Structure-based design of novel melanin-concentrating hormone receptor-1 ligands based on saturated nitrogen-containing heterocycles.

Melanin Concentrating Hormone (MCH) receptor is a G protein-coupled receptor (GPCR) with two subtypes R1 and R2. MCH-R1 is involved in the control of energy homeostasis, feeding behavior and body weight. Many studies have proved that administration of MCH-R1 antagonists significantly reduces food intake and causes weight loss in animal models. Herein, we report the optimization of our previously reported virtual screening hits into novel MCH-R1 ligands with chiral aliphatic nitrogen-containing scaffolds. The activity was improved from the micromolar range of the initial leads to 7 nM. We also disclose the first MCH-R1 ligands based on a diazaspiro[4.5]decane nucleus with sub-micromolar activity. A potent MCH-R1 antagonist with acceptable pharmacokinetic profile could represent a new hope for the management of obesity.

4-Azido-7-nitrobenzoxadiazole as innovative clickable fluorescence probe for trace and selective quantification of ethinylestradiol in human plasma.

Quantification of ethinylestradiol (EE) in biological matrices is challenging as it is a very potent drug with a very low Cmax (75 pg.ml-1 ). Despite the high sensitivity of fluorometric methods, the detection of EE was confined because its structure exhibited very limited fluorescence. Therefore, it must be derivatized first using a fluorogenic agent to produce a more potent fluorescence derivative to achieve the desired ultrasensitive bioanalysis. Here, for the first time, we proposed a promising click fluorescent probe, 4-azido-7-nitrobenzoxadiazole (NBD-AZ) to react with the alkyne group of EE, with the help of copper sulphate and l-ascorbic acid to give a highly fluorescent and stable 1,2,3-triazole derivative. Density functional theory calculation revealed how the triazole formation affects the quantum yield and fluorescence of click reaction product when compared with NBD-AZ. The resulting triazole exhibited a strong signal at a wavelength of 540 nm after excitation at 470 nm. Reaction parameters impacting the intensity of fluorescence were cautiously studied and optimized. The suggested approach has shown outstanding performance, high linearity (25-300 pg.ml-1 ) and a low detection limit of 7.5 pg.ml-1 . The enhanced sensitivity and selectivity were exploited for analyzing EE in plasma using liquid-liquid extraction for samples cleaning up without interference from any biological components and with a mean % recovery of 100.13 ± 0.39. Accuracy, sensitivity, selectivity, simplicity, and cost-effectiveness make this approach a convincing, promising, and appealing alternative to the reported analytical methods for EE bioanalysis in different matrices.

Evaluation of the Effect of Different Construction Techniques of CAD-CAM Milled, 3D-Printed, and Polyamide Denture Base Resins on Flexural Strength: An In Vitro Comparative Study.

PURPOSE: To compare the flexural strength of computer-aided design and computer-aided manufacturing (CAD-CAM) milled denture base resin (DBR), 3D-printed DBR, polyamide, and conventional compression-molded DBR. MATERIALS AND METHODS: Six denture base resins were used, one conventional heat-polymerized (Vertex), two milled CAD-CAM (AvaDent and Polident), two 3D-printed (Harz and NextDent), and one flexible polyamide (Polyamide). According to ISO 20795-1:2013, 60 specimens (65×10×3 mm) were constructed and divided into six groups (n = 10), according to DBR type. The flexural strength was measured using a universal testing machine and three-point loading test. Data were collected and analyzed using one-way ANOVA and Tukey's pair-wise post hoc tests (α = 0.05). RESULTS: One-way ANOVA results showed significant differences in flexural strengths between the tested DBRs (p˂0.001). Milled denture base resins (AvaDent and Polident) had significantly higher flexural strength values than the other groups (p˂0.001) and were followed by Vertex and NextDent, while Polyamide and Harz had the lowest values. Polyamide and Harz denture base resins had significantly lower flexural strength values than conventional denture base resin (p˂0.001). CONCLUSION: CAD-CAM milled DBRs showed the highest flexural strength when compared with conventional compression-molded or 3D-printed DBRs, while 3D-printed DBRs and polyamide showed the lowest flexural strengths.

