Mitochondrial targeting theranostic nanomedicine and molecular biomarkers for efficient cancer diagnosis and therapy.

Mitochondria play a crucial part in the cell's ability to adapt to the changing microenvironments and their dysfunction is associated with an extensive array of illnesses, including cancer. Mitochondrial dysfunction has been identified as a potential therapeutic target for cancer therapy. The objective of this article is to give an in-depth analysis of cancer treatment that targets the mitochondrial genome at the molecular level. Recent studies provide insights into nanomedicine techniques and theranostic nanomedicine for mitochondrial targeting. It also provides conceptual information on mitochondrial biomarkers for cancer treatment. Major drawbacks and challenges involved in mitochondrial targeting for advanced cancer therapy have also been discussed. There is a lot of evidence and reason to support using nanomedicine to focus on mitochondrial function. The development of a delivery system with increased selectivity and effectiveness is a prerequisite for a theranostic approach to cancer treatment. If given in large amounts, several new cancer-fighting medicines have been created that are toxic to healthy cells as well. For effective therapy, a new drug must be developed rather than an old one. When it comes to mitochondrial targeting therapy, theranostic techniques offer valuable insight

Modern Approaches in the Discovery and Development of Plant-Based Natural Products and Their Analogues as Potential Therapeutic Agents

Natural products represents an important source of new lead compounds in drug discovery research. Several drugs currently used as therapeutic agents have been developed from natural sources; plant sources are specifically important. In the past few decades, pharmaceutical companies demonstrated insignificant attention towards natural product drug discovery, mainly due to its intrinsic complexity. Recently, technological advancements greatly helped to address the challenges and resulted in the revived scientific interest in drug discovery from natural sources. This review provides a comprehensive overview of various approaches used in the selection, authentication, extraction/isolation, biological screening, and analogue development through the application of modern drug-development principles of plant-based natural products. Main focus is given to the bioactivity-guided fractionation approach along with associated challenges and major advancements. A brief outline of historical development in natural product drug discovery and a snapshot of the prominent natural drugs developed in the last few decades are also presented. The researcher’s opinions indicated that an integrated interdisciplinary approach utilizing technological advances is necessary for the successful development of natural products. These involve the application of efficient selection method, well-designed extraction/isolation procedure, advanced structure elucidation techniques, and bioassays with a high-throughput capacity to establish druggability and patentability of phyto-compounds. A number of modern approaches including molecular modeling, virtual screening, natural product library, and database mining are being used for improving natural product drug discovery research. Renewed scientific interest and recent research trends in natural product drug discovery clearly indicated that natural products will play important role in the future development of new therapeutic drugs and it is also anticipated that efficient application of new approaches will further improve the drug discovery campaign

