Steroid Derivatives Research Articles

Synthesis, In Vitro Biological Evaluation of Antiproliferative and Neuroprotective Effects and In Silico Studies of Novel 16E-Arylidene-5α,6α-epoxyepiandrosterone Derivatives

Steroids constitute an important class of pharmacologically active molecules, playing key roles in human physiology. Within this group, 16E-arylideneandrostane derivatives have been reported as potent anti-cancer agents for the treatment of leukemia, breast and prostate cancers, and brain tumors. Additionally, 5α,6α-epoxycholesterol is an oxysterol with several biological activities, including regulation of cell proliferation and cholesterol homeostasis. Interestingly, pregnenolone derivatives combining these two modifications were described as potential neuroprotective agents. In this research, novel 16E-arylidene-5α,6α-epoxyepiandrosterone derivatives were synthesized from dehydroepiandrosterone by aldol condensation with different aldehydes followed by a diastereoselective 5α,6α-epoxidation. Their cytotoxicity was evaluated on tumoral and non-tumoral cell lines by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Furthermore, the assessment of the neuroprotective activity of these derivatives was performed in a dopaminergic neuronal cell line (N27), at basal conditions, and in the presence of the neurotoxin 6-hydroxydopamine (6-OHDA). Interestingly, some of these steroids had selective cytotoxic effects in tumoral cell lines, with an IC50 of 3.47 µM for the 2,3-dichlorophenyl derivative in the breast cancer cell line (MCF-7). The effects of this functionalized epoxide on cell proliferation (Ki67 staining), cell necrosis (propidium iodide staining), as well as the analysis of the nuclear area and near neighbor distance in MCF-7 cells, were analyzed. From this set of biological studies, strong evidence of the activation of apoptosis was found. In contrast, no significant neuroprotection against 6-OHDA-induced neurotoxicity was observed for the less cytotoxic steroids in N27 cells. Lastly, molecular docking simulations were achieved to verify the potential affinity of these compounds against important targets of steroidal drugs (androgen receptor, estrogen receptor α, and 5α-reductase type 2, 17α-hydroxylase-17,20-lyase and aromatase enzymes). This in silico study predicted a strong affinity between most novel steroidal derivatives and 5α-reductase and 17α-hydroxylase-17,20-lyase enzymes.

Oxazolinyl derivatives of androst-16-ene as inhibitors of CYP17A1 activity and prostate carcinoma cells proliferation: Effects of substituents in oxazolinyl moiety

Steroid derivatives modified with nitrogen containing heterocycles are known to inhibit activity of steroidogenic enzymes, decrease proliferation of cancer cells and attract attention as promising anticancer agents. Specifically, 2′-(3β-hydroxyandrosta-5,16-dien-17-yl)-4′,5′-dihydro-1′,3′-oxazole 1a potently inhibited proliferation of prostate carcinoma cells. In this study we synthesized and investigated five new derivatives of 3β-hydroxyandrosta-5,16-diene comprising 4′-methyl or 4′-phenyl substituted oxazolinyl cycle 1 (b–f). Docking of compounds 1 (a–f) to CYP17A1 active site revealed that the presence of substitutents at C4′ atom in oxazoline cycle, as well as C4′ atom configuration, significantly affect docking poses of compounds in the complexes with enzyme. Testing of compounds 1 (a–f) as CYP17A1 inhibitors revealed that the only compound 1a, comprising unsubstituted oxazolinyl moiety, demonstrated strong inhibitory activity, while other compounds 1 (b–f) were slightly active or non active. Compounds 1 (a–f) efficiently decreased growth and proliferation of prostate carcinoma LNCaP and PC-3 cells at 96 h incubation; the effect of compound 1a was the most powerful. Compound 1a efficiently stimulated apoptosis and caused PC-3 cells death, that was demonstrated by a direct comparison of pro-apoptotic effects of compound 1a and abiraterone.

