- 1. New betulinic acid derivatives for bevirimat-resistant human immunodeficiency virus type-1.
Bevirimat (1, BVM) is an anti-HIV agent that blocks HIV-1 replication by interfering with HIV-1 Gag-SP1 processing at a late stage of viral maturation. However, clinical trials of 1 have revealed a high baseline drug resistance that is attributed to naturally occurring polymorphisms in HIV-1 Gag. To overcome the drug resistance, 28 new derivatives of 1 were synthesized and tested against compound 1-resistant (BVM-R) HIV-1 variants. Among them, compound 6 exhibited much improved activity against several HIV-1 strains carrying BVM-R polymorphisms. Compound 6 was at least 20-fold more potent than 1 against the replication of NL4-3/V370A, which carries the most prevalent clinical BVM-R polymorphism in HIV-1 Gag-SP1. Thus, compound 6 merits further development as a potential anti-AIDS clinical trial candidate....(more)
Dang Z, et al. J Med Chem 2013 Mar 14;56(5):2029-37.
Related Products: Betulinic Acid
- 2. Betulinic acid suppresses NGAL-induced epithelial-to-mesenchymal transition in melanoma.
Abstract Betulinic acid (BA) exhibits antitumoral activity by blocking proliferation, invasion, and angiogenesis. However, the impact of BA on epithelial-to-mesenchymal transition (EMT), a hallmark of cancer metastasis induced among others by neutrophil gelatinase-associated lipocalin (NGAL), remains unknown. The present study aimed at determining the effect of BA on NGAL-induced EMT. In A375 melanoma cells, BA downregulated mesenchymal markers, increased epithelial markers, and inhibited cytoskeletal reorganization. In addition, BA limited endogenous NGAL production and further suppressed EMT induced by exogenously added NGAL and the corresponding invasive cellular phenotype. In conclusion, BA interferes with EMT-associated changes, a mechanism to antagonize invasive melanoma cells....(more)
Gheorgheosu D, et al. Biol Chem 2013 Jun 1;394(6):773-81.
Related Products: Betulinic Acid
- 3. Synthesis of antitumor-active betulinic acid-derived hydroxypropargylamines by copper-catalyzend mannich reactions.
Several novel betulin derivates were prepared using Mannich reactions as a key step. Starting from 3-ethynyl-3-hydroxy-lup-20(29)-ene derivatives, copper-catalyzed Mannich reactions yielded hydroxypropargyl ammonium hydrochlorides or their corresponding methiodides. All compounds were screened in a sulforhodamine B assay for their antitumor activity using a panel of 9 human cancer cell lines. Some of these compounds showed significant cytotoxicity; they act by triggering apoptotic cell death as shown by additional acridine orange/propidium iodide assays, Trypan blue tests, DNA laddering experiments, and investigations of the cell cycle....(more)
Csuk R, et al. Arch Pharm (Weinheim) 2013 Mar;346(3):232-46.
Related Products: Betulinic Acid
- 4. Apoptosis of human breast cancer cells induced by microencapsulated betulinic acid from sour jujube fruits through the mitochondria transduction pathway.
Betulinic acid (BA), a natural pentacyclic triterpenoid, was isolated from sour jujube fruits for the first time. An inclusion complex comprising BA and β-cyclodextrin (β-CD) was formed to improve the dissolution of BA, but little is known about its anticancer effect. In this study, the anti-proliferative and apoptosis mechanisms of BA-β-CD on human breast cancer MCF-7 cells were further investigated. Experimental results confirmed that the complexation model inhibited the growth of MCF-7 cells in a dose-dependent manner, arrested cell cycle in the G2/M phase and induced apoptosis via the mitochondria transduction pathway. Gene and protein analyses showed that the complexation model significantly inhibited Bcl-2 expression and promoted Bax expression, causing caspase-3 and caspase-9 cascade activation. These findings corroborated evidence on microencapsulated BA as an apoptosis inducer in MCF-7 cells. Thus, sour jujube fruits may have potential use as a breast cancer chemotherapeutic agent....(more)
Sun YF, et al. Food Chem 2013 Jun 1;138(2-3):1998-2007.
Related Products: Betulinic Acid
- 5. Voltammetric determination of betulinic acid at lead film electrode after chromatographic separation in plant material.
The construction and application of a novel electrochemical detection system with an in situ plated lead film electrode for high-performance liquid chromatography (HPLC) is described. The working electrode of interest was tested as a potential sensor for the adsorptive stripping voltammetric determination of betulinic acid (BA). The high sensitivity of the proposed electrochemical detection results from the accumulation by adsorption of BA on the lead film surface before the proper electrode process. In a solution of sodium hydroxide, used as a supporting electrolyte for the proposed voltammetric method, the oxidation signal for BA was found to be proportional to the BA concentration in the range from 0.02 to 0.5μgL(-1) with a limit of detection equal to 0.009μgL(-1) (with preconcentration for 15s). The voltammetric detection was successfully applied to the determination of BA in the birch bark (Betula verrucosa) extracts after HPLC separation. The content of BA obtained by the proposed method was in close agreement with that obtained by HPLC coupled with photodiode array detector (HPLC-PAD). This appears to be the first application of electrochemical detection to the determination of pentacyclic triterpene....(more)
Tyszczuk-Rotko K, et al. Anal Biochem 2013 May 15;436(2):121-6.
Related Products: Betulinic Acid
- 6. Unusual Immuno-Modulatory Triterpene-Caffeates in the Skins of Russeted Varieties of Apples and Pears.
Three triterpene-caffeates have been isolated from skins of a russeted apple cultivar "Merton Russet" and identified by LC-MS and NMR as betulinic acid-3-cis-caffeate, betulinic acid-3-trans-caffeate, and oleanolic acid-3-trans-caffeate. Betulinic acid-3-trans-caffeate and oleanolic acid-3-trans-caffeate were also found in russeted pear skins. These compounds have not been previously reported in apples or pears, or in any other foods. Their presence was related to suberized tissue as they were only found in russet portions of the partially russeted apple cultivar "Cox's Orange Pippin" and were not detected in the waxy apple cultivar "Royal Gala". High concentrations of betulinic acid-3-trans-caffeate were found in the bark of both "Merton Russet" and "Royal Gala" trees. The three triterpene-caffeates showed anti-inflammatory activity in vitro, inhibiting NF-κB activation with IC50's of 6-9 μM. Betulinic acid-3-trans-caffeate, the predominant compound in the apples, was immuno-modulatory at around 10 μM in the in vitro and ex vivo bioassays, boosting production of the pro-inflammatory cytokine TNFα in cells stimulated with bacterial lipopolysaccharides....(more)
Andre CM, et al. J Agric Food Chem 2013 Mar 4.
