Current Pharmaceutical Biotechnology - Volume 24, Issue 10, 2023

Background: Small, non-coding microRNAs, usually of 20-25 nucleotides, are known to regulate the post-transcriptional gene expression, which has a significant role in human biological processes, including immune-biogenesis, homeostasis and infection control as differential expression of such miRNAs is responsible for fine-tuning the organismic development. Methods: A search of bibliographic databases was carried out with a focused question on microRNA- Disease Prediction. A deductive qualitative content analysis approach was employed to assess the research's overall outcomes, review articles on prediction tools in miRNA-Diseases, and analyse the interventions. Results: Diagnosis and therapeutics of diseases and miRNA prediction methods hold importance in identifying the regulatory mechanisms. Collections of efficient miRNA prediction methods to identify miRNA-mRNA-disease regulatory relationships have been presented through this review, consolidating the potential of miRNAs as a diagnostic and prognostic biomarker of multiple diseases, including COVID-19. Conclusion: The role of miRNA in the aetiology and pathogenesis of wide-range of pathologies, including viral, bacterial to chronic diseases such as cancer, is quite feasible through the modern tools in bioinformatics which has been elaborated focusing upon miRNA-disease prediction methods and their application potential establishing miRNAs as a robust and reliable biomarker in clinicomedical studies.

Terpenoids are naturally occurring secondary metabolites that consist of isoprene units (i.e., 2-methyl-1,3-butadiene). Terpenoids became recognized because of their diverse pharmacological benefits, such as anti-cancer, anti-inflammatory, antioxidant, analgesic, antibacterial, antifungal, hepatoprotective, antiviral, and antiparasitic activities. But most of these compounds have limited lipophilicity, dissolution rate, aqueous solubility, and drug permeability, so they are not used effectively. The low bioavailability significantly interferes with the performance of terpenoids to cure diseases, and the absorption process of terpenoids also becomes disrupted; therefore, their bioavailability in the blood becomes insufficient to achieve optimal treatment activity. Thus, to overcome this limitation, some strategies are used, such as nanotechnology (nanoparticles, carrier complexation), cocrystal, and glycosylation. Thus, this review summarizes the chemistry of terpenoids, factors that limit the bioavailability of terpenoids, and strategies employed to date with their design principles and outcomes possibly increasing their bioactivity.

The nature of phages was a matter of dispute, which was resolved in 1940, and it was continued to develop their activity and application in the Soviet Union and Eastern Europe. Bacteriophages were first employed in 1919 to treat bacterial illnesses caused by Citrobacter, Enterobacter, and Pseudomonas. Bacteriophages range in complexity from simple spherical viruses with genome sizes of less than 5 kbp to complicated viruses with genome sizes surpassing 280 kbp. They have two significant parts, head and tail, and are made up of numerous copies of more than 40 distinct proteins. Bacteriophages have been demonstrated to bind with receptors in the walls of both gram-positive and gram-negative bacteria, ranging from peptide sequences to polysaccharide moieties. Depending on the type of phage and the physiological state of the bacterium, the life cycle may diverge into the lytic cycle or lysogenic cycle. Lytic-lysogenic switch depends on a variety of inducing factors. Bacteriophage therapy can be administered via several routes, but parenteral routes are the most effective. Auto-dosing, single-dose potential, lack of cross-resistance with antibiotics, etc., are several advantages of phage therapy over antibiotic treatment. Bacteriophages are attracting much attention because of their potential advantages and wide applications as antibacterial agents, diagnostic technologies, phage-based products, and biocontrol agents. They also have several applications in the food industry, agriculture/crop, farm animal and bee protection, environmental, and biosensor development.

Antioxidant enzymes being an integral part of the defense mechanism have a crucial role in cellular metabolism, essential for healthy growth and living of the cells. The main function is to scavenge and degrade the free radicals, reactive oxygen species (ROS), and reactive nitrogen species (RNS). Endogenous antioxidant enzymes present in mitochondria, cytosol, and other cellular parts participate in capturing and repairing the oxidative damage to the system. Superoxide dismutase, catalase, and glutathione are antioxidant enzymes considered to be part of the first line of defense and are especially important in scavenging reactive oxygen species such as superoxide anion and hydrogen peroxide. Numerous studies in humans, as well as animal models, are correlated and reported about elevation in the enzymatic activity being involved in inhibiting oxidative damage and controlling the disease progression. Similarly, alterations due to enzymatic damage increase oxidative damage and have a key role in disease progression in diseases like cancer, atherosclerotic diseases, neurodegenerative diseases like Parkinson’s, Alzheimer’s, viral diseases, age-related ailments, etc. However, information about antioxidant enzymes, their specificity, free radicals involved in different diseases, and the oxidation process needs to be explored to a greater extent. This review focuses on our current understanding of the role of free radicals and the potential of various antioxidant enzymes, and their great scope in therapeutics against many dreadful diseases.

