ABSTRACT
Fungal infections are a public health problem that mainly affects immunosuppressed people, Candida spp. have been responsible for most sources of contamination and invasive fungal infections described around the world. The need arises to find new therapeutic approaches to combat growing infections. Plants and natural products have been considered a valuable source for discovering new molecules with active ingredients. Diosgenin is a sapogenin found in the families of Leguminosae and Dioscoreaceae, it is obtained mainly from the dioscin saponin through the hydrolysis method, it is a phytochemical that has been highlighted in the treatment of various diseases, as well as in combating microbial resistance. The present study aimed to evaluate the susceptibility of fungal strains to diosgenin, as well as verify the association with the reference drug and evaluate the inhibition of the virulence factor through morphological changes in the yeast state to the filamentous form of hyphae and pseudohyphae in strains of Candida albicans, Candida tropicalis and Candida krusei using the broth microdilution method and microculture technique. Antifungal assays revealed that diosgenin was not able to inhibit the growth of the tested strains. However, it was able to inhibit the fungal dimorphism of the strains evaluated, however further studies are recommended to verify its effectiveness against other virulence factors.
Subject(s)
Antifungal Agents , Candida , Diosgenin , Microbial Sensitivity Tests , Diosgenin/pharmacology , Diosgenin/chemistry , Diosgenin/analogs & derivatives , Antifungal Agents/pharmacology , Antifungal Agents/chemistry , Candida/drug effects , Virulence/drug effects , Candida albicans/drug effects , Candida albicans/pathogenicityABSTRACT
Diosgenin and its derivatives have proved a huge importance in diverse biological activities. The optimized production of the diastereoisomers of the epoxide of diosgenin acetate by means of mCPBA is reported herein. This transformation had a previous design of experiments using the application of a statistical factorial DoE of 4 parameters (nk), where one variable is varied at a time, while others stay constant. The temperature showed the greatest effect on the reaction yield; so, at 298 K the diastereomeric ratio 3:1 of α and ß-epoxides, normally found, was raised to 1:1. Time was the second significant variable, but due to its high correlation with temperature, 30 min were required for a global 90% conversion at least. These diastereoisomers were characterized both isolated and in the mixtures obtained, to determine their antioxidant, antimicrobial and antiproliferative activity, finding a low antioxidant capacity by DPPH, but antimicrobial activity at the level of penicillin in gram negative bacteria by 1ß better to 1α. The antiproliferative capacity was higher for diastereoisomer ß, agreeing with the proportion of the mixture obtained by different conditions, increasing this in relation to the amount of this diastereoisomer present in hormone-dependent cancer cell lines such as Hela, PC-3 and MCF-7, with 10.0 µM obtained values of viability at 21.8 %, 35.8 % and 12.3 % respectively. DoE optimization allows to manipulate the ratio between diastereoisomers with the minimum number of experiments, extending the analysis of the effect of the ratio between diastereoisomers and the in silico potential as well as the biological activity.
Subject(s)
Anti-Infective Agents , Diosgenin , Humans , Antioxidants/pharmacology , Antioxidants/chemistry , Diosgenin/chemistry , Cell Line, Tumor , Anti-Infective Agents/chemistry , HeLa CellsABSTRACT
Diosgenin is a phytoestrogen and a constituent of Dioscorea. It has several biological effects, and some of them are anti-inflammatory, antidiabetic, antitumor, and vasodilatory. The present study investigated both the vasorelaxing and antioxidant mechanisms of diosgenin in isolated rat aortic rings. Female rats weighing 200-220 g were subjected to sham or OVX operations at 8 weeks of age. Ovariectomy was performed for menopause induction after anesthesia. Diosgenin (10-9 M-3 × 10-4 M) produced a concentration-dependent relaxation in aortic rings precontracted with phenylephrine (1 µM), exhibiting Emax value of 55.34% ± 7.7% (in endothelium-intact rings) and Emax value of 30.30% ± 5.7% (in endothelium-denuded rings). In the endothelium-intact rings, the vasorelaxing effect of diosgenin was reduced by NG-nitro-l-arginine methyl ester (L-NAME) (100 µM), atropine (1 µM), indomethacin (10 µM), 1H-[1,2,4]oxadiazolo-[4,3-a]quinoxalin-1-one (ODQ) (10 µM), 4-aminopyridine (1 mM), tetraethylammonium (3 mM), glibenclamide (10 µM), apamin (10 µM), and Tiron (1 µM). Diosgenin (10-5 M) inhibited the contractions induced by cumulative addition of phenylephrine (10-9-10-5 M). The 28-days treatment with diosgenin (50 mg/kg, v.o.) did not imply changes in the myeloperoxidase parameter, but increased significantly, levels of glutathione, superoxide dismutase, and nitric oxide, as well as reduced the concentration of malondialdehyde related to lipid peroxidation. Our results suggest that diosgenin induced relaxation in aortic rings via an endothelium-dependent pathway, which involves the EDRF, the opening of potassium channels and antioxidant action.
