Betulonic Acid Research Articles

Paclitaxel and betulonic acid synergistically enhance antitumor efficacy by forming co-assembled nanoparticles

The side effects and low bioavailability of paclitaxel (PTX) limit its clinical application. The formation of self-assembled nanomedicines without structural modification is attractive for biomedical applications. Here, we constructed a supramolecular co-assembled nanoparticles (NPs), which is formed by betulonic acid (BTA) and PTX mainly through hydrogen bond interaction and hydrophobic interaction. It not only has the characteristics of NPs but also the activity of natural small molecules (NSMs). The results of in vitro and in vivo experiments showed that BTA-PTX NPs showed excellent synergistic enhancement of anti-tumor efficacy, because BTA and PTX have different anti-tumor mechanisms. What's more, BTA-PTX NPs showed excellent biosafety and low toxicity, because BTA has impressive biological activity and biosafety. This work provides an effective and simple method to construct high efficiency and minimize side effects of NPs, which provides more possibilities for the application of NSMs in drug delivery.

Betulinic Acid-Azaprostanoid Hybrids: Synthesis and Pharmacological Evaluation as Anti-inflammatory Agents

Background: Prevention and treatment of chronic inflammatory diseases require effective and low-toxic medicines. Molecular hybridization is an effective strategy to enhance the biological activity of new compounds. Triterpenoid scaffolds are in the focus of attention owing to their anti-inflammatory, antiviral, antiproliferative, and immunomodulatory activities. Heteroprostanoids have different pleiotropic effects in acute and chronic inflammatory processes.Objective: The study aimed to develop structurally new and low toxic anti-inflammatory agents via hybridization of betulinic acid with azaprostanoic acids.Methods: A series of betulinic acid-azaprostanoid hybrids was synthesized. The synthetic pathway included the transformation of betulin via Jones' oxidation into betulonic acid, reductive amination of the latter and coupling obtained by 3β-amino-3-deoxybetulinic acid with the 7- or 13-azaprostanoic acids and their homo analogues. The hybrids 1-9 were investigated in vivo on histamine-, formalin- and concanavalin A-induced mouse paw edema models and two models of pain - the acetic acid-induced abdominal writhing and the hot-plate test. The hybrids were in vitro evaluated for cytotoxic activity on cancer (MCF7, U-87 MG) and non-cancer humane cell lines.Results: In the immunogenic inflammation model, the substances showed a pronounced anti-inflammatory effect, which was comparable to that of indomethacin. In the models of the exudative inflammation, none of the compounds displayed a statistically significant effect. The hybrids produced weak or moderate analgesic effects. All the agents revealed low cytotoxicity on human immortalized fibroblasts and cancer cell lines compared with 3β-amino-3-deoxybetulinic acid and doxorubicin.Conclusion: The results indicate that the principal anti-inflammatory effect of hybrids is substantially provided with the triterpenoid scaffold and in some cases with the azaprostanoid scaffold, but the latter makes a significant contribution to reducing the toxicity of hybrids. Hybrid 1 is of interest as a potent low toxic agent against immune-mediated inflammation.

Bioactive Natural Small Molecule-Tuned Coassembly of Photosensitive Drugs for Highly Efficient Synergistic and Enhanced Type I Photochemotherapy.