Design, Synthesis, and Evaluation of GUNW-3 as a Brain Targeting Agent

The blood brain barrier (BBB) is a barrier in the brain that separates the peripheral blood circulation system from the central nerve system (CNS). The barrier effectively protects the brain from xenobiotics. The BBB serves as a physical barrier through the tight junction of endothelial cells that were found to be 50-100 times tighter than that of normal endothelial cells. Different drug efflux pumps such as P-glycoproteins and multidrug-resistance proteins are found to be overdistributed on the BBB. These drug efflux pumps help pump xenobiotics out if they enter the cells and serv as an additional mechanism to prevent xenobiotics from entering the CNS. The tight junction, drug efflux pumps, and other features of the BBB prevent almost 98% of small molecules, such as most therapeutics, and almost all large molecules such as biologics, recombinant genes and proteins from entering the brain. The inability to reach the therapeutic concentration caused by the barrier is often the major cause of treatment failure for brain diseases. Although the BBB blocks foreign compounds from entering the CNS, endogenous compounds, such as glucose, amino acids, peptides, neurotransmitters, and glutathione (GSH), enter the CNS readily through their corresponding receptors or transporters present in the BBB. Some of these receptors or transporters have been targeted for facilitating therapeutics, diagnostics, and other compounds to cross the BBB to reach the CNS. GSH is an endogenous three amino acid peptide. It plays various roles in the body: as a major antioxidant, a compound that removes toxic compounds, and a compound involved in other cellular functions. GSH crosses the BBB through a Nadependent GSH transporter. Recently, GSH transporters have been found effective in facilitating crossing of compounds through the BBB to reach the CNS. To achieve GSH transporter-mediated BBB crossing, GSH has been linked to a therapeutic agent (GSHDrug) to form a prodrug. The prodrug crosses the BBB by binding the GSH part to a GSH transporter followed by internalization of the prodrug. GSH has also been linked to polyethylene glycol (PEG) which is connected to a phospholipid (P) to form GSH-PEG-P or polyethylene glycol connected to vitamin E to form GSH-PEG-E. GSH-PEG-P and GSH-PEG-E have been coated on the surface of liposomes (GSH-PEGylated liposomes) to facilitate crossing of the liposomes through the BBB using the mechanism of binding the GSH moiety to a GSH transporter followed by internalization of the liposomes through endocytosis or transcytosis. The GSH-PEGylated liposomes have been shown to safely enhance the delivery to the brain by approximately 3-folds. We have developed GUNW-3 as a GSH-transporter dependent brain targeting agent. GUNW-3 was designed by connecting a hydrophilic GSH molecule to a hydrophobic cholesterol molecule through a two ehthylene glycol unit linker with a hope that the GSH part can serve as a brain-targeting structure through binding to the GSH transporter and facilitate the entry into the brain. This dissertation describes the design, synthesis, and fully characterization of GUNW-3. The dissertation also describes the ability of GUNW-3 to form micelles by itself (GUNW-3 micelles), the ability of GUNW- 3 micelles to cross the BBB to reach the brain, and the ability of GUNW-3 micelles to carry a dye (DiR) to the brain. Further, the dissertation shows the ability of GUNW-3 helps guide liposomes to the brain by forming GUNW-3 liposomes and the ability of GUNW-3 liposomes to deliver a dye (DiR) to the brain. Below is a brief description of the findings in this dissertation. GUNW-3 was synthesized in 4 steps from cholesterol and other commercially available reagents. GUNW-3 was found to be relatively stable. A cytotoxicity study of GUNW-3 revealed IC50 values of 0.65 mM and 0.47 mM for CV-1 cells (monkey kidney cells) and NCI-H226 cells (human lung cancer cells) respectively. GUNW-3 was found to form micelles by itself with a CMC value of 3.9 μM. CMC is a critical micelle parameter to reflect the stability of micelles and is also a parameter to determine if the micelles are stable enough to be used for a clinical application. The CMC of micelles need to be in μM concentration so that the micelles are stable enough to remain as micelles once being diluted in the blood stream. The CMC in low μM (3.9 μM) of GUNW-3 suggests that GUNW-3 micelles can be used for a therapeutic application. Further, the CMC of GUNW-3 is much lower than the IC50 values of the molecule indicating that GUNW-3 is not cytotoxic. For brain targeting, our data with ex-vivo imaging of the brains shows that the brain uptake of DiR, a dye, delivered by GUNW-3 micelles were 5 times higher than that of the control liposome and 12 times higher than that of free DiR at the first hour. After 48 h, the brain uptake of DiR delivered by GUNW-3 micelles was 6.5 times higher than that of the control liposome and 14 times higher than that of free DiR. GUNW-3 was also found to help deliver liposomes to the brain most likely by embedding the hydrophobic cholesterol part into the liposome double lipid layer and the hydrophilic GSH part floating on the surface of the liposomes for brain targeting. Our data from ex-vivo imaging of the brains demonstrate that GUNW-3 liposomes were able to significantly (\u3e3 folds) improve the delivery of DiR to the brain and retain in the brain well when compared with the control liposomes. Liposomes and micelles are known effective drug carriers that can be used to deliver various drugs or compounds such as small molecule therapeutics, DNA, RNA, and proteins (e.g., antibodies). Liposomes and micelles can encapsulate drugs and protect them from in vivo/in vitro degradation. They can also help reduce drug clearance, increase in vivo drug half-life, enhance the drug payload, control drug release, and improve drug solubility. The abilities to deliver DiR to the brain by GUNW-3 micelles and GUNW-3 liposomes warrantee further investigation of these two brain targeting delivery systems for delivering compounds to the brain for brain disease treatment or prevention. In summary, we have synthesized and characterized the rationally designed GUNW-3 as a brain targeting agent. GUNW-3 micelles and GUNW-3 liposomes showed promising brain targeting abilities. GUNW-3 micelles and GUNW-3 liposomes will be promising delivery systems for therapeutic and diagnostic molecules