Anabolic–androgenic steroid abuse: psychiatric manifestations and treatment

IntroductionAnabolic-androgenic steroids (AAS) are synthetic derivatives of steroids which are frequently utilized in order to increase muscle mass and strength. Estimates of AAS abuse vary according to different studies. However, a global lifetime prevalence of 3.3% has been reported. The increasing illicit use of AAS over the years has raised serious public health concerns.ObjectivesTo report a clinical case of AAS abuse and review the psychopathology associated with AAS abuse and its treatment.MethodsThe authors report a clinical case and conducted a non-systematic review on the Pubmed database with the terms “anabolic-androgenic steroids”, “abuse” and “psychiatric manifestations”.ResultsA 25-year-old male patient with history of body dysmorphic disorder was admitted to the emergency services following severe psychomotor agitation and verbal and physical aggression towards his family members. He explained he had begun using AAS years prior in order to improve his physical appearance. Recently he had been administering AAS injections on a more regular basis and restricting his food and water intake. His sleep-wake cycle was disrupted and he began experiencing increasing anxiety and persecutory ideation. Additionally, increased aggression was reported and numerous conflicts with his family members took place in the weeks previous to his psychiatric observation. AAS abuse has been associated with psychiatric symptoms such as aggression and violence, mania and less commonly psychosis and suicidal ideation. Moreover, its prolonged abuse can lead to symptoms of dependence and withdrawal following suspension. A biphasic model of steroid dependence has been described. First there is a brief hyperadrenergic state with opoid-like withdrawal symptoms which is then followed by a prolonged period of depression and craving. During this phase symptoms such as fatigue, muscle and joint pain, insomnia, anxiety and depression may occur. There are no established guidelines for treatment. Acute care of agitation should follow a similar course as the one utilized in other forms of substance induced-agitation: firstly, the least invasive interventions should be implemented. If medication is required, haloperidol has been reported to be effective, although evidence is scarce. Benzodiazepines may be considered, although its use with AAS has not been reported. In regards to long term care, AAS discontinuation is vital in conjunction with proper management of withdrawal symptoms. If a patient presents symptoms of opioid-like withdrawal, treatment with clonidine may be initiated. The use of medically prescribed steroids has been suggested in order to alleviate withdrawal symptoms. Other treatment options such as human chorionic gonadotrophin and clomiphene have also been proposed.ConclusionsAAS abuse is a serious public health concern. Clinicians should be aware of its serious psychiatric effects and possible treatment courses.Disclosure of InterestNone Declared

Improved 2α-Hydroxylation Efficiency of Steroids by CYP154C2 Using Structure-Guided Rational Design.

Cytochrome P450 enzymes are promising biocatalysts for industrial use because they catalyze site-selective C-H oxidation and have diverse catalytic reactions and a broad substrate range. In this study, the 2α-hydroxylation activity of CYP154C2 from Streptomyces avermitilis MA-4680T toward androstenedione (ASD) was identified by an in vitro conversion assay. The testosterone (TES)-bound structure of CYP154C2 was solved at 1.42 Å, and this structure was used to design eight mutants, including single, double, and triple mutants, to improve the conversion efficiency. Mutants L88F/M191F and M191F/V285L were found to enhance the conversion rates significantly (i.e., 8.9-fold and 7.4-fold for TES, 46.5-fold and 19.5-fold for ASD, respectively) compared with the wild-type (WT) enzyme while retaining high 2α-position selectivity. The substrate binding affinity of the L88F/M191F mutant toward TES and ASD was enhanced compared with that of WT CYP154C2, supporting the measured increase in the conversion efficiencies. Moreover, the total turnover number and kcat/Km of the L88F/M191F and M191F/V285L mutants increased significantly. Interestingly, all mutants containing L88F generated 16α-hydroxylation products, suggesting that L88 in CYP154C2 plays a vital role in substrate selectivity and that the amino acid corresponding to L88 in the 154C subfamily affects the orientation of steroid binding and substrate selectivity. IMPORTANCE Hydroxylated derivatives of steroids play essential roles in medicine. Cytochrome P450 enzymes selectively hydroxylate methyne groups on steroids, which can dramatically change their polarity, biological activity and toxicity. There is a paucity of reports on the 2α-hydroxylation of steroids, and documented 2α-hydroxylate P450s show extremely low conversion efficiency and/or low regio- and stereoselectivity. This study conducted crystal structure analysis and structure-guided rational engineering of CYP154C2 and efficiently enhanced the conversion efficiency of TES and ASD with high regio- and stereoselectivity. Our results provide an effective strategy and theoretical basis for the 2α-hydroxylation of steroids, and the structure-guided rational design of P450s should facilitate P450 applications in the biosynthesis of steroid drugs.