Subject(s)
Diosgenin/administration & dosage , Menopause/drug effects , Phytoestrogens/administration & dosage , Plant Extracts/administration & dosage , Vasodilator Agents/administration & dosage , Animals , Aorta/drug effects , Aorta/metabolism , Dioscorea/chemistry , Diosgenin/chemistry , Endothelium, Vascular/drug effects , Endothelium, Vascular/metabolism , Glutathione/metabolism , Humans , In Vitro Techniques , Male , Menopause/metabolism , Nitrites/metabolism , Ovariectomy , Oxidative Stress/drug effects , Phytoestrogens/chemistry , Plant Extracts/chemistry , Potassium Channels/metabolism , Rats , Rats, Wistar , Vasodilator Agents/chemistryABSTRACT
The stereoselective preparation of diosgenin-derived thio(seleno)ureas and glycomimetics bearing a 1,2,3-triazolyl tether on C-3 has been accomplished. The key steps in the synthetic pathway are the incorporation of an amino moiety and its further transformation into thio- and selenoureas, and also a click chemistry reaction involving a propargyl residue and an azido moiety to afford carbohydrate-derived 1,2,3-triazoles; subsequent BF3-promoted acetolysis of the spiranic moiety afforded the corresponding 22-oxocholestanic structure. The N-phenyl selenourea, an hitherto unknown steroidal derivative, turned out to be a potent ROS scavenger, in particular against free radicals (EC50 = 29.47 ± 2.33 µM, DPPH method), and as a glutathione peroxidase mimic in the elimination of H2O2 (t1/2 = 4.8 min, 1% molar ratio). 22-Oxocholestane structures bearing a C-3 azido, propargyl, thioureido, and particularly selenoureido moiety behaved as strong antiproliferative agents against HeLa cells (IC50 1.87-11.80 µM). N-phenyl selenourea also exhibited IC50 values lower than 6.50 µM for MDA-MB-231, MCF-7 and HepG2 cancer cells; apoptosis was found to be involved in its mode of action. Such compound was also capable of efficiently eliminating ROS endogenously produced by HeLa cells. Antiproliferative properties of thioxo and selenoxo derivatives were stronger than diosgenin.
Subject(s)
Diosgenin/chemistry , Diosgenin/pharmacology , Drug Design , Glycoconjugates/chemistry , Glycoconjugates/pharmacology , Organoselenium Compounds/chemistry , Triazoles/chemistry , Urea/analogs & derivatives , Animals , Antineoplastic Agents/chemistry , Antineoplastic Agents/metabolism , Antineoplastic Agents/pharmacology , Biomimetic Materials/chemistry , Biomimetic Materials/metabolism , Biomimetic Materials/pharmacology , Biphenyl Compounds/metabolism , Cell Line, Tumor , Cell Proliferation/drug effects , Diosgenin/metabolism , Drug Screening Assays, Antitumor , Free Radical Scavengers/chemistry , Free Radical Scavengers/pharmacology , Glutathione Peroxidase/metabolism , Glycoconjugates/metabolism , Humans , Mice , Picrates/metabolism , Reactive Oxygen Species/metabolism , Urea/chemistryABSTRACT
Brachiaria spp. are the most important grasses for ruminants in central-western Brazil. However, the use of these pastures is limited by their toxicity due to steroidal saponins. This experiment was conducted for 60 days to demonstrate the resistance of sheep raised on Brachiaria spp. pastures to steroidal saponin poisoning. The experiment was composed by 48 animals randomly divided into four groups (n = 12). Among them, 32 4- to 5-month-old castrated male crossbred Santa Inês sheep, originating from flocks that had been grazing on Brachiaria spp. for more than three consecutive years, and 16 were non-adapted (naïve) sheep from flocks that never had prior contact with pastures of Brachiaria spp. were randomly divided into four groups. Each of the four experimental groups was composed by eight adapted and four non-adapted animals. The four experimental groups were introduced into paddocks, each of which contained a single grass: either Brachiaria decumbens, Brachiaria brizantha, Panicum maximum, or Andropogon gayanus. The addition of the naïve sheep to the