Dušica P. Ilić

ALLICIN AND RELATED COMPOUNDS: BIOSYNTHESIS, SYNTHESIS AND PHARMACOLOGICAL ACTIVITY †

In this review, the biosynthesis of allicin (allyl thiosulfinate) by enzymatic transformation of alliin and various methods of its synthesis with detailed investigation of mechanisms and kinetics are summarized. A convenient method is also described for determination of allicin stability and the utility of the inclusion complexes of this pharmacologically active agent with β-cyclodextrins in increasing its stability. Allicin is the initial precursor for the production of ajoene ((E)- and (Z)-4,5,9-trithiadodeca- 1,6,11-triene 9-oxides) and vinyldithiin (2-vinyl-4H-1,3-dithiin, and 3-vinyl-4H-1,2- dithiin), which are more stable and show various pharmacological effects. The mechanisms of allicin transformations to these compounds are given in detail. Finally, the data on the pharmacological effects of allicin and its transformation products, ajoene and vinyldithiin, are presented.

Thermal degradation, antioxidant and antimicrobial activity of the synthesized allicin and allicin incorporated in gel

The main carriers of the pharmacological activity of garlic (Allium sativum L.) are organic sulfur compounds, the most important among them being allicin, a sulfenic acid thioester, or allylthiosulfonate. In this paper, the identification of synthesized and purified allicin was determined by using various spectroscopic methods (UV/VIS, FTIR, NMR). A HPLC method was developed for the detection and determination of the allicin content. The thermal degradation of allicin by using FTIR method was monitored. The method for the production of allicin gel based on Carbopol 940 (poly(acrylic acid)) was elaborated. The antimicrobial activity of pure allicin and allicin incorporated into gel by using a disk diffusion method was determined. In order to determine the antioxidant activity of allicin DPPH test was done and it was proved that with low concentrations (1 mgcm-3) a high DPPH radicals scavenging capacity (90%) was achieved.

Transformation of Synthetic Allicin: The Influence of Ultrasound, Microwaves, Different Solvents and Temperatures, and the Products Isolation

The transformation of the synthesized allicin, using conventional method, the influence of ultrasound and microwaves, in different organic solvents (acetonitrile, acetone, methanol, and chloroform), at various temperatures (room temperature, 45°C, and 55°C) was investigated. Allicin degradation kinetic was monitored by HPLC. Allicin transformation under the effect of microwaves is faster than transformations performed under the influence of ultrasound or by conventional method. Increase of the temperature accelerates allicin transformation. Pharmacologically active compounds of (E)-ajoene, (Z)-ajoene, 3-vinyl-4H-1,2-dithiin, 2-vinyl-4H-1,3-dithiin, and diallyl disulfide were isolated from the mixture of transformation products of allicin under the influence of microwaves in methanol at 55°C, which is according to kinetic parameters (highest values of the order of reaction and the lowest activation energy) the optimal method.

Inclusion complexes of amlodipine besylate and cyclodextrins

AbstractIn this paper the procedure for the preparation of inclusion complexes of amlodipine besylate with β-cyclodextrin (β-CD) and 2-hydrohypropyl-β-cyclodextrin (HPβ-CD) and their structural characterization was described. Molecular inclusion complexes of amlodipine besylate are prepared by the coprecipitation method and characterised by the application of spectroscopic methods FTIR, 1H-NMR and XRD. The photosensitivity of amlodipine besylate in the inclusion complexes was also determined with respect to uncomplexed agent. DSC curves indicate the loss of the clear peak due to melting of amlodipine besylate at about 200°C, while on XR diffractograms certain reflections are lost belonging to amlodipine besylate in complexes. This indicates its inclusion in the vacancies of the host. The inclusion of amlodipine besylate with cyclodextrins increases the stability, i.e. decreases the photosensitivity of amlodipine besylate.

Inclusion complexes of sulfanilamide with β-cyclodextrin and 2-hydroxypropyl-β-cyclodextrin

Sulfanilamide belongs to the group of drugs that have a bacteriostatic effect on different pathogenic microorganisms. This activity originates from the competitive antagonism with p-aminobenzoic acid, which is an integral part of folic acid. The safe use of sulfanilamide is limited due to poor solubility in the aqueous medium. Therefore, the aim of this paper is the synthesis of sulfanilamide, as well as preparing and structural characterization of its inclusion complexes with cyclodextrins. The crude sulfanilamide was obtained in the synthesis between acetanilide and chlorosulfonic acid according to the standard procedure. The synthesized sulfanilamide was recrystallized from water in order to obtain the satisfactory purity of the substance. Sufanilamide was complexed with β-cyclodextrin and 2-hydroxypropyl-β-cyclodextrin by the co-precipitation method. A molecular encapsulation of sulfanilamide was confirmed by using FTIR, 1H-NMR, XRD and DSC methods. Phase-solubility techniques were used to assess the formation of the inclusion complex between sulfanilamide and cyclodextrins. The photostability of sulfanilamide and its inclusion complexes was estimated by UVB irradiation in a photochemical reactor by applying the UV–Vis method. Based on the UV–Vis analysis, sulfanilamide:2-hydroxypropyl-β-cyclodextrin complex was presented as more photostable than sulfanilamide:β-cyclodextrin complex and sulfanilamide. The obtained results enable the potential use of these inclusion complexes for the preparation of oral formulations due to the enhanced solubility of sulfanilamide.

The effect of hydrodistillation techniques on yield, kinetics, composition and antimicrobial activity of essential oils from flowers of Lavandula officinalis L.