3D QSAR Studies of 3, 16 and 17 Position Modifications in Steroidal Derivatives for CNS Anticancer Activity

3D QSAR Studies of 3, 16 and 17 Position Modifications in Steroidal Derivatives for CNS Anticancer Activity

Synthesis, antiproliferative evaluation and in silico studies of a novel steroidal spiro morpholinone

Estrogens play a pivotal role in the development of estrogen-dependent breast cancer and other hormone-dependent disorders. A common strategy to overcome the pathological effects of estrogens is the use of aromatase inhibitors (AIs), which bind to the enzyme and prevent the union with the natural substrate, decreasing the amount of estrogens produced. Several AIs have been developed, including inhibitors with a steroidal backbone and a nitrogen heterocycle in their structure. Encouraged by the notable results presented by current and clinical steroidal drugs, herein we present the synthesis of a steroidal spiro morpholinone derivative as a plausible aromatase inhibitor.The morpholinone derivative was synthesized over a six-step methodology starting from estrone. The title compound and its hydroxychloroacetamide derivative precursor were evaluated for their antiproliferative profile against estrogen-dependent and independent solid tumor cell lines: A549, HBL-100, HeLa, SW1573, T-47D and WiDr. Both compounds exhibited a potent antiproliferative activity in the micromolar range against the six cancer cell lines, with the hydroxychloroacetamide derivative precursor being a more potent inhibitor (GI50 = 0.25–2.4 µM) than the morpholinone derivative (GI50 = 2.0–11 µM). Furthermore, both compounds showed, in almost all cases, better GI50 values than the steroidal anticancer drugs abiraterone and galeterone.Docking simulations of the derivatives were performed in order to explain the experimental biological activity. The results showed interactions with the iron heme (derivative 3) and important residues of the steroidal binding-site (Met374) for the inhibition of human aromatase. A correlation was found between in vitro assays and the score obtained from the molecular docking study.

Production of 11α-Hydroxysteroid Derivatives by Corynebacterium glutamicum Expressing the Rhizopus oryzae Hydroxylating System.

Hydroxylation of steroids has acquired special relevance for the pharmaceutical industry. Particularly, the 11α-hydroxylation of steroids is a process of biotechnological importance currently carried out at industrial scale for the production of contraceptive drugs and glucocorticoids. This process is performed by several fungal species including Rhizopus nigricans, Aspergillus ochraceus, Aspergillus niger, and Rhizopus oryzae that are used to produce by biotransformation hydroxylated steroids for pharmaceutical purposes (Wang et al., J Steroid Biochem Mol Biol 171:254-261, 2017). However, the development of more efficient biotransformation processes is essential since the steroidal derivatives obtained by the in vivo hydroxylation are often a mixture of hydroxylated compounds in different positions of the steroid molecule. This phenomenon is due to the large number of different CYPs contained in the fungal strains.The genes responsible for the 11α-hydroxylase activity in R. oryzae consisting in the cytochrome CYP509C12 and its redox partner, the reductase RoCPR1, have been chemically synthetized forming a synthetic operon named FUN optimized to be expressed in bacteria. To express this operon, we have selected the strain Corynebacterium glutamicum R31 that is a robust and GRAS bacterial strain widely used for industrial purposes. The synthetic operon has been cloned in the pECXK-99E vector, yielding pXKFUN plasmid, and transformed C. glutamicum R31 to generate C. glutamicum R31 (pXKFUN) strain. This strain is not a steroid degrader and can efficiently transport C19 and C21 steroids across the cytoplasmic membrane (García-Fernandez et al. Catalysts 316:1-12, 2017). C. glutamicum can be used as a clean host for steroid biotransformation, because it does not introduce additional undesired side reactions on the steroids, thus reducing the contamination of the final products (Felpeto-Santero et al., Microbiol Biotechnol 12:856-868, 2019). Here we show a proof of concept that C. glutamicum can be used as a suitable chassis to perform steroid biotransformation expressing eukaryotic cytochromes. The protocol below provides detailed information on steroid 11α-hydroxylations by Corynebacterium recombinant strain.

