Ursolic Acids Research Articles

Homo‐ and Cross Coupling of C‐2 Propargyl Substituted Triterpenoic Acids: Synthesis of Novel Symmetrical and Unsymmetrical Triterpene 1,3‐Diynes

AbstractA synthesis of the first representatives of symmetrical and unsymmetrical triterpene 1,3‐diynes in which one or two triterpene cores are linked by a diyne spacer at the C‐2 position of ring A is described. The new triterpenoids are prepared by Cu‐ and Pd‐catalyzed acetylenic homo‐ and cross coupling of accessible C‐2 propynyl derivatives of betulinic, ursolic, and oleanolic acids.

Both Oleanolic Acid and a Mixture of Oleanolic and Ursolic Acids Mimic the Effects of Fructus ligustri lucidi on Bone Properties and Circulating 1,25-Dihydroxycholecalciferol in Ovariectomized Rats

Oleanolic acid (OA) and ursolic acid (UA) are major chemical constituents found in Fructus ligustri lucidi (FLL), a Chinese herb previously shown to increase bone properties and modulate calcium-vitamin D metabolism in rats. OA and UA have been reported to exert osteoprotective effects in vitro. The present study was designed to determine whether OA or OA+UA mimicked the effects of FLL on bone and calcium homeostasis using ovariectomized rats. Three-month-old ovariectomized Sprague-Dawley rats were stabilized for 2 mo and randomly assigned to 4 groups offered the same amount (15-17 g/d) of a control diet or experimental diets containing FLL (18.8 g/kg), OA (0.67 g/kg), or OA (0.67 g/kg)+UA (0.22 g/kg) for 6 wk. Serum was obtained for measurement of 1,25-dihydroxycholecalciferol [1,25(OH)2D3] and bones were collected for micro-CT analysis. Calcium balance was measured at weeks 1 and 6. A calcium kinetic study using 45Ca was conducted at week 6 and modeled using WinSAAM software. Compared with the control group, rats fed the FLL-, OA-, and OA+UA-enriched diets had better bone properties and 51%, 31%, and 27% higher serum 1,25(OH)2D3 concentrations at week 6, respectively. These variables did not differ between the treatments. Calcium balance was not affected by diet at either week 1 or week 6. Kinetic modeling predicted that FLL and OA+UA diet-fed rats had 9% and 15% less endogenous excretion of calcium, respectively, compared with the control group. All 3 treatments resulted in a higher calcium mass of compartment 3 because of changes in transfer rate between compartments 2 and 3, and were positively associated with the serum 1,25(OH)2D3 concentration (R2=0.28; P<0.01). Similar to FLL, OA and OA+UA increase bone properties, serum 1,25(OH)2D3 concentration, and calcium use in ovariectomized rats, suggesting their potential role in management of osteoporosis.

Ursolic and Oleanolic Acids as Potential Anticancer Agents Acting in the Gastrointestinal Tract

Background:Cancer is one of the main causes of death worldwide. Contemporary therapies, including chemo- and radiotherapy, are burdened with severe side effects. Thus, there exists an urgent need to develop therapies that would be less devastating to the patient’s body. Such novel approaches can be based on the anti-tumorigenic activity of particular compounds or may involve sensitizing cells to chemotherapy and radiotherapy or reducing the side-effects of regular treatment.Objective:Natural-derived compounds are becoming more and more popular in cancer research. Examples of such substances are Ursolic Acid (UA) and Oleanolic Acid (OA), plant-derived pentacyclic triterpenoids which possess numerous beneficial properties, including anti-tumorigenic activity.Results:In recent years, ursolic and oleanolic acids have been demonstrated to exert a range of anticancer effects on various types of tumors. These compounds inhibit the viability and proliferation of cancer cells, prevent their migration and metastasis and induce their apoptosis. Both in vitro and in vivo studies indicate that UA and OA are promising anti-cancer agents that can prevent carcinogenesis at each step. Furthermore, cancers at all stages are susceptible to the activity of these compounds. &lt;/P&gt;&lt;P&gt; Neoplasms that are formed in the gastrointestinal tract, i.e. gastric, colorectal, pancreatic, and liver cancers, are among the most common and most lethal malignancies. Their localization in the digestive system, however, facilitates the action of orally-administered (potential) anti-cancer agents, making chemopreventive drugs more accessible.In this paper, the anti-tumorigenic effect of ursolic and oleanolic acids on gastric, colon, pancreatic, and liver cancers, as well as the mechanisms underlying this process, are presented.

