Selective detection of azelnidipine in pharmaceuticals via carbon dot mediated spectrofluorimetric method: A green approach.

A spectrofluorimetric method using fluorescent carbon dots (CDs) was developed for the selective detection of azelnidipine (AZEL) pharmaceutical in the presence of other drugs. In this study, N-doped CDs (N-CDs) were synthesized through a single-step hydrothermal process, using citric acid and urea as precursor materials. The prepared N-CDs showed a highly intense blue fluorescence emission at 447 nm, with a photoluminescence quantum yield of ~21.15% and a fluorescence lifetime of 0.47 ns. The N-CDs showed selective fluorescence quenching in the presence of all three antihypertensive drugs, which was used as a successful detection platform for the analysis of AZEL. The photophysical properties, UV-vis light absorbance, fluorescence emission, and lifetime measurements support the interaction between N-CDs and AZEL, leading to fluorescence quenching of N-CDs as a result of ground-state complex formation followed by a static fluorescence quenching phenomenon. The detection platform showed linearity in the range 10-200 µg/ml (R2 = 0.9837). The developed method was effectively utilized for the quantitative analysis of AZEL in commercially available pharmaceutical tablets, yielding results that closely align with those obtained from the standard method (UV spectroscopy). With a score of 0.76 on the 'Analytical GREEnness (AGREE)' scale, the developed analytical method, incorporating 12 distinct green analytical chemistry components, stands out as an important technique for estimating AZEL.

Magnetic nanoflowers: a hybrid platform for enzyme immobilization.

The use of organic-inorganic hybrid nanoflowers as a support material for enzyme immobilization has gained significant attention in recent years due to their high stability, ease of preparation, and enhanced catalytic activity. However, a major challenge in utilizing these hybrid nanoflowers for enzyme immobilization is the difficulty in handling and separating them due to their low density and high dispersion. To address this issue, magnetic nanoflowers have emerged as a promising alternative enzyme immobilization platform due to their easy separation, structural stability, and ability to enhance catalytic efficiency. This review focuses on different methods for designing magnetic nanoflowers, as well as future research directions. Additionally, it provides examples of enzymes immobilized in the form of magnetic nanoflowers and their applications in environmental remediation, biosensors, and food industries. Finally, the review discusses possible ways to improve the material for enhanced catalytic activity, structural stability, and scalability.

Magnetic nanoflowers can be used as a novel platform for enzyme immobilization.There are three different approaches to the synthesis of efficient magnetic nanoflower.The magnetic nanoflowers provides excellent stability and good reusability of enzymes.The hybrid biocatalyst was applied in biotransformation, environmental, and food applications.The challenges and their remedies of hybrid biocatalyst have been discussed.

Functionalized Graphene Quantum Dots (GQDs) based Label-Free Optical Fluorescence Sensor for CD59 Antigen Detection and Cellular Bioimaging.

Cluster of differentiation (CD59), a cell surface glycoprotein, regulates the complement system to prevent immune damage. In cancer, altered CD59 expression allows tumors to evade immune surveillance, promote growth, and resist certain immunotherapies. Targeting CD59 could enhance cancer treatment strategies by boosting the immune response against tumors. Herein, we present a one-step synthesis of Polyethyleneimine (PEI) functionalized graphene quantum dots (Lf-GQDs) from weathered lemon leaf extract. The fabricated Lf-GQDs were successfully used for the quantitative detection of the cluster of CD59 antigen that is reported for its expression in different types of cancer. In this work, we utilized orientation-based attachment of CD59 antibody (Anti-CD59). Our findings reveal that, instead of using random serial addition of antigen or antibody, oriented conjugation saves accumulated concentration offering greater sensitivity and selectivity. The Anti-CD59@Lf-GQDs immunosensor was fabricated using the oriented conjugation of antibodies onto the Lf-GQDs surface. Besides, the fabricated immunosensor demonstrated detection of CD59 in the range of 0.01 to 40.0 ng mL-1 with a low detection limit of 5.3 pg mL-1. Besides, the cellular uptake potential of the synthesized Lf-GQDs was also performed in A549 cells using a bioimaging study. The present approach represents the optimal utilization of Anti-CD59 and CD59 antigen. This approach could afford a pathway for constructing oriented conjugation of antibodies on the nanomaterials-based immunosensor for different biomarkers detection.

