23-Hydroxyursolic acid

23-Hydroxyursolic Acid Isolated from the Stem Bark of Cussonia bancoensis Induces Apoptosis through Fas/Caspase-8-Dependent Pathway in HL-60 Human Promyelocytic Leukemia Cells.[Pubmed:30551620]

Molecules. 2018 Dec 13;23(12). pii: molecules23123306.

The natural product 23-Hydroxyursolic acid (23-HUA) is a derivative of ursolic acid, which is known to induce cancer cell apoptosis. However, apoptotic effects and mechanisms of 23-HUA have not been well characterized yet. Herein, we investigated the molecular mechanisms of 23-HUA-induced apoptosis in HL-60 human promyelocytic leukemia cells. 23-HUA-treated HL-60 cells showed apoptotic features including internucleosomal DNA condensation and fragmentation as well as externalization of phosphatidylserine residues. 23-HUA induced a series of mitochondrial events including disruption of mitochondrial membrane potential (DeltaPsim), cytochrome c and Smac/DIABLO release and loss of balance between pro-apoptotic and anti-apoptotic Bcl-2 proteins in HL-60 cells. In addition, 23-HUA activated caspase-8, caspase-9 and caspase-3. Pretreatment with a broad caspase inhibitor (z-VAD-fmk), a caspase-3 inhibitor (z-DEVD-fmk), and a caspase-8 inhibitor (z-IETD-fmk) significantly attenuated 23-HUA-induced DNA fragmentation. After 23-HUA-induced apoptosis, proteins expression levels of FasL, Fas and FADD constituting the death-inducing signaling complex (DISC) were upregulated in HL-60 cells. Moreover, transfection with Fas or FADD siRNA significantly blocked 23-HUA-induced DNA fragmentation and caspases activation. Taken together, these findings indicate that 23-HUA induces apoptosis in HL-60 human promyelocytic leukemia cells through formation of DISC and caspase-8 activation leading to loss of DeltaPsim and caspase-3 activation.

Antimicrobial pentacyclic triterpenes and glycosides from the stem bark of Cussonia bancoensis.[Pubmed:30289000]

Nat Prod Res. 2020 Mar;34(6):859-862.

The stem bark of Cussonia bancoensis is used traditionally for the treatment of different types of infection and pain. A bioassay guided fractionation of the methanol stem bark extract led to the isolation of five pentacyclic triterpenes and glycosides identified based on spectroscopic data as 23-Hydroxyursolic acid (CB1), hederagenin (CB2), 3-O-alpha-L-arabinopyranosyl-echinocystic acid (CB3), 3-O-alpha-L-arabinopyranosyl- oleanolic acid (CB4) and 3-O-alpha-L-arabinopyranosyl-ursolic acid (CB5). CB2 - CB5 are being reported for the first time from this species. The compounds were evaluated for antimicrobial activity against ten microorganisms using the HT-SPOTi method. CB3 demonstrated remarkable antimicrobial activity against S. aureus, S. pyogens, E. faecalis, S. typhi and C. albicans at MICs between 3.12 and 12.5 microg/mL. Among the studied compounds, it was observed that hydroxylation of position C-16 of the oleanane skeleton may enhance antimicrobial activity. This study gives insight into the anti-infective constituents of the stem bark of C. bancoensis and justifies its use in ethnomedicine.[Formula: see text].

A Novel Multifunctional C-23 Oxidase, CYP714E19, is Involved in Asiaticoside Biosynthesis.[Pubmed:29579306]

Plant Cell Physiol. 2018 Jun 1;59(6):1200-1213.