groups was designed to detect pastures' toxicity to naïve sheep and to adjust the stocking rate to optimize the use of forage. The weight gains of sheep grazing on B. decumbens, B. brizantha, and P. maximum were similar; however, the A. gayanus group showed lower weight gains compared with the other groups (P < 0.05). The mean serum activities of γ-glutamyltransferase in the sheep grazing on B. decumbens were higher than those in the sheep from the other groups. No significant differences among the groups were found in aspartate aminotransferase, creatinine, albumin, or total protein serum concentrations. No clinical signs were observed in the adapted sheep in any of the pastures. Of the four non-adapted sheep introduced into the B. decumbens pasture, two showed clinical signs of steroidal saponin poisoning, and one died. No clinical signs were observed in the non-adapted sheep in the other pastures. The saponin (protodioscin) concentrations in the pastures varied from 3.3 to 12.2 g/kg DM in B. decumbens, from 2.8 to 9.1 g/kg DM in B. brizantha, and from 1 to 1.5 g/kg DM in A. gayanus. No saponins were found in P. maximum. It is concluded that sheep from flocks reared in pastures of B. decumbens and B. brizantha were resistant to steroidal saponin poisoning and showed similar weight gains to those of sheep grazing in other tropical pastures.
Subject(s)
Brachiaria/chemistry , Diosgenin/analogs & derivatives , Panicum/chemistry , Plant Poisoning/veterinary , Saponins/chemistry , Sheep Diseases/chemically induced , Andropogon , Animals , Brazil/epidemiology , Diosgenin/chemistry , Male , Plant Poisoning/epidemiology , Sheep , Sheep Diseases/epidemiologyABSTRACT
This report presents evidence that the following Solanum steroids: solasodine, diosgenin and solanine interact with human erythrocytes and molecular models of their membranes as follows: a) X-ray diffraction studies showed that the compounds at low molar ratios (0.1-10.0mol%) induced increasing structural perturbation to dimyristoylphosphatidylcholine bilayers and to a considerable lower extent to those of dimyristoylphosphatidylethanolamine; b) differential scanning calorimetry data showed that the compounds were able to alter the cooperativity of dimyristoylphosphatidylcholine, dimyristoylphosphatidylethanolamine and dimyristoylphosphatidylserine phase transitions in a concentration-dependent manner; c) in the presence of steroids, the fluorescence of Merocyanine 540 incorporated to the membranes decreased suggesting a fluidization of the lipid system; d) scanning electron microscopy observations showed that all steroids altered the normal shape of human erythrocytes inducing mainly echinocytosis, characterized by the formation of blebs in their surfaces, an indication that their molecules are located into the outer monolayer of the erythrocyte membrane.
Subject(s)
Diosgenin/chemistry , Erythrocyte Membrane/chemistry , Lipid Bilayers/chemistry , Solanaceous Alkaloids/chemistry , Solanine/chemistry , Calorimetry, Differential Scanning , Dimyristoylphosphatidylcholine/chemistry , Diosgenin/pharmacology , Erythrocyte Membrane/drug effects , Fluorescent Dyes/chemistry , Humans , Microscopy, Electron, Scanning , Phase Transition/drug effects , Phosphatidylethanolamines/chemistry , Phosphatidylserines/chemistry , Pyrimidinones/chemistry , Scattering, Small Angle , Solanaceous Alkaloids/pharmacology , Solanine/pharmacology , X-Ray DiffractionABSTRACT
Two unnatural steroid sapogenins bearing a furospirostane side chain were prepared starting from the readily available spirostane sapogenins, tigogenin and diosgenin following a synthetic protocol that included: (i) introduction of a carbonyl group at position C-23, (ii) diacetoxyiodobenzene-induced F-ring contraction and (iii) LiAlH4 reduction of the newly emerged methoxycarbonyl moiety. The structures of the new compounds were corroborated by NMR and X-ray studies.