Awareness, perspectives and satisfaction levels among patients with psoriatic arthritis: a multicentric cross-sectional survey.

Perception of the disease and its management impacts patients with Psoriatic arthritis (PsA) to a great degree. Studies examining patients' viewpoints and perception of their disease and its management are scarce. This multicentric cross-sectional survey was undertaken to understand the perspectives of patients with PsA. A survey questionnaire with items on demographics, awareness about their disease, treatment, physical therapy, quality of life and satisfaction with the care received was designed. After internal and external validation, a pilot survey was conducted, and the questionnaire was finalized. The final survey (with translations in local languages) was carried out at 17 centres across India. There were 262 respondents (56% males) with mean age of 45.14 ± 12.89 years. In 40%, the time lag between onset of symptoms and medical assessment for it was more than a year. In most of the patients, the diagnosis of PsA was made by a rheumatologist. Over 83% of patients were consulting their rheumatologist periodically as advised and fully compliant with the treatment. Lack of time and cost of therapy were the most common reasons for non-adherence to therapy. Eighty-eight patients (34%) were not fully satisfied with their current treatment. Over two-third of patients had never seen a physiotherapist due to barriers including a lack of time, pain, and fatigue. The daily activities and employment status were affected in nearly 50% of patients with PsA. The current survey has identified a gap in patients' awareness levels and helps healthcare providers in understanding the varied perceptions of patients with PsA. Addressing these issues in a systematic manner would potentially improve the treatment approaches, outcomes, and patient satisfaction levels.

A Bifurcated Multicomponent Synthesis Approach to Polycyclic Quinazolinones.

The rapid synthesis of diverse substituted polycyclic quinazolinones was achieved by two orthogonal Ugi four-component reaction (Ugi-4CR)-based protocols: the first two-step approach via an ammonia-Ugi-4CR followed by palladium-catalyzed annulation; in the second approach, cyanamide was used unprecedently as an amine component in Ugi-4CR followed by an AIBN/tributyltin hydride-induced radical reaction. Like no other method, MCR and cyclization could efficiently construct many biologically interesting compounds with tailored properties in very few steps.

Solvent-assisted cavitation for enhanced removal of organic pollutants - Degradation of 4-aminophenol.

A new approach of solvent-assisted cavitation process was proposed for degradation of organic pollutants. The process envisages the use of suitable solvent as an additive, (1-5% v/V), in the conventional cavitation process to enhance the pollutant removal efficiency. A proof of concept was provided for the removal of ammoniacal nitrogen with significantly improved efficiency using solvent-assisted hydrodynamic cavitation (HC) compared to conventional HC. The efficacy of the process was studied on a pilot plant scale (1 m3/h) and using vortex flow based vortex diode as a cavitating device. Degradation studies were carried out using a model pollutant, 4-aminophenol and four different solvents as additives, 1-octanol, cyclohexanol, 1-octane and toluene. Relatively polar solvents were found to increase the efficiency of the pollutant removal (>65%) and also increase the rates to an extent of more than 200%, compared to only HC. A very high removal of ammoniacal nitrogen, more than 90%, was obtained for solvents 1-octanol and cyclohexanol, indicating the importance of the selection of solvent. Per-pass degradation model showed 3 to 4 times increase in the per pass degradation for polar solvents compared to cavitation alone. The results confirm no role of conventional solvent extraction and no specific contamination of wastewater due to the use of solvent as an additive in the process. Further, the cost was 2-3 times lower as compared to the conventional HC. The interesting observations in the proposed process can fuel further research to provide possible improvements in existing methodologies of wastewater treatment, in general, and for removal of ammoniacal nitrogen, in particular.

