Base-Mediated, Chemo- and Regioselective (4+2) Annulation of Indole-2-carboxamides with 2,3-Epoxy Tosylates toward 1,2-Fused Indoles.

Base-mediated [4+2] annulation of indole-2-carboxamides with 2,3-epoxy tosylates has been explored. The protocol delivers 3-substituted pyrazino[1,2-a]indol-1-ones in high yields in diastereoselective fashion, and neither 4-substituted pyrazino[1,2-a]indol-1-ones nor tetrahydro-1H-[1,4]diazepino[1,2-a]indol-1-ones are generated, irrespective of whether the distal epoxide C3 substituent is alkyl or aryl, or the epoxide is cis- or trans-configured. This reaction proceeds in one pot via N-alkylation of the indole scaffold with 2,3-epoxy tosylates, concomitantly followed by 6-exo-selective epoxide-opening cyclization. Notably, the process is chemo- and regioselective with respect to both the starting materials. To our knowledge, the process represents the first successful example of one-pot annulation of indole-based diheteronucleophiles with epoxide-based dielectrophiles.

Diastereoselective Synthesis of Indoline- and Pyrrole-Embedded Tetracycles via an Unprecedented Dearomative Indole-C3-Alkylation/Aza-Friedel-Crafts Cascade Reaction.

Dearomative indole C3-alkylation─intramolecular iminium trapping cascade reaction of indole-C3-tethered nucleophiles is a well-known blueprint for accessing 2,3-fused indolines. In exploring this strategy, synthetic chemists have utilized diverse classes of electrophilic reagents. However, the tethered nucleophiles have mainly been limited to heteronucleophiles and enolates; exploitation of tethered arenes/heteroarenes remains unknown. We herein describe the first examples of pyrrole-intercepted dearomative indole C3-allylation and benzylation of indole-tethered pyrroles toward the synthesis of 2,3-cis-fused tetracyclic indolines featuring a C3 all-carbon quaternary stereocentre. Our methodology capitalizes on the capability of NaOtBu/Et3B combination to direct the intermolecular alkylation to take place regioselectively at the indole C3 position over the other reactive sites (indole N and C2 and pyrrole C2 positions) and leverages the high nucleophilicity of the pyrrole template for the concomitant aza-Friedel-Crafts ring closure that traditionally would require an additional acid-catalyzed synthetic step. This cascade reaction is accomplished with broad substrate scope and excellent yields and chemo-, regio-, and diastereoselectivities.

Synthesis of salicylic acid phenylethyl ester (SAPE) and its implication in immunomodulatory and anticancer roles.

Salicylic acid phenylethyl ester (SAPE) was synthesized by Zn(OTf)2-catalyzed selective esterification of salicylic acid and phenylethyl alcohol and studied for its role as an immunomodulatory and anticancer agent. Low toxicity and favorable physical, Lipinski-type, and solubility properties were elucidated by ADME-tox studies. Molecular docking of SAPE against COX-2 revealed favorable MolDockscore, rerank score, interaction energy, internal pose energy, and hydrogen bonding as compared to ibuprofen and indomethacin. An average RMSD of ~ 0.13 nm for the docked complex with stable dynamic equilibrium condition was noted during the 20 ns MD simulation. A low band gap predicting a strong binding affinity at the enzyme's active site was further predicted by DFT analysis. The ester caused a reduction in the percentage of erythrocyte hemolysis and was shown to be non-cytotoxic against human lymphocytes, CaCo-2, and HepG-2 cells by the MTT assay. Moreover, it's in vitro efficacy in inhibiting COX-2 enzyme under both LPS stimulated intestinal cells and direct sequestration assays was found to be higher than salicylic acid and indomethacin. The anticancer activity of SAPE was tested on the breast cancer cell line MCF-7, and potential efficacy was exhibited in terms of decreased cell viability. Flow cytometry analysis exhibited the arrest of the cell cycle at G1/G0 and S phases, during which induction of autophagic vesicle formation and decrease in mitochondrial membrane potential was observed owing to increased ROS production. Furthermore, at these phases, the onset of apoptosis along with DNA damage was also observed. Pre-treatment with SAPE in colitis-induced Wistar rats displayed low disease activity index and reduction in the extent of intestinal tissue disruption and lipid peroxidation. A marked increase of anti-oxidative enzymes viz., catalase, GGT, and GST, and a decrease of pro-inflammatory cytokines IL-6 and TNF-α in the intestinal tissue extracts of the treated groups was noted. The results of this study have sufficient credence to support that the synthesised ester (SAPE) be considered as an anti-oxidative and anti-inflammatory compound with therapeutic potential for the effective management of cancer.