Self-assembling natural small molecules (NSMs) with favorable anticancer activity are of increasing interest as novel drug delivery platforms without structural modification for biomedical applications. However, a lack of knowledge and practicability of NSMs as drug carriers limited their current biomedical application. Here, via a green and facile supramolecular coassembly strategy, we report and develop a series of carrier-free terpenoid natural small molecule-mediated coassembled photosensitive drugs for enhanced and synergistic chemo/photodynamic therapy. After screening 17 terpenoid NSMs, we identified 11 compounds that could form coassembled NSMs-Ce6 NPs with regulatable drug sizes. Analysis of the representative betulonic acid (BC)-mediated nano-coassemblies (BC-Ce6 NPs) reveals the high efficiency of the coassembly strategy and highlights the tremendous potential of NSMs as novel drug delivery platforms. Through molecular dynamics simulation and theoretical calculations, we elucidate the mystery of the coassembly process, indicating that the linear coplanar arrangement of BC dimeric units is primarily responsible for the formation of rod-like or spherical morphology. Meanwhile, we demonstrated that the reduced energy gap between the singlet and triplet excited states (ΔEST) facilitates efficient reactive oxygen species generation by promoting ·OH generation via a type I photoreaction mechanism. The assembled nanodrugs exhibit multiple favorable therapeutic features, ensuring a remarkably enhanced, synergistic, and secure combinatorial anticancer efficacy of 93.6% with highly efficient tumor ablation. This work not only expands the possibility of natural biodegradable materials for wide biological applications but also provides a new perspective for the construction of NSM-mediated nano-coassemblies for precision therapy.

Discovery and characterization of pentacyclic triterpenoid acids in Styrax as potent and reversible pancreatic lipase inhibitors

• Styrax extract was found with strong pancreatic lipase (PL) inhibition activity. • Bioactivity and LC-MS guided fractionation strategy was used to identify naturally occurring PL inhibitors in Styrax. • Oleanonic acid (ONA) and betulonic acid (BNA) potently inhibit PL, with IC 50 values less than 1.5 μM. • Inhibitory mechanisms of ONA and BNA against PL were investigated by inhibition kinetic analyses and in silico analysis. Pancreatic lipase (PL) inhibitor therapy is one of the most effective ways to treat obesity and obesity-associated metabolic disorders. Herein, bioactivity and liquid chromatograph-mass spectrometer (LC-MS) guided fractionation strategy was used to identify the naturally occurring PL inhibitors in Styrax, a popular folk medicine in Asia and Turkey. The results clearly showed that five pentacyclic triterpenoid acids in Styrax displayed strong inhibition on PL, with IC 50 values ranging from 0.49 µM to 6.35 µM. Further investigations demonstrated that oleanonic acid and betulonic acid (the most two potent PL inhibitors in Styrax) potently inhibited PL in a reversible and mixed inhibition manner, while molecular dynamics simulations showed that these two agents could stably bind at the entrance to the active site of PL. Collectively, our findings revealed the molecular basis for the anti-obesity effects of Styrax, while the newly identified PL inhibitors in this herb could be used for development of novel anti-obesity agents.

The New Pharmaceutical Compositions of Zinc Oxide Nanoparticles and Triterpenoids for the Burn Treatment.

We studied oleogels containing zinc oxide nanoparticles (ZnO NPs) and lupane triterpenoids in sunflower oil for the treatment of burns. The modification of ZnO was carried out by treatment with alcohol solutions of betulin, betulonic acid, betulin diacetate and betulin diphosphate. The properties of modified ZnO NPs were studied by powder XRD (average sizes of 10–20 nm), FTIR (νZnO 450 cm−1), UV–vis (345–360 nm), and blue–violet emission (380–420 nm). The identification and assay of modified ZnO NPs and triterpenoids were estimated. The treatment by oleogels of deep II-degree burns was studied on rats using histological studies, Doppler flowmetry and evaluation of enzymes activity and malonic dialdehyde (MDA) level. After the action of oleogels, burn wound area, and the necrosis decreased twice on the 10th day in comparison with the 1st day after burn. The microcirculation index in the near-wound zone by 20–30% improved compared with the group without treatment. Evaluation of the enzyme activity and the MDA level after treatment by oleogels during the course of 10 days showed them returning to normal. The improvement of antioxidant biochemical indexes, as well as wounds’ healing, was mainly determined by the influence of zinc oxide nanoparticles.

