A transmembrane scaffold from CD20 helps recombinant expression of a chimeric claudin 18.2 in an in vitro coupled transcription and translation system.

Cluster of differentiation 20 (CD20) is a nonglycosylated, multispanning transmembrane protein specifically integrated by B lymphocytes. Similar to CD20, another four-pass transmembrane protein, claudin 18.2, has attracted attention as an emerging therapeutic target for cancer. However, their poor solubility and toxic nature often hinder downstream applications, such as antibody drug development. Therefore, developing a cost-effective method for producing drug targets with multiple membrane-spanning domains is crucial. In this study, a high yield of recombinant CD20 was achieved through an E. coli-based in vitro coupled transcription-translation system. Surface plasmon resonance results showed that rituximab (an antileukemia drug) has nanomolar affinity with the CD20 protein, which aligns with published results. Notably, a previously hard-to-express claudin 18.2 recombinant protein was successfully expressed in the same reaction system by replacing its membrane-spanning domains with the transmembrane domains of CD20. The folding of the extracellular domain of the chimeric protein was verified using a commercial anti-claudin 18 antibody. This study provides a novel concept for promoting the expression of four-pass transmembrane proteins and lays the foundation for the large-scale industrial production of membrane-associated drug targets, similar to claudin 18.2.

Designing a novel drug-drug conjugate as a prodrug for breast cancer therapy: in silico insights.

Breast cancer (BC) poses a significant global health threat, necessitating innovative therapeutic approaches. The ribosomal s6 kinase 2 (RSK2) has emerged as a promising target due to its roles in cell proliferation and survival. This study proposes a drug-drug conjugate prodrug comprising Methotrexate (hydrophobic) and Capecitabine (hydrophilic) for BC treatment. In silico approaches, including Molecular Docking, Molecular Dynamics Simulations, MM-PBSA, ADME, and DFT calculations were employed to evaluate the prodrug's potential. The designed MET-CAP ligand exhibits a robust docking score (-8.980 kcal/mol), superior binding affinity (-53.16 kcal/mol), and stable dynamic behavior (0.62 nm) compared to native ligands. The DFT results reveal intramolecular charge transfer in MET-CAP (HLG = 0.09 eV), indicating its potential as a BC inhibitor. ADME analysis suggests satisfactory pharmaceutically relevant properties. The results indicate that the conjugated MET-CAP ligand exhibits favorable binding characteristics, stability, and pharmaceutically relevant properties, making it a potential RSK2 inhibitor for BC therapy. The multifaceted approach provides insights into binding interactions, stability, and pharmacokinetic properties, laying the foundation for further experimental validation and potential clinical development.

Bioengineered gold nanoparticles from marine seaweed Acanthophora spicifera for pharmaceutical uses: antioxidant, antibacterial, and anticancer activities.

In this study, we report a green synthesis of pharmaceutically active gold nanoparticles from marine red alga Acanthophora spicifera by the reduction of chloroauric acid. The formation of A. spicifera-mediated gold nanoparticles (As-AuNPs) was characterized by several analytical techniques. The crystalline and face-centered cubic (fcc) structure were confirmed by X-ray diffraction (XRD) analysis. Electron microscopy results confirmed that As-AuNPs were spherical and the average size of particles was < 20 nm. As-AuNPs hold a significant level of antioxidant activities than A. spicifera extract. As-AuNPs exhibited the highest antibacterial activity against Vibrio harveyi than Staphylococcus aureus at 100 µg/ml. Furthermore, As-AuNPs exhibited the utmost cytotoxicity against human colon adenocarcinoma (HT-29) cells and registered the half-maximal inhibitory concentration (IC50) at 21.86 µg/ml. These findings authenticated that the synthesized As-AuNPs possess a broad spectrum of biological activities, and it can be effectively applied in the field of aquaculture and biomedical application.

Evaluation of Rapanone and Nectandrin B as novel inhibitors for targeting the metastatic regulator protein BACH1 using breast cancer cell line Mcf-7.

Cancer formation is defined as the unrestrained proliferation of cells due to various factors acting as a causing agent. A limited number of over-expressed transcription factors are contributed to the development of numerous types of cancer. The metastatic regulator protein BTB And CNC Homology 1 (BACH1) is Cap 'N' Collar (CNC) and it belongs to a basic region leucine zipper (bZIP) family. The presence of the least level concentration of intracellular heme BACH1 forms heterodimers with musculo aponeurotic fibrosarcoma (sMAF) proteins and inhibits or induces the target gene expression. Based on the previous studies, BACH1 plays a critical player in the conditions of senescence and oxidative stress, cycling of cell life, heme degradation pathway and cancer, especially in metastasis. Discovering new anti-cancer drugs (identification of bioactive compounds) stages finally needs to inhibit the target protein. This present study is aimed to screen and identify stability, binding affinity and analysis of pharmacokinetics of selected compounds through structural screening, ADMET, DFT and MESP. From this study, it is revealed that Rapanone and Nectandrin B have the potential to alter the degree of gene expression via binding with the BACH1 allosteric region which will further change the degree of expression of BACH1 downstream target genes involved in the regulation of cancer progression particularly in metastasis. The two plant origin compounds Rapanone and Nectandrin B might be novel candidates for developing anti-cancer drugs. The predicted compounds were further validated through in-vitro experimental approaches.Communicated by Ramaswamy H. Sarma.