A unified multivariate statistical approach for the assessment of deep groundwater quality of rapidly growing city of Maharashtra Province, India, with potential health risk.

The main aim of this research is to assess the consequences of natural and anthropogenic processes on the groundwater quality of 65 deep aquifers of Nagpur city, Maharashtra Province, India, using a unified multivariate statistical approach. The dominant groundwater type recognized is Ca-HCO3 (recharge waters) in 43.1 and 38.5% of groundwater samples of pre- and post-monsoon seasons, followed by mixed water types. The seasonal distribution of physicochemical parameters shows increase in the concentration of EC, TDS, TH, Mg2+, SO42-, and NO3- signifying the high mineralization and anthropogenic loading from pre- and post-monsoon season respectively. The entropy-weight water quality index categorizes the 84.6% and 75.4% of total samples from pre- and post-monsoon seasons into moderate quality. The multiple linear regression and PCA analysis reveal the masking of rock weathering mechanism by anthropogenic activities. The % of PCA Variance varies from 79 to 83.7% from pre- to post-monsoon season. The high contributions of EC (0.76, 0.72), TDS (0.79, 0.73), TH (0.97, 0.962), Ca2+ (0.84, 0.78), Mg2+ (0.79, 0.83), Cl- (0.73, 0.75), and NO3- (0.78, 0.68) in PC1 components expose high salinity and hardness in urban groundwater that signifies the consequences of urbanization on the groundwater regime. About 55.4 and 70.8% of children population as compared to the adult female (53.8%, 69.2%) and male (32.3%, 46.1%) population in PRM and POM respectively were at high non-carcinogenic health threat of NO3--enriched groundwater. The study is beneficial for understanding the variation in groundwater composition due to unplanned urbanization and is very useful for protecting groundwater resources in urban areas.

Enzyme embedded microfluidic paper-based analytic device (µPAD): a comprehensive review.

Low-cost paper-based analytical devices are the latest generation of portable lab-on-chip designs that offers an innovative platform for the on/off-site analysis (biosensing) of target analytes, especially in rural and remote areas. Recently, microfluidic paper-based analytical devices (µPADs) have attained significant recognition owing to their exciting fundamental features such as: ease of fabrication, rapid operation, and precise interpretations. The incorporation of enzymes with paper-based analytical devices significantly improves analytical performance while exhibiting excellent chemical and storage stability. In addition to that, these devices are highly compact, portable, easy-to-use, and do not require any additional sophisticated equipment for the detection and quantification of target analytes. This review provides a holistic insight into design, fabrication, and enzyme immobilization strategies for the development of enzyme-µPADs, which enables them to be widely implemented for in-field analysis. It also highlights the recent application of enzyme-µPADs in the area of: biomedical, food safety, and environmental monitoring while exploring the mechanisms of detection involved. Further, in order to improve the accuracy of analysis, researchers have designed a smartphone-based scanning tool for multi-variant point-of-care devices, which is summarized in the latter part of the review. Finally, the future perspectives and outlook of major challenges associated with enzyme-µPADs are discussed with their possible solutions. The development of enzyme integrated µPADs will open a new avenue as an exceptional analytical tool to explore various applications.HIGHLIGHTSEnzyme embedded paper-based analytical devices are a revolution in the field of biosensing.The design, fabrication, and enzyme immobilization on µPADs have been comprehensively discussed.The application of enzyme-µPADs food safety, environmental monitoring, and clinical diagnostic have been reviewed.Smartphones can be used as an on-site, user-friendly, and compact next-gen scanning tool for biosensing.

Novel use of fused cardiac computed tomography and transesophageal echocardiography for left atrial appendage closure.

The use of the Watchman left atrial appendage occlusion device (Boston Scientific Inc.) is becoming increasingly frequent in patients with atrial fibrillation. Cardiac computed tomography (CT) for device sizing pre-procedure can help facilitate more accurate device selection compared with transesophageal echo (TEE) alone. CT can also help identify minor lobes and trabeculations that may not be apparent on TEE. We report a series of three cases to highlight the utility of a novel application of CT-TEE fusion imaging to provide procedural guidance during Watchman implant and to assess for peri-device leak post-implant.

