High-throughput screening of small-molecules libraries identified antibacterials against clinically relevant multidrug-resistant A. baumannii and K. pneumoniae.

BACKGROUND: The current pipeline for new antibiotics fails to fully address the significant threat posed by drug-resistant Gram-negative bacteria that have been identified by the World Health Organization (WHO) as a global health priority. New antibacterials acting through novel mechanisms of action are urgently needed. We aimed to identify new chemical entities (NCEs) with activity against Klebsiella pneumoniae and Acinetobacter baumannii that could be developed into a new treatment for drug-resistant infections. METHODS: We developed a high-throughput phenotypic screen and selection cascade for generation of hit compounds active against multidrug-resistant (MDR) strains of K. pneumoniae and A. baumannii. We screened compound libraries selected from the proprietary collections of three pharmaceutical companies that had exited antibacterial drug discovery but continued to accumulate new compounds to their collection. Compounds from two out of three libraries were selected using "eNTRy rules" criteria associated with increased likelihood of intracellular accumulation in Escherichia coli. FINDINGS: We identified 72 compounds with confirmed activity against K. pneumoniae and/or drug-resistant A. baumannii. Two new chemical series with activity against XDR A. baumannii were identified meeting our criteria of potency (EC50 ≤50 µM) and absence of cytotoxicity (HepG2 CC50 ≥100 µM and red blood cell lysis HC50 ≥100 µM). The activity of close analogues of the two chemical series was also determined against A. baumannii clinical isolates. INTERPRETATION: This work provides proof of principle for the screening strategy developed to identify NCEs with antibacterial activity against multidrug-resistant critical priority pathogens such as K. pneumoniae and A. baumannii. The screening and hit selection cascade established here provide an excellent foundation for further screening of new compound libraries to identify high quality starting points for new antibacterial lead generation projects. FUNDING: BMBF and GARDP.

Novel detection method to rapidly quantify toxic cucurbitacin in Lagenaria siceraria (bottle gourd).

Bottle gourd (Lagenaria siceraria) is widely used in India for its medicinal properties. Sometimes its consumption as a fruit or juice can lead to serious health issues due to the presence of toxic cucurbitacins (Cuc) like Cuc B at high concentration. In the present study, a molecularly imprinted polymer (MIP) based method was developed to quantify Cuc B in commercial bottle gourd juice. Cuc B quantification was also done in peel and pulp of fresh bottle gourd fruit. Developed MIP was superior to NIP (non-imprinted polymer) in terms of equilibrium binding, solid-phase extraction, selectivity, and loading capacity of Cuc B. Scatchard plot analysis, adsorption kinetics, and surface area study further strengthen the fact that prepared imprinted polymer was better than NIP for Cuc extraction. TLC and high-resolution LC-MS based analysis revealed that MIP selectively extracts Cuc B from all the studied samples. It was further observed that gamma irradiation and ß-glucosidase treatment did not have any significant (p > 0.05) effect on Cuc concentration. The quantity of Cuc in 100 g of juice, peel, and pulp was 2.81 ± 0.26, 3.37 ± 0.31, and 2.39 ± 0.21 mg, respectively. Analytical method validation indicates that the MIP based method was highly linear, precise, and accurate to rapidly quantify toxic Cuc B in bottle gourd consequently assuring its quality. Supplementary Information: The online version contains supplementary material available at 10.1007/s13197-022-05600-3.

Collaborative Virtual Screening Identifies a 2-Aryl-4-aminoquinazoline Series with Efficacy in an In Vivo Model of Trypanosoma cruzi Infection.

Probing multiple proprietary pharmaceutical libraries in parallel via virtual screening allowed rapid expansion of the structure-activity relationship (SAR) around hit compounds with moderate efficacy against Trypanosoma cruzi, the causative agent of Chagas Disease. A potency-improving scaffold hop, followed by elaboration of the SAR via design guided by the output of the phenotypic virtual screening efforts, identified two promising hit compounds 54 and 85, which were profiled further in pharmacokinetic studies and in an in vivo model of T. cruzi infection. Compound 85 demonstrated clear reduction of parasitemia in the in vivo setting, confirming the interest in this series of 2-(pyridin-2-yl)quinazolines as potential anti-trypanosome treatments.

A quantitative and qualitative investigation of the impact of irradiation on the physico-chemical of pointed gourd during storage.