Nanoemulsion Strategy for Ursolic and Oleanic Acids Isolates from Plumeria Obtusa Improves Antioxidant and Cytotoxic Activity in Melanoma Cells.

Triterpenoids are an important class of natural bioactive products present in many medicinal plants. The aim of present study is to investigate the antioxidant and anticarcinogenic potential of Oleanolic Acid (OA) and Ursolic Acid (UA) on B16 murine melanoma cell line isolated from Plumeria obtusa, free and loaded in a nanoemulsion (NEm) system. The nanoemulsion was characterized by dynamic light scattering, transmission electron microscopy. The viscosity was also evaluated. The antioxidant activity was determined by the reduction of 2,2-diphenyl-2- picrylhydrazyl (DPPH) free radical. In vitro proliferation studies were determined using the sulforhodamine-B method. OA/UA natural mixture exhibited high percentage of inhibition of DPPH (86.06% and 85.12%, with and without irradiation). Percentages of inhibition higher than 85% in samples with and without ultraviolet irradiation were recorded when loaded in the NEm system. The natural mixture incorporated into the NEm showed cytotoxic activity from 2.9 µM, whereas the free compounds from 17.4 µM. We conclude that these pentacyclic triterpenes loaded in a NEm system could be considered as a new potential tool for further investigation as anticancer agents.

Chemical composition of lipophilic extractives from six Eucalyptus barks

Barks show a large diversity in chemical composition and are generally characterized by a substantial content of extractives. Today, these components are viewed with a renewed interest as valuable source of chemicals and bioactive compounds. The barks of six eucalypt species, Eucalyptus botryoides, E. camaldulensis, E. maculata, E. rudis, E. sideroxylon and E. viminalis, were studied in relation to their lipophilic extractives that represented between 0.1% (E. rudis) and 1.8% (E. viminalis) of the oven-dry bark. It is the first time that a detailed chemical composition is given for the lipophilic extracts from the barks of these eucalypt species. They showed a high content in triterpenes (between 3623.1 and 13,616.5 mg/kg of bark), especially triterpenic acids such as betulinic, ursolic, oleanolic and arjunolic acids. Sterols were also identified in considerable amounts (174.0–1736.7 mg/kg of bark). β-Sitosterol was the major sterol, with 1350.1 mg/kg of bark in E. maculata. Fatty acids C16–C28 existed in smaller amounts. A saponification treatment (MeOH/KOH) showed that fatty acids, sterols and triterpenes existed in both esterified and non-esterified forms. The presence of triterpenes represents a potential added value for the valorization of bark residues by production of bioactive or high-value compounds.

Free Solvent Amidation of Ursolic and Oleanolic Acids of Fagraea Fragrans Fruits: Their P-388 Antitumor Activity

This is an our laboratory effort in developing major compounds, ursolic acid (UA) and its isomer oleanolic acid (OA) of Fagraea Fragrans Roxb fruits, namely buah tembesu, Loganiaceae by free solvent amidation of the acids with n-buthylamine and phenylamine. The reactions have been conducted by using thionyl chloride as a chlorination reagent to make their acid chlorides respectively. In this reaction, the secondary hydroxyl group at C-3 position was preferably converted to be olefinic while the carboxyl group at C-28 position was normally amidated to give N-buthyl-urs-2,12-dien-28-amide [N-buthyl-olean-2,12-dien-28-amide] with yield 45.5% % and N-phenyl-urs-2,12-dien-28-amide [N-phenyl-olean-2,12-dien-28-amide] with yield 39%. Cytotoxic activity of amidation products toward P-388 murine leukemia cells was decreasing about half fold compared to starting material ursolic acid [oleanolic acid]. The IC50 values of N-butyl-urs-2,12-dien-28-amide [N-butyl-olean-2,12-dien-28-amide] and N-phenyl-urs-2,12-dien-28-amide [N-phenyl-olean-2,12-dien-28-amide] were 81, 4 μg/mL and 83.6 μg/mL respectively, whereas the IC50 value of ursolic acid [oleanolic acid] was 53.4 µg/mL.

Fragmentation of the main triterpene acids of apple by LC-APCI-MSn.