Bacterial infections continue to jeopardize human and animal health, impacting millions of lives by causing significant deaths every year. The use of antibiotics remains the primary choice of therapy and has only been partly successful in reducing the disease burden due to the evolving nature of resistant microbes. Widespread and inappropriate use of antibiotics resulted in the development of antibiotic-resistant microbial species provoking substantial economic burdens. The most promising way to resolve the issue of antibiotic resistance is the use of bacterial viruses called bacteriophages to treat microbial infections. Earlier reports on experimental bacteriophage therapy showed successful patient outcomes, and many clinical trials of such clinical bacteriophages have already been investigated in many western countries. In this review, we are focusing on the advantages as well as drawbacks of bacteriophage therapy to use it as an alternative to antibiotics for microbial infections, together with its current success status. There is also a need to extensively study the past, present, and future outlook of phage therapy in comparison to presently available antimicrobial agents and especially immunological response by the host after phage administration. Our aim is to highlight the fast-promoting field of bacteriophage therapy and provocations that lie ahead as the world is gradually moving aside from complete dependence on antimicrobial agents.

Background: Chrysin (CH) is one of the important natural flavonoids with antioxidant and anti-inflammatory activity. The aim of this study was to assess the protective effects of CH on biochemical indexes and histopathological changes in the liver of male Wistar rats exposed to chlorpyrifos (CPF). Methods: We induced sub-chronic toxicity in rats using CPF (10 mg/kg/day, orally) and administrated CH at 12.5, 25, and 50 mg/kg/day for 45 days. Results: In this study, CPF increased liver enzyme activities compared with the control group (p < 0.05), and co-treated CH with CPF reduced them compared with the non-treated CPF group (p < 0.05). A significant reduction in the liver GSH concentration along with a significant elevation in the concentrations of MDA and NO in the CPF group was observed compared with the control group (p < 0.001). However, CH at a dose of 50 mg could reverse them nearly to the control group (p < 0.001). In the CPF, CPF + CH1, and CPF + CH2 groups, a marked (p < 0.05) increase was found in the serum concentration of IL-6 compared with the control animals. No significant changes were found in the IL-6 concentration of the CPF + CH3 compared with the controls. Moreover, the coadministration of CH plus CPF induced histopathological alterations in liver. Conclusion: These results suggest that CH attenuates hepatic enzymes and histopathological alterations induced by CPF via modulating oxidative stress and inflammatory indices in rats.

Background: In recent years, the delivery of drugs by nanocomposites has emerged as an exciting field of research for bio-imaging tools and targeted cancer treatment. The large surface area and porous volume of mesoporous silica nanocomposites (MSN’s) have gained a lot of interest for their application in the delivery of drugs and the magnetic properties of iron oxide (IO) nanocomposites play a key role in the targeted delivery system. Methods: In this study, mesoporous silica encapsulated IO nanocomposites loaded with doxorubicin (DOX) were synthesized for the magnetically guided delivery of anticancer drugs. The synthesis of IO nanocomposites was done through the precipitation method, and then silica encapsulation and drug loading were done by the StÖber method. Results: The magnetically driven delivery of the drug is produced by the encapsulation of magnetically active IO in the mesoporous silica shell. The controlled release of DOX is possible because of the MSN’s. TEM images show that the nanocomposites have a spherical morphology and average diameter in the range of 120 nm. Power-XRD data confirm the crystalline nature of nanocomposites. The strong absorption peak was observed in UV-Visible spectroscopy at 490 nm and quenching in fluorescence spectra confirms the encapsulation of DOX in the mesoporous silica shell. VSM data showed the magnetic nature of nanocomposites, with large magnetic susceptibility (74.88 emu/g). The use of DOX/IO@Silica nanocomposites as a sustainable drug release and targeted drug delivery vehicle has been reported here. The pH dependent release of DOX was studied and significant release was observed at lower pH. In-vitro cell viability assay and fluorescence imaging assay have demonstrated that these nanocomposites show significant dose-dependent toxicity to cancer cells in the presence of a magnetic field. Conclusion: In-vitro studies via the MTT assay showed that these synthesized nanocomposites in culture are non-toxic to healthy cells compared to DOX-induced cytotoxicity due its controlled release and can be further strengthened by magnetic guidance. Therefore, due to its optical properties and potential for guided delivery of drug to the targeted site, these nanocomposites are ideal as an anticancer agent and bio-imaging prob.

