In-Silico Aided Screening and Characterization Results in Stability Enhanced Novel Roxadustat Co-Crystal.

Roxadustat (RXD) is an approved drug substances for the treatment of renal anemia. It has poor aqueous solubility and photochemical stability. This study employs a comprehensive approach to enhance the stability and physicochemical properties RXD through coformer selection and characterization. The investigation integrates delta pKa analysis, molecular complementary assessment, molecular electrostatic potential surface analysis, and machine learning techniques to predict potential co-crystal formation and binding interactions between drug molecules and coformers. The co-crystal screening which lead to in a novel RXD-nicotinamide co-crystal (RXD-NA). Experimental characterization underscores the physical and chemical stability of the co-crystals. To elucidate the supramolecular synthons and understand the intermolecular interactions in the RXD-NA co-crystal, Hirshfeld surfaces analysis, quantum theory of atoms in molecules (QTAIM) analysis and non-covalent interaction (NCI) analysis were performed. Computational analysis of photo-isomer formation aligns with experimental observations, further enhancing our understanding of RXD-coformer interactions. RXD-NA co-crystal was found photo-chemically stable as compared to free base API drug substance. This integrated methodology provides a systematic framework for informed co-crystal design, holding promise for optimizing RXD formulations based on molecular interactions and stability considerations. Consequently, this study contributes valuable insights to the field of rational drug design and formulation optimization.

Transaminase-mediated chiral selective synthesis of (1R)-(3-methylphenyl)ethan-1-amine from 1-(3-methylphenyl)ethan-1-one: process minutiae, optimization, characterization and 'What If studies'.

Transaminases capable of carrying out chiral selective transamination of 1-(3-methylphenyl)ethan-1-one to (1R)-(3-methylphenyl)ethan-1-amine were screened, and ATA-025 was the best enzyme, while dimethylsulfoxide (10% V/V) was the best co-solvent for said bioconversion. The variables such as enzyme loading, substrate loading, temperature, and pH for development of process displaying maximum conversion with good product formation and higher yield were optimized. The ambient processing conditions were 10% enzyme loading/50 g/L substrate loading/45 °C/pH 8.0, and 5% enzyme loading/36.78 g/L substrate loading/42.66 °C/pH 8.2 displaying maximum conversion 99.01 ± 2.47% and 96.115 ± 1.97%, and 76.93 ± 1.05% and 73.12 ± 1.04% yield with one factor at a time approach and numerical optimization with Box Behnken Design, respectively. In the final optimized reaction, ATA-025 showed the highest 99.22 ± 2.61% conversion, 49.55 g/L product formation, with an actual product recovery of 38.16 g corresponding to a product yield 77.03 ± 1.01% with respect to the product formed after reaction. The purity of recovered product (1R)-(3-methylphenyl)ethan-1-amine formed was ≥ 99% (RP-HPLC), and chiral purity ≥ 98.5% (Chiral-GC), and it was also confirmed and characterized with instrumental methods using boiling point, LC-MS, ATR-FTIR, and 1H NMR. The findings of 'What If' studies performed by investigating timely progress of reaction on gram scale by drastically changing the process parameters revealed a substantial modification in process variables to achieve desired results. (1R)-(3-methylphenyl)ethan-1-amine synthesized by green, facile and novel enzymatic approach with an optimized process could be used for synthesis of different active pharma entities.

Preparation, characterization and stability of cross linked nitrilase aggregates (nitrilase-CLEAs) for hydroxylation of 2-chloroisonicotinonitrile to 2-chloroisonicotinic acid.