Related Products: Betulinic Acid
- 7. Betulinic acid selectively increases protein degradation and enhances prostate cancer-specific apoptosis: possible role for inhibition of deubiquitinase activity.
Inhibition of the ubiquitin-proteasome system (UPS) of protein degradation is a valid anti-cancer strategy and has led to the approval of bortezomib for the treatment of multiple myeloma. However, the alternative approach of enhancing the degradation of oncoproteins that are frequently overexpressed in cancers is less developed. Betulinic acid (BA) is a plant-derived small molecule that can increase apoptosis specifically in cancer but not in normal cells, making it an attractive anti-cancer agent. Our results in prostate cancer suggested that BA inhibited multiple deubiquitinases (DUBs), which resulted in the accumulation of poly-ubiquitinated proteins, decreased levels of oncoproteins, and increased apoptotic cell death. In normal fibroblasts, however, BA did not inhibit DUB activity nor increased total poly-ubiquitinated proteins, which was associated with a lack of effect on cell death. In the TRAMP transgenic mouse model of prostate cancer, treatment with BA (10 mg/kg) inhibited primary tumors, increased apoptosis, decreased angiogenesis and proliferation, and lowered androgen receptor and cyclin D1 protein. BA treatment also inhibited DUB activity and increased ubiquitinated proteins in TRAMP prostate cancer but had no effect on apoptosis or ubiquitination in normal mouse tissues. Overall, our data suggests that BA-mediated inhibition of DUBs and induction of apoptotic cell death specifically in prostate cancer but not in normal cells and tissues may provide an effective non-toxic and clinically selective agent for chemotherapy....(more)
Reiner T, et al. PLoS One 2013;8(2):e56234.
Related Products: Betulinic Acid
- 8. Betulinic acid alleviates non-alcoholic fatty liver by inhibiting SREBP1 activity via the AMPK-mTOR-SREBP signaling pathway.
Non-alcoholic fatty liver disease (NAFLD) is emerging as the most common liver disease in industrialized countries. The discovery of food components that can ameliorate NAFLD is therefore of interest. Betulinic acid (BA) is a triterpenoid with many pharmacological activities, but the effect of BA on fatty liver is as yet unknown. To explore the possible anti-fatty liver effects and their underlying mechanisms, we used insulin-resistant HepG2 cells, primary rat hepatocytes and liver tissue from ICR mice fed a high-fat diet (HFD). Oil Red O staining revealed that BA significantly suppressed excessive triglyceride accumulation in HepG2 cells and in the livers of mice fed a HFD. Ca(+2)-calmodulin dependent protein kinase kinase (CAMKK) and AMP-activated protein kinase (AMPK) were both activated by BA treatment. In contrast, the protein levels of sterol regulatory element-binding protein 1 (SREBP1), mammalian target of rapamycin (mTOR) and S6 kinase (S6K) were all reduced when hepatocytes were treated with BA for up to 24h. We found that BA activates AMPK via phosphorylation, suppresses SREBP1 mRNA expression, nuclear translocation and repressed SREBP1 target gene expression in HepG2 cells and primary hepatocytes, leading to reduced lipogenesis and lipid accumulation. These effects were completely abolished in the presence of STO-609 (a CAMKK inhibitor) or compound C (an AMPK inhibitor), indicating that the BA-induced reduction in hepatic steatosis was mediated via the CAMKK-AMPK-SREBP1 signaling pathway. Taken together, our results suggest that BA effectively ameliorates intracellular lipid accumulation in liver cells and thus is a potential therapeutic agent for the prevention of fatty liver disease....(more)
Quan HY, et al. Biochem Pharmacol 2013 May 1;85(9):1330-40.
Related Products: Betulinic Acid
- 9. Effect of fermented bilberry extracts on visual outcomes in eyes with myopia: a prospective, randomized, placebo-controlled study.
Abstract Purpose: To investigate clinically the effects of yeast-fermented bilberry extract on visual outcomes in myopic eyes. Methods: In a prospective, randomized, placebo-controlled, cross-over study, we examined 30 eyes of 30 middle-aged healthy volunteers (mean age±standard deviation, 39.5±7.2 years) with myopia [manifest spherical equivalent, -2.40±1.88 diopters (D)], who were randomly assigned to 1 of 2 oral regimens: fermented bilberry extract (400 mg/day) or placebo. We quantitatively assessed visual acuity, refraction, pupil constriction rate, accommodation, and mesopic contrast sensitivity (CS), before and 1 month after treatment. Only the right eyes were tested. The amplitude of accommodation and CS were measured with an accommodometer (D'ACOMO; WOC) and a CS unit (VCTS-6500; Vistech), respectively. From the CS, the area under the log contrast sensitivity function (AULCSF) was calculated. Results: The mean amplitude of accommodation increased significantly, from 4.62±1.88 D before treatment, to 5.33±2.03 D after treatment in the study group (Wilcoxon signed-rank test, P=0.002). Moreover, the mesopic AULCSF was significantly increased, from 1.04±0.16 before, to 1.13±0.17 after, treatment (P=0.009). However, we found no significant changes in accommodation or AULCSF in the control group (P>0.05), or any significant changes in any other parameters in either group (P>0.05). Conclusions: The present data show that fermented bilberry extract is effective in causing increases in subjective accommodation and in mesopic CS in myopic eyes....(more)
Kamiya K, et al. J Ocul Pharmacol Ther 2013 Apr;29(3):356-9.
Related Products: Bilberry Extract
- 10. Degradation Index for quality evaluation of commercial dietary supplements of bilberry extract.