Herb-drug interaction between Styrax and warfarin: Molecular basis and mechanism

BackgroundStyrax, one of the most famous folk medicines, has been frequently used for the treatment of cardiovascular diseases and skin problems in Asia and Africa. It is unclear whether Styrax or Styrax-related herbal medicines may trigger clinically relevant herb-drug interactions. PurposeThis study was carried out to investigate the inhibitory effects of Styrax on human cytochrome P450 enzymes (CYPs) and to clarify whether this herb may modulate the pharmacokinetic behavior of the CYP-substrate drug warfarin when co-administered. Study DesignThe inhibitory effects of Styrax on CYPs were assayed in human liver microsomes (HLM), while the pharmacokinetic interactions between Styrax and warfarin were investigated in rats. The bioactive constituents in Styrax with strong CYP3A inhibitory activity were identified and their inhibitory mechanisms were carefully investigated. MethodsThe inhibitory effects of Styrax on human CYPs were assayed in vitro, while the pharmacokinetic interactions between Styrax and warfarin were studied in rats. Fingerprinting analysis of Styrax coupled with LC-TOF-MS/MS profiling and CYP inhibition assays were used to identify the constituents with strong CYP3A inhibitory activity. The inhibitory mechanism of oleanonic acid (the most potent CYP3A inhibitor occurring in Styrax) against CYP3A4 was investigated by a panel of inhibition kinetics analyses and in silico analysis. ResultsIn vitro assays demonstrated that Styrax extract strongly inhibited human CYP3A and moderately inhibited six other tested human CYPs, as well as potently inhibited warfarin 10-hydroxylation in liver microsomes from both humans and rats. In vivo assays demonstrated that compared with warfarin given individually in rats, Styrax (100 mg/kg) significantly prolonged the plasma half-life of warfarin by 2.3-fold and increased the AUC(0-inf) of warfarin by 2.7-fold when this herb was co-administrated with warfarin (2 mg/kg) in rats. Two LC fractions were found with strong CYP3A inhibitory activity and the major constituents in these fractions were characterized by LC-TOF-MS/MS. Five pentacyclic triterpenoid acids (including epibetulinic acid, betulinic acid, betulonic acid, oleanonic acid and maslinic acid) present in Styrax were potent CYP3A inhibitors, and oleanonic acid was a competitive inhibitor against CYP3A-mediated testosterone 6β-hydroxylation. ConclusionStyrax and the pentacyclic triterpenoid acids occurring in this herb strongly modulate the pharmacokinetic behavior of warfarin via inhibition of CYP3A.

Effect of betulin and betulonic acid on isolated rat liver mitochondria and liposomes

The paper considers the effects of plant triterpenoid betulin and its derivative betulonic acid on rat liver mitochondria and liposomes. It was found that betulonic acid and, to a lesser extent, betulin, activate mitochondrial respiration in states 2 and 4 and inhibit ADP- and DNP-stimulated (uncoupled) respiration. The effect of betulonic acid resulted in a significant decrease of the respiratory control and ADP/O ratios and decrease in Δψ. The effects of both compounds were most pronounced in the case of succinate-fueled mitochondrial respiration. This may include both the possible protonophore effect of betulonic acid and the inhibition of respiratory chain complexes by both compounds. Both agents enhanced H2O2 production in succinate-fueled mitochondria, while betulonic acid exerted an antioxidant effect with NAD-dependent substrates. Betulin was found to induce mitochondrial aggregation, but had no effect on membrane permeability. A similar pattern was found on liposomes. As revealed by the laurdan generalized polarization (GP) technique, betulin increased laurdan GP in lecithin liposomes, indicating a decrease in membrane fluidity. Measurements of GP as a function of fluorescence excitation wavelength gave an ascending line for high concentrations of betulin, which can be interpreted as phase heterogeneity of the lipid/betulin system. High concentrations of betulin (> 60 mol%) was also demonstrated to cause permeabilization of lecithin liposomes. Betulonic acid was much less effective in inducing the aggregation of mitochondria and liposomes and had no effect on membrane permeability. The possible mechanisms of betulin and betulonic acid effect on rat liver mitochondria and liposomes are discussed.

Synthesis, Anticancer, and Antibacterial Activity of Betulinic and Betulonic Acid C-28-Triphenylphosphonium Conjugates with Variable Alkyl Linker Length.