Centella asiatica is widely used as a medicinal plant due to accumulation of the ursane-type triterpene saponins asiaticoside and madecassoside. The molecular structure of both compounds suggests that they are biosynthesized from alpha-amyrin via three hydroxylations, and the respective Cyt P450-dependent monooxygenases (P450 enzymes) oxidizing the C-28 and C-2alpha positions have been reported. However, a third enzyme hydroxylating C-23 remained elusive. We previously identified 40,064 unique sequences in the transcriptome of C. asiatica elicited by methyl jasmonate, and among them we have now found 149 unigenes encoding putative P450 enzymes. In this set, 23 full-length cDNAs were recognized, 13 of which belonged to P450 subfamilies previously implicated in secondary metabolism. Four of these genes were highly expressed in response to jasmonate treatment, especially in leaves, in accordance with the accumulation patterns of asiaticoside. The functions of these candidate genes were tested using heterologous expression in yeast cells. Gas chromatography-mass spectrometry (GC-MS) analysis revealed that yeast expressing only the oxidosqualene synthase CaDDS produced the asiaticoside precursor alpha-amyrin (along with its isomer beta-amyrin), while yeast co-expressing CaDDS and CYP716A83 also contained ursolic acid along with oleanolic acid. This P450 enzyme thus acts as a multifunctional triterpenoid C-28 oxidase converting amyrins into corresponding triterpenoid acids. Finally, yeast strains co-expressing CaDDS, CYP716A83 and CYP714E19 produced hederagenin and 23-Hydroxyursolic acid, showing that CYP714E19 is a multifunctional triterpenoid oxidase catalyzing the C-23 hydroxylation of oleanolic acid and ursolic acid. Overall, our results demonstrate that CaDDS, CYP716A83 and CYP714E19 are C. asiatica enzymes catalyzing consecutive steps in asiaticoside biosynthesis.

[Triterpenoids from Leaves of Ilex latifolia].[Pubmed:26983238]

Zhong Yao Cai. 2015 Aug;38(8):1653-5.

OBJECTIVE: To investigate the chemical constituents from the leaves of Ilex latifolia. METHODS: The constituents were iso- lated and purified by various chromatographic techniques. Their structures were identified by UV, IR, MS and NMR spectroscopic analy- sis. RESULTS: Ten compounds were isolated from the leaves of flex latifolia. Their structures were respectively identified as 3beta,13beta-Di- hydroxy-urs-11-en-28-oic acid-13-lactone (1), beta-sitosterol (2) 3-hydroxy-11-oxours-12-ene (3), (20S,24S)-epoxydammarane-3beta,25- diol (4), 25-deuteriostigmasterol (5), alpha-amyrin (6), foliasalacin A4 (7), 23-Hydroxyursolic acid (8), p-coumaric acid (9) and beta-dau- costerol (10). CONCLUSION: Compounds 1,3-5,7 and 9 are isolated from this plant for the first time.

Arboreasides A-E, triterpene saponins from the bark of Cussonia arborea.[Pubmed:19456116]

J Nat Prod. 2009 Jun;72(6):1081-6.

Five new triterpene saponins, arboreasides A-E (1-5), and two known saponins, ciwujianoside C(3) and 23-Hydroxyursolic acid 28-O-alpha-L-rhamnopyranosyl-(1-->4)-beta-D-glucopyranosyl-(1-->6)-beta-D-glucopy ranosyl ester, were isolated from the bark of Cussonia arborea. The structures were established using extensive 1D and 2D NMR spectroscopic analyses and mass spectrometry.

23-hydroxyursolic acid causes cell growth-inhibition by inducing caspase-dependent apoptosis in human cervical squamous carcinoma HeLa cells.[Pubmed:19414337]

Anticancer Res. 2009 Apr;29(4):995-1000.

BACKGROUND: There are few reports on the biological activities of 23-Hydroxyursolic acid (23-HUA). The mechanism of growth-inhibition induced by 23-HUA, isolated from Cussonia bancoensis, in human cervical squamous carcinoma HeLa cells is hereby investigated. MATERIALS AND METHODS: The growth-inhibitory activity was measured by MTS assay. Caspases activation and expression of apoptosis-related proteins were detected by Western blotting. Apoptotic cells were observed by morphological analysis with Hoechst 33342. RESULTS: 23-HUA inhibited the growth of HeLa cells in a concentration dependent manner. Proteolytically generated fragments of caspase-3, -8 and -9 were observed in HeLa cells treated with 60 microM 23-HUA. The expression of Bcl-X(L), an anti-apoptotic protein, was markedly decreased by 60 microM 23-HUA. Morphological analysis showed that apoptotic changes occurred after treatment with 60 microM 23-HUA, and the changes were inhibited by a pan-caspase inhibitor, Z-VAD-FMK. CONCLUSION: These results indicate that 23-HUA causes potent growth-inhibition by the induction of apoptosis via activation of caspases in HeLa cells.

Triterpene acids isolated from Lagerstroemia speciosa leaves as alpha-glucosidase inhibitors.[Pubmed:19107840]

Phytother Res. 2009 May;23(5):614-8.