Anticancer Effects of New Ceramides Isolated from the Red Sea Red Algae Hypnea musciformis in a Model of Ehrlich Ascites Carcinoma: LC-HRMS Analysis Profile and Molecular Modeling.

Different classes of phytochemicals were previously isolated from the Red Sea algae Hypnea musciformis as sterols, ketosteroids, fatty acids, and terpenoids. Herein, we report the isolation of three fatty acids-docosanoic acid 4, hexadecenoic acid 5, and alpha hydroxy octadecanoic acid 6-as well as three ceramides-A (1), B (2), and C (3)-with 9-methyl-sphinga-4,8-dienes and phytosphingosine bases. Additionally, different phytochemicals were determined using the liquid chromatography coupled with electrospray ionization high-resolution mass spectrometry (LC-ESI-HRMS) technique. Ceramides A (1) and B (2) exhibited promising in vitro cytotoxic activity against the human breast adenocarcinoma (MCF-7) cell line when compared with doxorubicin as a positive control. Further in vivo study and biochemical estimation in a mouse model of Ehrlich ascites carcinoma (EAC) revealed that both ceramides A (1) and B (2) at doses of 1 and 2 mg/kg, respectively, significantly decreased the tumor size in mice inoculated with EAC cells. The higher dose (2 mg/kg) of ceramide B (2) particularly expressed the most pronounced decrease in serum levels of vascular endothelial growth factor -B (VEGF-B) and tumor necrosis factor-α (TNF-α) markers, as well as the expression levels of the growth factor midkine in tumor tissue relative to the EAC control group. The highest expression of apoptotic factors, p53, Bax, and caspase 3 was observed in the same group that received 2 mg/kg of ceramide B (2). Molecular docking simulations suggested that ceramides A (1) and B (2) could bind in the deep grove between the H2 helix and the Ser240-P250 loop of p53, preventing its interaction with MDM2 and leading to its accumulation. In conclusion, this study reports the cytotoxic, apoptotic, and antiangiogenic effects of ceramides isolated from the Red Sea algae Hypnea musciformis in an experimental model of EAC.

Comparative Effect of Different Surface Treatments on the Shear Bond Strength of Two Types of Artificial Teeth Bonded To Two Types of Denture Base Resins.

PURPOSE: This in vitro study aims to assess the impact of various surface treatments on the shear bond strength (SBS) of two types of artificial teeth and denture base resins (DBRs). MATERIALS AND METHODS: Two types of DBRs (CAD/CAM-milled and heat-polymerized) and two types of denture teeth (acrylic and composite) were investigated. Teeth were cut into slices (5 × 5 × 2 mm) and divided according to surface treatment into four subgroups (n = 10): no treatment (control), air abrasion (Alumina-blasting; AB), bur roughening, and dichloromethane (DCM) subgroups. According to manufacturer recommendations, the treated tooth slices were bonded to the acrylic disk of DBRs. The SBS test was performed using a universal testing machine. ANOVA was used for results analysis followed by Tukey's post hoc tests (α = 0.05). RESULTS: DCM and AB increased the SBS of acrylic teeth to heat-polymerized DBR compared with other groups (p < 0.001). All surface treatments showed no significant difference in CAD/CAM DBR with acrylic teeth (p = 0.059; AB, p = 0.319; bur roughening, p = 0.895; DCM), while there was a significant decrease in SBS with composite teeth (p ˂ 0.001). Between teeth, acrylic teeth showed a statistically significant increase in SBS compared to composite teeth (p < 0.001). CONCLUSION: AB and DCM application improved the SBS for acrylic teeth with the heat-polymerized DBR when compared with the untreated group, but none of the surface treatment agents showed significant improvement with CAD/CAM DBR. All surface treatment agents reduced the SBS for composite teeth with CAD/CAM DBR while AB only increased the SBS with heat-polymerized DBR.