Effects of long-term intake of whole wheat and aleurone-enriched Chinese steamed bread on gut microbiome and liver metabolome in mice fed high-fat diet

The protective effects of whole wheat flour steamed bread (WWF) and aleurone-enriched wheat flour steamed bread (AWF) on gut microbiota and metabolomic disorders were investigated in long-term high-fat diet (HFD) mice. The results showed that AWF and WWF significantly ameliorated dyslipidemia, hepatic oxidative stress, and damage in HFD-fed mice. The fecal microbiome analysis showed that AWF and WWF significantly increased diversity in the gut microbiota and altered the structure of the gut microbiota. An increase in Bacteroidetes (by 31% and 34%) and decrease in Firmicutes (by 17% and 17%) were observed in the AWF and WWF groups, respectively. Liver metabolomics demonstrated that AWF and WWF upregulated carboxylic acids and derivatives and organooxygen compounds, and downregulated fatty acyl compounds. Moreover, WWF significantly modulated the concentrations of steroids and steroid derivatives, and glycerophospholipids. Overall, our findings suggest that AWF and WWF dramatically altered hepatic metabolism concurrent with shifts in specific gut bacteria in C57BL/6J mice, exhibiting potential as functional foods for the prevention of metabolic disorders induced by a long-term HFD. Despite some differences, the AWF is generally able to achieve the same level of health benefits as the WWF.

Protective effect of biogenic selenium nanoparticles against diquat-induced acute toxicity via regulation of gut microbiota and its metabolites.

Selenium nanoparticles (SeNPs) with unique biological properties have been suggested as a safer and more effective platform for delivering of Selenium for biological needs. In this study, we investigated the association between gut microbiota altered by SeNPs supplementation and its metabolites under oxidative stress conditions through 16S rDNA gene sequencing analysis and untargeted metabolomics. The results showed that dietary supplementation with SeNPs attenuated diquat-induced acute toxicity in mice, as demonstrated by lower levels of inflammatory effector cells, and biochemical markers in serum such as aspartate aminotransferase (AST), alanine aminotransferase (ALT), blood urea nitrogen (BUN) and lactate dehydrogenase (LDH). SeNPs also reversed the perturbed gut microbiota composition induced by diquat, decreased the Firmicutes/Bacteroidetes ratio, and increased the abundance of beneficial bacteria such as Akkermansia, Muribaculaceae, Bacteroides and Parabacteroides. Untargeted fecal metabolomics showed that SeNPs can regulate the production of steroids and steroid derivatives, organonitrogen compounds, pyridines and derivatives and other metabolites. Microbiome-metabolome correlation analysis suggested that Parabacteroides was the key bacteria for the SeNPs intervention, which might upregulate the levels of metabolites such as trimethaphan, emedastine, berberine, desoxycortone, tetrahydrocortisone. This study demonstrated that dietary SeNPs supplementation can extensively modulate the gut microbiota and its metabolism, thereby alleviating diquat-induced acute toxicity.

Two new antimicrobial steroids and other constituents from the whole plant of Ludwigia abyssinica

The genus Ludwigia belongs to the Onagraceae family and it encompasses seventy-five species of aquatic plants. The chemistry of this genus is scarcely investigated, although some studies have demonstrated the potential of Ludwigia leptocarpa to produce important bioactive compounds. Herein, we describe the phytochemical investigation of Ludwigia abyssinica A. Rich. Two new steroids named 3 β -formyloxy-5 α ,6α-dihydroxysitostane (Ludwigiaformyl A, 1 ) and 3 β ,6 α -diformyloxy-5 α -hydroxysitostane (Ludwigiaformyl B, 2 ), along with six known compounds, 3 β -formyloxysitost-5-en ( 3 ), 5 α ,6 β -dihydroxysitosterol ( 4 ), maslinic acid ( 5 ), oleanolic acid ( 6 ) and a mixture of two iridoids: linearin ( 7 ) and 1-epilinearin ( 8 ) were obtained from whole plant of L. abyssinica . The structures of the isolated compounds were established by extensive analysis of their spectroscopic and spectrometric data, which included HR-TOF-ESIMS, 1 D NMR ( 1 H, 13 C) and 2 D NMR ( 1 H– 1 H COSY, HSQC, HMBC and ROESY) and by comparison with data reported in the literature. The antimicrobial activities of extracts, fractions, and new compounds ( 1) and ( 2 ) were evaluated using broth microdilution method against fungi and bacteria strains. The MeOH extract and the ethyl acetate fraction displayed different degrees of antibacterial and antifungal activities (MIC = 32 – 512 µg/mL; MMC = 64 – 512 µg/mL) whereas compounds 1 and 2 showed moderate antimicrobial activities against Pseudomonas aeruginosa , Staphylococcus aureus , Escherichia coli , Shigella flexneri and Cryptococcus neoformans (MIC = 8 – 32 µg/mL; MMC = 8 – 64 µg/mL). • Two new steroid derivatives trivially named Ludwigiaformyl A and B, were isolated from Ludwigia abyssinica. • Their structures were characterized by extensive 2 D NMR studies. • Six known compounds were also isolated from this plant. • Antimicrobial activities were evaluated.