Synthesis, characterization and biological activity of a Schiff base derived from o-amino benzoic acid and its Co(II), Cd(II) and Ni(II) complexes

Equimolar amounts of imidazoleacetophenone and 2-aminobenzoic acid were combined together and the Schiff base 4(1H-imidazole-yl)acetophenoneanthranilic acid was prepared as a new bidentate complexing agent. The synthesized ligand was reacted with cobalt(II), cadmium(II), and nickel(II) ions yielding air stable complexes. For quantification and characterization purposes, elemental analysis, infrared spectra, electronic spectra, proton nuclear magnetic resonance spectra and mass spectra studies were carried out on the obtained complexes and ligand. Thermogravimetric analysis and magnetic susceptibility measurements were also used for characterization. The ligand IR spectrum showed that the ligand acts as a bidentate coordinates to the metal ions through the nitrogen and oxygen atoms.Measurements of magnetic susceptibility for Ni(II) and Co(II) complexes were found to be 3.4 and 3.8 B.M., respectively, in the range normal for the octahedral geometry. The conductivity measurements revealed that the chelates are non-electrolytes. An in vitro antimicrobial investigation was also carried out for the free ligand and its metal complexes against a number of bacterial and fungal strains, to assess their antimicrobial properties by diffusion technique. Antimicrobial activity of the prepared complexes showed higher activity than the free ligand

Optimization of Chromatographic Conditions with QbD for Method Development and Validation of Bosutinib by HPLC: Applications in Dosage Forms and Rat Plasma Analysis

Aim: Bosutinib (BST) is an anti-cancer medicine that is used to treat a variety of different types of cancer. Using the HPLC method of analysis and the Quality by Design (QbD) strategy, the study aimed to precisely quantify the drug in tablet form and in rat plasma. Methodology: For the developed method’s validation, the chromatographic settings were fine-tuned by making use of the Box–Behnken Design (BBD). In the BBD, two dependent variables and three independent variables were selected. Isocratically, samples were eluted, having eluent phase composition of ammonium acetate (CH3COONH4) buffer pH 3.0 and acetonitrile (CH3CN) (60:40% v/v), in Raptor C-18 column at temperature 25 ∘C with a flow rate of 1 mL/min for 5 min. The wavelength of detection was set at 260 nm. In this study, encorafenib (ENC) was employed as an internal standard. Result: A sharp and resolved peak of BST and ENC at a retention time of 1.92 min and 4.01 min, respectively, was observed by the developed method. The limits of quantification and detection of the newly established method were found to be 1.503 μg/mL−1 and 0.496 μg/mL−1. The calibration curve’s observed linearity range was between 2 and 20 μg/mL−1, with an r2 of 0.999. The developed and optimized method was verified in compliance with the ICH guidelines. The results of all validation parameters were within the acceptable range, for example, % RSD of system suitability (0.63–4.46), % RSD of linear regression (1.659), interday and intraday precision % RSD value (1.723–1.892), and (1.762–1.923), respectively, and accuracy (1.476–1.982). Conclusion: The quantity of BST in tablet dosage form and in rat plasma samples was determined using a simple, quick, and robust method that was devised and validated

Development of screen- printed carbon electrode-based immunosensors for the electrochemical detection of dengue virus antigen