Irradiation effect on the physico-chemical characteristics and shelf stability of pointed gourd was investigated in research. It was estimated that moisture content, total phenol and total carotenoids was significantly (p≤0.05) decreased while physiological loss in weight, total soluble solids, lutein and α-cryptoxanthin were increased during the completion of study. The three carotenoid pigments (ß-carotene, lutein, α-cryptoxanthin) were identified by Thin Layer Chromatography (TLC) and spectrophotometric method and its changes reported during storage in control and irradiated sample. The principal component analysis showed that b* value, Physiological Loss Weight (PLW), adhesiveness and acidity have negative correlation with rest of the parameters during storage. The research will present a progressive strategy for increasing the storage stability of pointed gourd during transportation and storage in India's government and private mandis. Supplementary Information: The online version contains supplementary material available at 10.1007/s13197-022-05395-3.

Cyclic peptides nanospheres: A '2-in-1' self-assembled delivery system for targeting nucleus and cytoplasm.

Vascular endothelial growth factor (VEGF) is considered as one of the vital growth factors for angiogenesis, which is primarily responsible for the progress and maintenance of new vascular network in tumor. Numerous studies report that inhibition of VEGF-induced angiogenesis is a potent technique for cancer suppression. Recently, RNA interference, especially small interfering RNA (siRNA) signified a promising approach to suppress the gene expression. However, the clinical implementation of biological macromolecules such as siRNA is significantly limited because of stability and bioavailability issues. Herein, self-assembled peptide nanospheres have been generated from L,L-cyclic peptides using hydrophobic (Trp), positively charged (Arg) and cysteine (Cys) amino acid residues and demonstrated as vehicles for intracellular delivery of VEGF siRNA and VEGF antisense oligonucleotide. Formation of peptide nanostructures is confirmed by HR-TEM, AFM, SEM and DLS analysis. Possible mechanism of self-assembly of the cyclic peptides and their binding with macromolecules are demonstrated by in-silico analysis. Gel electrophoresis reveals that the newly generated peptide based organic materials exhibit strong binding affinity toward siRNAs / antisense oligonucleotides (ASOs) at optimum concentration. Flow cytometry and confocal microscopy results confirm the efficiency of the new biomaterials toward the intracellular delivery of fluorescent labeled siRNA / ASOs. Furthermore, VEGF expression evaluated by western blot and RT-PCR upon the delivery of functional VEGF siRNA/ASOs suggests that very low concentrations of VEGF siRNA/ASOs cause significant gene knockdown at protein and mRNA levels, respectively.

Carbon nano-dot for cancer studies as dual nano-sensor for imaging intracellular temperature or pH variation.

Cellular temperature and pH govern many cellular physiologies, especially of cancer cells. Besides, attaining higher cellular temperature plays key role in therapeutic efficacy of hyperthermia treatment of cancer. This requires bio-compatible, non-toxic and sensitive probe with dual sensing ability to detect temperature and pH variations. In this regard, fluorescence based nano-sensors for cancer studies play an important role. Therefore, a facile green synthesis of orange carbon nano-dots (CND) with high quantum yield of 90% was achieved and its application as dual nano-sensor for imaging intracellular temperature and pH was explored. CND was synthesized from readily available, bio-compatible citric acid and rhodamine 6G hydrazide using solvent-free and simple heating technique requiring purification by dialysis. Although the particle size of 19 nm (which is quite large for CND) was observed yet CND exhibits no surface defects leading to decrease in photoluminescence (PL). On the contrary, very high fluorescence was observed along with good photo-stability. Temperature and pH dependent fluorescence studies show linearity in fluorescence intensity which was replicated in breast cancer cells. In addition, molecular nature of PL of CND was established using pH dependent fluorescence study. Together, the current investigation showed synthesis of highly fluorescent orange CND, which acts as a sensitive bio-imaging probe: an optical nano-thermal or nano-pH sensor for cancer-related studies.

Targeting RECQL5 Functions, by a Small Molecule, Selectively Kills Breast Cancer in Vitro and in Vivo.

Clinical and preclinical data reveal that RECQL5 protein overexpression in breast cancer was strongly correlated with poor prognosis, survival, and therapeutic resistance. In the current investigation, we report design, synthesis, and specificity of a small molecule, 4a, which can preferentially kill RECQL5-expressing breast cancers but not RECQL5 knockout. Our stringent analysis showed that compound 4a specifically sensitizes RECQL5-expressing cancers, while it did not have any effect on other members of DNA RECQL-helicases. Integrated approaches of organic synthesis, biochemical, in silico molecular simulation, knockouts, functional mutation, and rescue experiments showed that 4a potently inhibits RECQL5-helicase activity and stabilizes RECQL5-RAD51 physical interaction, leading to impaired HRR and preferential killing of RECQL5-expressing breast cancer. Moreover, 4a treatment led to the efficient sensitization of cisplatin-resistant breast cancers but not normal mammary epithelial cells. Pharmacologically, compound 4a was orally effective in reducing the growth of RECQL5-expressing breast tumors (human xenograft) in NUDE-mice with no appreciable toxicity to the vital organs.