Screening of inhibitors as potential remedial against Ebolavirus infection: pharmacophore-based approach.

Ebola virus disease (EVD) has been recognized as a major threat for humans and primates. Till now, therapeutic solution is a challenging for the treatment of Ebola virus disease. Therefore, new, novel with suitable effective antiviral drug against EVD is essential. In the present study pharmacophore modeling, 3DQSAR, molecular docking, DFT and molecular dynamic simulations were used to identify the new, novel with suitable effective potential inhibitors against Ebola virus through the computational methods. From the pharmacophore PHASE modeling, the best five hypothesis pharmacophore features such as three hydrogen bond acceptor, one positive ion and one aromatic ring are taken to study the 3D-QSAR structural model. The designed with correlation co-efficient is found to be R2 = 0.92 and the excellent predictive power with correlation co- efficient is found to be Q2 = 0.82. The 3D-QSAR model is used to study the virtual screening against the chemical libraries compounds (NCI, ZINC, Asinex, LifeChemical, ChemBridge, MayBridge, Enamine and specs) to identify the novel scaffolds. The best binding free energy (39.368769 kcal/mol) and the best docking score (12.419 kcal/mol) for NCI database are obtained from molecular docking and MM-GBSA respectively. The good electronic features of ligands are observed from DFT analysis. Finally, molecular dynamics simulations revealed the stability of ligand protein complexes ranging from 1 nm to 1.5 nm. We anticipated that, this ligand protein complex could be supportive to improve the potent inhibitor against the Ebola viral treatment. Communicated by Ramaswamy H. Sarma.

Induction of Caspase-Mediated Apoptosis in HepG2 Liver Carcinoma Cells Using Mutagen-Antioxidant Conjugated Self-Assembled Novel Carbazole Nanoparticles and In Silico Modeling Studies.

In this study, novel self-assembled carbazole-thiooctanoic acid nanoparticles (CTNs) were synthesized from amino carbazole (a mutagen) and thiooctanoic acid (an antioxidant). The nanoparticles were characterized using hyperspectral techniques. Then, the antiproliferative potential of CTNs was determined in HepG2 liver carcinoma cells. This study employed a solvent-antisolvent interaction method to synthesize a spherical CTN of size less than 50 nm. Moreover, CT was subsequently capped to gold nanoparticles (AuNPs) in the additional comparative studies. The CT derivative was synthesized from carbazole and lipoic acid by the amide bond formation reaction using a coupling agent. Furthermore, it was characterized using infrared (IR), 1H nuclear magnetic resonance, dynamic light scattering (DLS), and transmission electron microscopy techniques. The CT-capped gold nanoparticles (CTAuNPs) were prepared from CT, chloroauric acid, and NaBH4. The CTAuNPs were characterized using ultraviolet-visible, high-resolution TEM, DLS, and Fourier transform IR techniques. The cytotoxicity and apoptosis-inducing ability of both nanoparticles were determined in HepG2 cells. The results demonstrate that CTNs exhibit antiproliferative activity in the cancerous HepG2 cells. Moreover, molecular docking and molecular dynamics studies were conducted to explore the therapeutic potential of CT against human EGFR suppressor protein to gain more insights into the binding mode of the CT, which may show a significant role in anticancer therapy.

In silico Screening of Natural Phytocompounds Towards Identification of Potential Lead Compounds to Treat COVID-19.

COVID-19 is one of the members of the coronavirus family that can easily assail humans. As of now, 10 million people are infected and above two million people have died from COVID-19 globally. Over the past year, several researchers have made essential advances in discovering potential drugs. Up to now, no efficient drugs are available on the market. The present study aims to identify the potent phytocompounds from different medicinal plants (Zingiber officinale, Cuminum cyminum, Piper nigrum, Curcuma longa, and Allium sativum). In total, 227 phytocompounds were identified and screened against the proteins S-ACE2 and M pro through structure-based virtual screening approaches. Based on the binding affinity score, 30 active phytocompounds were selected. Amongst, the binding affinity for beta-sitosterol and beta-elemene against S-ACE2 showed -12.0 and -10.9 kcal/mol, respectively. Meanwhile, the binding affinity for beta-sitosterol and beta-chlorogenin against M pro was found to be -9.7 and -8.4 kcal/mol, respectively. Further, the selected compounds proceeded with molecular dynamics simulation, prime MM-GBSA analysis, and ADME/T property checks to understand the stability, interaction, conformational changes, binding free energy, and pharmaceutical relevant parameters. Moreover, the hotspot residues such as Lys31 and Lys353 for S-ACE2 and catalytic dyad His41 and Cys145 for M pro were actively involved in the inhibition of viral entry. From the in silico analyses, we anticipate that this work could be valuable to ongoing novel drug discovery with potential treatment for COVID-19.