The potential antidiabetic activity of ethyl acetate extract of the leaves of Lagerstroemia speciosa (LSL) was investigated by alpha-amylase and alpha-glucosidase inhibition assay. Six pentacyclic triterpenes (oleanolic acid, arjunolic acid, asiatic acid, maslinic acid, corosolic acid and 23-Hydroxyursolic acid) were isolated from LSL. Their structures were determined by spectroscopic analysis and their alpha-glycosidase and alpha-amylase inhibitory activities were investigated. They exhibited no or weak inhibitory activity against alpha-amylase and middle alpha-glucosidase inhibitory activities. Corosolic acid, which shows best bioactivity against alpha-glucosidase (IC(50) = 3.53 microg/mL), contributes most to the alpha-glucosidase inhibitory activity of EtOAc extract. The kinetics of inhibition of corosolic acid was also discussed. Results from this study might provide the scientific evidence for LSL for the treatment of diabetes in traditional medicine.

A new pancreatic lipase inhibitor isolated from the roots of Actinidia arguta.[Pubmed:18481026]

Arch Pharm Res. 2008 May;31(5):666-70.

A new coumaroyl triterpene, 3-O-trans-p-coumaroyl actinidic acid (1), as well as five known triterpenes, ursolic acid (2), 23-Hydroxyursolic acid (3), corosolic acid (4), asiatic acid (5) and betulinic acid (6) were isolated from an EtOAc-soluble extract of the roots of Actinidia arguta. The structure of compound 1 was elucidated from interpretation of the spectroscopic data, particularly by extensive 1D and 2D NMR studies. All the isolates (1-6) were evaluated in vitro for their inhibitory activities on pancreatic lipase (PL). Of the isolates, the new compound 1 possessed the highest inhibitory activity on PL, with an IC(50) of 14.95 microM, followed by ursolic acid (2, IC(50) = 15.83 microM). The other four triterpenes (3-6) also showed significant PL inhibitory activity, with IC(50) values ranging from 20.42 to 76.45 microM.

Heme oxygenase-1 inducing constituent of Prunella vulgaris in HepG2 cells.[Pubmed:18310925]

Biol Pharm Bull. 2008 Mar;31(3):531-3.

Inducible heme oxygenase (HO)-1 is known to play a major role in the pathogenesis of several diseases, and it protects cells against oxidant-mediated injury. The bioassay-guided fractionation of the EtOH extract of the flowered fruit-spike of Prunella vulgaris L. (Labiatae) yielded two ursane-type triterpenes, 3beta,23-dihydroxyurs-12-en-28-oic acid (23-Hydroxyursolic acid) (1) and 3beta-hydroxyurs-12-en-28-oic acid (ursolic acid) (2). Western blotting demonstrated that treatment with compound 1 increased the expression of HO-1 in a dose-dependent manner in human liver-derived HepG2 cells. Investigation of structure-related HO-1 inducing activity suggested that the hydroxyl group at the C-23 position in the ursane skeleton is important for this activity.

[Chemical constituents from root of Actinidia chinensis].[Pubmed:18027663]

Zhongguo Zhong Yao Za Zhi. 2007 Aug;32(16):1663-5.

OBJECTIVE: To study the chemical constituents from the root of Actinidia chinensis. METHOD: The roots of A. chinensis were extracted with 95% EtOH. The EtOH extract was suspended in H2O and extracted with petroleum ether, EtOAc and BuOH successively. The compounds were isolated with column chromatography from the EtOAc fraction, and elucidated on the basis of spectral analysis(MS, IR, 1H-NMR, 13C-NMR). RESULT: Seven compounds were isolated from the root of A. chinensis, and the structures were identified as 2alpha-hydroxyoleanolic acid (1), 2alpha-hydroxyursolic acid (2), euscaphic acid (3), 23-Hydroxyursolic acid (4), 3beta-O-acetylursolic acid (5), ergosta4, 6, 8, (14), 22-tetraen-3-one (6), beta-steriol (7). CONCLUSION: All the compounds were obtained from the root of A. chinensis for the first time.

Triterpenoids from the flower of Campsis grandiflora K. Schum. as human acyl-CoA: cholesterol acyltransferase inhibitors.[Pubmed:15974441]

Arch Pharm Res. 2005 May;28(5):550-6.