'BhAVI-23'-A spice-herb based dietary infusion possessing in-vitro anti-viral potential.

BACKGROUND: Viruses cause many life threatening human diseases. Recently, COVID-19 pandemic has challenged the health care systems worldwide. As a disease preventive approach and to bring relief to the severity of the symptoms, a infusion termed as Bhabha Anti-Viral Infusion-23 ('BhAVI-23') was conceptualized and formulated which comprised of 23 selected spices and herbals. OBJECTIVE: The present study was conducted to assess the in vitro antiviral potential of the formulation, BhaAVI-23. MATERIAL AND METHODS: The in-vitro anti-viral potential of BhAVI-23 was assessed through inhibition of HIV1 reverse transcriptase (RT) as well as through a novel P1 (virulent) bacteriphage based screening assay system. Anti-diabetic potential was assessed by non-enzymatic glycosylation of haemoglobin and the bioactive volatile components were detected through headspace gas chromatography followed by molecular docking analysis. RESULTS: The infusion displayed prominent anti-viral activity as evident from significant (57%) inhibition of the HIV1-RT as well as through reduction in the infectivity of P1 (virulent) bacteriophage. The infusion also exerted profound protection (∼64%) to non-enzymatic glycosylation of haemoglobin. Headspace gas chromatography and mass spectrometric analysis confirmed the presence of at least 47 major compounds. Docking analysis indicated possible interaction of α-pinene and eugenol with SARS-CoV spike protein. CONCLUSION: This 'BhAVI-23' infusion displayed prominent in-vitro anti-viral and anti-diabetic potential in different model systems. These attributes have relevance as diabetic patients are more prone to COVID-19 morbidity. 'BhAVI-23' opens the avenue for its potential inclusion as a supportive health care system upon due regulatory approval during the current pandemic.

Intussusception in children aged under two years in India: Retrospective surveillance at nineteen tertiary care hospitals.

OBJECTIVE: Intussusception has been linked with rotavirus vaccine (RVV) as a rare adverse reaction. In view of limited background data on intussusception in India and in preparation for RVV introduction, a surveillance network was established to document the epidemiology of intussusception cases in Indian children. METHODS: Intussusception in children 2-23 months were documented at 19 nationally representative sentinel hospitals through a retrospective surveillance for 69 months (July 2010 to March 2016). For each case clinical, hospital course, treatment and outcome data were collected. RESULTS: Among the 1588 intussusception cases, 54.5% were from South India and 66.3% were boys. The median age was 8 months (IQR 6, 12) with 34.6% aged 2-6 months. Seasonal variation with higher cases were documented during March-June period. The most common symptoms and signs were vomiting (63.4%), bloody stool (49.1%), abdominal pain (46.9%) and excessive crying (42.8%). The classical triad (vomiting, abdominal pain, and blood in stools) was observed in 25.6% cases. 96.4% cases were diagnosed by ultrasound with ileocolic location as the commonest (85.3%). Management was done by reduction (50.8%) and surgery (41.1%) and only 1% of the patients' died. 91.1% cases met Brighton criteria level 1 and 3.3% Level 2. Between 2010 and 2015, the case load and case ratio increased across all regions. CONCLUSION: Intussusception cases have occurred in children across all parts of the country, with low case fatality in the settings studied. The progressive rise cases could indicate an increasing awareness and availability of diagnostic facilities.

Ag(I)-Fesulphos-Catalyzed Enantioselective Synthesis of 3-Silylproline Derivatives.

An efficient catalytic asymmetric 1,3-dipolar cycloaddition of N-benzylidineiminoglycinate-derived azomethine ylides to ß-silylmethylene malonates catalyzed by a Ag(I)-Fesulphos complex has been developed, affording fully substituted 3-silylproline derivatives with an all carbon quaternary center. The silylproline derivatives were obtained in moderate-to-good yields (up to 81%) in high diastereoselectivities and enantioselectivities (dr up to 95:5; er up to 96:4). Tamao-Fleming oxidation of selected 3-silylproline derivatives provided not only an efficient route but also the shortest route to 3-hydroxyproline derivatives, which are not accessible by direct 1,3-dipolar cycloadditions of azomethine ylide with frequently used arylidene/alkylidene malonates.