Effects of N-terminus modified Hx-amides on DNA binding affinity, sequence specificity, cellular uptake, and gene expression.

Five X-HxIP (Hx-amides) 6a-e, in which the N-terminus p-anisyl moiety is modified, were designed and synthesised with the purpose of optimising DNA binding, improving cellular uptake/nuclear penetration, and enhancing the modulation of the topoisomerase IIα (TOP2A) gene expression. The modifications include a fluorophenyl group and other heterocycles bearing different molecular shapes, size, and polarity. Like their parent compound HxIP 3, all five X-HxIP analogues bind preferentially to their cognate sequence 5'-TACGAT-3', which is found embedded on the 5' flank of the inverted CCAAT box-2 (ICB2) site in the TOP2A gene promoter, and inhibit protein complex binding. Interestingly, the 4-pyridyl analog 6a exhibits greater binding affinity for the target DNA sequence and abolishes the protein:ICB2 interaction in vitro, at a lower concentration, compared to the prototypical compound HxIP 3. Analogues 6b-e, display improved DNA sequence specificity, but reduced binding affinity for the cognate sequence, relative to the unmodified HxIP 3, with polyamides 6b and 6e being the most sequence selective. However, unlike 3 and 6b, 6a was unable to enter cells, access the nucleus and thereby affect TOP2A gene expression in confluent human lung cancer cells. These results show that while DNA binding affinity and sequence selectivity are important, consideration of cellular uptake and concentration in the nucleus are critical when exerting biological activity is the desired outcome. By characterising the DNA binding, cellular uptake and gene regulatory properties of these small molecules, we can elucidate the determinants of the elicited biological activity, which can be impacted by even small structural modifications in the polyamide molecular design.

Silk industry waste protein: isolation, purification and fabrication of electrospun silk protein nanofibers as a possible nanocarrier for floating drug delivery.

Amongst assorted regio-selective and targeted oral drug delivery strategies accepted for the gastro-retentive drug delivery system (GRDDS), the floating drug delivery system (FDDS) holds a major share as clinically accepted formulations. The major objective of the present investigation was to explore the silk industry waste protein, silk fibroin (SF) as a possible electrospun nanocarrier for the FDDS. In a nutshell, electrospinning (ES) is one of the flexible and astonishing strategies for the fabrication of porous electrospun nanofibers (NFs), which offers the potential to amend the floating profile, dissolution rate, solubility, and release patterns of the drug, etc as per compendial requirements. Looking at the prospects of floating SF-NFs preparation, we have isolated and lyophilized the SF from industrial waste cocoons and prepared drug-loaded SF single polymer nanofibers (SPN). Lafutidine (LF) being a good candidate for GRDDS selected as a model drug, which is an excellent proton pump inhibitor, mainly used in the treatment of gastric ulcers. Finally, the obtained LF loaded SF-NFs (LF-SF-NFs) were successfully analyzed for physicochemical characteristics, porosity, swelling index, antioxidant activity, mucoadhesion strength, floating properties, enzymatic degradation, and accelerated stability study, etc. Further, these LF-SF-NFs were evaluated for percent drug content, weight variation, in-vitro dissolution in 0.1 N hydrochloric acid (HCl, pH:1.2) and fasted state simulated gastric fluid (FSSGF), and accelerated stability study. It has shown significant floating time >18 h, about 99% ± 0.58% floating buoyancy with sustained release up to 24 h. LF-SF-NFs showed good compatibility, entrapment efficiency, antioxidant activity, mucoadhesion strength, enzymatic degradation, and long term stability. Soon, the essential floating and drug release profiles can claim single polymer (SF) based electrospun protein NFs as a possible novel oral nanocarrier for FDDS.

Racial and ethnic differences in left atrial appendage occlusion wait time, complications, and periprocedural management.

