Triterpenoid Acids Research Articles

ANALYSIS OF TRITERPENOID ACIDS IN CHINESE JUJUBE

Chinese jujube (Ziziphus jujuba Mill.) is a well-known tradotional Chinese medicinal plant and fruit tree. Triterpenoid acid is one of its main bioactive components. The HPLC analysis method for three kinds of triterpenoid acids of Chinese jujube, i.e., betulinic acid, oleanolic acid and ursolic acid, were firstly established. Furthermore, the contents of the triterpenoid acids in the fruits of 27 excellent cultivars, different organs of cultivar 'Jinsixiaozao' as well as in the fruits at different ripening stages of cultivar 'Dongzao' and 'Jinsixiaozao' were analyzed. Significant differences were found among cultivars and organs. The oleanolic acid content was highest in cultivar 'Tongbaidazao' and 'Xuanchengjianzao', and the highest contents of betulinic acid and ursolic acid were detected in 'Neihuangbianhesuan' and 'Yuanlingzao', respectively. Leaf contains much more triterpenoid acids than flower and fruit. The contents of betulinic acid, oleanolic acid and ursolic acid in leaf are 5.38 and 3.58 times, 21.71 and 15.03 times, and 41.68 and 15.33 times of those in flower and fruit. Betulinic acid was most dominant in fruit during ripening, followed by oleanolic acid and ursolic acid. All the three kinds of triterpenoid acids showed higher contents in white mature fruit than in full red fruit.

Oleanolic acid derivative Dex-OA has potent anti-tumor and anti-metastatic activity on osteosarcoma cells in vitro and in vivo

This study was undertaken to investigate the inhibitory effects of triterpenoid compound oleanolic acid and its synthetic derivatives on osteosarcoma cells in order to identify new therapeutic candidates for the treatment of this disease. We used the 3-(4,5-dimethyl-2 thiazolyl)-2,5-diphenyl tetrazolium bromide assay to assess the effect of oleanolic acid compounds on the proliferation of osteosarcoma cells. The effect of dextrose-oleanolic acid (the most potent oleanolic acid derivative) on apoptosis of osteosarcoma cells was evaluated using the Annexin-V method. The cell cycle of dextrose-oleanolic acid-treated cells was examined by flow cytometry, and the in vivo effects of dextrose-oleanolic acid were evaluated in a mouse osteosarcoma model. Oleanolic acid compounds had an overall inhibitory effect on the proliferation of osteosarcoma cells. Our in vitro data showed that the dextrose-oleanolic acid derivative brought about maximal inhibition of proliferation of osteosarcoma cells while inducing apoptosis. It could also inhibit the growth of osteosarcoma and decreased the rate of lung metastasis in vivo. Of the oleanolic acid derivatives, dextrose-oleanolic acid exhibited the most potent anti-osteosarcoma activity; it may represent a new frontier in the treatment of osteosarcoma.

Lignans and Triterpenoids from the Stems of Kadsura induta

Abstractmagnified imageTwo new lignans, kadsurindutins C and H (1 and 2, resp.), and a new 18(13→12)‐abeo‐lanostane triterpenoid acid, kadindutic acid (3), were isolated from the stems of Kadsura induta, together with known lignans and triterpenoids. Their structures and configurations were elucidated by spectroscopic methods, including 2D‐NMR techniques.

High-performance liquid chromatography—Two wavelength detection of triterpenoid acids from the fruits of Ziziphus jujuba containing various cultivars in different regions and classification using chemometric analysis

A simple and sensitive HPLC–DAD method has been developed for the first time to simultaneously determine 10 triterpenoid acids (ceanothic acid, alphitolic acid, zizyberanal acid, zizyberanalic acid, epiceanothic acid, ceanothenic acid, betulinic acid, oleanolic acid, ursonic acid and zizyberenalic acid) in the dried fruit of Ziziphus jujuba (called Dazao) which has been widely used as one of the traditional Chinese medicines (TCMs). This HPLC assay was performed on a reversed-phase C18 column (250mm×4.6mm, 5μm) with the column temperature at 35°C. The mobile phase was composed of A (acetonitrile) and B (0.05% aqueous phosphoric acid, v/v). The flow rate was 1.0ml/min and the detection wave length was set at 205nm for reference compounds 1–9 and 238nm for reference compound 10. All calibration curves showed good linear regression (r2>0.9999) within the test range. The established method showed good precision and accuracy with overall intra-day and inter-day variations of 0.43–1.72% and 0.53–2.45%, respectively, and overall recoveries of 94.98–104.09% for the 10 compounds analyzed. The validated method was successfully applied for the simultaneous determination of the 10 triterpenoid acids in 42 batches of Dazao which contained 36 cultivars from 22 cultivation regions, and were investigated and authenticated as Z. jujuba. Hierarchical clustering analysis (HCA) and principal components analysis (PCA) were performed to differentiate and classify the samples based on the contents of the 10 triterpenoid acids. The presented HPLC–DAD method conjugated with chemometrics approach was demonstrated to be very helpful in using Dazao resources, and was possibly useful in chemotaxonomic characterization.