MMP-9 responsive dipeptide-tempted natural protein hydrogel-based wound dressings for accelerated healing action of infected diabetic wound.

The infected diabetic wound ulcer is a significant problem for the diabetic patients, which leads to removal of affected foot site due to its delayed/non-healing tissues. The poly-microbial infections and active matrix metalloproteinases (MMP) are the significant influencing factors to delayed healing in diabetic mice. The main purposes of present investigation are to evaluation the targeted inactivation of MMP and avoid polymicrobial infections by using a combined therapeutic effect of metal chelating dipeptide (L-carnosine) and curcumin with the biocompatible silk protein hydrogel (L-car@cur/SF) dressing in the infected diabetic wound ulcer. The in vitro biological assay methods, such as, cell viability, anti-oxidant activity, anti-inflammatory macrophage cells and inhibition collagenase exhibited that the designed hydrogel matrix to be human cell compatible and could be accelerate for significant diabetic healing potential. The activation of cur/SF matrix by the L-carnosine was persuading the inactivation of matrix metalloproteinase-9 (MMP-9) through its potent chelating effects of Zn2+ ions from the MMP-9 active center. The L-car@cur/SF hydrogel was demonstrated for the effective MMP-9 inactivation and bacterial inhibition via in vivo mice wound site, which indorsed the diabetic wound healing efficiency in streptozotocin-tempted diabetic mice.

Photoluminescent reduced graphene oxide quantum dots from latex of Calotropis gigantea for metal sensing, radical scavenging, cytotoxicity, and bioimaging in Artemia salina: A greener route.

In this work, we report the fabrication of green fluorescent reduced graphene oxide quantum dots (rGOQDs) from the latex of Calotropis gigantea by simple one-step microwave assisted greener route. The latex of Calotropis gigantea calcined at 300°C and its ethanolic extract is used for the synthesis of QDs, The rGOQDs showed particle size ranging from 2 to 8nm and it exhibited green fluorescent in longer UV region at 360-520nm. The rGOQDs graphitic nature was confirmed by RAMAN and XRD analysis. The FTIR, XPS demonstrate that presence of functional groups such as CO, COC, -OH, hence it's addressing them as rGOQDs. It is used to design the greener and economically adopted fluorescent probe for the detection of Pb2+ ions. It provides simple and appropriate for the selective and sensitive detection of Pb2+ ions in water purification process. It also trapped the free radicals and neutralized that and act as an excellent radical scavenger in DPPH radical scavenging assessment. These rGOQDs showed excellent biocompatibility on brine shrimp nauplii (Artemia salina) up to 160µg/mL for 24h incubation. Furthermore, rGOQDS were demonstrated as fluorescent bioimaging probe selectively in the inner digestion part of Artemia salina. In summary, stable, economically viable, highly biocompatible, greener method based rGOQDs were prepared for heavy metal ion detecting, radical scavenging, bioimaging applications which can play a vital role in the future nanotechnology-based biomedical field.

Phyto-mediated synthesis of silver nanoparticles using fucoidan isolated from Spatoglossum asperum and assessment of antibacterial activities.

The present study was aimed to investigate the antibacterial efficacy of fucoidan mediated silver nanoparticles (Fu-AgNPs) synthesized from Spatoglossum asperum. The synthesized Fu-AgNPs were characterized by UV-visible, Field emission - scanning electron microscope (FE-SEM), Tranmission electron microscope (TEM), X-ray diffraction (XRD), Selected area electron diffraction (SAED) pattern, Energy-dispersive X-ray spectroscopy (EDAX), Fourier transform infrared spectroscopy (FT-IR), Dynamic light scattering (DLS) and Zeta potential analysis. The UV-visible spectrum of Fu-AgNPs exhibited a characteristic surface plasmon resonance (SPR) peak at 440 nm. The electron microscopic results revealed that the nanoparticles were spherical to oval in shape and are found to be 20 to 46 nm. Altogether the X-ray diffraction analysis showed that the Fu-AgNPs were crystalline in nature. The FT-IR spectrum confirmed the existence of CC stretching vibration of aromatic compounds and sulfated groups of fucoidan plays a major role in the synthesis of Fu-AgNPs. The biosynthesized Fu-AgNPs shows potential antibacterial activity against Klebsiella pneumoniae in agar bioassay, disk diffusion, reactive oxygen species, protein leakage and confocal laser scanning microscopy assays. Furthermore, Artemia toxicity assay results showed less mortality (3.3 ±â€¯0.8%) even at higher concentration of Fu-AgNPs. Therefore, Fu-AgNPs can be effectively used as an antibacterial agent in the pharmaceutical fields.