In this paper, we investigated the fragmentation of the main triterpene acids of apple using an liquid chromatography atmospheric pressure chemical ionization mass spectrometry (LC-APCI-MSn ) approach and high-resolution mass spectrometry (HR-MS) (Q-TOF). Triterpenes were isolated using semipreparative high-performance liquid chromatography, and chemical structures were elucidated by HR-MS and nuclear magnetic resonance spectroscopy. Finally, compounds were used to study MSn behavior in ion trap. Isolated triterpenes present similar structures, bearing carboxyl group linked to C-17 and different substitutions. We observed significant changes in MS2 spectra, which were useful for further compound identification. The observed fragments allowed the discrimination of different derivatives, namely, pomaceic, annurcoic, euscaphic, pomolic, corosolic, maslinic, betulinic, oleanolic, and ursolic acids. The proposed method allows a rapid identification of triterpene acids, and it could be useful for the analysis of these compounds in apple fruits and in other natural sources.

Deodorization by Solar Steam Distillation of Rosemary Leaves Prior to Solvent Extraction of Rosmarinic, Carnosic, and Ursolic Acids

This study aims total valorization of rosemary for food applications, moving toward developing an original biorefinery concept using solar steam distillation (SSD) to extract essential oil while preserving active compounds in the raw material, mainly antioxidants, for a further extraction. The deodorization process is important to avoid the extraction of antioxidants with strong aromas which limit their utilization for food preservation. The SSD process was compared with conventional steam distillation (CSD) for essential oil extraction as well as deodorization of dried rosemary leaves. The treated leaves were recovered afterward for the extraction of antioxidants by solvent extraction to recover rosmarinic, carnosic, and ursolic acids. With standard protocol extraction and analysis, the antioxidants seem to be mostly preserved by SSD rather than by CSD. The Folin–Ciocalteu reagent was used to determine the polyphenol content, whereas diphenylpicrylhydrazyl (DPPH) was used to analyze the antiradical prope...

New Targets for Growth Inhibition of Mycobacterium tuberculosis: Why Do Natural Terpenoids Exhibit Antitubercular Activity?

The article draws the attention of chemists to the literature data reporting the discovery of new targets for growth inhibition of Mycobacterium tuberculosis, namely, diterpene cyclase (Rv3377c) and tuberculosinol phosphatase (Rv3378c), which produce diterpenoids of tuberculosinols in the cell membrane of M. tuberculosis, and these diterpenoids ensure the pathogenicity and the virulence of M. tuberculosis. For the first time, by the example of diterpenoid of isosteviol, its binuclear derivatives, triterpenoid betulinic, oleanolic, and ursolic acids, it has been shown by the molecular docking method that the antitubercular activity of natural terpenoids is caused by their ability to bind to the active site of tuberculosinol phosphatase (Rv3378c) of M. tuberculosis. It is suggested that natural and semisynthetic terpenoids represent a promising platform for design of a new generation of antitubercular agents that affect this enzyme.

Identification and component analysis of triterpenoids in Monarda fistulosa L. and Ocimum americanum L. (Lamiaceae) aerial parts

Despite the progress in the creation of synthetic drugs, herbal medicines do not lose their relevance nowadays. Triterpenoids are quite common in the plant world and exhibit a wide range of biological activity. Medicinal plants of the Lamiaceae family can accumulate mainly pentacyclic triterpenoids which demonstrate the anti-inflammatory, antioxidant, hepatoprotective, anticancer and antimicrobial properties. Nowadays a number of new, poorly-studied species of the family have been introduced in Ukraine, including Monarda fistulosa L. and Ocimum americanum L.The aim of this study was to identify the triterpenoids and to analyze the contents of their components in ethanolic extracts of M. fistulosa and O. americanum (Lamiaceae) aerial parts under the condition of their farming in West Podillya (Ukraine).Methods: foam test, sedimentation and color reaction were used to determine the presence of triterpenoids in plant material; high-performance liquid chromatography (HPLC) method was used to determine their components' content; determination was carried out on a liquid chromatograph Shimadzu LC 20 Prominence.Results. The presence of triterpenoids in M. fistulosa and O. americanum ethanolic extracts was proved by the reactions of identification and HPLC. Effective extraction of triterpene compounds from the investigated plants was achieved using 96 % and 70 % ethanol. The comparative HPLC-analysis showed significant differences in content of ursolic, euscaphic, tormentic and oleanolic acids, betulin and lupeol in the aerial parts of investigated species. The most abundant components of M. fistulosa and O. americanum herbs were the ursolic and euscaphic acids.Conclusions: It was identified the presence and evaluated the contents of 6 triterpenoids in M. fistulosa and O. americanum aerial parts. The obtained data can be used in the planning of pharmacological studies and in the chemotaxonomy of Lamiaceae family

α-Glucosidase enzyme inhibitory effects and ursolic and oleanolic acid contents of fourteen Anatolian Salvia species