Bioassay-Guided Isolation of Antiproliferative Compounds from Grape (Vitis vinifera) Stems

The fractionation, guided by cell-growth inhibition assay, of the EtOAc crude extract from grape stems of the Sicilian Vitis vinifera variety 'Nerello Mascalese' allowed identification often constituents, isolated either as pure compounds (1, 3-5, 7-10) or inseparable mixtures (2a-d and 6a-e). The pure constituents were: two triterpenoid acids, oleanolic (1) and betulinic acids (5); daucosterol (7); a stilbenoid, E-resveratrol (3) and its dimer E-epsilon-viniferin (4); the simple phenol gallic acid (8); and the flavanols catechin (9) and gallocatechin (10). Four 6'-O-acyldaucosterols (2a-d) and five 1,2-di-O-acyl-3-O-beta-D-galactopyranosyl glycerols (6a-e) were also identified as inseparable mixtures. All the isolated compounds were subjected to spectroscopic analysis and MTT bioassay on MCF-7 human breast cancer cells. The majority showed growth-inhibitory activity, 5 being the most active (GI50 = 0.57 microM). Compounds 3-5 were also tested on HT-29, U-87-MG and U-373-MG cell lines.

Asiatic Acid Induces Colon Cancer Cell Growth Inhibition and Apoptosis through Mitochondrial Death Cascade

Cancer is one of the leading causes of death in the world. The triterpenoid compound asiatic acid derived from the tropical medicinal plant Centella asiatica displays cytotoxic activity on fibroblast cells and several other kinds of cells. The present work studies asiatic acid-mediated growth inhibition of cancer cells and the underlying mechanism. Asiatic acid markedly inhibited cancer cell proliferation. Apoptosis of SW480 human colon cancer cells was induced by asiatic acid as shown by flow cytometry, DNA fragmentation and nuclear chromatin condensation experiments. Through increasing mitochondrial membrane permeability and cytochrome c release from mitochondria into cytosol, asiatic acid induced caspase-9 activity, which further activated caspase-3 and poly(ADP-ribose) polymerase cleavage resulting in irreversible apoptotic death in the tumor cells. Taken together, these results suggest that mitochondrial death apoptosis cascade plays very important roles in asiatic acid-induced cancer apoptosis.

Enhancing the divergent activities of betulinic acid via neoglycosylation.

Using neoglycosylation, the impact of differential glycosylation upon the divergent anticancer and anti-HIV properties of the triterpenoid betulinic acid (BA) was examined. Each member from a library of 37 differentially glycosylated BA variants was tested for anticancer and anti-HIV activities. Enhanced analogs for both desired activities were discovered with the corresponding antitumor or antiviral enhancements diverging, on the basis of the appended sugar, into two distinct compound subsets.

Inhibition of PGHS-1 and PGHS-2 by Triterpenoid Acids from Chaenomelis Fructus

The dichloromethane extract of the dried fruits of Chaenomeles speciosa (Sweet) Nakai (Rosaceae) showed strong inhibitory activity against both prostaglandin-H-synthase isoenzymes [IC50 (PGHS-1) = 5.1 μg/mL; IC50 (PGHS-2) = 2.3 μg/mL]. The lipophilic portion of the extract was mainly responsible for the inhibitory effect. Several triterpenoid acids were isolated and identified as contributing to this inhibitory activity (oleanolic, pomolic, 3β-O-acetylursolic and 3β-O-acetylpomolic acids). Comparison of their inhibitory potential with their selectivity to PGHS-2 showed that 3β-O-acetylursolic acid had the highest potency in the inhibition of PGHS-1 and PGHS-2 enzymes, whilst pomolic and 3β-O-acetylpomolic acid, with a hydroxyl group at position 19α, showed selectivity for PGHS-2. The inhibitory effect of the extract seems to be the result of the activity of the mixture of these different triterpenoid acids.

