SYNTHESIS OF COCRYSTALS OF BETULIN WITH SUBERIC ACID AND STUDY OF THEIR PROPERTIES

Lupane triterpenoids, betulin and its derivatives, are attracting great interest due to their wide range of biological and pharmacological activities. However, the poor solubility of betulin and its derivatives in aqueous media significantly reduces their bioavailability. Obtaining cocrystals, that is, multicomponent crystal systems containing active pharmaceutical ingredients and non-toxic partner molecules in their structure, is used in pharmacy to change the physicochemical properties of drugs, including the rate of dissolution and solubility. In this study, cocrystals of betulin with suberic acid were obtained by liquid-assisted grinding method using organic solvents of different polarity: ethanol, acetone, ethyl acetate, chloroform, toluene, dioxane. The formation of cocrystals was confirmed by X-ray diffraction analysis, IR spectroscopy, and thermal analysis. It has been shown that cocrystals of betulin with suberic acid contain water molecules in their structure; anhydrous cocrystals can be obtained by heating a physical mixture of reagents until the acid melts. The results of experiments on the dissolution of cocrystals of betulin with suberic acid in comparison with the data for cocrystals of betulin with adipic acid showed that an increase in the length of the aliphatic acid chain leads to a decrease in the rate of betulin release into solution.

Biocatalysis in the Chemistry of Lupane Triterpenoids

Pentacyclic triterpenes are important representatives of natural products that exhibit a wide variety of biological activities. These activities suggest that these compounds may represent potential medicines for the treatment of cancer and viral, bacterial, or protozoal infections. Naturally occurring triterpenes usually have several drawbacks, such as limited activity and insufficient solubility and bioavailability; therefore, they need to be modified to obtain compounds suitable for drug development. Modifications can be achieved either by methods of standard organic synthesis or with the use of biocatalysts, such as enzymes or enzyme systems within living organisms. In most cases, these modifications result in the preparation of esters, amides, saponins, or sugar conjugates. Notably, while standard organic synthesis has been heavily used and developed, the use of the latter methodology has been rather limited, but it appears that biocatalysis has recently sparked considerably wider interest within the scientific community. Among triterpenes, derivatives of lupane play important roles. This review therefore summarizes the natural occurrence and sources of lupane triterpenoids, their biosynthesis, and semisynthetic methods that may be used for the production of betulinic acid from abundant and inexpensive betulin. Most importantly, this article compares chemical transformations of lupane triterpenoids with analogous reactions performed by biocatalysts and highlights a large space for the future development of biocatalysis in this field. The results of this study may serve as a summary of the current state of research and demonstrate the potential of the method in future applications.

Determining Stability Constants of Complexes Between Ester Betulin Derivatives and β-Cyclodextrin by Affinity Capillary Electrophoresis

Inclusion complexes of ester betulin derivatives (pentacyclic lupane triterpenoids) with β-cyclodextrin (β-CD) were studied by affinity capillary electrophoresis. Stability constants were calculated from dependencies of viscosity-corrected effective electrophoretic mobilities on β-CD concentration (0-10 mM) in background electrolytes using nonlinear regression fitting. Logarithms of stability constants at 25 °C for 1:1 and 1:2 β-CD complexes of betulin 3,28-diphthalate, betulin 3,28-disuccinate and betulin 3,28-disulfate (95 % confidence interval) are 4.25 (4.16-4.32) and 7.27 (6.73-7.50), 4.38 (4.26-4.48) and 7.58 (6.90-7.84), 4.04 (4.00-4.08) and 5.91 (4.60-6.20), respectively

Design, Synthesis, and Photophysical Properties of BODIPY-Labeled Lupane Triterpenoids

Novel boron-dipyrromethene difluoride (4,4-difluoro-4-bora-3α,4α-diaza-s-indacene) (BODIPY)-lupane triterpenoid conjugates bearing a fluorescent marker at the C-2 position of ring A of the triterpene core were obtained via the Sonogashira reaction as a key step. The starting compounds in the cross-coupling reaction were C-2 propynyl derivatives of betulinic or betulonic acids and fluorescent dyes with an iodo-group at C-2 or meso position of BODIPY-platform. The newly elaborated coupling procedure might have applicability in the synthesis of fluorescently-labeled triterpenoid conjugates suitable for biological assays.

Evaluation of Cytotoxicity and α-Glucosidase Inhibitory Activity of Amide and Polyamino-Derivatives of Lupane Triterpenoids

A series of two new and twenty earlier synthesized branched extra-amino-triterpenoids obtained by the direct coupling of betulinic/betulonic acids with polymethylenpolyamines, or by the cyanoethylation of lupane type alcohols, oximes, amines, and amides with the following reduction were evaluated for cytotoxicity toward the NCI-60 cancer cell line panel, α-glucosidase inhibitory, and antimicrobial activities. Lupane carboxamides, conjugates with diaminopropane, triethylenetetramine, and branched C3-cyanoethylated polyamine methyl betulonate showed high cytotoxic activity against most of the tested cancer cell lines with GI50 that ranged from 1.09 to 54.40 µM. Betulonic acid C28-conjugate with triethylenetetramine and C3,C28-bis-aminopropoxy-betulin were found to be potent micromolar inhibitors of yeast α-glucosidase and to simultaneously inhibit the endosomal reticulum α-glucosidase, rendering them as potentially capable to suppress tumor invasiveness and neovascularization, in addition to the direct cytotoxicity. Plausible mechanisms of cytotoxic action and underlying disrupted molecular pathways were elucidated with CellMinner pattern analysis and Gene Ontology enrichment analysis, according to which the lead compounds exert multi-target antiproliferative activity associated with oxidative stress induction and chromatin structure alteration. The betulonic acid diethylentriamine conjugate showed partial activity against methicillin-resistant S. aureus and the fungi C. neoformans. These results show that triterpenic polyamines, being analogs of steroidal squalamine and trodusquemine, are important substances for the search of new drugs with anticancer, antidiabetic, and antimicrobial activities.

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.

Design, Synthesis, and Cytotoxic Evaluation of Novel Lupane Triterpenoid Derived Hydroxamates

A series of new hydroxamate derivatives of lupane triterpenoids has been designed and successfully synthesized. The synthesized compounds were evaluated for their in vitro antitumor activity using the 3-[4,5-dimethylthiazol-2-yl]−2,5-diphenyltetrazolium bromide-based assay against the human cancer cell lines KB and HepG2. Most of these derivatives possess at least moderate cytotoxic activity and the hydroxamate derivative compounds 3c, 3e, 7a, and 15b could be lead compounds for further optimization to develop novel anticancer agents.