Background: Attention Deficit Hyperactivity Disorder (ADHD) can be comorbid with depression, often leading to the prescription of both methylphenidate (MP) and selective serotonin reuptake inhibitor (SSRI) antidepressants, such as fluoxetine (FLX). Moreover, these drugs are often misused as cognitive enhancers. This study examined the effects of chronic oral co-administration of MP and FLX on depressive- and anxiety-like behaviors. Methods: Adolescent rats received daily either water (control), MP, FLX, or the combination of MP plus FLX in their drinking water over the course of 4 weeks. Results: Data analysis shows a decrease in food consumption and body weight for rats exposed to FLX or the combination of MP and FLX. Sucrose consumption was significantly greater in FLX or MP+FLX groups compared to controls. FLX-treated rats showed no effect in the elevated plus maze (EPM; open arm time) and forced swim test (FST; latency to immobility). However, rats treated with the combination (MP+FLX) showed significant anxiolytic-like and anti-depressive-like behaviors (as measured by EPM and FST), as well as significant increases in overall activity (distance traveled in open field test). Finally, the combined MP+FLX treatment induced a decrease in anxiety and depressive- like behaviors significantly greater than the response from either of these drugs alone. Conclusion: These behavioral results characterize the long-term effects of these drugs (orally administered) that are widely co-administered and co-misused and provide important insight into the potential neurobiological and neurochemical effects. Future research will determine the potential risks of the long-term use of MP and FLX together.

Background: Peptide-Fc fusion proteins are inherently heterogeneous and complex molecules. Protein post-translational modifications (PTMs) or truncation can arise during manufacturing or product storage. Some of these product attributes could potentially impact the efficacy or safety of the bio-molecule and are thus classified as critical quality attributes (CQAs). These CQAs should be controlled in order to ensure manufacturing and quality consistency. Methods: A subunit UPLC-ToF MS based MAM method was developed for identity test and quantitatively monitored two critical quality attributes (CQAs) resulting from two truncations of that fusion protein (fragment 1 and 2). Three independent laboratories are involved in the method validation according to ICH Q2(R1), ICH Q6B, FDA and NMPA guidance. Results: This developed method fully meets the pre-defined analytical target profile (ATP), including specificity, accuracy, precision, quantitation limit, linearity, range and robustness. Three independent labs co-validate a UPLC-ToF MS based MAM method for protein drug QC release and stability testing. Conclusion: The experimental design of method validation can be a reference for LC-HRMS-based subunit MAM methods that have been widely used in the characterization of antibodies, ADCs and other protein-based biologics. This work paves the way for implementing MAM in QC with more targeted control of product quality.

Background: Streptokinase, one of the most widely used thrombolytic medicines, is a favorable protein for site-specific PEGylation as it lacks any cysteine residues in its amino acid sequence; however, any changes in the protein’s structure should be carefully planned to avoid undesired changes in its function. Objectives: This study aimed to design and produce novel di/tri-cysteine variants of streptokinase from previously developed cysteine analogues, Arg45, Glu263, and Arg319, as candidates for multiple site-specific PEGylation. Methods: Using bioinformatics tools and site-directed mutagenesis, we incorporated concurrent mutations at Arg45, Glu263, and Arg319 (carried out in our previous study) to create di/tri-cysteine variants of streptokinase proteins (SK45-319cys, SK263-319cys, and SK45-263-319cys) and evaluated their kinetic activity parameters by a colorimetric method, using H-D-Val-Leu-Lys-pNA.2HCl (S2251) as substrate. Results: Based on the kinetic results, SK263-319cys with 44% enzyme efficiency increment compared to wild-type SK was the superior protein in terms of activity; as well, SK45-319cys and SK45-263-319cys showed 17 and 22% activity enhancement, respectively. Docking of the mutant streptokinase proteins with μ-plasmin demonstrated that changes in intermolecular interactions caused by amino acid substitution could be the reason for activity difference. Conclusion: The novel mutant proteins created in this study exhibit remarkable biological activity and may be uniquely suitable for simultaneous PEGylation on two/three domains. As well, PEGylated derivates of these variants might prove to be more proficient proteins, compared to the singlecysteine analogs of streptokinase; because of their more surface coverage and increased molecular weight.

Background: Lung cancer is a major threat to public health and remains difficult to treat. Repositioning of existing drugs has emerged as a therapeutic strategy in lung cancer. Clinically, low-dose montelukast has been used to treat asthma. Objective: We evaluated the potential of using montelukast to treat lung cancer. Methods: Migration was detected using wound-healing and Transwell assays, the expression of CysLT1 using western blotting, and subcellular localization of CysLT1 using immunofluorescence. CRISPR/Cas9 technology was used to further investigate the function of CysLT1. Results: Subcellular localization staining showed that the CysLT1 distribution varied in murine and human lung cancer cell lines. Furthermore, montelukast suppressed CysLT1 expression in lung cancer cells. The treated cells also showed weaker migration ability compared with control cells. Knockout of CysLT1 using CRISPR/Cas9 editing in A549 cells further impaired the cell migration ability. Conclusion: Montelukast inhibits the migration of lung cancer cells by suppressing CysLT1 expression, demonstrating the potential of using CysLT1 as a therapeutic target in lung cancer.