A New Cytotoxic Friedelane Acid – Pluricostatic Acid – and Other Compounds from the Leaves of Marila pluricostata

Bioassay-guided fractionation of the dichloromethane extract of the leaves of Marila pluricostata led to the isolation of 2α,3β-dihydroxy-D:A-friedoolean-28-oic acid (pluricostatic acid), a new friedelane triterpenoid, (1), ten known triterpenoids and three sterols. Their chemical structures were elucidated through spectroscopic analysis. The less polar fractions, on GC/MS analysis and comparison with a MS library, resulted in the identification of twenty four sesquiterpenoids. The new triterpenoid acid 1 showed cytotoxicity against the MCF-7, H-460, and SF-268 human cancer cell lines with GI50 values from 1.2 to 3.3 μg/mL.

Characterisation of two phenotypes of Centella asiatica in Southern Africa through the composition of four triterpenoids in callus, cell suspensions and leaves

Two morphologically distinct phenotypes of Centella asiatica (Type-1 and Type-2) in South Africa were compared in relation to the levels of triterpenoid saponins with the aim of assessing their potential for biotechnological manipulation of triterpenoid synthesis. The metabolites investigated included madecassoside and asiaticoside and their sapogenins madecassic—and asiatic acid; produced in cultured undifferentiated cells (cell suspensions and calli) and leaves. Weight determination in plant cell suspensions and the accumulation of secondary metabolites after 16 days for Type-1 and 20 days for Type-2 were investigated since these secondary metabolites accumulate during the period that follows the active growth phase. The four triterpenoids of interest were analysed and quantified by HPLC in crude ethanolic extracts. A difference in bioactive triterpenoids was exhibited that was tissue specific and varied between the two phenotypes. The triterpenoids from leaf tissue were more easily quantifiable in each phenotype than in the case of the undifferentiated cells (callus and cell suspensions), which had lower, but still quantifiable, levels of these targeted secondary metabolites. Leaves contained the highest triterpenoid levels (ranging from 1.8 to 5% dry weight for the triterpenoid acids and their glycosides, respectively), with the free acids occurring in a ratio of approximately 1:2.5 in relation to the glycoside content.

Structure-dependent inhibition of bladder and pancreatic cancer cell growth by 2-substituted glycyrrhetinic and ursolic acid derivatives

Derivatives of oleanolic acid, ursolic acid and glycyrrhetinic acid substituted with electron-withdrawing groups at the 2-position in the A-ring which also contains a 1-en-3-one structure are potent inhibitors of cancer cell growth. In this study, we have compared the effects of several 2-substituted analogs of triterpenoid acid methyl esters derived from ursolic and glycyrrhetinic acid on proliferation of KU7 and 253JB-V bladder and Panc-1 and Panc-28 pancreatic cancer cells. The results show that the 2-cyano and 2-trifluoromethyl derivatives were the most active compounds. The glycyrrhetinic acid derivatives with the rearranged C-ring containing the 9(11)-en-12-one structure were generally more active than the corresponding 12-en-11-one isomers. However, differences in growth inhibitory IC(50) values were highly variable and dependent on the 2-substituent (CN vs CF(3)) and cancer cell context.

In Vitro. Inhibition of Tumor Cell Growth by Miconia fallax.

In vitro. tumor growth inhibition by the ethanol extract of Miconia fallax. DC. (Melastomataceae) was evaluated in culture media containing HeLa cells at the following concentrations: 5, 15, 25, 35, and 45 μ g/mL. Bioassay-guided fractionation of this extract furnished a mixture of ursolic and oleanolic acids. Both the ethanol extract and the mixture of the triterpenoid acids produced dose-dependent tumor growth inhibition. This antitumor activity was more pronounced when a mixture of the triterpene acids at a concentration of 45 μ g/mL was added to a HeLa cell culture medium.

Methyl 2-Cyano-3,11-dioxo-18β-olean-1,12-dien-30-oate Is a Peroxisome Proliferator-Activated Receptor-γ Agonist That Induces Receptor-Independent Apoptosis in LNCaP Prostate Cancer Cells