Conjugation of triterpenoids such as betulinic acid 1 with the Triphenylphosphonium (TPP) group is a powerful approach to generating medicinal compounds. Their development proposes structure optimization in respect of availability and activity towards target cells and organelles. Selection of 1 or its precursor betulonic acid 2 and the optimal linker is of particular importance for drug candidate identification among the TPP-triterpenoid conjugates. In this study, new C-28-TPP conjugated derivatives of 1 and 2 with the alkyl/alkoxyalkyl linkers of variable length were synthesized and compared regarding their anticancer, antibacterial, and mitochondriatargeted effects. The TPP conjugates of 1 and 2 [6a-f, 7a-f] were synthesized by the reaction of halogenalkyl esters [3a-f, 4a-f, 5] with triphenylphosphine in acetonitrile upon heating. Cytotoxicity (MTT assay), antibacterial activity (microdilution assay), and mitochondrial effects (flow cytofluorometry) were studied. Conjugation with the TPP group greatly increased the cytotoxicity of the triterpenoids up to 30 times. The conjugates were up to 10-17 times more active against MCF-7 (IC50 = 0.17μM, 72h, 6c) and PC-3 (IC50 = 0.14μM, 72h, 6a) cancer cells than for human skin fibroblasts. The enhanced antibacterial (bactericidal) activity of the TPP-triterpenoid conjugates with MIC for Gram-positive bacteria as low as 2μM (6a, 7a) was for the first time revealed. The conjugates were found to effectively inhibit fluorescence of 2′,7′-dichlorofluorescin probe in the cytosol upon oxidation, decrease transmembrane potential, and increase superoxide radical level in mitochondria. Relationships between the effects and structure of the TPP-triterpenoid conjugates were evaluated and discussed. Based on the results, 6a can be selected for further preclinical investigation as a potential anticancer compound.

SYNTHESIS OF TRITERPENE POLYMER CONSTRUCTIONS

The pentacyclic triterpenoids betulin and betulonic acid are promising sources of new biologically active compounds. Grafting them onto polymer matrices leads to the formation of effective dosage forms compared to the original drug. On the basis of betulin and betulonic acid, the synthesis of polymer structures with a fragment of triterpene in the side chain was carried out. New triterpene-containing polymer ensembles were obtained by the method of polymer-analogous transformations of copolymers of N-vinylpyrrolidone with N-(n-carboxy) phenylmaleimide and p-aminostyrene. By crosslinking the carboxyl groups of the copolymer of N-vinylpyrrolidone with N-(n-carboxy) phenylmaleimide with betulin hydroxyl groups and the reaction of the amino groups of the copolymer of N-vinyl pyrrolidone with p-aminopyrol with carboxyl groups of betulonic acid, polymer structures with polycyclic triterpene fragments are obtained. The resulting polymer ensembles have higher activity against melanoma compared to the original copolymers. A polymer based on a copolymer of N-vinylpyrrolidone with N-(n-carboxy) phenylmaleimide and betulin at a concentration of 40.48 μM inhibits 50% of MS cells, while 50% of MS cells die under the influence of 68.29 μM betulin. New triterpene-containing polymer ensembles are promising for the development of new biologically active polymer bioconjugates.

A Study of Plant Coumarins. 18. Conjugates of Coumarins with Lupane Triterpenoids and 1,2,3-Triazoles: Synthesis and Anti-Inflammatory Activity

In the reaction of coumarin peuruthenicin with α,ω-dibromoalkanes, the corresponding 7-(ω-bromoalkyloxy)coumarins have been synthesized. The interaction of the resulting compounds with sodium azide has yielded 7-(ω-azidoalkyl)-substituted peuruthenicin derivatives, which exhibited high activity in the Cu(I)-catalyzed Huisgen cycloaddition reaction with the propargyl ester of betulonic acid. As a result of the reaction, 28-O-(chromenoalkyl-triazolylmethyl)lupenones have been synthesized. The reaction of betulonic acid propargyl ester with 2-azidooreoselones in the presence of aqueous copper sulfate and sodium ascorbate has led to the corresponding (furochromen-triazolyl)-20(29)-lupen-3-ones. The newly synthesized hybrid compound triterpenoid furocoumarin containing a triazole linker exhibits anti-inflammatory activity in a model of histamine inflammation.