In silico ADMET analysis of the A-, B- and D-modified androstane derivatives with potential anticancer effects

The major challenge in the fight against cancer is to design new drugs that will be more selective for cancer cells, with fewer side effects. Synthetic steroids such as cyproterone, fulvestrant, exemestane and abiraterone are approved powerful drugs for the treatment of hormone-dependent diseases such as breast and prostate cancers. Therefore, androstane derivatives in 17-substituted, 17a-homo lactone and 16,17-seco series, with potent anticancer activity, were selected for pharmacokinetic and druglike predictions from the absorption, distribution, metabolism and excretion (ADME) models. In silico determination of physico-chemical and ADMET properties was performed using SwissADME and ProTox-II web tools. The possibility of gastrointestinal absorption and brain penetration was analyzed using the BOILED-Egg model, while the in silico evaluation of the similarities between selected steroid derivatives and FDA-approved drugs was carried out using the SwissSimilarity tool. Of all tested, two compounds that showed good in silico ADMET results, in addition to promising cytotoxicity and molecular docking results, could potentially be evaluated in in vivo tests.

L‐Proline Catalyzed Synthesis of Steroidal Pyridones And Their DFT Studies

AbstractA steroidal‐based synthetic protocol has been developed for the synthesis of substituted steroidal pyridones in the presence of L‐proline. A wide variety of steroidal pyridones were synthesized in better yields. Initially, steroidal benzylidenes were prepared by the aldol condensation of pregnenolone with various substituted aromatic aldehydes under basic conditions. After that heterocyclization of resulting benzylidenes with cyanoacetamide in the presence of L‐proline gives the desired products. With the mild reaction conditions, good conversions, operational simplicity, and inexpensive and readily available catalyst, it is a practical and appealing approach for pyridone synthesis. Structural and quantum chemical insights of synthesized steroidal pyridone derivatives have been also presented using the density functional theory (DFT) approach.

Metabolic Variations in Brown Rice Fertilised with Different Levels of Nitrogen

Nitrogen is a necessary element for plant growth; therefore, it is important to study the influence of N fertilisers on crop metabolites. In this study, we investigate the variability of endogenous metabolites in brown rice fertilised with different amounts of nitrogen. We identified 489 metabolites in brown rice. Compared to non-nitrogen fertilised groups, there were 59 differentially activated metabolic pathways in the nitrogen-fertilised groups. Additionally, there were significantly differential secondary metabolites, especially flavonoids, between groups treated with moderate (210 kg N/hm2) and excessive amounts of nitrogen (420 kg N/hm2). Nitrogen fertilisation upregulated linoleic acid metabolism and most steroids, steroid derivatives, and flavonoid compounds, which have antioxidant activity. The DPPH, ABTS, and hydroxyl radical scavenging rates were higher in fertilised groups than in the non-fertilised group. These findings provide a theoretical basis to enhance the health benefits of brown rice by improving fertilisation.

Multivariate Analysis Revealed Ultrasonic-Assisted Extraction Improves Anti-Melanoma Activity of Non-Flavonoid Compounds in Indonesian Brown Algae Ethanol Extract.

Indonesia has high biodiversity of algae that are under-utilised due to limitations in the processing techniques. Here, we observed the effect of two different extraction methods (cold maceration and ultrasonic-assisted extraction (UAE)) on multiple variables of Indonesian brown algae ethanol extracts (Sargassum polycystum, Sargassum cristaefolium, Sargassum aquifolium and Turbinaria ornata). The variables observed included metabolites screening by untargeted metabolomics liquid chromatography-high-resolution mass spectrometry (LC-HRMS), observation of total phenolic content (TPC), total flavonoid content (TFC), anti-oxidant and B16-F10 melanoma cells cytotoxicity. UAE extracts had higher extraction yield and TPC, but no TFC difference was observed. UAE extract had more lipophilic compounds, such as fatty acids (Palmitic acid, Oleamide, Palmitoleic acid, Eicosapentaenoic acid, α-Linolenic acid, Arachidonic acid), lipid-derived mediators (11,12-Epoxyeicosatrienoic acid ((±)11(12)-EET)), steroid derivatives (Ergosterol peroxide), lipophilic metabolite (Fucoxanthin), and lipid-soluble vitamins (all-trans-retinol). Principle component analysis (PCA) revealed that TPC, not TFC, in the UAE extracts was correlated with the anti-oxidant activities and cytotoxicity of the extracts towards B16-F10 melanoma cells. This means other non-flavonoid phenolic and lipophilic compounds may have contributed to its bioactivity. These results suggest that out of the two methods investigated, UAE could be a chosen method to extract natural anti-melanogenic agents from brown algae.