Methyl 2-cyano-3,11-dioxo-18beta-olean-1,12-diene-30-oate (beta-CDODA-Me) is a synthetic analog of the naturally occurring triterpenoid glycyrrhetinic acid, which contains a 2-cyano substituent in the A-ring. beta-CDODA-Me was a potent inhibitor of LNCaP prostate cancer cell growth (IC(50) approximately 1 muM) and activated peroxisome proliferator-activated receptor gamma (PPARgamma), whereas analogs without the cyano group were weakly cytotoxic and did not activate PPARgamma. beta-CDODA-Me induced p21 and p27, down-regulated cyclin D1 protein expression, and induced two other proapoptotic proteins, namely nonsteroidal anti-inflammatory drug-activated gene-1 and activating transcription factor-3. However, induction of these responses by beta-CDODA-Me was PPARgamma-independent and due to activation of phosphatidylinositol-3-kinase, mitogen-activated protein kinase, and jun N-terminal kinase pathways by this compound. In contrast, beta-CDODA-Me also decreased androgen receptor (AR) and prostate-specific antigen (PSA) mRNA and protein levels through kinase-independent pathways. beta-CDODA-Me repressed AR mRNA transcription, whereas decreased PSA mRNA levels were dependent on protein synthesis and were reversed by cycloheximide. Thus, potent inhibition of LNCaP cell survival by beta-CDODA-Me is due to PPARgamma-independent activation of multiple pathways that selectively activate growth-inhibitory and proapoptotic responses.

2-Cyano-lup-1-en-3-oxo-20-oic acid, a cyano derivative of betulinic acid, activates peroxisome proliferator-activated receptor in colon and pancreatic cancer cells

Betulinic acid (BA) is a phytochemical triterpenoid acid from bark extracts and is cytotoxic to cancer cells and tumors. We modified the A-ring of BA to give a 2-cyano-1-en-3-one moiety and the effects of the 2-cyano-lup-1-en-3-oxo-20-oic acid (CN-BA), 2-cyano derivative of BA, and its methyl ester (CN-BA-Me) were investigated in colon and pancreatic cancer cells. Both CN-BA and CN-BA-Me were highly cytotoxic to Panc-28 pancreatic and SW480 colon cancer cells. CN-BA and CN-BA-Me also induced differentiation in 3T3-L1 adipocytes, which exhibited a characteristic fat droplet accumulation induced by peroxisome proliferator-activated receptor gamma (PPARgamma) agonists. Based on these results, we investigated the activities of CN-BA and CN-BA-Me as PPARgamma agonists using several receptor-mediated responses including activation of transfected PPARgamma-responsive constructs, induction of p21 in Panc-28 cells and induction of caveolin-1 and Krüppel-like factor 4 in colon cancer cells. The results clearly demonstrated that both CN-BA and CN-BA-Me activated PPARgamma-dependent responses in colon (caveolin-1) and pancreatic (p21) cancer cells, whereas induction of KLF4 by these compounds in colon cancer cells was PPARgamma independent and also dependent on cell context. The PPARgamma agonist activities of CN-BA and CN-BA-Me were structure-, response/gene- and cell context-dependent suggesting that these compounds are a novel class of selective PPARgamma modulators with potential for clinical treatment of colon and pancreatic cancer.

Synthetic triterpenoids inhibit growth and induce apoptosis in human glioblastoma and neuroblastoma cells through inhibition of prosurvival Akt, NF-κB and Notch1 signaling

Glioblastomas are high-risk primary brain tumors that are generally unresponsive or only weakly responsive to the currently available antineoplastic agents. Thus novel therapeutic strategies and agents are urgently needed to treat these incurable cancers. Oleanolic acid and ursolic acid are naturally occurring triterpenoids that have been used in traditional Asian medicine as anti-inflammatory and anti-cancer agents. Recently, synthetic oleanolic acid triterpenoid 2-cyano-3,12-dioxooleana-1,9(11)-dien-28-oic acid (CDDO) and its C-28 methyl ester (CDDO-Me) and C-28 imidazole (CDDO-Im) derivatives have been shown to exhibit potent antitumor activity against diverse types of tumor cell lines, including leukemia, multiple myeloma, osteosarcoma, breast, lung, and pancreatic cancer cell lines; however, the anticancer activity of these agents for brain tumors has not been reported. In the present study, we investigated the apoptosis-inducing activity of CDDOs in glioblastoma (U87MG, U251MG) and neuroblastoma (SK-N-MC) cell lines. Cell growth/viability (MTS) and cytotoxicity (LDH release) assays demonstrated that glioblastoma cell lines are least sensitive to CDDO, but are highly sensitive to CDDO-Me and CDDO-Im at concentrations of 2.5-10 muM. CDDO-Im and CDDO-Me were equipotenent in their growth inhibitory activity. The primary mode of tumor cell destruction was apoptosis as demonstrated by significant increase in the number of hypo-diploid (sub-G0) cells and annexin V-FITC binding. Induction of apoptosis was associated with the activation of procaspases-3, -8, and -9, mitochondrial depolarization and the release of cytochrome c from mitochondria. Furthermore, CDDO-Me inhibited the levels of anti-apoptotic and prosurvival p-Akt, NF-kappaB (p65) and Notch1 signaling molecules. These studies provide rationale for clinical evaluation of these novel agents for the management of lethal brain neoplasms.