Bioassay guided isolation of mosquito biting deterrent compounds from Strumpfia maritima.

In our natural products screening program against mosquitoes, we screened ethanolic extract of Strumpfia maritima Jacq. which showed high biting deterrent activity in Klun and Debboun (K&D) bioassays against Aedes aegypti L. Through bioguided fractionation we isolated six compounds from the active fractions. The extract and pure compounds were evaluated for larvicidal and biting deterrent activity against Aedes aegypti. Ethanolic extract with proportion not biting (PNB) value of 0.8 ± 0.08 provided biting deterrent activity similar to N,N-dimethyl-m-toluamide (DEET) (PNB = 0.9 ± 0.05) against Ae. aegypti. Five out of eight fractions, prepared through bioassay-guided fractionation, showed biting deterrent activity similar to DEET. Six compounds were isolated from active fractions and characterized as squalene, ursolic acid, betulonic acid, cabraleone, ursolic acid acetate, and 27-p-E-coumaroyloxyursolic acid. The biting deterrent activity of ursolic acid acetate (biting deterrence index, BDI = 0.85), cabraleone (BDI = 0.79), and ursolic acid (BDI = 0.78) was similar to DEET against Ae. aegypti. Strumpfia maritima extract did not show larvicidal activity at the highest screening dose of 125 ppm in the larvicidal bioassay. The ethanolic extract and pure compounds showed biting deterrent activity against Ae. aegypti. The high biting deterrent activity of ursolic acid acetate and cabraleone indicated a great potential for these natural compounds to be developed as mosquito repellents. © 2020 Society of Chemical Industry.

Ultrastructure of the Liver in Response to Cyclophosphamide and Triterpenoids.

Ultrastructural reorganization of liver cells after isolated injections of cyclophosphamide, betulonic acid or its β-alanylamide, and combined treatment with the cytostatic and each of the triterpenoids is studied. Cyclophosphamide causes significant ultrastructural changes in all intracellular compartments of hepatocytes. Both triterpenoids cause moderate cytotoxic and stimulatory effects on the liver cell populations (hepatocytes, sinusoidal endotheliocytes, and Kupffer cells), when used alone. The cytotoxic effect of betulonic acid manifests in modification of the fine structure of hepatocyte mitochondria, sequestration of glycogen, intensification of autophagic processes, emergence of necrobiotic changes in hepatocytes and endotheliocytes; betulonic acid amide actively modifies the mitochondrial fine structure (hypertrophic organelles, matrix rarefaction, uneven dilatation of cristae). The effects of combinations of cyclophosphamide with betulonic acid or its amide on liver are polytarget: the cytotoxic activity of the cytostatic is potentiated towards some cells, while in other cells the regeneratory reactions are stimulated. The common cytological cytoprotective effects of betulonic acid and its amide used alone and in combination with cytostatics include stimulation of the endocytotic (pinocytotic) activities of the cells and stimulation of intracellular regeneration processes in them.

Promising Terpenes as Natural Antagonists of Cancer: An In-Silico Approach.

Overexpression of murine double minute 2 (MDM2) results in the inactivation of p53 and causes cancer which is a leading cause of death in recent era. In recent decades, much attention has been paid to discover potential inhibitors against MDM2 in order to cure cancer. Outcomes from studies proposes that the MDM2 is a hot target to screen potent antagonists. Thus, this study aims at discovering natural compounds using several computational approaches to inhibit the MDM2 and to eliminate p53-MDM2 interaction, which would result in the reactivation of p53 activity. A library of 500 terpenes was prepared and several virtual screening approaches were employed to find out the best hits which could serve as p53-MDM2 antagonists. On the basis of the designed protocol, three terpenes were selected. In the present study, for the stability and validation of selected three protein-ligand complexes 20 ns molecular dynamics simulations and principal component analyses (PCA) were performed. Results found that the selected terpenes hits (3-trans-p-coumaroyl maslinic acid, Silvestrol and Betulonic acid) are potential inhibitors of p53–MDM2 interaction and could serve as potent antagonists.