During the last decade, ursolic and oleanolic acids have been of considerable interest because of their α-glucosidase inhibitory activities and potential effects for treatment of type 2 diabetes. A simple and sensitive reversed-phase HPLC method was developed for the simultaneous determination of ursolic acid and oleanolic acid. The optimal mobile phase was selected as 85% acetonitrile solution. The limit of detection of the method for ursolic acid and oleanolic acid were 14 ng mL−1 and 13 ng mL−1, respectively. The method showed good precision and accuracy with intra-day and inter-day variations of 0.54% and 7.33% for ursolic acid, intra-day and inter-day variations of 0.51% and 5.26% for oleanolic acid, and overall recoveries of 97.8% and 98.5% for ursolic acid and oleanolic acid, respectively. Application of the method to determine the ursolic acid and oleanolic acid contents in the Salvia species revealed both compounds, with varying amounts between 0.21–9.76 mg g−1 ursolic acid and 0.20–12.7 mg g−1 oleanolic acid, respectively, among 14 Salvia species analyzed. Additionally, the plant extracts were analyzed for their inhibitory activities on α-glucosidase. According to the results of this assay, the extracts showed considerable activity on α-glucosidase with IC50 values from 17.6 to 173 μg mL−1. A strong negative correlation was detected between the amounts of both acids and IC50 values of extracts. Anatolian Salvia species have great potential as functional plants in the management of diabetes.

Known Triterpenes and their Derivatives as Scaffolds for the Development of New Therapeutic Agents for Cancer.

Plants produce several bioactive secondary metabolites whose are used as therapeutic agents to treat several diseases, among whom cancer. Triterpenes are secondary metabolites that exert inhibitory activity against multiple intracellular and extracellular targets in euchariotic cells. These targets are proteins involved in apoptosis, cell development and differentiation, angiogenesis, metastasis and inflammatory processes. The inhibition of their functions leads to decreased cellular growth, differentiation and migration, resulting in antitumor activity, as shown by several authors. Furthermore, during recent years synthetic triterpenoid derivatives have also been developed to implement potency and efficacy of starting compounds, allowing the obtainment of new agents having promising anticancer activity. In this review we report the latest results regarding anticancer activity of some of the most studied triterpenes in the field, as well as of their semi-synthetic derivatives, with the aim to summarize the role of triterpenes as molecular leads for the development of new classes of antitumor agents. We focused on the most recent literature regarding triterpenes in cancer treatment, highlighting the potential of developing new drugs starting from these natural compounds. Several "old" triterpenes as ursolic, betulinic and oleanolic acids were recently reconsidered as model compounds for the development of innovative anticancer agents. Their activity against proteins involved in tumor development enhances the opportunity to exploit these compounds as new multi-target therapeutic agents. Furthermore, the possibility to synthetize new compounds from their natural-occurring structures could be an alternative to overcome cellular resistance to drugs and could improve their therapeutic efficacy.

Synthesis of Water-soluble, Polyester-based Dendrimer Prodrugs for Exploiting Therapeutic Properties of Two Triterpenoid Acids

Dendrimers are macromolecules characterized by high controlled size, shape and architecture, presence of inner cavities able to accommodate small molecules and many peripheral functional groups to bind target entities. They are of eminent interest for biomedical applications, including gene transfection, tissue engineering, imaging, and drug delivery. The well-known pharmacological activities of ursolic and oleanolic acids are limited by their small water solubility, non-specific cell distribution, low bioavailability, poor pharmacokinetics, and their direct administration could result in the release of thrombi. To overcome such problems, in this paper we described their physical incorporation inside amino acids-modified polyester-based dendrimers which made them highly water-soluble. IR, NMR, zeta potential, mean size of particles, buffer capacity and drug release profiles of prepared materials were reported. The achieved water-soluble complexes harmonize a polycationic character and a buffer capacity which presuppose efficient cell penetration and increased residence time with a biodegradable cell respectful scaffold, thus appearing as a promising team of not toxic prodrugs for safe administration of ursolic and oleanolic acids.

Chemical characterization and bioactive properties of decoctions and hydroethanolic extracts of Thymus carnosus Boiss.