First Report on the Separation of Trivalent Lanthanides from Trivalent Actinides Using an Aqueous Soluble Multiple N-Donor Ligand, 2,6-bis(1 H-tetrazol-5-yl)pyridine: Extraction, Spectroscopic, Structural, and Computational Studies.

A terdentate multiple N donor ligand, 2,6-bis(1 H-tetrazol-5-yl)pyridine (H2BTzP), was synthesized, and its complexation with trivalent americium, neodymium, and europium was studied using single-crystal X-ray diffraction, attenuated total reflectance-fourrier transform infrared spectroscopy, time-resolved fluorescence spectroscopy, UV-vis absorption spectrophotometry. Higher complexation strength of BTzP toward trivalent actinide over lanthanides as observed from UV-vis spectrophotometric study resulted in an effective separation of Am3+ and Eu3+ in liquid-liquid extraction studies employing N,N, N',N'-tetra- n-octyl diglycolamide in the presence of BTzP as the aqueous complexant. The selectivity of BTzP toward Am3+ over Eu3+ was further investigated by DFT computations, which indicated higher metal-ligand overlap in the Am3+ complex as indicated from the metal-nitrogen bond order and frontier molecular orbital analysis of the BTzP complexes of Am3+ and Eu3+.

Superparamagnetic graphene oxide-magnetite nanoparticle composites for uptake of actinide ions from mildly acidic feeds.

Super paramagnetic graphene oxide (GO) - Fe3O4 nanoparticle composites were prepared and characterized by conventional techniques such as XRD, SEM, EDX, FT-IR, Raman, XPS, DLS and zeta potential, etc. TEM studies have confirmed nanoparticle nature of the composites. The GO-magnetic nanoparticle composites can be dispersed in mildly acidic aqueous solutions and get concentrated in a small volume under application of an external magnetic field. The composites were evaluated for the uptake of actinide ions such as Am3+, UO22+, Th4+ and Pu4+ from mildly acidic aqueous solutions. Am3+ sorption sharply increased with pH as the Kd values increased from about 10 at pH 1 to 105 at pH 3 beyond which a plateau in the Kd values was seen. Eu3+ displayed nearly comparable uptake behaviour to that of Am3+ while the uptake of other metal ions followed the trend: Pu(IV)>Th(IV)>>UO22+. The adsorption behaviour of Am3+ onto the graphene oxide - Fe3O4 nanoparticle composites fitted very well to the Langmuir as well as Temkin isotherm models. The desorption rate (using 1M HNO3) was fast and reusability study results were highly encouraging. The very high uptake values suggest possible application of the magnetic nanoparticles in radioactive waste remediation in natural ground water.

Organocatalytic decarboxylative aldol reaction of ß-ketoacids with α-ketophosphonates en route to the enantioselective synthesis of tertiary α-hydroxyphosphonates.

The first example of an asymmetric organocatalyzed decarboxylative aldol reaction of ß-ketoacids (aroylacetic acids) with α-ketophosphonates that produces a quaternary chiral centre has been developed. A quinidine based bifunctional urea derivative was identified as the preferred catalyst affording γ-aroyl tertiary α-hydroxyphosphonates in good yield and enantioselectivity. The 31P NMR spectroscopic study was performed to shed light on the reaction mechanism.

Tuning the Stability and the Reactivity of Substituted [3]Dendralenes for Quick Access to Diverse Copiously Functionalized Fused Polycycles with Step and Atom Economy.

This is the first comprehensive study that details the synthesis of stable acyclic trisubstituted [3]dendralenes and deciphers their structural requisite for a successful diene transmissive Diels-Alder (DTDA) reaction by employing two different dienophiles and eventually generating a small repository of complex molecules, thus exemplifying how substituted [3]dendralenes could be deployed in diversity-oriented synthesis with high selectivities. A balance of reactivity and stability was struck by prudent selection of the position and nature of functional groups on these [3]dendralenes. Upon tandem Diels-Alder reactions with several symmetrical as well as unsymmetrical dienophiles, these dendralenes afforded diversity-oriented quick access to many polycyclic complex motifs possessing several functional groups and multiple stereogenic centers. Thus, the full potential of the dendralenes could be harnessed. The reactions proceeded under mild conditions with step and atom economy and were highly regio- and stereoselective besides being excellent yielding. The DTDA sequence resulted in the generation of four new carbon-carbon bonds, two new rings, and 3-7 stereogenic centers. The key feature of the method is that we could rapidly generate complexity along with functional and structural diversity from a trivial acyclic substrate with no stereogenic centers.