β- Sitosterol and Betulonic Acid from n-Hexane Extract the Stem Bark of Tetracera indica

The Tetraceraindicahasbeen used traditionally to treat various diseases including skin medications, lower blood pressure, fever, headache, wound andgout medicine. The objective of the research was isolatedand characterizedthe β-sitosterol (1) and betulonicacid (2) fromn-hexaneextract of the stem bark of Tetraceraindica. The dry powder thestembark (550 g)wasmaceration with n-hexane for 24 hrs.Thisextraction process was repeated three times. The n-hexane extract was separated and purified by chromatographytechnique. The structure of isolatedcompound wasdetermined by spectroscopy data UV, IR, NMR and compared with reported data. Two compounds have been isolated and base on spectroscopy data thecompounds is β-sitosterol (1) andbetulonic acid (2). Betulonic acid is being reported for the first time from T.indica,meanwhile theβ-sitosterolpreviously reported from MeOHextractof leaves of T. indicabut is being reported for the first time from the stembarkof T.indica.

Synthesis and biological evaluation of triterpenoid thiazoles derived from betulonic acid, dihydrobetulonic acid, and ursonic acid

In this work, 35 new derivatives of betulonic, dihydrobetulonic and ursonic acid were prepared including 30 aminothiazoles and all of them were tested for their in vitro cytotoxic activity in eight cancer cell lines and two non-cancer fibroblasts. Compounds with the IC50 below 5 μM in CCRF-CEM cells and low toxicity in non-cancer fibroblasts (4m, 5c, 5m, 6c, 6m, 7b, and 7c) were further subjected to tests of pharmacological parameters yielding the final set for advanced biological evaluation (4m, 5m, 6m, and 7b). It was proved by several methods, that all of them trigger apoptosis via the intrinsic pathway and derivatives 5m and 7b are the most effective (IC50 2.4 μM and 3.6 μM). They are the best candidates to become potentially new anticancer drugs and will be subjected to in vivo tests in mice. In addition, compounds 6b and 6c deserve more attention because their activity is not limited only to chemosensitive CCRF-CEM cell line. Specifically, compound 6b is highly active against K562 leukemic cell line (0.7 μM) and its IC50 activity in colon cancer HCT116 cell line is 1.0 μM. Compound 6c is active in both normal K562 and resistant K562-TAX cell lines (IC50 3.4 μM and 5.4 μM) and both colon cancer cell lines (HCT116 and HCT116p53-/-, IC50 3.5 μM and 3.4 μM).

Chemical profiles of birch and alder bark by ambient mass spectrometry

Desorption atmospheric pressure photoionization (DAPPI) is an ambient mass spectrometry (MS) technique that allows the analysis of both polar and nonpolar compounds directly from the surfaces of various sample types. Here, DAPPI was used to study the chemical profiles in different parts of birch and alder tree barks. Four distinct fractions of Betula pendula (silver birch) bark were collected from three different developmental stages of the stem, after which the chemical profiles of the different tissue types were measured. Of special interest were triterpenoids, a class of important defensive substances, which are found in the bark of the silver birch. Additionally, the chemical profiles of lenticels and the surrounding surfaces in the phellem of B. pendula (silver birch), Alnus glutinosa (black alder), and Alnus incana (gray alder) were screened with DAPPI. Another ambient MS technique, laser ablation atmospheric pressure photoionization (LAAPPI), was further used for the mass spectrometry imaging of lenticels on the B. pendula phellem. All the studied birch bark fractions showed individual chemical profiles in DAPPI. The mass spectra from the young apical stem and the transition zone resembled each other more than the mature stem. Instead, the phellem was found to contain a high amount of triterpenoids in all the developmental stages of the stem. The most intense peaks in the DAPPI mass spectra of the birch bark fractions were those of betulin and lupeol. Betulinic and betulonic acid peaks were intense as well, and these compounds were detected especially in the lenticels of the tree samples.Graphical abstract

Synthesis of betulonic acid S,S′-bis(carboxymethyl) dithioketal

Betulonic acid S,S′-bis(carboxymethyl) dithioketal was obtained by condensation of betulonic and thioglycolic acids.