In recent years, many anthocyanin-containing dietary supplements of various dosages and formulations have been sold through advertising their large number of beneficial effects. On the other hand, there is an increased risk of distributing deteriorated supplements to consumers due to lax regulations, because in Japan these supplements are classified as food. Spectrophotometric methods are commonly used to control the quality of supplements. However, these methods have limitations with regard to assessing deterioration. In this study, we evaluated a new index for detection of deteriorated products. The stability of 3 formulations and the quality of 20 supplements were investigated by ultra-high performance liquid chromatography, which is superior to spectrophotometry for identifying and quantifying individual anthocyanins. The stability was not only affected by storage temperature but also by formulation. We defined "Degradation Index" (DI) as an indicator of the deterioration of supplements. Of 20 supplements investigated, the DI of 5 supplements was more than 3-fold the value of Bilberon-25, and the worst one was 12.7-fold. These results suggest that DI could be a useful quality control index for detecting deteriorated supplements....(more)
Yamamoto M, et al. J Food Sci 2013 Mar;78(3):S477-83.
Related Products: Bilberry Extract
- 11. Bilberry anthocyanin-rich extract alters expression of genes related to atherosclerosis development in aorta of apo E-deficient mice.
Intake of anthocyanin-rich foods has been associated with a reduced risk of cardiovascular diseases. We recently reported that a nutritional supplementation with a bilberry anthocyanin-rich extract (BE) attenuates atherosclerotic lesion development in apolipoprotein E-deficient (apoE/) mice. However, the mechanism(s) of their preventive action are not completely understood. Anthocyanins may alter mRNA levels of genes related to atherosclerosis in cultured macrophages and endothelial cells, but in vivo studies remain scarce. The aim of the present study was to explore the in vivo mechanisms of action of the same bilberry extract, administered by supplementation at a nutritional level, in the aorta of apo E/ mice using a global transcriptomic approach. This study focused on the early stage of atherosclerosis development for better assessment of BE action on initiation mechanisms of this pathology. After a two week period, plasma lipid and antioxidant capacity were evaluated and the global genomic analysis was carried out using pangenomic microarrays. BE supplementation significantly improved hypercholesterolemia whereas the plasmatic antioxidant status remained unchanged. Nutrigenomic analysis identified 1261 genes which expression was modulated by BE in the aorta. Bioinformatic analysis revealed that these genes are implicated in different cellular processes such as oxidative stress, inflammation, transendothelial migration and angiogenesis, processes associated with atherosclerosis development/protection. Some of the most significantly down-regulated genes included genes coding for AOX1, CYP2E1 or TXNIP implicated in the regulation of oxidative stress, JAM-A coding for adhesion molecules or VEGFR2 implicate in regulation of angiogenesis. Other genes were up-regulated, such as CRB3, CLDN14 or CDH4 potentially associated with increased cell-cell adhesion and decreased paracellular permeability. These results provide a global integrated view of the mechanisms involved in the preventive action of bilberry anthocyanin-rich extract against atherosclerosis....(more)
Mauray A, et al. Nutr Metab Cardiovasc Dis 2012 Jan;22(1):72-80.
Related Products: Bilberry Extract
- 12. Vision preservation during retinal inflammation by anthocyanin-rich bilberry extract: cellular and molecular mechanism.
Anthocyanin-rich bilberry extract, a plant-derived antioxidant, has been utilized as a popular supplement for ocular health worldwide. However, it is unclear whether this extract has any biological effect on visual function, and the mechanism for such an effect is completely unknown. In this study, we generated a mouse model of endotoxin-induced uveitis (EIU) that shows retinal inflammation, as well as uveitis, by injecting lipopolysaccharide. We pretreated the mice with anthocyanin-rich bilberry extract and analyzed the effect on the retina. Anthocyanin-rich bilberry extract prevented the impairment of photoreceptor cell function, as measured by electroretinogram. At the cellular level, we found that the EIU-associated rhodopsin decreased and the shortening of outer segments in photoreceptor cells were suppressed in the bilberry-extract-treated animals. Moreover, the extract prevented both STAT3 activation, which induces inflammation-related rhodopsin decrease, and the increase in interleukin-6 expression, which activates STAT3. In addition to its anti-inflammatory effect, the anthocyanin-rich bilberry extract ameliorated the intracellular elevation of reactive oxygen species and activated NF-κB, a redox-sensitive transcription factor, in the inflamed retina. Our findings indicate that anthocyanin-rich bilberry extract has a protective effect on visual function during retinal inflammation....(more)
Miyake S, et al. Lab Invest 2012 Jan;92(1):102-9.
Related Products: Bilberry Extract
- 13. Sealing liquid-filled pectinate capsules with a shellac coating.
Liquid-filled pectinate capsules have a large potential for the controlled and site-specific delivery of liquid drugs. Earlier studies have shown that pure pectinate capsules can store drugs only for a few minutes. Here, we show that the retention time can be extended to several hours by coating the capsules with the natural resin shellac. A bilberry extract containing anthocyanins with promising therapeutic properties was used as model drug to characterize the permeability of the capsules by in vitro drug release measurements. Characterizing the structure of the double-layered capsule membranes by NMR microscopy, we optimized the capsule production by adjusting the pH-value in the coating process and the gelation time of the pectinate hydrogel layer. A comparison of the layer thicknesses with drug release measurements reveals that capsules with the thinnest shellac layers provide the best entrapment. Additional squeezing experiments show that the shellac layer makes the capsules also mechanically more stable....(more)
Henning S, et al. J Microencapsul 2012;29(2):147-55.
Related Products: Bilberry Extract
- 14. Preparation and comparative release characteristics of three anthocyanin encapsulation systems.
Bilberries (Vaccinium myrtillus L.) and their major polyphenolic constituents, anthocyanins, have preventive activities inter alia against colon cancer and inflammatory bowel diseases. However, anthocyanins are sensitive to environmental conditions; thus their bioavailability in the gastrointestinal tract is an important determinant of their in vivo activity. In the study reported here, the potential benefits of encapsulating an anthocyanin rich bilberry extract (BE) on anthocyanin stability were investigated. Nonencapsulated BE and three different BE loaded microcapsule systems were incubated in simulated gastric fluid (SGF) and fed state simulated intestinal fluid (FeSSIF). After exposure to these media, released anthocyanins were identified and quantified by HPLC with UV/Vis detection. Although a rapid release of anthocyanins was observed within the first 20 min, encapsulation of anthocyanins doubled the amount of available anthocyanins after 150 min of incubation. These results illustrate the ability of encapsulation to inhibit early degradation of anthocyanins in the intestinal system....(more)
Oidtmann J, et al. J Agric Food Chem 2012 Jan 25;60(3):844-51.
Related Products: Bilberry Extract
- 15. Vaccinium myrtillus ameliorates unpredictable chronic mild stress induced depression: possible involvement of nitric oxide pathway.