The flower of Campsis grandiflora K. Schum. was extracted with 80% aqueous MeOH, and the concentrated extract was partitioned with EtOAc, n-BuOH and H2O. From the EtOAc fraction, seven triterpenoids were isolated through the repeated silica gel, ODS column chromatographies and preparative HPLC. From the result of physico-chemical data including NMR, MS and IR, the chemical structures of the compounds were determined as 3beta-hydroxyolean-12-en-28-oic acid (oleanolic acid, 1), 3beta-hydroxyurs-12-en-28-oic acid (ursolic acid, 2), 3beta-hydroxyurs-12-en-28-al (ursolic aldehyde, 3), 2alpha,3beta-dihydroxyolean-12-en-28-oic acid (maslinic acid, 4), 2alpha,3beta-dihydroxyurs-12-en-28-oic acid (corosolic acid, 5), 3beta,23-dihydroxyurs-12-en-28-oic acid (23-Hydroxyursolic acid, 6) and 2alpha,3beta,23-trihydroxyolean-12-en-28-oic acid (arjunolic acid, 7). These teriterpenoids were isolated for the first time from this plant. Also, compounds 4, 5, 6, and 7 revealed relatively high hACAT-1 inhibitory activity with the value of 46.2+/-1.1, 46.7+/-0.9, 41.5+/-1.3 and 60.8+/-1.1% at the concentration of 100 microg/mL, respectively.

In vitro anti-inflammatory activity of 23-hydroxyursolic acid isolated from Cussonia bancoensis in murine macrophage RAW 264.7 cells.[Pubmed:15386188]

Planta Med. 2004 Sep;70(9):803-7.

In the present study, the effects of various triterpenoids isolated from the stem bark of Cussonia bancoensis, namely, ursolic acid ( 1), 23-Hydroxyursolic acid ( 2), 3-O-alpha- L-arabinopyranosyl-23-Hydroxyursolic acid (3), and 3-O-beta-D-glucopyranosyl-23-Hydroxyursolic acid ( 4) were evaluated on lipopolysaccharide (LPS)-induced nitric oxide (NO) and prostaglandin E (2) (PGE (2)) release by the macrophage cell line RAW 264.7. Of the tested triterpenoids, 23-Hydroxyursolic acid ( 2) was found to be the most potent inhibitor of NO production, and also significantly reduced PGE (2) release. Consistent with these observations, the protein and mRNA expression levels of inducible NO synthase (iNOS) and cyclooxygenase (COX)-2 enzymes were inhibited by 23-Hydroxyursolic acid ( 2) in a concentration-dependent manner. Furthermore, 23-Hydroxyursolic acid ( 2) inhibited the LPS-induced DNA binding activity of nuclear factor- kappaB (NF- kappaB), which was associated with a decrease of p65 protein levels in the nucleus. These results suggest that the 23-Hydroxyursolic acid-mediated inhibition of iNOS and COX-2 expression, via blocking NF- kappaB activation, may mechanistically responsible for the anti-inflammatory effects of Cussonia bancoensis stem bark in vitro.

Antitubercular constituents of Valeriana laxiflora.[Pubmed:15229801]

Planta Med. 2004 Jun;70(6):509-14.

Antitubercular bioassay-guided fractionation of the n-hexane- and CH (2)Cl (2)-soluble extracts of the above-ground biomass and roots of Valeriana laxiflora led to the isolation of a new iridolactone, (4R,5R,7S,8S,9S)-7-hydroxy-8-hydroxymethyl-4-methyl-perhydrocyclopenta[ c]pyran-1-one ( 1), and a new lignan, (+)-1-hydroxy-2,6-bis- epi-pinoresinol ( 2), along with eleven known compounds, betulin ( 3), betulinic acid ( 4), 5,7-dihydroxy-3,6,4'-trimethoxyflavone ( 5), 23-Hydroxyursolic acid ( 6), oleanolic acid ( 7), tricin ( 8), ursolic acid ( 9), ferulic acid, (+)-1-hydroxypinoresinol, prinsepiol, and 5,7,3'-trihydroxy-4'-methoxyflavone. The structures of compounds 1 and 2 were elucidated on the basis of spectroscopic evidence. The absolute stereochemistry of 1 was determined by chemical transformations and Mosher ester procedures. In a microplate alamar blue assay against Mycobacterium tuberculosis, compounds 2 - 9 exhibited minimum inhibitory concentrations (MIC) of 15.5 - 127 microg/mL, while the other isolates were regarded as inactive (MIC > 128 microg/mL). In addition, all the isolates were tested for cytotoxicity against African green monkey Vero cells in order to evaluate their selectivity potential.