Lupan‐type triterpenoids from the stems of Salacia chinensis L. (Celastraceae) and their α‐glucosidase inhibitory activities

AbstractSix lupan‐type triterpenoids: 29‐norlupan‐3,20‐dione (1), 3β‐hydroxy‐29‐norlupan‐20‐one (2), betulonic acid (3), salacianol (4), 30‐hydroxylup‐20(29)‐en‐3‐one (5), and lup‐20(29)‐ene‐3β,30‐diol (6) have been isolated from the CHCl3‐soluble fraction of the stem of Salacia chinensis L. (Celastraceae). Their chemical structures were elucidated based on the NMR spectroscopic analysis and comparison with the literatures. Compounds (5) and (6) showed very weak α‐glucosidase inhibitory activity with the IC50 values of 211.5 and 246.8 μM, respectively.

MOLECULAR DOCKING OF ANTITRYPANOSOMAL INHIBITORS FROM EUCALYPTUS TERETICORNIS FOR SLEEPING SICKNESS

Objectives: This study aims to investigate the antitrypanosomal inhibitors of Eucalyptus tereticornis for sleeping sickness through molecular docking and studies on Absorption distribution metabolism excursion and toxicology (ADMET).
 Methods: In silico molecular docking in ArgusLab software and ADMET analysis in AdmetSAR software was performed for the antitrypanosomal inhibitors of E. tereticornis for sleeping sickness.
 Results: Interactions were studied for the ten proteins responsible for sleeping sickness with the 50 antitrypanosomal inhibitors of E. tereticornis. Docking was performed to see the interaction and the best binding energy of compounds with the proteins involved in sleeping sickness. The docking scores were highest for betulonic acid with −15.66 kcal/mol followed by euglobal with −12.24 kcal/mol, B-pinene with −10.313 kcal/mol, A-pinene with −10.3418 kcal/mol, and the least docking score for P-cymene with −10.6045 kcal/mol. Docking results showed that only betulonic acid and euglobal showed that hydrogen bond interaction was as b-pinene, a-pinene, and p-cymene yielded no hydrogen bond interactions so we will be taking the former docking results for further studies. The best docking result was shown by betulonic acid with trypanothione reductase giving binding energy of −15.66 kcal/mol with hydrogen bond interaction of 2.9, so this result was taken for further analysis.
 Conclusion: The results of the compound extracted from E. tereticornis will become physiological relevant only when (i) the pure compounds of this plant is available in large quantities; (ii) the Eucalyptus is biochemically stabilized to avoid degradation and enhance absorption in the gastrointestinal tract; and (iii) special delivery methods for this drug to reach the areas of treatment. In this work, the efficacy of E. tereticornis to act against trypanosomal protein was initiated and thus further research in this process would help us to take full advantage of the remedial effects of the compounds extracted from this plant.

Lupane-type conjugates with aminoacids, 1,3,4- oxadiazole and 1,2,5-oxadiazole-2-oxide derivatives: Synthesis, anti-inflammatory activity and in silico evaluation of target affinity

With the purpose to improve anti-inflammatory activity, the impact of introduction of 1,2,5- and 1,3,4-oxadiazole fragments to betulonic acid core as well as hybrids tethered with short ω-amino acids has been studied. The anti-inflammatory activity of synthesized compounds was tested in vivo using models of inflammation induced by concanavalin A and histamine. The majority of new compounds demonstrated higher anti-inflammatory activity compared with starting betulonic acid. To confirm the molecular targets of new derivatives in NRf2 and NFκB pathways the docking at Kelch and BTB active sites of Keap1 as well as IKK was done. The novelty of the present work is the development of new class of low toxic anti-inflammatory substances consisting of amino acid-linked betulonic acid - oxadiazole conjugates. These compounds can be considered as prospective chemopreventive agents.