Extraction and TLC Desitometric Determination of Triterpenoid Acids (Arjungenin, Arjunolic Acid) from Terminalia arjuna Stem Bark Without Interference of Tannins

The stem bark of Terminalia arjuna Linn. (fam: Combretaceae), commonly known as Arjuna in Indian systems of medicine, is a reputed drug used for various cardiac disorders. T. arjuna stem bark is reported to contain different groups of chemical constituents including phenolics, tannins, saponins and triterpenoid acids. From our earlier experience with tannin containing herbal drugs, we are aware that tannins interfere in the extraction of certain compounds and hence in their quantification. In the present experiment, we report a sample preparation method to overcome the interference of the tannins by adsorbing them with carboxy methyl cellulose, which facilitates the efficient extraction of the triterpenoid acids. Further we established TLC densitometric methods for the quantification of two of the triterpenoid acids of T. arjuna stem bark viz., arjungenin and arjunolic acid using HPTLC. The methods were validated in terms of accuracy, precision and repeatability. The calibration curve showed linearity in the range of 160–480 ng spot−1 and 160–560 ng spot−1 for arjungenin and arjunolic acid respectively. The percentage of arjungenin and arjunolic acid were found to be 0.324% w/w and 0.524% w/w in the stem bark by this modified method of extraction, which was many times higher than when compared to that using the extraction method without CMC (0.018% and 0.049% respectively). The study reiterates the importance of sample preparation in the quantification of non polar phytochemicals from herbal raw materials, such that the compounds of interest are extracted efficiently, overcoming the interference of other compounds like tannins in the matrix of plant material.

Triterpenoids and ellagic acid derivatives from in vitro cultures of Camptotheca acuminata Decaisne

The metabolic profile of secondary products in calli and cell suspension cultures of Camptotheca acuminata Decaisne was investigated and compared to that of the leaves and roots taken from the plant. Neither in vitro system produced the anticancer quinoline alkaloid camptothecin (CPT); they accumulated discrete quantities of polyhydroxylated triterpenoids, different from those found in the plant organs, and ellagic acid derivatives. Nine ellagic acid derivatives ( 1a– 1d and 2a– 2e) and eight triterpenoid acids ( 3a– 3e and 4a– 4c) were isolated and characterised. All the identified triterpenes were related to ursane- or oleanane-type skeletons and their concentrations rose to 4.5% in suspended cells.

Study of the oleanolic and ursolic acid contents of some species of the Lamiaceae

The oleanolic and ursolic acid contents from 88 taxa of Lamiaceae (19 genera, 66 species, 8 subspecies, 9 varieties and 5 hybrids) were investigated using gas chromatography. Both triterpenoid acids were present in all of the investigated taxa, but the plants belonging in the subfamily Nepetoideae produced significantly higher amounts than those in the subfamily Lamioideae. The oleanolic acid content ranged from traces to 1.840% dry weight, and that of ursolic acid from traces to 4.019% dry weight.

CHEMICAL CONSTITUENTS FROM THE LEAVES OF SANDORICUM KOETJAPE

CHEMICAL CONSTITUENTS FROM THE LEAVES OF SANDORICUM KOETJAPE

Triterpenoid and caffeic acid derivatives in the leaves of ragweed, Ambrosia artemisiifolia L. (Asterales: Asteraceae), as feeding stimulants of Ophraella communa LeSage (Coleoptera: Chrysomelidae)

The leaf beetle Ophraella communa infests almost exclusively Ambrosia artemisiifolia in the fields of Japan, even though it normally feeds on several Asteraceous plants. A filter paper bioassay showed that the feeding of O. communa is strongly stimulated by methanolic extracts of A. artemisiifolia. The feeding stimulants for O. communa have been isolated from methanolic extracts of A. artemisiifolia. α-Amyrin acetate, β-amyrin acetate, 5-caffeoylquinic acid (chlorogenic acid) and 3,5-dicaffeoylquinic acid from A. artemisiifolia have been identified as feeding stimulants for O. communa. Triterpenoid derivatives (α-amyrin acetate or β-amyrin acetate) and caffeic acid derivatives (3, 5-dicaffoylquinic acid or 5-caffeoylquinic acid) showed feeding stimulant activity when mixed together.