL RELEVANCE: Hyptis suaveolens is used by the traditional population in several parts of the world to treat inflammation, gastric ulcer and infection and is used as a crude drug to relieve symptoms related with gastric ulcer or gastritis in northeaster and central region of Brazil. MATERIALS AND METHODS: the standardized ethanolic extract (Hs-EtOHE) and hexanic fraction (Hs-HexF) of Hyptis suaveolens (62,5, 125, 250 and 500 mg/kg) was evaluated in several models of acute gastric ulcers. The participation of NO was evaluated by pretreatment with L-NAME and non-protein sulfyhydryls by NEM in the gastroprotective effect. RESULTS: Hs-EtOHE and Hs-HexF markedly reduced the gastric lesions induced by all ulcerogenic agents (HCl/ethanol, ethanol, NSAIDs and hypothermic restraint-stress). Gastric ulcerations were exacerbated by administration of NEM suggesting that the gastroprotective mechanism of action of Hs-EtOHE and Hs-HexF involves sulfhydryl groups. CONCLUSION: Ours results show that an extract of Hyptis suaveolens, administered orally to rodents, present gastro protective activity in different models of acute of gastric ulcer and give some support to the reported claims on the use of this plant as a gastro protective agent.

Abietane diterpenes from Hyptis suaveolens.

Investigation of the constituents of the whole plant of Hyptis suaveolens led to the isolation of three new abietane diterpenes, isosuaveolic acid (1), 8α,9α-epoxysuaveolic acid (2), and 14-O-methylsuaveolic acid (3), together with eleven known compounds. The structures of 1-3 were established by spectroscopic methods and chemical correlations. Some isolates were tested for their antimycobacterial and cytotoxic activities.

Production of podophyllotoxin from roots and plantlets of Hyptis suaveolens cultivated in vitro.

BACKGROUND: Hyptis suaveolens was an important source of food and medicines in pre-hispanic Mιxico and is actually used popularly to treat respiratory and skin diseases, fever, pain, and cramps, between other ailments. In 2008 the presence of podophyllotoxin (PTOX) was reported in this plant. OBJECTIVE: To establish in vitro cultures of H. suaveolens able to produce PTOX. MATERIALS AND METHODS: Explants of H. suaveolens were cultivated in Murashige and Skoog (MS) medium supplemented with different concentrations of the phytohormones 6-benzylaminopurine (6-BAP), 2,4-dichlorophenoxyacetic acid (2,4-D), 1-naphthaleneacetic acid (NAA) and kinetin (Kin), in order to induce the production of podophyllotoxin. Root cultures without hormones were also established and the quantification of PTOX was performed by HPLC analysis. RESULTS: The presence of growth regulators during in vitro cultivation of H. suaveolens, provoked morphological variations in explants, and induced the accumulation of different levels of PTOX. Roots grown without phytohormones accumulated PTOX at 0.013% dry weight (DW), while in three of the callus cultures cell lines growing together with roots, PTOX accumulated at concentrations of 0.003, 0.005 and 0.006% DW when NAA was combined with either Kin or BAP. In wild plant material PTOX was present in trace amounts in the aerial parts, while in the roots it was found at 0.005% DW. CONCLUSION: This study demonstrated that although it is possible to obtain PTOX in a variety of in vitro cultures of H. suaveolens, in vitro roots grown without the addition of growth regulators were better producers of PTOX.