Chronic unpredictable stressors can produce a situation similar to clinical depression and such animal models can be used for the preclinical evaluation of antidepressants. Nitric oxide, a secondary messenger molecule, has been implicated in neurotransmission, synaptic plasticity, learning, aggression and depression. Vaccinium myrtillus (bilberry) extract is a potent inhibitor of reactive oxygen/nitrogen species and cytokine production. The present study investigated the role of nitric oxide in the antidepressant action of Vaccinium myrtillus in unpredictable chronic mild stress-induced depression in mice. Animals were subjected to different stress paradigms daily for a period of 21 days to induce depressive-like behavior. Pretreatment with L-arginine significantly reversed the protective effect of bilberry (500 mg/kg) on chronic stress-induced behavioral (immobility period, sucrose preference) and biochemical (lipid peroxidation and nitrite levels; endogenous antioxidant activities) in stressed mice. Furthermore, L-NAME (10 mg/kg) pretreatment with a sub-effective dose of bilberry (250 mg/kg) significantly potentiated the protective effect of bilberry extract. The study revealed that modulation of the nitric oxide pathway might be involved in antidepressant-like effects of Vaccinium myrtillus in stressed mice....(more)
Kumar B, et al. Phytother Res 2012 Apr;26(4):488-97.
Related Products: Bilberry Extract
- 16. Anthocyanins, but not anthocyanidins, from bilberry (Vaccinium myrtillus L.) alleviate pruritus via inhibition of mast cell degranulation.
We have previously reported that bilberry anthocyanins exhibit an anti-pruritic effect in a mouse model of allergic contact dermatitis. It has been reported that anthocyanins are particularly sensitive to thermal treatment and are easily hydrolyzed to anthocyanidins when exposed to high temperatures. The objective of this study was to compare the anti-pruritic effect of anthocyanin-rich quality-controlled bilberry extract and anthocyanidin-rich degraded extract using a mouse model of allergic contact dermatitis. BALB/c mice with allergic contact dermatitis induced by 4 weeks of repeated application of 2,4,6-trinitro-1-chlorobenzene (TNCB) were administered Bilberon-25 orally for 4 weeks after sensitization with TNCB. The effect of Bilberon-25 on pruritus was evaluated by measurement of scratching behavior. RBL-2H3 mast cells were used to investigate the effect of Bilberon-25 on degranulation in 48/80-stimulated mast cells. Compared with nonheated Bilberon-25, the proportion of anthocyanins in heated Bilberon-25 decreased, and the proportion of anthocyanidins was increased in heated-time dependent manner. Treatment with non-heated Bilberon-25 significantly attenuated the TNCB-induced increase in scratching behavior, whereas treatment with 2 h-heated Bilberon-25 did not. Moreover, 300 μg/mL nonheated Bilberon-25 showed significant inhibition of degranulation in RBL-2H3 mast cells, whereas 2 h-heated Bilberon-25 had no effect at any concentration studied. It is assumed that the inhibitory effect of bilberry anthocyanins on pruritus might be mediated, at least in part, by its inhibitory effect on mast cell degranulation. In conclusion, the anthocyanin-rich but not anthocyanidin-rich bilberry extract may be a useful dietary supplement for skin diseases involving pruritic symptoms, such as chronic allergic contact dermatitis, atopic dermatitis, and rhinitis....(more)
Yamaura K, et al. J Food Sci 2012 Dec;77(12):H262-7.
Related Products: Bilberry Extract
- 17. Stability of anthocyanin-rich w/o/w-emulsions designed for intestinal release in gastrointestinal environment.
Anthocyanins belong to the most important hydrophilic plant pigments. Outside their natural environment, these molecules are extremely unstable. Encapsulating them in submicron-sized containers is one possibility to stabilize them for the use in bioactivity studies or functional foods. The containers have to be designed for a target release in the human gastrointestinal system. In this contribution, an anthocyanin-rich bilberry extract was encapsulated in the inner aqueous phase of water-in-oil-in-water-double emulsions. The physical stability as well as the release of free fatty acids and encapsulated, bioactive substances from the emulsions during an in vitro gastrointestinal passage were investigated. The focus was on the influence of emulsion microstructural parameters (for example, inner and outer droplet size, disperse phase content) and required additives (emulsifier systems), respectively. It could be shown that it is possible to stabilize anthocyanins in the inner phase of double emulsions. The release rate of free fatty acids during incubation was independent of the emulsifier used. However, the exterior (O/W)-emulsifier has an impact on the stability of multiple emulsions in gastrointestinal environment and, thus, the location of release. Long-chained emulsifiers like whey proteins are most suitable to transport a maximum amount of bioactive substances to the effective location, being the small intestine for anthocyanins. In addition, it was shown that the dominating release mechanism for entrapped matter was coalescence of the interior W(1) -droplets with the surrounding W(2) -phase....(more)
Frank K, et al. J Food Sci 2012 Dec;77(12):N50-7.
Related Products: Bilberry Extract
- 18. Bilberry and blueberry anthocyanins act as powerful intracellular antioxidants in mammalian cells.
Berry anthocyanins have pronounced health effects, even though they have a low bioavailability. The common mechanism underlying health protection is believed to relate to antioxidant activity. Berry extracts, chemically characterised for their phenolic content, were prepared from bilberries (Vaccinium myrtillusL.) and blueberries (Vaccinium corymbosumL.); the bilberry extract was further purified to obtain the anthocyanin fraction. The antioxidant activity of each extract was examined at the cellular level. For this purpose a specific assay, known as cellular antioxidant activity assay (CAA), was implemented in different cell lines: human colon cancer (Caco-2), human hepatocarcinoma (HepG2), human endothelial (EA.hy926) and rat vascular smooth muscle (A7r5). Here we show for the first time that anthocyanins had intracellular antioxidant activity if applied at very low concentrations (<1 μg/l; nM range), thereby providing a long-sought rationale for their health protecting effects in spite of their unfavorable pharmacokinetic properties....(more)
Bornsek SM, et al. Food Chem 2012 Oct 15;134(4):1878-84.
Related Products: Bilberry Extract
- 19. Distribution of isoflavones and coumestrol in neglected tropical and subtropical legumes.