Subject(s)
Acetates/chemistry , Diosgenin/chemistry , Iodobenzenes/chemistry , Sapogenins/chemical synthesis , Spirostans/chemistry , Crystallography, X-Ray , Hydrolysis , Magnetic Resonance Spectroscopy , Molecular ConformationABSTRACT
The naturally occurring dinorcholanic lactone vespertilin and two other non-natural derivatives bearing the 5α-hydroxy-6-oxo moiety were synthesized starting from the readily available steroid sapogenin diosgenin. The obtained compounds showed plant growth promoting activity in the bean's second internode elongation assay.
Subject(s)
Fabaceae/growth & development , Lactones/chemistry , Lactones/metabolism , Plant Growth Regulators/chemistry , Plant Growth Regulators/metabolism , Diosgenin/chemistry , Fabaceae/chemistry , Fabaceae/metabolism , Lactones/chemical synthesis , Plant Growth Regulators/chemical synthesis , Sapogenins/chemistry , Seeds/chemistry , Seeds/growth & development , Seeds/metabolismABSTRACT
The regioselective opening of the F ring of 22-oxo-23-spiroketals using a saturated solution of HCl in acetic anhydride yielded novel cholestanic frameworks with pyranone or cyclopentenone E rings. The structures of the new derivatives of sarsasapogenin, diosgenin and hecogenin thus obtained were established using one and two dimensional (1)H, (13)C experiments (DEPT, COSY, HETCOR, HMBC, ROESY, and NOESY). The X-ray analysis for compound 11b confirmed the 23R configuration for the new stereogenic center.
Subject(s)
Cholestanes/chemistry , Cyclopentanes/chemistry , Furans/chemistry , Pyrones/chemistry , Spiro Compounds/chemistry , Acetic Anhydrides/chemistry , Crystallography, X-Ray , Diosgenin/chemistry , Hydrochloric Acid/chemistry , Magnetic Resonance Spectroscopy/methods , Models, Chemical , Molecular Structure , Sapogenins/chemistry , Spirostans/chemistry , StereoisomerismABSTRACT
Certain steroidal compounds have demonstrated an antiproliferative effect against several tumor cell lines; however, their complete role on cancer cells is not currently established. Herein, we report the synthesis and evaluation of two new 26-hydroxy-22-oxocholestanic steroids on cervical cancer CaSki cells. The title compounds were prepared from diosgenin and hecogenin in excellent yields. We determined their effect on cell proliferation, cell cycle, and cell death. The cytotoxic effect of the title compounds on CaSki and human lymphocytes was also evaluated, indicating that the main cell death process is not necrosis; the null effect on lymphocytes implies that they are not cytotoxic. The observation of apoptotic bodies as well as the increase in the expression of active caspase-3 along with the fragmentation of DNA confirmed that such new cholestanic frameworks induced apoptosis in tumor cells. Significantly, their antiproliferative activity on tumor cells did not affect the proliferative potential of normal fibroblasts from cervix and peripheral blood lymphocytes. The title compounds show selective antitumor activity and therefore serve as promising lead candidates for further optimization.
Subject(s)
Antineoplastic Agents, Phytogenic/chemical synthesis , Antineoplastic Agents, Phytogenic/pharmacology , Apoptosis/drug effects , Cholestanols/chemistry , Cholestanols/chemical synthesis , Cholestanols/pharmacology , Diosgenin/chemistry , Sapogenins/chemistry , Steroids/chemistry , Steroids/chemical synthesis , Steroids/pharmacology , Carbohydrate Sequence , Caspase 3/metabolism , Cell Cycle/drug effects , Cell Line, Tumor , Cell Proliferation/drug effects , DNA Fragmentation/drug effects , Female , Fibroblasts/drug effects , Fluorescent Dyes , Humans , In Situ Nick-End Labeling , Indoles , Lymphocytes/drug effects , Magnetic Resonance Spectroscopy , Molecular Sequence Data , Structure-Activity Relationship , Uterine Cervical Neoplasms/drug therapy , Uterine Cervical Neoplasms/pathologyABSTRACT
We report the deacylation of (20R)-20-acetyl-23,24-dinorcholanic lactones by hydrazine hydrate, under microwave irradiation in high yields. The elimination of the 20-acetyl group proceeded with retention of configuration which contrast with other proved deacylation methods that yield a mixture of diastereoisomers. In this way, unnatural (20R)-23,24-dinorcholanic lactones can be produced rapidly on a large scale. Both (20R)- and (20S)-lactones were prepared starting from diosgenin, hecogenin and sarsasapogenin, in 72-80% overall yields.