PURPOSE: Non-white patients are underrepresented in left atrial appendage occlusion (LAAO) trials, and racial disparities in LAAO periprocedural management are unknown. METHODS: We assessed sociodemographics and comorbidities of consecutive patients at our institution undergoing LAAO between 2015 and 2020, then in adjusted analyses, compared procedural wait time, procedural complications, and post-procedure oral anticoagulation (OAC) use in whites versus non-whites. RESULTS: Among 109 patients undergoing LAAO (45% white), whites had lower CHA2 DS2 VASc scores, on average, than non-whites (4.0 vs. 4.8, p = .006). There was no difference in median time from index event (IE) or initial outpatient cardiology encounter to LAAO procedure (whites 10.5 vs. non-whites 13.7 months, p = .9; 1.9 vs. 1.8 months, p = .6, respectively), and there was no difference in procedural complications (whites 4% vs. non-whites 5%, p = .33). After adjusting for CHA2 DS2 VASc score, OAC use at discharge tended to be higher in whites (OR 2.4, 95% CI [0.9-6.0], p = .07). When restricting the analysis to those with prior gastrointestinal (GI) bleed, adjusting for CHA2 DS2 VASc score and GI bleed severity, whites had a nearly five-fold odds of being discharged on OAC (OR 4.6, 95% CI [1-21.8], p = 0.05). The association between race and discharge OAC was not mediated through income category (total mediation effect 19% 95% CI [-.04-0.11], p = .38). CONCLUSION: Despite an increased prevalence of comorbidities amongst non-whites, wait time for LAAO and procedural complications were similar in whites versus non-whites. Among those with prior GI bleed, whites were nearly five-fold more likely to be discharged on OAC than non-whites, independent of income.

Design and evaluation of hyaluronic acid-coated PLGA nanoparticles of raloxifene hydrochloride for treatment of breast cancer.

CONTEXT: In the present study, hyaluronic acid (HA)-coated raloxifene-loaded poly(l-lactic-co-glycolic acid) (PLGA) nanoparticles have been developed to improve the anticancer potential and reduce side effects associated with the drug. AIM AND OBJECTIVES: The investigation was aimed to formulate and optimize raloxifene hydrochloride (RALH)-loaded PLGA nanoparticles with surface modification using HA as a targeting moiety. To perform physicochemical characterization, in vitro cytotoxicity study (using MCF-7), in vitro drug release study and in vivo pharmacodynamic study of optimized formulation. METHODOLOGY: Raloxifene hydrochloride-loaded PLGA nanoparticles were prepared by nanoprecipitation technique, followed by surface modification with HA. Formulation was optimized by using 23 factorial design and characterized by physicochemical, in vitro drug release, in vitro cytotoxicity studies, and in vivo pharmacokinetics. RESULTS AND DISCUSSION: The particle size, PDI, zeta potential, entrapment efficiency, and loading capacity of spherically shaped RALH-loaded nanoparticles were 207.3 ± 4.2 d.nm, 0.218 ± 0.127, -.127 mV, 43.75 ± 1.2%, and 7.55 ± 1.14%, respectively. The in vitro drug release showed sustained release and followed Korsmeyer-Peppas model with non-Fickian release pattern. The in vitro cytotoxicity study of drug-loaded NPs by MTT assay on MCF-7 breast carcinoma cell showed anti-cancer activity after 48 h of treatment. CONCLUSION: The results of the present investigation suggested that RALH-loaded HA-modified PLGA nanoparticles showed sustained drug release with anticancer activity and can be a promising approach for treatment of breast cancer.

Combining High-Throughput Synthesis and High-Throughput Protein Crystallography for Accelerated Hit Identification.