BACKGROUND:
Isoflavones and coumestrol from dietary legumes are plant constituents showing multiple beneficial effects on humans. Owing to their ability to bind with mammalian estrogenic receptors and thereby intervention in several kinds of hormone-related cancers, they have received much attention. Soybean (Glycine max) is currently the major source of isoflavonoids in human diet. However, dozens of tropical and subtropical leguminous species remain unexplored for their isoflavonoids content.
RESULTS:
We have analyzed 55 extracts from 41 tropical and subtropical legume species used either in human or animal diet by high-performance liquid chromatography for the content of soy isoflavones, biochanin A, daidzein, daidzin, formononetin, genistein, genistin, sissotrin, ononin and the coumestan coumestrol. Genistein and biochanin A were the most abundant compounds. The highest content of genistein was found in aerial parts of Andira macrothyrsa, seeds of Pachyrhizus tuberosus and aerial parts of Calopogonium mucunoides (598, 250 and 184 µg g(-1), respectively) and biochanin A in aerial parts of Cratylia argentea, C. mucunoides and flowers of A. macrothyrsa (76, 53 and 40 µg g(-1), respectively).
CONCLUSION:
None of the samples tested was richer overall source of soy isoflavones and coumestrol than soybean; nevertheless several species (C. mucunoides or A. macrothyrsa) may serve as a promising source of individual compounds.
Copyright © 2012 Society of Chemical Industry....(more)
Leuner O, et al. J Sci Food Agric 2013 Feb;93(3):575-9.
Related Products: Biochanin A
- 20. Differential antiviral and anti-inflammatory mechanisms of the flavonoids biochanin A and baicalein in H5N1 influenza A virus-infected cells.
From a panel of 22 flavonoids, we identified six compounds (apigenin, baicalein, biochanin A, kaempferol, luteolin, naringenin) that inhibited influenza A nucleoprotein production in human lung epithelial (A549) cells infected with the highly pathogenic avian influenza H5N1 virus strain A/Thailand/Kan-1/04 in non-toxic concentrations. Baicalein (IC(50): 18.79±1.17μM, selectivity index 5.82) and biochanin A (IC(50) 8.92±1.87μM, selectivity index 5.60) were selected for further experiments. Both compounds reduced H5N1 infectious titres (baicalein 40μM: 29-fold reduction, biochanin A 40μM: 55-fold reduction after infection at MOI 0.01), virus-induced caspase 3 cleavage, nuclear export of viral RNP complexes, and enhanced the effects of the neuraminidase inhibitor zanamivir. Biochanin A and baicalein also inhibited the replication of the H5N1 strain A/Vietnam/1203/04. Time of addition experiments indicated that both compounds interfere with H5N1 replication after the adsorption period. Further mechanistic investigations revealed clear differences between these two flavonoids. Only baicalein interfered with the viral neuraminidase activity (39±7% inhibition at 100μM, the maximum concentration tested). In contrast to baicalein, biochanin A affected cellular signalling pathways resulting in reduced virus-induced activation of AKT, ERK 1/2, and NF-kB. Moreover, biochanin A inhibited the virus-induced production of IL-6, IL-8, and IP-10 while baicalein inhibited IL-6 and IL-8 production without affecting IP-10 levels. In primary human monocyte-derived macrophages, only baicalein but not biochanin A impaired H5N1 virus replication. Both flavonoids interfered with the H5N1-induced production of IL-6, IP-10, and TNF-α but not of IL-8 in macrophages. These findings indicate that closely related flavonoids can exert anti-H5N1 effects by different molecular mechanisms....(more)
Sithisarn P, et al. Antiviral Res 2013 Jan;97(1):41-8.
Related Products: Biochanin A
- 21. Neuroprotective effects of biochanin a against glutamate-induced cytotoxicity in PC12 cells via apoptosis inhibition.
L-Glutamate plays a crucial role in neuronal cell death, which is known to be associated with various neurodegenerative diseases, such as Alzheimer's, Parkinson's, and Huntington's diseases. In this study, we investigated the protective effects of biochanin A, a phytoestrogen compound found mainly in Trifolium pratense, against L-glutamate-induced cytotoxicity in a PC12 cell line. Exposure of the cells to 10 mM L-glutamate was found to significantly increase cell viability loss and apoptosis, whereas pretreatment with various concentrations of biochanin A attenuated the cytotoxic effects of L-glutamate. Specifically, the pretreatment led to not only decreases in the release of lactate dehydrogenase, the number of apoptotic cells, and the activity of caspase-3 but also an increase in the total glutathione level in the L-glutamate-treated PC12 cells. These results indicate that biochanin A may be able to exert neuroprotective effects against L-glutamate-induced cytotoxicity. Furthermore, our findings also imply that biochanin A may act as an antiapoptotic agent in order to perform its protective function....(more)
Tan JW, et al. Neurochem Res 2013 Mar;38(3):512-8.
Related Products: Biochanin A
- 22. Development of a food compositional database for the estimation of dietary intake of phyto-oestrogens in a group of postmenopausal women previously treated for breast cancer and validation with urinary excretion.
The scientific literature contains evidence suggesting that women who have been treated for breast cancer may, as a result of their diagnosis, increase their phyto-oestrogen (PE) intake. In the present paper, we describe the creation of a dietary analysis database (based on Dietplan6) for the determination of dietary intakes of specific PE (daidzein, genistein, glycitein, formononetin, biochanin A, coumestrol, matairesinol and secoisolariciresinol), in a group of women previously diagnosed and treated for postmenopausal breast cancer. The design of the database, data evaluation criteria, literature data entry for 551 foods and primary analysis by LC-MS/MS of an additional thirty-four foods for which there were no published data are described. The dietary intake of 316 women previously treated for postmenopausal breast cancer informed the identification of potential food and beverage sources of PE and the bespoke dietary analysis database was created to, ultimately, quantify their PE intake. In order that PE exposure could be comprehensively described, fifty-four of the 316 subjects completed a 24 h urine collection, and their urinary excretion results allowed for the description of exposure to include those identified as 'equol producers'....(more)
Clarke DB, et al. Br J Nutr 2013 Jan 4:1-8.
Related Products: Biochanin A
- 23. Methylation of genistein and kaempferol improves their affinities for proteins.