Nitrilases capable of performing hydroxylation of 2-chloroisonicotinonitrile to 2-chloroisonicotinic acid were screened, and ES-NIT-102 was the best nitrilase for said biotransformation. Nitrilase was immobilized as cross linked enzyme aggregates (nitrilase-CLEAs) by fractional precipitation with iso-propanol, and cross linked with glutaraldehyde. The nitrilase-CLEAs prepared with optimized 35 mM glutaraldehyde for 120 min cross linking time had 82.36 ± 4.45% residual activity, and displayed type-II structural CLEAs formation as confirmed by particle size, SEM, FTIR, and SDS-PAGE analysis. Nitrilase-CLEAs had superior pH and temperature stability, showed a shift in optimal temperature by 5 °C, and retained nearly 1.5 to 1.7 folds activity over free nitrilase at 50 °C and 55 °C after more than 9 h incubation. Nitrilase-CLEAs showed reduced affinity and decreased conversion of substrate as indicated by slightly higher Km values by 5.19% and reduced Vmax by 17%. Furthermore, these nitrilase-CLEAs showed 98% conversion, 94.72 g/L product formation, and 83.30% recovery after 24 h when used for hydroxylation of 2-chloroisonicotinonitrile to 2-chloroisonicotinic acid. Nitrilase-CLEAs were catalytically active for 3 cycles showcasing 81% conversion, 75.53 g/L product formation and 66.42% yield. The recovered product was confirmed by HPLC, FTIR, LC-MS, and 1H NMR, and displayed > 99% purity.

Substituent Orchestration in Dimethylquinoxaline Derivatives: A Tool for Fishing Out Appropriate CDK5 Inhibitors as Potential Therapeutics for Alzheimer's.

A set of new heterocyclic analogs (Compounds I-IX), comprising of 6,7 dimethyl Quinoxalines were found to be active against the receptor GSK3ß (Compounds IV-V) (Chem. Biodiversity 2021, 18, e2100364). In an effort to modulate effective CDK5 inhibitors herein our hypothesis underpinned to fish out an appropriate derivative from the same quinoxaline series, as these two targets GSK3ß and CDK5 shared structural resemblance with each other. Aligned to the goal we have synthesized Compounds I-IX, characterized them using a combination of spectroscopic techniques and evaluated their activities against CDK5. Our analysis reflected that the adjacently located alkoxy/hydroxy functionality derivatives namely Compounds III and VI, to be the most potent (micromolar) amongst others in the series, backed by Density Functional Theory (DFT) calculations and molecular modelling studies. Also, the efficacy of the Compounds I-IX, were monitored in few other members of the CMGC family namely DYRK1A, CLK1and CK1δ that have been known to be directly involved in hyperphosphorylation of Tau. But unfortunately in none of the targets, our quinoxaline series were active. In a nut shell further optimisation of these intelligent nucleus, would not only lead to the discovery of novel pharmacophores, but also marked selectivity against a pool of kinases, thereby implementing a distinct roadmap towards the design of potential therapeutics against Alzheimer's.

Rapid chiral separation of potential antibiotics using supercritical fluid chromatography.

WHO is searching most active antibiotics due to the bacterial resistance problem. The activities of the racemic antibiotics may be augment by preparing optically active antibiotics by the chiral separation. Chiral separation of potential antibiotics such as cefotaxime and ofloxacin was studied using amylose-based packing chiral stationary phases (CSPs) such as Chiralpak IA and Chiralpak IG. Supercritical fluid chromatography (SFC) was employed to carry out this study. Both immobilized CSPs such as Chiralpak IA and IG have sown remarkable selectivity for the reported drugs by using SFC. The values of retention factor (k) for ofloxacin enantiomers were 9.63 and 11.81, followed by 2.94 and 5.96 for cefotaxime enantiomers. The values of separation factor (α) for both the reported drugs were 1.22 and 2.03, respectively Similarly, the values of resolution factor (Rs) for both the enantio-selective drugs were 1.49 and 2.06, separately and respectively. The chiral recognition mechanism was developed and it was observed that the π-π interactions are playing a major role. The developed method is effective, reproducible, eco-friendly, and may be used to discriminate the enantiomers of the reported drugs in any sample.

APuML: An Efficient Approach to Detect Mobile Phishing Webpages using Machine Learning.