Protein crystallography (PX) is widely used to drive advanced stages of drug optimization or to discover medicinal chemistry starting points by fragment soaking. However, recent progress in PX could allow for a more integrated role into early drug discovery. Here, we demonstrate for the first time the interplay of high throughput synthesis and high throughput PX. We describe a practical multicomponent reaction approach to acrylamides and -esters from diverse building blocks suitable for mmol scale synthesis on 96-well format and on a high-throughput nanoscale format in a highly automated fashion. High-throughput PX of our libraries efficiently yielded potent covalent inhibitors of the main protease of the COVID-19 causing agent, SARS-CoV-2. Our results demonstrate, that the marriage of in situ HT synthesis of (covalent) libraires and HT PX has the potential to accelerate hit finding and to provide meaningful strategies for medicinal chemistry projects.

Novel isoniazid embedded triazole derivatives: Synthesis, antitubercular and antimicrobial activity evaluation.

In the present study, a series of new isoniazid embedded triazole derivatives have been synthesized. These compounds were evaluated for their in vitro antitubercular and antimicrobial activities. Among the screened compounds, six have exhibited potent antitubercular activity against Mycobacterium tuberculosis H37Rv strain with MIC value 0.78 µg/mL, whereas, three compounds have displayed activity with MIC value ranging from 1.56 to 3.125 µg/mL. The cytotoxicity of the active compounds was studied against RAW 264.7 cell line by MTT assay and no toxicity was observed even at 25 µg/mL concentration. The five compounds have displayed good antimicrobial activities. Molecular docking have been performed against mycobacterial InhA enzyme to gain an insight into the plausible mechanism of action which could pave the way for our endeavor to identify potent antitubercular candidates. We believe that further optimization of these molecules may lead to potent antitubercular agents.

Recent Advancement in Bio-precursor derived graphene quantum dots: Synthesis, Characterization and Toxicological Perspective.

Graphene quantum dots (GQDs), impressive materials with enormous future potential, are reviewed from their inception, including different precursors. Considering the increasing burden of industrial and ecological bio-waste, there is an urgency to develop techniques which will convert biowaste into active moieties of interest. Amongst the various materials explored, we selectively highlight the use of potential carbon containing bioprecursors (e.g. plant-based, amino acids, carbohydrates), and industrial waste and its conversion into GQDs with negligible use of chemicals. This review focuses on the effects of different processing parameters that affect the properties of GQDs, including the surface functionalization, paradigmatic characterization, toxicity and biocompatibility issues of bioprecursor derived GQDs. This review also examines current challenges and s the ongoing exploration of potential bioprecursors for ecofriendly GQD synthesis for future applications. This review sheds further light on the electronic and optical properties of GQDs along with the effects of doping on the same. This review may aid in future design approaches and applications of GQDs in the biomedical and materials design fields.

In Vitro and In Vivo Activity of Peptidomimetic Compounds That Target the Periodontal Pathogen Porphyromonas gingivalis.

The interaction of the periodontal pathogen Porphyromonas gingivalis with oral streptococci is important for initial colonization of the oral cavity by P. gingivalis and is mediated by a discrete motif of the streptococcal antigen I/II protein. A synthetic peptide encompassing this motif functions as a potent inhibitor of P. gingivalis adherence, but the use of peptides as topically applied therapeutic agents in the oral cavity has limitations arising from the relatively high cost of peptide synthesis and their susceptibility to degradation by proteases expressed by oral organisms. In this study, we demonstrate the in vitro and in vivo activity of five small-molecule mimetic compounds of the streptococcal peptide. Using a three-species biofilm model, all five compounds were shown to effectively inhibit the incorporation of P. gingivalis into in vitro biofilms and exhibited 50% inhibitory concentrations (IC50s) of 10 to 20 µM. Four of the five compounds also significantly reduced maxillary alveolar bone resorption induced by P. gingivalis infection in a mouse model of periodontitis. All of the compounds were nontoxic toward a human telomerase immortalized gingival keratinocyte cell line. Three compounds exhibited slight toxicity against the murine macrophage J774A.1 cell line at the highest concentration tested. Compound PCP-III-201 was nontoxic to both cell lines and the most potent inhibitor of P. gingivalis virulence and thus may represent a novel potential therapeutic agent that targets P. gingivalis by preventing its colonization of the oral cavity.