Methylation of flavonoids appears to be a simple and effective way to improve metabolic resistance and transport of flavonoids. Serum albumins are major soluble proteins serving as transport proteins for many exogenous compounds. This work in here mainly concerns about the effect of methylation of flavonoids on the affinity for human serum albumin (HSA) and ovalbumin. One isoflavone (genistein) and one flavonol (kaempferol) and their monomethylated derivatives at position 4' (biochanin A and kaempferide) were studied for their affinities for ovalbumin and HSA. The methylation of flavonoids significantly affects the binding process. In general, the methylation of flavonoids improved the affinities for proteins by 2-16 times. This result supports that the methylation of genistein and kaempferol enhanced the transporting ability, which leads to facilitated absorption and greatly increased bioavailability. The methylation increases the hydrophobicity of genistein and kaempferol, and the hydrophobic interaction plays an important role in binding flavonoids to HSA and ovoalbumin....(more)
Cao H, et al. Int J Food Sci Nutr 2013 Jun;64(4):437-43.
Related Products: Biochanin A
- 24. A new O-prenylated flavonol from the roots of Sophora interrupta.
A new O-prenylated flavonol, 3',4'-dimethoxy-7-(γ,γ-dimethylallyloxy)flavonol (1), together with three known compounds, 2'-hydroxy-3,4-dimethoxychalcone (2), biochanin A (3) and kaempferol-3-O-β-d-glucopyranoside (4), were isolated from the roots of Sophora interrupta Bedd. The structure of compound 1 was elucidated by extensive 1D and 2D NMR spectral studies....(more)
Munikishore R, et al. Nat Prod Res 2013 Feb 13.
Related Products: Biochanin A
- 25. The preventive effect of biochanin a on bone loss in ovariectomized rats: involvement in regulation of growth and activity of osteoblasts and osteoclasts.
Biochanin A (BCA) is a major isoflavone abundant in red clover (Trifolium pretense). The protective effect of BCA on bone loss in an ovariectomized (OVX) animal model has never been clarified. The objective of this study was to investigate the biological effects of BCA on bone loss in OVX rats in vivo and on the development of osteoblasts and osteoclasts in vitro. Ovariectomy resulted in a marked increase in body weight and a decrease in femoral bone mineral density and trabecular bone volume that was prevented by BCA or 17 β -estradiol (E2) treatment. However, an increase in uterine weight was observed in E2-treated OVX rats, but not in response to BCA treatment. Treatment with BCA increased the mRNA expression of osterix, collagen type I, alkaline phosphatase (ALP), and osteocalcin and decreased the mRNA expression of tartrate-resistant acid phosphatase (TRAP) and the receptor activator of nuclear factor- κ B ligand (RANKL)/osteoprotegerin (OPG) ratio in the femur of OVX rats. Treatment with BCA or E2 prevented the OVX-induced increase in urinary deoxypyridinoline (DPD) and serum tumor necrosis factor α (TNF- α ) and interleukin-1 β (IL-1 β ). In vitro, BCA induced preosteoblasts to differentiate into osteoblasts and increased osteoblast mineralization. BCA inhibited preosteoclasts and osteoclast proliferation and decreased osteoclast bone resorption. These findings suggest that BCA treatment can effectively prevent the OVX-induced increase in bone loss and bone turnover possibly by increasing osteoblastic activities and decreasing osteoclastic activities....(more)
Su SJ, et al. Evid Based Complement Alternat Med 2013;2013:594857.
Related Products: Biochanin A
- 26. Determination of seventeen endocrine disruptor compounds and their spatial and seasonal distribution in Ria Formosa Lagoon (Portugal).
In spite of its outstanding ecological and touristic importance the Ria Formosa Lagoon shows signs of anthropogenic pollution. Nonetheless, until the present survey no studies had ever documented the measurement of natural and pharmaceutical estrogens (17β-estradiol, estrone, and 17α-ethynylestradiol), xenoestrogenic industrial pollutants (4-octylphenol, 4-nonylphenol, and their mono and diethoxylates and bisphenol A), phytoestrogens (formononetin, biochanin A, daidzein, genistein), and sitosterol in this area. The 17 compounds measured herein are known as endocrine disrupters (EDCs) and act over the endocrine system even in few amounts (ng L<sup>-1</sup>-μg L<sup>-1</sup>). Thus to conclude about the influx of EDCs in the lagoon, water samples were taken every 2 months, during 1 year (2010), in low tide at nine sites distributed along the coastline. Water samples (1 L) were preconcentrated in the Oasis HLB cartridges and cleaned in silica cartridges before their analysis by GC-MS. Data showed the ubiquitous presence of potentially hazardous amounts of estrogens (particularly of ethynylestradiol, up to 24.3 ng L<sup>-1</sup>), nonylphenol (up to 547 ng L<sup>-1</sup>), and sitosterol (up to 12,300 ng L<sup>-1</sup>), mainly in summer, suggesting that the increase of the local number of inhabitants (tourists), the rise of the water temperature (up to 26 °C), and the blooming of local flora may interfere with the water quality parameters. This makes the lagoon a potential model to study. Taking into account the data, it was concluded that there are conditions for the occurrence of endocrine disruption in aquatic animals, even in areas included in the natural park of the Formosa. Besides, both the high amounts of un-ionized ammonia (up to 0.3 mg L<sup>-1</sup>) and phosphates (up to 1.6 mg L<sup>-1</sup>) my pose risks for local fauna and humans....(more)
Rocha MJ, et al. Environ Monit Assess 2013 Apr 18.
Related Products: Biochanin A
- 27. Functional Characterization of a Flavonoid Glycosyltransferase Gene from Withania somnifera (Ashwagandha).
Glycosylation of flavonoids is mediated by family 1 uridine diphosphate (UDP)-dependent glycosyltransferases (UGTs). Until date, there are few reports on functionally characterized flavonoid glycosyltransferases from Withania somnifera. In this study, we cloned the glycosyltransferase gene from W. somnifera (UGT73A16) showing 85-92 % homology with UGTs from other plants. UGT73A16 was expressed as a His6-tagged fusion protein in Escherichia coli. Several compounds, including flavonoids, were screened as potential substrates for UGT73A16. HPLC analysis and hypsochromic shift indicated that UGT73A16 transfers a glucose molecule to several different flavonoids. Based on kinetic parameters, UGT73A16 shows more catalytic efficiency towards naringenin. Here, we explored UGT73A16 of W. somnifera as whole cell catalyst in E. coli. We used flavonoids (genistein, apigenin, kaempferol, naringenin, biochanin A, and daidzein) as substrates for this study. More than 95 % of the glucoside products were released into the medium, facilitating their isolation. Glycosylation of substrates occurred on the 7- and 3-hydroxyl group of the aglycone. UGT73A16 also displayed regiospecific glucosyl transfer activity towards 3-hydroxy flavone compound, which is the backbone of all flavonols and also for a chemically synthesized compound, not found naturally. The present study generates essential knowledge and molecular as well as biochemical tools that allow the verification of UGT73A16 in glycosylation....(more)
Singh S, et al. Appl Biochem Biotechnol 2013 Apr 23.