Thymus carnosus Boiss. is an endangered species endemic to Portugal. Studies on its phytochemical composition and bioactivity would increase its use and protection, as no data is available. LC-MS analysis of T. carnosus decoction and hydroethanolic extracts allowed the identification of a salvianolic A isomer, which has not been described in Thymus genus before, rosmarinic and salvianolic K acids as the main phenolics. Luteolin, apigenin and eriodictyol glycosides were identified in lower amounts. Oleanolic and ursolic acids were present only in hydroethanolic extracts. Decoction extracted 32.4% of the phenolic compounds obtained by exhaustive hydroethanolic extraction. Both extracts reduced the viability of three cancer cell lines (human breast: MCF-7, BT-474; murine macrophage: Raw 264.7), with hydroethanolic extracts showing a higher activity. T. carnosus extracts possess significant antioxidant, anti-inflammatory and anti-proliferative activities and may be suggested as a new potential source of natural antioxidants and their decoctions as a novel functional food.

Antitumor effect of Japanese apricot extract (MK615) on human cancer cells in vitro and in vivo through a reactive oxygen species-dependent mechanism.

MK615 is produced from Japanese apricot and contains several cyclic triterpenes, such as oleanolic and ursolic acids. MK615 was shown to strongly suppress cutaneous in-transit metastasis in a patient with malignant melanoma. The present investigation was undertaken to clarify the antitumor effects of MK615 in vitro and in vivo. Several human cancer cell lines were exposed to MK615 for 7 days to examine its antiproliferative effects. The effect of MK615 on in vivo growth of human pancreatic cancer MIAPaCa-2 cells was also examined. MK615 inhibited the growth of several human cancer cell lines in a concentration-dependent way. Pancreatic cancer MIAPaCa-2 cells were highly sensitive to the growth-inhibiting effects of MK615. Treatment with MK615 preferentially induced cell death in human cancer cells while sparing normal cells such as human umbilical vein endothelial cells (HUVEC) and mouse bone marrow cells. When MIAPaCa-2 cells were incubated with MK615 in the presence of antioxidant, growth-inhibition was significantly reduced, and MK615 induced the accumulation of reactive oxygen species in cancer cells but not in HUVEC. MK615, in both the presence and absence of gemcitabine, significantly inhibited the growth of human pancreatic cancer cells as xenografts without apparent adverse effects. MK615, a supplement produced from Japanese apricot, may have therapeutic value in treating human cancers through a reactive oxygen species-dependent mechanism.

Recent progress in the antiviral activity and mechanism study of pentacyclic triterpenoids and their derivatives.

Viral infections cause many serious human diseases with high mortality rates. New drug‐resistant strains are continually emerging due to the high viral mutation rate, which makes it necessary to develop new antiviral agents. Compounds of plant origin are particularly interesting. The pentacyclic triterpenoids (PTs) are a diverse class of natural products from plants composed of three terpene units. They exhibit antitumor, anti‐inflammatory, and antiviral activities. Oleanolic, betulinic, and ursolic acids are representative PTs widely present in nature with a broad antiviral spectrum. This review focuses on the recent literatures in the antiviral efficacy of this class of phytochemicals and their derivatives. In addition, their modes of action are also summarized.

Synergy between Ursolic and Oleanolic Acids from Vitellaria paradoxa Leaf Extract and β-Lactams against Methicillin-Resistant Staphylococcus aureus: In Vitro and In Vivo Activity and Underlying Mechanisms.

Combining antibiotics with resistance reversing agents is a key strategy to overcome bacterial resistance. Upon screening antimicrobial activities of plants used in traditional medicine, we found that a leaf dichloromethane extract from the shea butter tree (Vitellaria paradoxa) had antimicrobial activity against methicillin-resistant Staphylococcus aureus (MRSA) with further evidence of synergy when combined with β-lactams. Using HPLC-MS, we identified ursolic (UA) and oleanolic acids (OA) in leaves and twigs of this species, and quantified them by HPLC-UV as the major constituents in leaf extracts (21% and 6% respectively). Both pure triterpenic acids showed antimicrobial activity against reference and clinical strains of MRSA, with MICs ranging from 8–16 mg/L for UA to 32–128 mg/L for OA. They were highly synergistic with β-lactams (ampicillin and oxacillin) at subMIC concentrations. Reversion of MRSA phenotype was attributed to their capacity to delocalize PBP2 from the septal division site, as observed by fluorescence microscopy, and to disturb thereby peptidoglycan synthesis. Moreover, both compounds also inhibited β-lactamases activity of living bacteria (as assessed by inhibition of nitrocefin hydrolysis), but not in bacterial lysates, suggesting an indirect mechanism for this inhibition. In a murine model of subcutaneous MRSA infection, local administration of UA was synergistic with nafcillin to reduce lesion size and inflammatory cytokine (IL-1β) production. Thus, these data highlight the potential interest of triterpenic acids as resistance reversing agents in combination with β-lactams against MRSA.