Subject(s)
Diacetyl/chemistry , Lactones/chemistry , Acylation , Antineoplastic Agents/chemical synthesis , Antineoplastic Agents/chemistry , Diacetyl/chemical synthesis , Diosgenin/chemistry , Hydrazines/chemistry , Lactones/chemical synthesis , Microwaves , Models, Molecular , Molecular Structure , Sapogenins/chemistry , Spirostans/chemistry , Stereoisomerism , X-Ray DiffractionABSTRACT
As part of an experimental study, crystal-associated cholangiopathy was induced in 9 sheep by grazing pure pastures of Brachiaria decumbens in Brazil. One of these sheep showed characteristic lesions of photosensitization. The analysis of the B decumbens samples by acidic hydrolysis followed by TLC and infrared spectrum revealed diosgenin as the principal sapogenin present in the plant. In the rumen contents samples from the B decumbens-grazing group were identified by TLC, 1H and 13C NMR and EIMS as epismilagenin, episarsasapogenin, and a mixture of smilagenin and sarsasapogenin. In the bile samples from the B decumbens-grazing group, TLC analysis demonstrated 2 compounds similar to epismilagenin and episarsasapogenin. However, by this same method, those compounds were not observed in the rumen contents and bile from 2 sheep which served as control animals. The P chartarum spore counts remained very low during the experimental period.
Subject(s)
Cholangitis/veterinary , Liver Diseases/veterinary , Photosensitivity Disorders/veterinary , Poaceae/chemistry , Sapogenins/isolation & purification , Sheep Diseases/etiology , Animals , Ascomycota/growth & development , Bile/chemistry , Brazil , Cholangitis/chemically induced , Cholangitis/etiology , Chromatography, Thin Layer/veterinary , Colony Count, Microbial/veterinary , Diosgenin/chemistry , Diosgenin/isolation & purification , Diosgenin/toxicity , Histocytochemistry , Liver/pathology , Liver Diseases/etiology , Liver Diseases/pathology , Magnetic Resonance Spectroscopy , Mass Spectrometry/veterinary , Photosensitivity Disorders/chemically induced , Photosensitivity Disorders/etiology , Poaceae/microbiology , Poaceae/toxicity , Rumen/chemistry , Sapogenins/chemistry , Sapogenins/toxicity , Sheep , Sheep Diseases/microbiology , Sheep Diseases/pathology , Spectrophotometry, Infrared/veterinary , Spirostans/chemistry , Spirostans/isolation & purification , Spirostans/toxicity , Spores, Fungal/growth & developmentABSTRACT
The period from late 1949 through 1951 was an extraordinarily productive one in steroid chemistry and especially so at Syntex S.A. in Mexico City. Two of the most important Syntex contributions--the synthesis of 19-nor-17 alpha-ethynyltestosterone (norethindrone) and of cortisone from diosgenin--are described from a historical perspective.
PIP: The pharmaceutical industry contributed more to the published record of steroid research during the 1950s than any industry has ever contributed before to any chemical subdiscipline. Syntex, a research-oriented company in Mexico city, contributed much of the publication of industrial research of steroids. Dr. Djerassi arrived at Syntex in 1949 as associate director of chemical research. He conducted partial aromatization studies leading him to the 1st synthesis of an oral contraceptive (OC) on October 15, 1951. This steroid was 19-nor-17 alpha-ethynyltestosterone, later called norethistrone or norethindrone. Syntex submitted the product to a commercial laboratory in Madison, Wisconsin, for biological evaluation. It was indeed the most active, orally effective progestational hormone at the time. Syntex applied for a patent in November 1951. In November 1954, clinical results of norethindrone used to treat various menstrual disorders and fertility problems was presented. G.D. Searle & Co. filed for a patent for the synthesis of the double bond isomer 13 of norethindrone called norethynodrel in August 1953. Acid or human gastric juice converts norethynodrel into norethindrone. Had it not been for Searle using norethindrone in its antimotion sickness drug, Dramamine, Syntex would have filed suit against Searle. Syntex sponsored contraceptive trials with norethindrone. Various incidents prevented Syntex from obtaining US Food and Drug Administration approval to use norethindrone for contraceptive indications before Searle obtained approval to use norethynodrel. By 1964, 3 companies including Syntex were marketing 2 mg doses of Syntex's norethindrone, the most widely used active ingredient in OCs. Dr. Djerassi also played a key role in the synthesis of cortisone from diosgenin, a chemical derived from Mexican yams. This synthesis was a more economical industrial route to cortisone than previous routes.