Related Products: Biochanin A
- 28. Biochanin A Protects against Acute Carbon Tetrachloride-Induced Hepatotoxicity in Rats.
Biochanin A (BCA) is an isoflavone found in red clover possessing multiple pharmacological activities including antimicrobial, antioxidant, and anticancer ones. The present study aimed to assess its hepatoprotective potential at different doses in a carbon tetrachloride (CCl4)-induced hepatotoxicity model in rats. The effects on hepatic injury were explored by measuring serum levels of alanine aminotransferase, aspartate aminotransferase, and alkaline phosphatase. Furthermore, the serum levels of glucose, urea, creatinine, total bilirubin, total proteins, triglycerides, and total cholesterol were determined. The metabolic capacity of the liver was assessed by measuring changes in cytochrome P450 2E1 activity. The underlying mechanisms were substantiated by measuring oxidative stress markers as catalase, superoxide dismutase, glutathione peroxidase, glutathione transferase, glutathione reductase, reduced glutathione, total antioxidant capacity, and lipid peroxidation, as well as inflammation markers such as nitric oxide, inducible nitric oxide synthase, cyclooxygenase2, tumor necrosis factor-α, and leukocyte-common antigen. The results were confirmed by histopathological examination, and the median lethal dose was determined to confirm the safety of the drug. BCA successively protected against CCl4-induced damage, normalizing many parameters to that of the control group. The study indicates that BCA possesses multimechanistic hepatoprotective activity that can be attributed to its antioxidant, anti-inflammatory, and immunomodulatory activity....(more)
Breikaa RM, et al. Biosci Biotechnol Biochem 2013 May 7.
Related Products: Biochanin A
- 29. Bitter melon extract inhibits proliferation of Trypanosoma brucei bloodstream forms in vitro.
Trypanosoma brucei is the causative agent of sleeping sickness, a fatal disease prevalent in sub-Saharan Africa. The few currently available drug treatments are dated and face problems with toxicity and resistance. For these reasons, there is an urgent need for the development of new chemotherapies for the treatment of sleeping sickness. In this study, we investigated the trypanocidal activity of bitter melon extract. Recently, it has been shown that bitter melon extracts display cytotoxic activity towards different cancer cell lines. However, agents exhibiting anti-tumour activity are usually also inhibiting the growth of T. brucei. Treatment of bloodstream forms of T. brucei with extracts prepared from Chinese and Indian bitter melon varieties resulted in a decrease in cell proliferation. In contrast, human myeloid leukaemia HL-60 cells were 3-6 times less sensitive to the extracts than trypanosomes. Initial fractionation of bitter melon extracts indicated that the trypanocidal activity of the extract is associated with at least two different classes of substances: one class of larger molecular weight compounds (>3 kDa) causing rapid lysis of trypanosomes and one class of smaller molecular weight compounds (<3 kDa) inducing accumulation of the parasites in the G(2)-M phase of the cell cycle. Together, the results suggest that bitter melon is a promising source for trypanocidal agents which could be used as lead compounds for the development of novel anti-sleeping sickness drugs....(more)
Phillips EA, et al. Exp Parasitol 2013 Mar;133(3):353-6.
Related Products: Bitter Melon Extract
- 30. Bitter melon (Momordica charantia) extract suppresses adrenocortical cancer cell proliferation through modulation of the apoptotic pathway, steroidogenesis, and insulin-like growth factor type 1 receptor/RAC-α serine/threonine-protein kinase signali
Adrenocortical carcinomas are rare but present with extremely poor prognosis. One of the approaches to control cancer progression and reduce cancer risk is prevention through diet. Bitter melon is widely consumed as a vegetable and especially as a traditional medicine in many countries. In this study, we have used human and mouse adrenocortical cancer cells as an in vitro model to assess the efficacy of bitter melon extract (BME) as an anticancer agent. The protein concentrations of BME and other extracts were measured before use. First, BME treatment of adrenocortical cancer cells resulted in a significantly dose-dependent decrease in cell proliferation. However, we did not observe an antiproliferative effect in adrenocortical cancer cells treated with extracts from blueberry, zucchini, and acorn squash. Second, apoptosis of adrenocortical cancer cells was accompanied by increased caspase-3 activation and poly(ADP-ribose) polymerase cleavage. BME treatment enhanced cellular tumor antigen p53, cyclin-dependent kinase inhibitor 1A (also called p21), and cyclic AMP-dependent transcription factor-3 levels and inhibited G1/S-specific cyclin D1, D2, and D3, and mitogen-activated protein kinase 8 (also called Janus kinase) expression, suggesting an additional mechanism involving cell cycle regulation and cell survival. Third, BME treatment decreased the key proteins involved in steroidogenesis in adrenocortical cancer cells. BME treatment decreased the level of phosphorylation of cyclin-dependent kinase 7, which is required, at least in part, for steroidogenic factor 1 activation. Finally, we observed that BME treatment significantly reduced the level of insulin-like growth factor 1 receptor and its downstream signaling pathway as evidenced by lower levels of phosphorylated RAC-α serine/threonine-protein kinase. Taken together, these data illustrate the inhibitory effect of bitter melon on cell proliferation of adrenocortical cancer through modulation of diverse mechanisms....(more)
Brennan VC, et al. J Med Food 2012 Apr;15(4):325-34.
Related Products: Bitter Melon Extract
- 31. Mechanisms underlying decreased hepatic triacylglycerol and cholesterol by dietary bitter melon extract in the rat.