Gastroprotective activity of the methanol extract from peels of Plinia edulis (Vell.) Sobral fruits and its isolated triterpenes: maslinic and ursolic acids.

According to the Brazilian folk medicine, the leaves of Plinia edulis (Vell.) Sobral (Myrtaceae), known as cambuca, are indicated in the treatment of gastric disorders. Infusions of P. edulis leaves were previously demonstrated to contain both maslinic (MA) and ursolic acids (UA). Both triterpenes have also been identified in the methanolic extract of peels from P. edulis fruit (MEPPE); however, the antiulcer effects of MEPPE have not yet been studied. This study therefore evaluates the gastroprotective potential of MEPPE, MA, and UA using ethanol/HCl- and indomethacin-induced gastric ulcers in mice. In addition, the in vitro effects of these compounds on the H+, K+-ATPase activity and on the free radical DPPH were measured. When used at concentration of 100μg/mL, both MEPPE and UA were found to reduce the DPPH radical levels by 78.66 and 60.14%, respectively. However, MA did not reduce DPPH radical levels. Our results illustrated the antiulcer effects of MEPPE, MA, and UA against experimental ulcer models when administered by either the oral or the intraperitoneal routes. In addition, MEPPE reduced the size of ethanol/HCl-induced ulcers in a dose-dependent manner (log half-maximal effective oral dose, LogED50 = 1.09). Interestingly, UA promoted gastroprotection at lower doses than MA by increasing the production of mucin levels at 692%; however, it does not alter the activity of H+, K+-ATPase. In contrast, both MEPPE and MA, when incubated at concentrations of 10 and 100μg/mL, inhibited H+, K+-ATPase activity in 61.81, 68.37, 54.04, and 70.45%, respectively. These results confirm that MEPPE, MA, and UA display gastroprotective activity through different modes of action; MA inhibits H+, K+-ATPase activity, whereas UA favour the mucus barrier.

Betulinic, oleanolic and ursolic acids inhibit the enzymatic and biological effects induced by a P-I snake venom metalloproteinase

Betulinic acid (BA), Oleanolic acid (OA) and Ursolic acid (UA), are pentacyclic triterpenoids with widespread occurrence throughout the plant kingdom, these compounds are widely recognized by their pharmacological and biological properties, such as, anti-tumoral, anti-inflammatory, anti-microbial and hepatoprotective activity. In this work we determined the inhibitory ability of these compounds on the enzymatic, hemorrhagic, myotoxic and edema-inducing activities of Batx-I, a P-I metalloproteinase isolated from Bothrops atrox venom. BA, UA and OA inhibited the proteolytic activity of Batx-I on gelatin with IC50 values of 115.3, 223.0 and 357.3 μM, respectively. Additionally, these compounds showed inhibition of the hemorrhagic activity of Batx-I in skin with IC50 345.7, 643.5 and 1077.0 μM for BA, UA and OA in preincubation experiments. In studies with independent-injection, in which Batx-I was injected and then, at the same site, a concentration of 600 μM of each compound were administered at either 0, 5 or 10 min, BA showed a significant reduction of hemorrhage at 0 and 5 min. In addition, these compounds inhibited myotoxicity and edema-forming activity of Batx-I at 600 μM concentration. Molecular docking studies suggested that these compounds could occupy part of the substrate binding cleft of the enzyme affecting its catalytic cycle. In this manner, triterpenic acids are candidates for the development of inhibitors for the prevention of local tissue damage in snakebite envenomation.

Determination of Oleanolic and Ursolic Acids in Sambuci flos Using HPLC with a New Reversed-phase Column Packed with Naphthalene Bounded Silica

Elder flowers have traditionally been used for their anti-inflammatory properties. However little is known about triterpenoid components in this Pharmacopoeial herbal drug. Herein, we report the quantification of ursolic and oleanolic acid in Sambuci flos by HPLC method. These biologically active triterpenoids were effectively separated and quantified using the elaborated HPLC chromatographic method with a naphthalene bounded silica reversed phase column. In this study, a very simple mobile phase for HPLC was developed to simultaneously determine both acids and the factors affecting separation were also discussed. The average content of oleanolic acid and ursolic acid in the elder flowers determined by proposed method was 2.06 mg/g and 11.56 mg/g respectively. This study showed both successful applications of the new type of reversed phase column and important contribution of mentioned triterpenoids to elder flower composition presumably resulting in its pharmacological action.