Molecular Origin and Self-Assembly of Fluorescent Carbon Nanodots in Polar Solvents.

Despite numerous efforts, there are several fundamental ambiguities regarding the photoluminescence of carbon dots (CDs). Spectral shift measurements display characteristic of both π-π* and n-π* transitions for the main absorption or excitation band at ∼350 nm, contrary to common assignment of exclusive n-π* transition. Additionally, the generally perceived core-state transition at ∼250 nm, involving sp2-networked carbogenic domains shielded from external environments, needs to be reassessed because it fails to explain the observed fluorescence quenching and spectral shift. These results have been explained based on the molecular origin of PL in CDs invoking the similarity between CD and citrazinic acid. Fluorescent derivatives of the latter are recognized to be produced during citric-acid-based CD synthesis. Concentration-dependent spectral splitting of the main excitation band in combination with the temperature-dependent PL results has been envisioned assuming self-assembly of CDs into various H-aggregates.

Organocatalyzed enantioselective Michael addition/cyclization cascade reaction of 3-isothiocyanato oxindoles with arylidene malonates.

An organocatalyzed Michael addition-cyclization reaction between 3-isothiocyanatooxindoles and arylidene malonates has been developed for the synthesis of highly functionalized 3,2'-pyrrolidinyl spirooxindole derivatives. The reaction was catalyzed by a quinine derived tertiary amino-thiourea based bifunctional catalyst or its pseudo-enantiomeric quinidine derived catalyst providing both the enantiomers of the desired product. The products were obtained in high yields and with excellent diastereo- (up to 99 : 1 dr) and enantioselectivities (up to >99% ee).

Origin of Excitation Dependent Fluorescence in Carbon Nanodots.

The fascinating aspect of excitation dependent fluorescence in carbon nanodots has led to several hypotheses, starting from particle size distribution to the presence of different emissive states and even to sluggish solvent relaxation around nanodot. In this contribution we provide definitive evidence for the involvement of discrete multiple electronic states for the excitation dependent emission in carbon nanodots. The presence of different types of aggregates even at very dilute solutions used in ensemble fluorescence spectroscopy, where fluorescence intensity shows linear dependence with absorbance, is the origin of these multiple electronic states. Inhomogeneous broadening due to slow solvent relaxation leading to excitation dependent spectral shift has negligible influence in conventional solvents.

Synthesis and extraction studies with a rationally designed diamide ligand selective to actinide(iv) pertinent to the plutonium uranium redox extraction process.

A new class of conformationally constrained oxa-bridged tricyclo-dicarboxamide (OTDA) ligand was rationally designed for the selective extraction of tetravalent actinides pertinent to the Plutonium Uranium Redox EXtraction (PUREX) process. Two of the designed diamide ligands were synthesized and extraction studies were performed for Pu(iv) from HNO3 medium. The mechanism of extraction was investigated by studying various parameters such as feed HNO3, NaNO3 and OTDA concentrations. The nature of the extracted species was found to be [Pu(NO3)4(OTDA)]. One of the OTDA ligands was elaborately tested and showed the selective extraction of Pu(iv) and Np(iv) over other actinide species, viz., U(vi), Np(v), Am(iii), lanthanides and fission products contained in a nuclear waste from the PUREX process. DFT calculations predicted the charge density on each of the coordinating 'O' atoms of OTDA supporting its high Pu(iv) selectivity over other ions studied and also provided the energy optimized structure of OTDA and its Pu(iv) complex.

"On Water" Organocatalyzed [4 + 2] Cycloaddition of Enones and Nitro Dienes for the Enantioselective Synthesis of Densely Substituted Cyclohexanones.

An "on water" hydroquinine-based primary amine-benzoic acid organocatalyst system was found to be best suited to produce 3,4,5-trisubstituted cyclohexanones with a nitro group in the 4-position from enones and nitro dienes under ambient conditions in good yield, with good diastereoselectivity, and with excellent enantioselectivity. An appreciable rate enhancement by water was observed compared to organic solvents. Mechanistic analysis of the reaction suggests that it followed an endo [4 + 2] cycloaddition with enamine of enone as diene and nitro diene as dienophile.