In these studies, we focused on finding the mechanism(s) underlying the bitter melon (Momordica charantia L.) methanol fraction (MF)-dependent reduction in the concentration of hepatic triacylglycerol (TAG) and cholesterol in the rat. Rats were fed diets containing low (5 %) fat for 2 weeks (experiment 1), or low (5 %) and high (15 %) fat for a longer period of 8 weeks (experiment 2). MF was supplemented at 1 % level in both experiments. After feeding, rats were sacrificed, and their livers were prepared as slices and hepatocytes, followed by incubation with [1(2)-¹C] acetate or [1-¹C] oleic acid (18:1 n-6). Under these conditions, we found that rats fed diets containing MF, as compared to those without MF, showed: (1) no adverse effects on food intake and growth, (2) a decreased hepatic TAG and total cholesterol, irrespective of the difference in dietary fat level or feeding period, and (3) a decreased incorporation of [1(2)-(¹C] acetate and [1-¹C] oleic acid into TAG of liver slices and hepatocytes. MF-supplemented rats also showed no altered incorporation of labeled acetate into cholesterol and cholesterol ester, an increased fecal excretion of neutral steroids, but not of acidic steroids, and an enhanced mRNA abundance of carnitine palmitoylacyltransferase I, which is the rate-limiting enzyme for fatty acid oxidation. These results suggest that dietary MF decreases hepatic TAG synthesis while enhancing fatty acid oxidation, thereby reducing the concentration of hepatic TAG. The liver cholesterol-lowering effect of MF, however, is probably mediated through an increased fecal excretion of neutral steroids, without an effect on cholesterogenesis....(more)
Senanayake GV, et al. Lipids 2012 May;47(5):495-503.
Related Products: Bitter Melon Extract
- 32. Bitter melon extract impairs prostate cancer cell-cycle progression and delays prostatic intraepithelial neoplasia in TRAMP model.
Prostate cancer remains the second leading cause of cancer deaths among American men. Earlier diagnosis increases survival rate in patients. However, treatments for advanced disease are limited to hormone ablation techniques and palliative care. Thus, new methods of treatment and prevention are necessary for inhibiting disease progression to a hormone refractory state. One of the approaches to control prostate cancer is prevention through diet, which inhibits one or more neoplastic events and reduces the cancer risk. For centuries, Ayurveda has recommended the use of bitter melon (Momordica charantia) as a functional food to prevent and treat human health related issues. In this study, we have initially used human prostate cancer cells, PC3 and LNCaP, as an in vitro model to assess the efficacy of bitter melon extract (BME) as an anticancer agent. We observed that prostate cancer cells treated with BME accumulate during the S phase of the cell cycle and modulate cyclin D1, cyclin E, and p21 expression. Treatment of prostate cancer cells with BME enhanced Bax expression and induced PARP cleavage. Oral gavage of BME, as a dietary compound, delayed the progression to high-grade prostatic intraepithelial neoplasia in TRAMP (transgenic adenocarcinoma of mouse prostate) mice (31%). Prostate tissue from BME-fed mice displayed approximately 51% reduction of proliferating cell nuclear antigen expression. Together, our results suggest for the first time that oral administration of BME inhibits prostate cancer progression in TRAMP mice by interfering cell-cycle progression and proliferation....(more)
Ru P, et al. Cancer Prev Res (Phila) 2011 Dec;4(12):2122-30.
Related Products: Bitter Melon Extract
- 33. Bitter melon: antagonist to cancer.
The incidence of cancer is increasing worldwide, in spite of substantial progress in the development of anti-cancer therapies. One approach to control cancer could be its prevention by diet, which inhibits one or more neoplastic events and reduces cancer risk. Dietary compounds offer great potential in the fight against cancer by inhibiting the carcinogenesis process through the regulation of cell homeostasis and cell-death machineries. For centuries, Ayurveda (Indian traditional medicine) has recommended the use of bitter melon (Momordica charantia) as a functional food to prevent and treat diabetes and associated complications. It is noteworthy to mention that bitter melon extract has no-to-low side effects in animals as well as in humans. The anti-tumor activity of bitter melon has recently begun to emerge. This review focuses on recent advancements in cancer chemopreventive and anti-cancer efficacy of bitter melon and its active constituents. Several groups of investigators have reported that treatment of bitter-melon-related products in a number of cancer cell lines induces cell cycle arrest and apoptosis without affecting normal cell growth. Therefore, the effect of bitter melon should be beneficial for health, and use of the non-modified dietary product is cost effective....(more)
Nerurkar P, et al. Pharm Res 2010 Jun;27(6):1049-53.
Related Products: Bitter Melon Extract
- 34. Vasorelaxant prenylated flavonoids from the roots of Sophora flavescens.
Bioassay-guided fractionation of the methanol extract from the root of Sophora flavescens led to the isolation of eight known prenylated flavonoids responsible for the vasorelaxation activity in porcine coronary arteries. Among them, kushenol N and 5-methylsophoraflavanone B strongly induced the relaxation of porcine coronary arteries with respective ED(50) values of 8.6 and 12.4 µM. This activity and the results of a high-performance liquid chromatographic analysis suggest that kushenol N and 5-methylsophoraflavanone B could be active markers in the S. flavescens extract for vasorelaxation activity....(more)
Kim CY, et al. Biosci Biotechnol Biochem 2013;77(2):395-7.
Related Products: Bitter Sophora Root Extract
- 35. [Study on anti-inflammatory efficacy accompanied by side effects of different components of Sophorae Tonkinensis Radix et Rhizoma].
OBJECTIVE:
To investigate the anti-inflammatory efficacy accompanied by side effects of water extract and alcohol extract of Sophorae Tonkinensis Radix et Rhizome (STRR), their molecular mechanism, and interpret the relationship of "toxicity-effect" of toxic medicine.
METHOD:
The ear swelling by croton oil and granuloma by agar test models were used, water extract and alcohol extract of STRR of different dosages were administrated ig to mice to observe the assident toxicity, at the same time the activities of ALT, AST and the content of SOD, MDA,PEG2, NO, NOS, Cr, BUN, GSH, TG and Gn in serum were tested.
RESULT:
Both water extract and alcohol extract of STRR have a strong inhibitory effect on ear swelling by croton oil and granuloma by agar. The activities of ALT, AST in serum were higher than that of normal group. SOD, MDA, PEG2, NO, NOS, GSH, TG and Gn had obvious changes.
CONCLUSION:
Both water extract and alcohol extract of STRR had an anti-inflammatory effect on acute and chronic inflammation. At the same time, side effects and liver toxicity. The anti-inflammatory effect of STRR in probable relation to the reduced inflammatory mediators release. Oxidative damnification might be one of the liver injury mechanism....(more)
Li X, et al. Zhongguo Zhong Yao Za Zhi 2012 Aug;37(15):2232-7. Chinese.
Related Products: Bitter Sophora Root Extract