Strength and Surface Properties of a 3D-Printed Denture Base Polymer.

PURPOSE: This in vitro study evaluated the flexural strength, impact strength, hardness, and surface roughness of 3D-printed denture base resin subjected to thermal cycling treatment. MATERIALS AND METHODS: According to ISO 20795-1:2013 standards, 120 acrylic resin specimens (40/flexural strength test, 40/impact strength, and 40/surface roughness and hardness test, n = 10) were fabricated and distributed into two groups: heat-polymerized; (Major.Base.20) as control and 3D-printed (NextDent) as experimental group. Half of the specimens of each group were subjected to 10,000 thermal cycles of 5 to 55°C simulating 1 year of clinical use. Flexural strength (MPa), impact strength (KJ/m2 ), hardness (VHN), and surface roughness (µm) were measured using universal testing machine, Charpy's impact tester, Vickers hardness tester, and profilometer, respectively. Data were analyzed by ANOVA and Tukey honestly significant difference (HSD) test (α = 0.05). RESULTS: The values of flexural strength (MPa) were 86.63 ± 1.0 and 69.15 ± 0.88; impact strength (KJ/m2 )-6.32 ± 0.50 and 2.44 ± 0.31; hardness (VHN)-41.63 ± 2.03 and 34.62 ± 2.1; and surface roughness (µm)-0.18 ± 0.01 and 0.12 ± 0.02 for heat-polymerized and 3D-printed denture base materials, respectively. Significant differences in all tested properties were recorded between heat-polymerized and 3D-printed denture base materials (P < 0.001). Thermal cycling significantly lowered the flexural strength (63.93 ± 1.54 MPa), impact strength (2.40 ± 0.35 KJ/m2 ), and hardness (30.17 ± 1.38 VHN) of 3D-printed resin in comparison to thermal cycled heat-polymerized resin, but surface roughness showed non-significant difference (p = 0.262). CONCLUSION: 3D-printed resin had inferior flexural strength, impact strength, and hardness values than heat-polymerized resin, but showed superior surface roughness. Temperature changes (thermal cycling) significantly reduced the hardness and flexural strength and increased surface roughness, but did not affect the impact strength.

Metal dichalcogenide nanomeshes: structural, electronic and magnetic properties.

Motivated by the successful preparation of two-dimensional transition metal dichalcogenide (2D-TMD) nanomeshes in the last three years, we use density functional theory (DFT) to study the structural stability, mechanical, magnetic, and electronic properties of porous 2H-MoX2 (X = S, Se and Te) without and with pore passivation. We consider structures with multiple, systematically created pores. The molecular dynamics simulations and cohesive energy calculations showed the stability of the 2D-TMD nanomeshes, with larger stability for those with smaller pores. The lattice undergoes some deformations to accommodate the pore energetically, and as the pore size increases Young's modulus decreases. In most cases, the missing metal atoms disrupt the spin states' even population, resulting in some nanomeshes becoming magnetic. The electronic gaps of the MoX2 nanomesh systems are diminished because of the emergence of pore-edge localized mid-gap metal 4d states in the spin-polarized spectrum, making some systems half-metallic. The oxygen passivation of the pore edges of 2D-TMD nanomeshes restores the even population of spin states, and makes those systems metallic. Our results can be used in different applications such as spintronics, ion chelation, and molecular sensing applications.

Nifuroxazide Mitigates Angiogenesis in Ehlrich's Solid Carcinoma: Molecular Docking, Bioinformatic and Experimental Studies on Inhibition of Il-6/Jak2/Stat3 Signaling.