One-Pot Double Intramolecular Cyclization Approach to Tetralin-Based Cis-Fused Tetracyclic Compounds.

Presented herein is a BF3·OEt2-mediated, diastereoselective one-pot double cyclization of 4-aryl-2-[(arylthio)methyl]butanals leading to the formation of cis-tetrahydro-6H-naphtho[2,1-c]thiochromenes for the first time. Mechanistically, the formation of the title products involves the one-pot intramolecular Friedel-Crafts hydroxyalkylation/intramolecular Friedel-Crafts alkylation cascade. This synthetic methodology is featured by its high atom economy, broad substrate scope, mild transition-metal-free reaction conditions, capability to assemble two new rings in one pot, and moderate to high yields (up to 94% yield). It was then applied in the synthesis of a thia analogue of brazilane and a chromeno[3,4-c]chromene derivative. Moreover, the methodology was successfully extended to the synthesis of cis-hexahydrobenzo[c]phenanthrenes. Specifically, 1,5-diarylpentan-3-ones were first subjected to the Corey-Chaykovsky reaction, and the resulting epoxides, without being chromatographically isolated, were treated with BF3·OEt2 to afford the cyclized products in high yields (up to 84% yield over two steps).

Hexafluoroisopropanol-Mediated Intramolecular Ring-Opening Cyclization of Indolyl-N-Tethered Epoxides: Tether-Length-Controlled Synthesis of 1,7- and 1,2-Fused Indoles.

Despite having the capability to construct benzo-fused heterocycles in complete atom economy and high chemo-, regio-, enantio-, and diastereoselectivities, intramolecular Friedel-Crafts epoxide arene cyclization (IFCEAC) remains underutilized in organic synthesis. The wide adaptation of this powerful Csp2-Csp3 bond-forming reaction, therefore, requires a broad understanding of the substrate scope to better impact heterocycle synthesis. Along this line, we investigated the applicability of IFCEAC for the synthesis of 1,7- and 1,2-fused indoles. In this article, we report the results of our systematic investigation into the scope and limitations of the first examples of the hexafluoro-2-propanol (HFIP)-mediated IFCEAC of readily accessible indolyl-N-tethered epoxides. We observed that the nature and position of the indole and epoxide substituents and the tether length separating these two reacting moieties have strong effects on the cyclization. This mild and transition-metal-free protocol delivered pyrrolo[3,2,1-ij]quinolin-5-ols in moderate to good yields from substrates bearing both a methylene linker that connects the indole and epoxide moieties and an electron-rich indole carbocyclic ring. Notably, the reactions required the presence of a π-activating aryl substituent on the reacting epoxide carbon atom. Interestingly, replacing the methylene tether with an ethylene unit resulted in regioswitching, which delivered the corresponding tetrahydropyrido[1,2-a]indol-8-ols in good to high yields. We could also successfully extend this methodology to pyrrolyl-N-tethered epoxides for a very high-yielding synthesis of tetrahydroindolizin-7-ols.

Switching of regioselectivity in base-mediated diastereoselective annulation of 2,3-epoxy tosylates and their N-tosylaziridine analogs with 2-mercaptobenzimidazole.