The essential oils from flowers of Lavandula officinalis L. (L. officinalis L.= L. angustifolia Mill.) were obtained by two hydrodistillation techniques: standard (technique I) and modified (technique II) Clevenger hydrodistillation. The yield, hydrodistillation kinetics, composition and antimicrobial activity of essential oils were investigated. The higher yield of oil was obtained by hydrodistillation technique II. The new analytical hydrodistillation kinetics model of essential oil from Lavandulae flowers was defined. The essential oil composition was analyzed by gas chromatography–mass spectroscopy (GC–MS). There were differences in chemical compositions of the oils obtained by different techniques. Antimicrobial activity of essential oil obtained by technique I and II against Salmonella enteritidis, Klebsiella pneumoniae, Staphylococcus aureus, Enterococcus faecalis, Candida albicans and Aspergillus niger was the same, while the activity against Escherichia coli and Pseudomonas aeruginosa was slightly different.

The protection of Nifedipin from photodegradation due to complex formation with β-cyclodextrin

AbstractThe inclusion complex β-cyclodextrin:nifedipin was prepared in solid state by coprecipitation with 1:1 mol ratio. The structure of the obtained complex and nifedipin was characterized by use of X-ray diffraction (XR), infrared spectroscopy (FTIR), nuclear magnetic resonance (NMR), and differential scanning calorimetry (DSC) methods. The photodegradation of nifedipin and the β-cyclodextrin:nifedipin inclusion complex in solid state was monitored under natural daylight by infrared spectroscopy, whereby the free nifedipin degraded four to five times faster than the complexed nifedipin. The photodegradation products of both free and complexed nifedipin, formed during irradiation at 350 nm (with corresponding energy flux of 18 W m−2) were monitored by liquid chromatography during various time intervals. The speed of formation of nitroso- and nitro-phenyl derivatives by nifedipin irradiation was significantly higher than those of complexed nifedipin irradiation, which indicates its increased photostability in the inclusion complex. The effect on this property is significant because it contributes both to the improvement of the therapeutic effect of nifedipin and to the safer application thereof.

The effect of extraction techniques on yield, extraction kinetics, and antioxidant activity of aqueous-methanolic extracts from nettle (Urtica dioica L.) leaves

ABSTRACT Five extraction techniques, maceration, reflux, Soxhlet, Tillepape, and ultrasonic extraction, were used to obtain the extractive matter from nettle leaves. The antioxidant activity of extracts was assessed by DPPH, FRAP, and H2O2 test, while the total phenolic and total flavonoid content was determined according to the Folin–Ciocalteu and aluminium chloride methods, respectively. Model Ponomarev and a non-stationary diffusion model through the plant material were used for modelling extraction process. The extract obtained by Soxhlet extraction, containing higher amounts of extractive matter as well as phenolic and flavonoid compounds, showed better antioxidant activity than those obtained by other extraction techniques.

Biological evaluation of synthesized allicin and its transformation products obtained by microwaves in methanol: antioxidant activity and effect on cell growth.

Allicin is the most biologically active substance present in garlic. It can be synthesized or obtained by extraction of fresh garlic. Transformation products of allicin are also biologically active. The aim of this study was to examine the antioxidant activity of synthesized allicin and its transformation products obtained using microwaves in methanol at 55 °C as well as their effect on HeLa cells growth. The antioxidant activity was determined by DPPH (2,2-diphenyl-1-picrylhydrazyl radical) test. The effect on HeLa cells growth was determined by MTT (3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyl-2H-tetrazolium bromide) test. For MTT test, allicin and its transformation products were dispersed in carmellose sodium solution and examined in concentrations ranging from 0.3 μg/mL to 3 mg/mL. Allicin showed stronger antioxidant activity than the transformation products. A maximum degree of neutralization of DPPH radicals, about 90%, was reached when the concentration of allicin was 2 mg/mL, with an EC50 (concentration of sample which is required for reduction of the initial concentration DPPH radicals to 50%) value of 0.37 mg/mL. In our study, allicin and its transformation products were not cytotoxic to HeLa cells under the examined conditions. The highest concentration of allicin and its transformation products had a slight antiproliferative effect, with a more pronounced effect of allicin, which reflected on the morphology of HeLa cells. The examined substances are safe to use on epithelial cells at concentrations up to 3 mg/mL when applied in carmellose sodium solution. Using carmellose sodium as a dispersing agent could be recommended as a good approach for testing liposoluble substances in liquid cell cultures.

Synthesis of polyrotaxanes from acetyl-β-cyclodextrin

Polyrotaxanes are intermediary products in the synthesis of topological gels. They are created by inclusion complex formation of hydrophobic linear macromolecules with cyclodextrins or their derivatives. Then, pairs of cyclodextrin molecules with covalently linkage were practically forming the nodes of the semi-flexible polymer network. Such gels are called topological gels and they can absorb huge quantities of water due to the net flexibility allowing the poly(ethylene oxide) chains to slide through the cyclodextrin cavities, without being pulled out altogether. For polyrotaxane formation poly(ethylene oxide) was used like linear macromolecules. There are hydroxyl groups at poly(ethylene oxide) chains, whereby the linking of the voluminous molecules should be made. To avoid the reaction of cyclodextrin OH groups with stoppers, they should be protected by, e.g., acetylation. In this work, the acetylation of the OH groups of β-cyclodextrin was performed by acetic acid anhydride with iodine as the catalyst. The acetylation reaction was assessed by the FTIR and HPLC method. By the HPLC analysis was found that the acetylation was completed in 20 minutes. Inserting of poly(ethylene oxide) with 4000 g/mol molecule mass into acetyl-β-cyclodextrin with 2:1 poly(ethylene oxide) monomer unit to acetyl-β-cyclodextrin ratio was also monitored by FTIR, and it was found that the process was completed in 12 h at the temperature of 10°C. If the process is performed at temperatures above 10°C, or for periods longer than 12 hours, the process of uncontrolled hydrolysis of acetate groups was initiated.