Nifuroxazide is an antidiarrheal medication that has promising anticancer activity against diverse types of tumors. The present study tested the anticancer activity of nifuroxazide against Ehrlich's mammary carcinoma grown in vivo. Furthermore, we investigated the effect of nifuroxazide on IL-6/jak2/STAT3 signaling and the possible impact on tumor angiogenesis. The biological study was supported by molecular docking and bioinformatic predictions for the possible effect of nifuroxazide on this signaling pathway. Female albino mice were injected with Ehrlich carcinoma cells to produce Ehrlich's solid tumors (ESTs). The experimental groups were as follows: EST control, EST + nifuroxazide (5 mg/kg), and EST + nifuroxazide (10 mg/kg). Nifuroxazide was found to reduce tumor masses (730.83 ± 73.19 and 381.42 ± 109.69 mg vs. 1099.5 ± 310.83) and lessen tumor pathologies. Furthermore, nifuroxazide downregulated IL-6, TNF-α, NFk-ß, angiostatin, and Jak2 proteins, and it also reduced tumoral VEGF, as indicated by ELISA and immunohistochemical analysis. Furthermore, nifuroxazide dose-dependently downregulated STAT3 phosphorylation (60% and 30% reductions, respectively). Collectively, the current experiment shed light on the antitumor activity of nifuroxazide against mammary solid carcinoma grown in vivo. The antitumor activity was at least partly mediated by inhibition of IL-6/Jak2/STAT3 signaling that affected angiogenesis (low VEGF and high angiostatin) in the EST. Therefore, nifuroxazide might be a promising antitumor medication if appropriate human studies will be conducted.

The impacts of seasonal variation on the immune status of Nile tilapia larvae and their response to different immunostimulants feed additives.

Few data are available on the thermal tolerance of Nile tilapia fish larvae in relation to their immune status and survival. The aims of this work were to evaluate the immune status of one day old Nile tilapia (Oreochromis niloticus) larval stage collected at the beginning (March), middle (August) and at the end (October) of hatching season through morphometric assessment of the larvae parameters including yolk sac diameter, body length and width as well as the expression of some immune-related genes (rag, sacs and tlr), inflammatory (il1b and il8) and stress related genes (hsp27, hsp70). Also, to compare the effect of three different immunostimulants (ß-glucan, Vitamin C, and methionine/lysine amino acids mix) on the expression of the studied genes at two variant temperatures (23 ± 1 °C and 30 ± 1 °C) in experimental study for 21 days. The immune status of Nile tilapia is affected by thermal fluctuation throughout the hatching season reflected by altered yolk sac size, length, and expression of the immune and stress related genes of the larvae, the best performances was observed at the beginning of the hatching season (March). High temperature (30 °C) suppress immune and stress responses throughout downregulation of all the genes under study, mask any effects for the immunostimulants, increased mortality in fish larvae suggesting narrow thermal tolerance range for the larvae compared with the adult fish. We recommend the use of amino acid mix as immunostimulant for Nile tilapia larvae, it reduces the mortality percentage and improve cellular response. Also, the use of ß-glucan should be prohibited during this developmental stage of larvae, it induced the highest mortality percentage.

Synthesis and Molecular Modelling Studies of New 1,3-Diaryl-5-Oxo-Proline Derivatives as Endothelin Receptor Ligands.

The synthesis of seventeen new 1,3-diaryl-5-oxo-proline derivatives as endothelin receptor (ETR) ligands is described. The structural configuration of the new molecules was determined by analyzing selected signals in proton NMR spectra. In vitro binding assays of the human ETA and ETB receptors allowed us to identify compound 31h as a selective ETAR ligand. The molecular docking of the selected compounds and the ETA antagonist atrasentan in the ETAR homology model provided insight into the structural elements required for the affinity and the selectivity of the ETAR subtype.

Identification of a new small molecule chemotype of Melanin Concentrating Hormone Receptor-1 antagonists using pharmacophore-based virtual screening.