A base-mediated dinucleophilic cyclization of readily accessible 2,3-epoxy tosylates with 2-mercaptobenzimidazole has been developed for the one-pot diastereoselective synthesis of benzimidazole-based tricyclic compounds equipped with two stereogenic centres. With trans-substrates bearing an aryl or alkyl substituent at the C3 position, the reaction involves an initial S-C1 bond-forming intermolecular alkylation followed by an N-C3 bond-forming, endo-selective intramolecular epoxide ring-opening cyclization reaction. A spectacular regioselectivity switching (tandem S-C3 and N-C1 bond formation reactions) was observed with related trans-N-tosylaziridine substrates. Wide substrate scope, complete diastereoselectivity, high to complete regioselectivity and mild transition metal-free conditions render this protocol particularly efficient and practical.

Studies on in vitro bioavailability and starch hydrolysis in zinc fortified ready-to-eat parboiled rice (komal chawal).

Zinc fortified parboiled rice (komal chawal) was produced from a low amylose variety of rice by applying 'brown rice parboiling' method. In addition to the effect of milling on fortification, the effectiveness of fortification upon the amount of bioaccessible (in vitro digest) and bioavailable (cellular uptake) form of Zn was tested. The effect on glycaemic index was also assessed by employing an in vitro starch hydrolysis assay. The bioaccessible form of Zn in the unmilled fortified rice were ranged in between 4.24 and 11.07 mg/100 g, which was significantly higher (p < 0.05) than the milled and unfortified parboiled rice. Similarly, the % absorption of bioavailable Zn was negligible in the unfortified parboiled rice as compared to the fortified rice (14.5-24.5%). The estimated GI of fortified parboiled rice samples was in the range of 50.97-59.79, which was lower than the unfortified parboiled rice (58.80-62.53) and raw rice (78.71-84.64). The results thus demonstrated that Zn fortified komal chawal can be a novel and rapidly produced micronutrient enhanced ready-to-eat rice.

Effect of iron and folic acid fortification on in vitro bioavailability and starch hydrolysis in ready-to-eat parboiled rice.

Iron (Fe) and folic acid (FA) fortified parboiled rice was produced by applying 'brown rice parboiling' method. The effect of milling and the effectiveness of fortification were tested in relation to the amount of bioaccessible and bioavailable form of Fe and FA. An in vitro starch hydrolysis assay was employed to assess the effect on simulated glycaemic index (GI). The % bioaccessiblity of Fe and FA in the unmilled fortified rice were in the range of 57.6-65.8%, and 55.1-91.9%, respectively. The % bioavailability in the unfortified parboiled rice was negligible as compared to Fe (14.7-32.1%) and FA (13.5-27.5%) fortified rice. The GI of unfortified and fortified parboiled rice samples was in the range of 56-69, which was lower than the raw rice. The results demonstrated that this approach can be a novel and rapid method to produce micronutrient enhanced ready-to-eat rice.

Growth and metabolic characterization of four lactic acid bacteria species isolated from rice beer prepared in Assam, India.

Isolation and identification of lactic acid bacteria (LAB) from rice beer prepared in Assam, India was performed and their growth associated and functional properties were studied. LAB strains were identified as Lactobacillus casei , Pediococcus pentosaceus , Lactobacillus pentosus and Lactobacillus plantarum based on 16 s rRNA sequencing. Their growth characteristics at different pH, NaCl concentration, temperature and presence of carbohydrates were profiled. High tolerance against acid and bile salts was shown by all the strains, particularly L. pentosus TEZU174 and P. pentosaceus TEZU199 up to a pH of 1.5, and L. pentosus TEZU174 up to 14  % bile concentration. They were susceptible towards the common antibiotics, wherein erythromycin, chloramphenicol and linezolid were the most effective. The strains displayed antibiosis activity against Escherichia coli and Staphylococcus aureus and antioxidant activity in terms of resistance to H2O2, scavenging of ·OH and DPPH free radicals was also displayed, wherein L. casei TEZU374 and P. pentosaceus TEZU482 were the most effective with above 70  % scavenging activity. The strains displayed cellular aggregation and L. casei TEZU262 and L. casei TEZU309 were highly aggregated, which attained 100  % autoaggregation within a period of 5 h. High cell surface hydrophobicity was shown by L. casei TEZU309 towards xylene and chloroform, and P. pentosaceus TEZU427 towards ethyl acetate. The strains evinced good gut tolerance capacity, antioxidant activity and adherence properties, which are characteristics of probiotic bacteria and thus are candidates for therapeutic uses and also to be used as starter cultures.