DNA-Binding Properties of New Fluorescent AzaHx Amides: Methoxypyridylazabenzimidazolepyrroleimidazole/pyrrole.

DNA minor groove binding polyamides have been extensively developed to control abnormal gene expression. The establishment of novel, inherently fluorescent 2-(p-anisyl)benzimidazole (Hx) amides has provided an alternative path for studying DNA binding in cells by direct observation of cell localization. Because of the 2:1 antiparallel stacking homodimer binding mode of these molecules to DNA, modification of Hx amides to 2-(p-anisyl)-4-azabenzimidazole (AzaHx) amides has successfully extended the DNA-recognition repertoire from central CG [recognized by Hx-I (I=N-methylimidazole)] to central GC [recognized by AzaHx-P (P=N-methylpyrrole)] recognition. For potential targeting of two consecutive GG bases, modification of the AzaHx moiety to 2- and 3-pyridyl-aza-benzimidazole (Pyr-AzaHx) moieties was explored. The newly designed molecules are also small-sized, fluorescent amides with the Pyr-AzaHx moiety connected to two conventional five-membered heterocycles. Complementary biophysical methods were performed to investigate the DNA-binding properties of these molecules. The results showed that neither 3-Pyr-AzaHx nor 2-Pyr-AzaHx was able to mimic I-I=N-methylimidazole-N-methylimidazole to target GG dinucleotides specifically. Rather, 3-Pyr-AzaHx was found to function like AzaHx, f-I (f=formamide), or P-I as an antiparallel stacked dimer. 3-Pyr-AzaHx-PI (2) binds 5'-ACGCGT'-3' with improved binding affinity and high sequence specificity in comparison to its parent molecule AzaHx-PI (1). However, 2-Pyr-AzaHx is detrimental to DNA binding because of an unfavorable steric clash upon stacking in the minor groove.

Design, synthesis, nuclear localization, and biological activity of a fluorescent duocarmycin analog, HxTfA.

HxTfA 4 is a fluorescent analog of a potent cytotoxic and antimalarial agent, TfA 3, which is currently being investigated for the development of an antimalarial vaccine, PlasProtect®. HxTfA contains a p-anisylbenzimidazole or Hx moiety, which is endowed with a blue emission upon excitation at 318 nm; thus enabling it to be used as a surrogate for probing the cellular fate of TfA using confocal microscopy, and addressing the question of nuclear localization. HxTfA exhibits similar selectivity to TfA for A-tract sequences of DNA, alkylating adenine-N3, albeit at 10-fold higher concentrations. It also possesses in vitro cytotoxicity against A549 human lung carcinoma cells and Plasmodium falciparum. Confocal microscopy studies showed for the first time that HxTfA, and by inference TfA, entered A549 cells and localized in the nucleus to exert its biological activity. At biologically relevant concentrations, HxTfA elicits DNA damage response as evidenced by a marked increase in the levels of γH2AX observed by confocal microscopy and immunoblotting studies, and ultimately induces apoptosis.

Alternate pathway for the click reaction of 2-(2-azidophenyl)-4,5-diaryloxazoles.

The CuSO4/ascorbate-mediated 'click' reaction of 2-(2-azidophenyl)-4,5-diaryloxazoles and arylacetylenes proceeded through an alternate pathway whereby reduction of the azide predominated over formation of the 1,2,3-triazole-forming cycloaddition. The unimolecular product, 2-(2-aminophenyl)-4,5-diphenyloxazole, was isolated which appears to be a formal reduction of the arylazide to the corresponding arylamine. A series of oxazoles which possessed various substituents (F, Cl, Br, OCH3) on the 4,5-diaryl rings and having the 2-azido group on the 2-oxazolylphenyl position were submitted to the same 'click' conditions and gave the corresponding arylamine products (73-99%). The reaction appears to be specific toward the ortho-azido substitution of the polycyclic system, as the corresponding azidomethyl-substituted phenyl oxazoles do not give the 'reduction' products but gave the expected click products with the acetylenic co-reactants.