MCH receptor is a G protein-coupled receptor with two subtypes R1 and R2. Many studies have demonstrated the role of MCH-R1 in feeding and energy homeostasis. It has been proven that oral administration of small molecule MCH-R1 antagonists significantly reduces food intake and causes a dose-dependent weight loss. In this study, two ligand-based pharmacophores were developed and validated based on recently published MCH-R1 antagonists with diverse structures. Successful pharmacophores had one hydrogen bond acceptor, one positive ionizable, one ring aromatic and two or three hydrophobic groups. These 3D-QSAR models were used for virtual screening of the ZINC chemical database resulting in the identification of nine compounds with more than 50% displacement of radiolabeled MCH at a 20 µM concentration. Moreover, four of these compounds showed antagonistic activities in Aequorin functional assay, including MH-3 which is the first MCH-R1 antagonist based on a diazaspiro[4.5]decane scaffold. The most active compounds were also docked into our previously published MCH-R1 homology model to gain insights into their binding determinants. These compounds could represent a viable starting scaffold for the design of potent MCH-R1 antagonists with improved pharmacokinetic properties as an effective treatment for obesity.

Synthesis and biological evaluation of novel cYY analogues targeting Mycobacterium tuberculosis CYP121A1.

The rise in multidrug resistant (MDR) cases of tuberculosis (TB) has led to the need for the development of TB drugs with different mechanisms of action. The genome sequence of Mycobacterium tuberculosis (Mtb) revealed twenty different genes coding for cytochrome P450s. CYP121A1 catalyzes a CC crosslinking reaction of dicyclotyrosine (cYY) producing mycocyclosin and current research suggests that either mycocyclosin is essential or the overproduction of cYY is toxic to Mtb. A series of 1,4-dibenzyl-2-imidazol-1-yl-methylpiperazine derivatives were designed and synthesised as cYY mimics. The derivatives substituted in the 4-position of the phenyl rings with halides or alkyl group showed promising antimycobacterial activity (MIC 6.25 µg/mL), with the more lipophilic branched alkyl derivatives displaying optimal binding affinity with CYP121A1 (iPr KD = 1.6 µM; tBu KD = 1.2 µM). Computational studies revealed two possible binding modes within the CYP121A1 active site both of which would effectively block cYY from binding.

Synthesis, biological evaluation, and molecular docking investigation of benzhydrol- and indole-based dual PPAR-γ/FFAR1 agonists.

Type-2 diabetes mellitus is a progressive cluster of metabolic disorders, representing a global public health burden affecting more than 366 million people worldwide. We recently reported the discovery of three series of novel agents showing balanced activity on two metabolic receptors, peroxisome proliferator activated receptor-γ (PPAR-γ) and free fatty acid receptor 1 (FFAR1), also known as GPCR40. Our designing strategy relied on linking the thiazolidinedione head with known GPCR privilege structures. To further investigate this concept, two new scaffolds, the benzhydrol- and indole-based chemotypes, were introduced here in. Our optimization campaign resulted in three compounds; 15a, 15c, and 15d, with affinities in the low micromolar range on both targets. In vivo study of selected test compounds, revealed that 15c possesses a significant anti-hyperglycemic and anti-hyperlipidemic activities superior to rosiglitazone in fat-fed animal models. Molecular docking analysis was conducted to explain the binding modes of both series. These compounds could lead to the development of the unique antidiabetic agent acting as insulin sensitizer as well as insulin secretagogue.

Novel pyrazoles and pyrazolo[1,2-a]pyridazines as selective COX-2 inhibitors; Ultrasound-assisted synthesis, biological evaluation, and DFT calculations.

COX-2 is an inducible enzyme mediating inflammatory responses. Selective targeting of COX-2 is useful for developing anti-inflammatory agents devoid of ulcerogenic activity. Herein, we report the design and synthesis of a series of pyrazoles and pyrazolo[1,2-a]pyridazines with selective COX-2 inhibitory activity and in vivo anti-inflammatory effect. Both series were accessed through acid-catalyzed ultrasound-assisted reactions. The most active compounds in this study are two novel molecules, 11 and 16, showing promising selectivity and decent IC50 of 16.2 and 20.1nM, respectively. These compounds were also docked into the crystal structure of COX-2 enzyme (PDB ID: 3LN1) to understand their mode of binding. Finally, Mulliken charges and electrostatic surface potential were calculated for both compound 11 and celecoxib using DFT method to get insights into the molecular determinants of activity of this compound. These results could lead to the development of novel COX-2 inhibitors with improved selectivity.