Optimization of ultrasound and microwave assisted extractions of polyphenols from black rice (Oryza sativa cv. Poireton) husk.

Phenolic compounds were extracted from the husk of milled black rice (cv. Poireton) by using a combination of ultrasound assisted extraction and microwave assisted extraction. Extraction parameters were optimized by response surface methodology according to a three levels, five variables Box-Behnken design. The appropriate process variables (extraction temperature and extraction time) to maximize the ethanolic extraction of total phenolic compounds, flavonoids, anthocyanins and antioxidant activity of the extracts were obtained. Extraction of functional components with varying ethanol concentration and microwave time were significantly affected by the process variables. The best possible conditions obtained by RSM for all the factors included 10.02 min sonication time, 49.46 °C sonication temperature, 1:40.79 (w/v) solute solvent ratio, 67.34% ethanol concentration, and 31.11 s microwave time. Under the given solutions, the maximum extraction of phenolics (1.65 mg/g GAE), flavonoids (3.04 mg/100 g), anthocyanins (3.39 mg/100 g) and antioxidants (100%) were predicted, while the experimental values included 1.72 mg/g GAE of total phenolics, 3.01 mg/100 g of flavonoids, 3.36 mg/100 g of anthocyanins and 100% antioxidant activity. The overall results indicated positive impact of co-application of microwave and ultrasound assisted extractions of phenolic compounds from black rice husk.

Amylolytic fungi in starter cakes for rice beer production.

Two types of starter cakes, viz. amou and perok-kushi, used in the production of rice beer in Assam, India, by the Bodo and Deori communities, respectively, were used for the isolation of amylolytic fungi. Based on the sequencing of their internal transcribed spacer (ITS) regions the fungi were identified as Amylomyces rouxii and Rhizopus oryzae, and given the strain names TU460 and TU465, respectively. Both the strains showed the ability to degrade and saccharify starch. The glucoamylase activity was considerably high in A. rouxii TU460 (14.92 µmol/min) as compared to R. oryzae TU465 (1.41 µmol/min), whereas α-amylase activity was found to be closely related, i.e. 7.02 and 6.09 unit mL-1, respectively. SDS PAGE for the determination of the molecular size of the glucoamylase enzymes revealed the production of two distinct units of 59 kDa and 31 kDa by A. rouxii TU460, and one unit of 72 kDa by R. oryzae TU465. LC MS/MS analysis revealed that no mycotoxins were produced by either of the strains. The overall study indicated a good amylolytic property of both strains and a potential for application in the starch processing industries.

Optimization of the extraction of phenolic compounds from Cyclosorus extensa with solvents of varying polarities.

The leaves of Cyclosorus extensa are used in the preparation of rice beer in Assam, India. The optimal conditions of time and temperature of fermentation for extraction of bioactive compounds from the dried leaves were obtained using response surface methodology. The central composite rotatable design was used and 13 experimental runs based on two-factor-five-level design were generated and performed for each of the solvents. The independent variables were extraction time (12 and 48 h) and temperature (25 and 55°C). The responses studied were total polyphenol content, radical scavenging activity, antibacterial activity, and antifungal activity. The analysis of variance of the test data was performed and the sequential sum of squares, F-value, R2, and adjusted R2 were deduced. The predicted models for all the response variables were adequately fitted to the observed experimental data (p ≤ 0.001). The maximum extraction of bioactive compounds under the optimum conditions of extraction temperature and time for hexane, ethyl acetate, methanol, and distilled water were found to be 25°C for 29.43 h, 28.28°C for 41.27 h, 43.95°C for 29.61 h, and 55.00°C for 48.00 h, respectively. It was also observed that the solubility of the polyphenols was higher in methanol, followed by ethyl acetate, and the highest antibacterial activity against Escherichia coli was shown by the ethyl acetate extracts.