Nowadays, the growth of mobile phones users has gained a significant increase because of the features offered by them in abundant amounts. These devices are being used rapidly for accessing the web and many online services. However, the security mechanisms that are available in smartphones are not yet mature. Therefore, smartphones are vulnerable to various types of attacks, such as phishing. The browsers on smartphones are very trivial and the smartphones security abilities have been lessened, to match the smartphone's capabilities. Therefore, detection of the malicious website is different from the previously known technique, which is used on the desktop. Many anti-phishing techniques for mobile devices have been developed but still, there is a lack of a full-fledged solution. Therefore, this paper presents an efficient approach to detect malicious mobile webpages. The proposed approach APuML (Anti Phishing using Machine Learning) extracts all the static and site popularity features from the given URL to create a feature vector. An appropriate machine learning classification algorithm is then applied on the feature set to obtain the result and update the database accordingly. In our approach, the Random Forest classifier outperforms over other classifiers and achieved detection accuracy of 93.85%. We have also created an endpoint application for the users to interact with our system using his/her mobile devices. Moreover, the proposed approach can identify drive-by downloads attack, zero-day attack and clickjacking attack with high accuracy.

A comparative study of chiral separation of proton pump inhibitors by supercritical fluid chromatography and high-performance liquid chromatography.

A comparative study of chiral separation of pantoprazole and rabeprazole is carried out using supercritical fluid chromatography and high-performance liquid chromatography. The columns used were Chiralpak IA and Chiralpak IE. The best mobile phase in supercritical fluid chromatography was carbon dioxide-0.2% triethylamine in methanol (60:40) and 0.1% triethylamine in n-hexane-ethanol (50:50) in high-performance liquid chromatography. For supercritical fluid chromatography, values of the retention factor of pantoprazole enantiomers were 3.97 and 4.88. These values for rabeprazole enantiomers were 6.10 and 7.52. The values of separation and resolution factor for pantoprazole and rabeprazole were 1.23 and 1.23 and 2.20 and 3.36, respectively. Similarly, for high-performance liquid chromatography, the values of retention factor for enantiomers of pantoprazole were 4.02 and 7.32. These values for rabeprazole enantiomers were 5.32 and 7.88, respectively. The values of separation and resolution factor for pantoprazole and rabeprazole were 1.82 and 1.48 and 9.22 and 6.58, respectively. A comparison was carried out, which confirmed supercritical fluid chromatography as the best method due to its fastness, eco-friendly, and inexpensiveness. The reported methods are effective, efficient, and reproducible and may be used to separate and identify pantoprazole and rabeprazole in any unknown samples.

Human CDC2-like kinase 1 (CLK1): a novel target for Alzheimer's disease.

The cdc2-like kinases (CLKs) are an evolutionarily conserved group of dual specificity kinases belonging to the CMGC (cyclin-dependent kinases (CDKs), mitogen-activated protein kinases (MAP kinases), glycogen synthase kinases (GSK) and CDK-like kinases). The CLK family consists of four isoforms namely CLK1, CLK2, CLK3 and CLK4. The human CLK1 encoded protein comprises 454 amino acids and the catalytic domain of CLK1 exhibits the typical protein kinase fold. CLK1 has been shown to autophosphorylate on serine, threonine and tyrosine residues and phosphorylate exogenous substrates on serine and threonine residues. CLK1 plays an important role in the regulation of RNA splicing through phosphorylation of members of the serine and arginine-rich (SR) family of splicing factors. CLK1 is involved in the pathophysiology of Alzheimer's disease by phosphorylating the serine residue in SR proteins. Nuclear speckles of the nucleoplasm contain the stored form of SR proteins and are moderately responsible for the choice of splicing sites during pre-mRNA splicing. Hence, the inhibition of CLK1 can be used as a therapeutic strategy for Alzheimer's disease. Many natural and synthetic molecules are reported to possess CLK1 inhibitory activity. Some specific examples are Marine alkaloid Leucettamine B and KH-CB19. Leucettamine B is a potent inhibitor of CLK1 (15 nM), Dyrk1A (40 nM), and Dyrk2 (35 nM) and a moderate inhibitor of CLK3 (4.5 µM) whereas KH-CB19 is a highly specific and potent inhibitor of the CLK1/CLK4. X-ray crystallographic studies have revealed the binding mode of marine sponge metabolite hymenialdisine and a dichloroindolyl enamino nitrile (KH-CB19) to CLK1. This review focuses on the role of CLKs in the pathophysiology of Alzheimer's disease and therapeutic potential of targeting CLK1 in Alzheimer's disease drug discovery and development. In addition, the recent developments in drug discovery efforts targeting human CLK1 are also highlighted.