Homology modeling and explicit membrane molecular dynamics simulation to delineate the mode of binding of thiazolidinediones into FFAR1 and the mechanism of receptor activation.

Free fatty acid receptor 1 (FFAR1) is a member of a previously characterized cluster of orphan G protein-coupled receptors (GPCRs). Later, this orphan receptor was identified as a target of medium- to long-chain free fatty acids in ß-cells of the pancreas. Administration of FFAR1 agonists has been proved to potentiate glucose-stimulated insulin secretion from pancreatic ß-cells. It was reported that some thiazolidinediones (TZDs), the best studied PPARγ agonists, are also able to stimulate FFAR1 in a dose-dependent manner. In the present study, a homology model of the human FFAR1 was constructed and inserted into a pre-equilibrated DPPC/TIP3P membrane system. This system was then simulated for 20 ns in complex with the FFAR1 agonist GW9085, as well as rosiglitazone and pioglitazone. We noticed that the salt bridge between Glu172 and Arg258 and the H bond between Glu145 and His153 could be responsible for the stabilization of the receptor in the inactive state. Moreover, we described for the first time the binding mode of TZDs in the binding site of FFAR1. The thiazolidinedione head forms a hydrogen bonding network with the critical polar residues in the binding site, Arg258 and Asn244, while the rest of the molecule is embedded into the receptor hydrophobic pocket. Based on this modeling study, we arrived at a proposal of the pharmacophore required for binding to both PPARγ and FFAR1. Insights gained from this investigation should provide future directions for the design of novel dual acting antidiabetic agents.

Design, synthesis, and enzyme kinetics of novel benzimidazole and quinoxaline derivatives as methionine synthase inhibitors.

Methionine synthase catalyzes the transfer of a methyl group from 5-methyltetrahydrofolate to homocysteine, producing methionine and tetrahydrofolate. Benzimidazole and deazatetrahydrofolates derivatives have been shown to inhibit methionine synthase by competing with the substrate 5-methyltetrahydrofolate. In this study, a novel series of substituted benzimidazoles and quinoxalines were designed and assessed for inhibitory activity against purified rat liver methionine synthase using a radiometric enzyme assay. Compounds 3g, 3j, and 5c showed the highest activity against methionine synthase (IC50: 20 µM, 18 µM, 9 µM, respectively). Kinetic analysis of these compounds using Lineweaver-Burk plots revealed characteristics of mixed inhibition for 3g and 5c; and uncompetitive inhibition for 3j. Docking study into a homology model of the rat methionine synthase gave insights into the molecular determinants of the activity of this class of compounds. The identification of these drug-like inhibitors could lead the design of the next generation modulators of methionine synthase.

Effect of clasp design on retention at different intervals using different abutment materials and in a simulated oral condition.

PURPOSE: The purpose of this study was to compare the retention of circlet (E) clasps and back-action clasps against three abutment surface materials during long-term simulation of attachment and detachment. MATERIAL AND METHODS: Forty-eight test models were constructed by placing premolars (natural abutments or metal dies) inside metal blocks to test different abutment retention surface materials (sound enamel, composite resin, and glass-ceramic; 16 each). The models were duplicated into investment models for construction of circlet (E) and back-action clasps. Removal and insertion cycling of clasps was carried out for 250, 500, 1000, 2000, 4000, 8000, and 16,000 cycles. The retention of each clasp was measured before cycling and after each interval. Data were analyzed using 1-way-ANOVA, 2-way-ANOVA, and Mann-Whitney U tests. RESULTS: No significant differences in retention of either clasp were found between the three abutment material surfaces; however, there was a significant decrease in retention force of the circlet (E) clasp between 1000 and 2000 cycles but not of the back-action clasp. CONCLUSIONS: (1) The back-action clasp maintains its retention force for a longer period than the circlet (E) clasp. (2) Composite resin contouring of teeth provided retention comparable to enamel and a ceramic material.

Differential binding of latrunculins to G-actin: a molecular dynamics study.