Integrated analysis of gene expressions and metabolite features unravel immunometabolic interplay in hybrid fish (Carassius cuvieri ♀ × Carassius auratus red var ♂) infected with Aeromonas hydrophila

Aeromonas hydrophila is one of important pathogenic bacteria that can imperil fish survival. Current study suggested that hybrid crucian carp (WR) receiving acute A. hydrophila challenge exerted a molecular regulation linking peroxisomal fatty acid β-oxidation to immune response. A. hydrophila infection may alleviate antioxidant status along with the significant alternation of immune-, apoptosis- and metabolic-related gene expressions in kidney, whereas most portions of differential metabolites (DMs) were enriched in steroid derivatives and phospholipids. In addition, mevalonate (MVA) pathway and phospholipid hydrolysis were identified as crucial cellular processes in WR after A. hydrophila challenge. These results may provide an insight into immunometabolic strategy against A. hydrophila infection.

Functional Characterization and Synthetic Application of Is2-SDR, a Novel Thermostable and Promiscuous Ketoreductase from a Hot Spring Metagenome.

In a metagenome mining-based search of novel thermostable hydroxysteroid dehydrogenases (HSDHs), enzymes that are able to selectively oxidize/reduce steroidal compounds, a novel short-chain dehydrogenase/reductase (SDR), named Is2-SDR, was recently discovered. This enzyme, found in an Icelandic hot spring metagenome, shared a high sequence similarity with HSDHs, but, unexpectedly, showed no activity in the oxidation of the tested steroid substrates, e.g., cholic acid. Despite that, Is2-SDR proved to be a very active and versatile ketoreductase, being able to regio- and stereoselectively reduce a diversified panel of carbonylic substrates, including bulky ketones, α- and β-ketoesters, and α-diketones of pharmaceutical relevance. Further investigations showed that Is2-SDR was indeed active in the regio- and stereoselective reduction of oxidized steroid derivatives, and this outcome was rationalized by docking analysis in the active site model. Moreover, Is2-SDR showed remarkable thermostability, with an apparent melting temperature (TM) around 75 °C, as determined by circular dichroism analysis, and no significant decrease in catalytic activity, even after 5 h at 80 °C. A broad tolerance to both water-miscible and water-immiscible organic solvents was demonstrated as well, thus, confirming the potential of this new biocatalyst for its synthetic application.

Investigation of Potential In Vitro Anticancer and Antimicrobial Activities of Balanites aegyptiaca (L.) Delile Fruit Extract and Its Phytochemical Components.

The therapeutic importance of Balanites aegyptiaca in folk medicine for the treatment of several common human diseases has led researchers to conduct phytochemical and pharmacological studies on extracts from various parts of the plant. In the current study, the phytochemical composition of the B. aegyptiaca methanolic fruit extract was characterized, and its antimicrobial activity was evaluated together with the cytotoxic activity against MCF-7, PC-3, and Caco-2, compared with normal Vero cells. Further, its effects on cell cycle arrest, apoptosis induction and expression of apoptosis-related genes were assessed. The phytochemical screening revealed the presence of fatty acids and their esters in addition to phytosterols, steroid derivatives, and bioflavonoid glycosides with oleic and palmitic acids being the prevalent components (24.12 and 21.56%, respectively). The results showed considerable cytotoxic activity of the extract against the three cancer cell lines (MCF-7, PC-3, and Caco-2) with a selectivity index ranging from 5.07 to 6.52. This effect was further confirmed with the accompanied increased total apoptosis of treated PC-3 cells (19.22% of the total number of cells) compared to the control cells (0.64% of the total number of cells) with cell cycle arrest at G1 phase and the increased transcription of pro-apoptotic genes including P53 (3.69) and BAX (3.33) expressed as fold change (2^ ΔΔCT). The calculated minimum inhibitory concentration (MIC) was similar (62.5 µg/mL) against the three tested bacterial strains (Acinetobacter johnsonii, Serratia marcescens and Agrobacterium tumefaciens), while it was higher than 1000 µg/mL for the fungal species (Rhizoctonia solani, Penicillium italicum, and Fusarium oxysporium). Our findings suggest a promising anticancer activity for B. aegyptiaca, which paves the way for more detailed future studies.