Latrunculins are unique macrolides containing a thiazolidinone moiety. Latrunculin A (1), latrunculin B (2), 16-epi-latrunculin B (3), and latrunculin T (4) were isolated from the Red Sea sponge Negombata magnifica. In the present study, after testing compounds 2-4 for cytotoxic activity, they were docked into the crystal structure of G-actin and subjected to binding energy calculation and a 20 ns MD simulation. The modeling study shows that latrunculins binding depends on both hydrophobic interaction of the macrocycle as well as H bonding of the thiazolidinone ring with Asp157 and Thr186. It was noticed that epimerization at C16 of latrunculin B was well tolerated as it could form an alternative H bonding network. However, opening of the macrocyclic ring deteriorates the actin binding due to reduced hydrophobicity. MD simulation showed that latrunculin B (2) possesses a more significant stabilizing effect on G-actin than latrunculin T (4) and could efficiently hinder the flattening transition of G-actin into F-actin. These findings could explain, at the molecular level, the impact of epimerization and macrolide ring-opening on latrunculins activity, an issue that has not been addressed before. Also, the study gives insights into the mechanism of cytotoxicity of diverse latrunculins and provides direction for future lead optimization studies.

Structural basis for the selectivity of 3rd generation EGFR inhibitors: a molecular dynamics study.

Activating mutations in the EGFR kinase domain are known to be a common cause of Non-Small Cell Lung Cancer (NSCLC) and are thus targeted for treatment. First generation Tyrosine Kinase Inhibitors (TKIs) were used to treat NSCLC patients with the known activating mutations L858R and exon 19 deletion but were resisted by a second mutation T790M in the active site of the kinase domain. Second generation members of TKIs have an electrophilic moiety that can form a covalent bond with Cys797 and are effective against T790M EGFR but are toxic because they inhibit WT EGFR as well. Third generation TKIs, like Osimertinib, can bind to and irreversibly inhibit T790M mutants selectively, while sparing the wild-type enzyme. Thus, they possess a better safety profile and a wider therapeutic window. However, the reason behind their selectivity is still not well understood. In this study, computational MD simulations were carried out on Osimertinib in complex with both WT and L858R/T790M Double Mutant (DM) EGFR to provide an insight into the selectivity of Osimertinib and its molecular interactions within the active site. A high-resolution trajectory analysis showed that the key selectivity residues are Val726, met793, and Cys797. Interaction of Osimertinib with these residues is improved due to the T790M mutation which optimizes the ligand orientation for binding, as evident from the RMSD and the distances monitored. These results can provide guidance for the development of more selective 3rd generation EGFR TKIs.Communicated by Ramaswamy H. Sarma.

Novel fluorescence approach for trace quantification of levonorgestrel in breast milk based on click reaction with benzonitrofurazan azide (NBD-AZ).

Although the great importance of oral contraceptive agents in birth control, their existence in breast milk became a cause for concern, since infant exposure to these hormones is associated with many health problems. Consequentially, developing a sensitive bioanalytical method for monitoring their concentrations in breast milk is an urgent demand to examine the safety or the risk of these compounds on infants. Levonorgestrel is one of the most common contraceptive hormones under concern. Despite the high sensitivity of the fluorometric methods, detection of Levonorgestrel by them is confined because its structure does not exhibit any fluorescence. For the first time, we proposed a promising click fluorescent probe, 4-azido-7-nitrobenzoxadiazole to react with the alkyne group of Levonorgestrel, to give a highly fluorescent triazole derivative that exhibited strong signal at wavelength of 544 nm after excitation at 470 nm. Reaction parameters impacting the fluorescence were cautiously studied and optimized. The suggested approach has been successfully applied in Levonorgestrel estimation in breast milk samples with linearity of (0.4-80 ng.ml-1) and low detection limit of 0.12 ng.ml-1without interferences from any biological components and with mean % recovery of 97.84 ± 2.73. Accuracy, sensitivity, selectivity, simplicity, and low-cost makes this approach a convincing, promising, and appealing alternative over reported analytical methods for Levonorgestrel bioanalysis in different matrices.