Design and synthesis of hetero-steroids via ring-closing metathesis: Biological studies towards in vitro anticancer activity

Here, we report a synthetic approach to hetero-steroids and also studied their biological activities as anticancer agents. A novel class of oxacycles containing estrone moiety were synthesized in this report. Allyl ether derived from estrone underwent Claisen rearrangement (CR) and again O-allylation and subsequent ring-closure gave A-ring-furan and oxepine fused derivatives in high yields. We used double bond isomerization and ring-closing metathesis (RCM) as key steps to assemble hetero steroids containing a mixture of regio isomers like benzofurans and benzoxepine moieties. The novel benzofuran and benzoxepine-based hybrid steroid derivatives were subjected to in vitro cytotoxicity analysis and were found to exert cancer cell-specific activity.

Trypsin Inhibitor Isolated From Glycine max (Soya Bean) Extraction, Purification, and Characterization.

The current study aims to isolate, purify, and characterize the trypsin inhibitor protein from seeds of soya beans, scientifically known as Glycine max. Its seeds were ground, and the powder was soaked several times using n-hexane. It was added to phosphate buffer saline (PBS) followed by filtration and centrifugation of the PBS dissolved extract. The supernatant was subjected to ammonium sulfate precipitation and about six fractions, 30% to 80% were prepared. The centrifuged pellets obtained from each fraction were dialyzed and run on SDS-PAGE. The trypsin inhibitor protein was precipitated and characterized in 30% pellet and molecular weight was 21.5 kDa compared to protein ladder (ThermoFisher 10-170 kDa). GC-MS analysis revealed the steroid derivatives such as stigmasterol, campesterol, beta-sitosterol, and gamma-tocopherol. Glycine max trypsin inhibitor could be used as a plant-derived drug to overcome the over-activation of trypsin without its real substrate (proteins) becoming activated and start auto digestion leading to pancreatitis.

3D-QSAR, Molecular Docking and Pharmacokinetic Studies: In-Silico Approach to Search Novel Inhibitors of 5-Alpha Reductase for Treatment of Benign Prostatic Hyperplasia

Aim:To identify novel steroidal 5-alpha reductase (5AR) inhibitors using computational approaches.Objectives:To exploit the steroidal nuclei for possible modifications by creating a library of 17-oximino-5-androsten-3-carboxamide derivatives and identify potent 5AR inhibitors based on docking and pharmacokinetic parameters.Background:Benign prostatic hyperplasia (BPH), is a condition of aged men, that is characterized by lower urinary tract symptoms. Excessive production of dihydrostestosterone (DHT) from testosterone has been found to play a major role in its pathophysiology. Studies targeting the 5AR enzyme have so far resulted in the development of two clinical approved 5AR inhibitors.Methods:Atom-based three dimensional-quantitative structure activity relationship (3D-QSAR) models have been developed using a selected series of steroidal derivatives as 5AR inhibitors, to elucidate the structural properties required for 5AR inhibitory activities. Further In‒silico studies (molecular docking and pharmacokinetic properties like adsorption, distribution, metabolism, and excretion) of 17-oximino-5-androsten-3-carboxamide derivatives have also been carried out to identify the binding orientation and protein-ligand interactions responsible for the exhibited activity and drug like properties.Results:The best 3D-QSAR model was generated using Partial Least Square method with an excellent correlation coefficient (R², training set) of 0.882, standard deviation (SD) of 0.09, and a predicted coefficient (Q², test set) of 0.814. Docking analysis indicated that the designed series of compounds have comparable binding affinity from -8.961 to -8.017 to the protein and suggested that hydrophobic and electrostatic moieties can have a key role in the inhibition mechanism.Conclusion:3D-QSAR, molecular docking and pharmacokinetic studies indicated that 17-oximino-5-androsten-3-carboxamide derivatives could be studied further to provide a new strategy for the treatment of BPH.