Objective: This study describes the development of a cost-effective and sensitive immunosensor for rapid detection of the dengue virus (DENV) antigen in human plasma. Methods: Screen-printed carbon electrodes (SPCEs) were used to fabricate the immunosensor by immobilizing the DENV antibodies to its surface using 1-ethyl-3-(3-dimethylamino propyl)carbodiimide hydrochloride/N-hydroxysulfosuccinimide (EDC/s-NHS) as linker. The detection of the antigen–antibody interaction was achieved by linear sweep voltammetry (LSV) using potentiostat instrument and a sensor connector. The developed method was validated as per the ICH guidelines for linearity, sensitivity and accuracy. Results: The developed immunosensors showed excellent sensitivities in detecting the DENV antigens in both phosphate buffer saline (PBS) maintained at pH 7.4 as well as diluted human plasma. The limit of detection (LOD) values obtained in PBS was 0.11 nM; whereas in human plasma, it was calculated to be 0.16 nM. Good linearity was obtained for both PBS and human plasma over a wide range of concentrations showing promising applications of the immunosensor in both qualitative and quantitative detection of dengue virus antigen. Also, the developed immunosensor was able to detect the dengue antigen in 5 min showing rapid detection. Conclusion: A sensitive, reliable and cost-effective biosensor was developed which was able to detect the dengue virus antigen selectively and rapidly

Natural coumarins from Murraya paniculata as mixed-type inhibitors of cholinesterases: In vitro and in silico investigations

Currently, acetylcholinesterase (AChE) inhibiting drugs in clinical use, such as tacrine, donepezil, rivastigmine, and galanthamine, are associated with serious side effects and short half-lives. In recent years, numerous phytochemicals have been identified as inhibitors of cholinesterases with potential applications in the management of Alzheimer\u27s disease (AD). In this study three natural coumarins, 2\u27-O-ethylmurrangatin (1), murranganone (2), and paniculatin (3) isolated previously by our group from the leaves of Murraya paniculata, were tested against the two cholinesterases (ChE) enzymes, AChE and butyrylcholinesterase (BChE) using in vitro assay. Molecular docking was performed to highlight the structural properties that contribute to the molecular recognition pattern in the inhibition of ChE and the structural differences resulting in the selectivity of these compounds toward AChE. Classical enzyme inhibition kinetics data suggested that compounds 2 and 3 were potent inhibitors of AChE and BChE, while 1 was found inactive against both enzymes. The findings from molecular docking studies revealed the competitive and non-competitive inhibition mechanisms of compounds 2 and 3 against both enzymes. Molecular docking and simulations have revealed that hydrogen bonding, mediated by ketone and hydroxyl functionalities in various positions, significantly contributes to the binding of the inhibitor to the receptor. According to MD simulation studies, the stability of the ligand-AChE complex for the most active compound (3) is found to be comparable to that of the widely used drug Tacrine. In addition, to evaluate the drug-likeness of compounds, in silico ADME evaluation was performed, and the compounds presented good ADME profiles. Data suggested that the coumarin nucleus having diverse side chains at the C-8 position can serve as a potential inhibitor of cholinesterases and can act as a lead to develop a new semisynthetic drug for the treatment of A

Alkaloidal Phytoconstituents for Diabetes Management: Exploring the Unrevealed Potential

The main characteristic feature of diabetes mellitus is the disturbance of carbohydrate, lipid, and protein metabolism, which results in insulin insufficiency and can also lead to insulin resistance. Both the acute and chronic diabetic cases are increasing at an exponential rate, which is also flagged by the World Health Organization (WHO) and the International Diabetes Federation (IDF). Treatment of diabetes mellitus with synthetic drugs often fails to provide desired results and limits its use to symptomatic treatment only. This has resulted in the exploration of alternative medicine, of which herbal treatment is gaining popularity these days. Owing to their safety benefits, treatment compliance, and ability to exhibit effects without disturbing internal homeostasis, research in the field of herbal and ayurvedic treatments has gained importance. Medicinal phytoconstituents include micronutrients, amino acids, proteins, mucilage, critical oils, triterpenoids, saponins, carotenoids, alkaloids, flavonoids, phenolic acids, tannins, and coumarins, which play a dynamic function in the prevention and treatment of diabetes mellitus. Alkaloids found in medicinal plants represent an intriguing potential for the inception of novel approaches to diabetes mellitus therapies. Thus, this review article highlights detailed information on alkaloidal phytoconstituents, which includes sources and structures of alkaloids along with the associated mechanism involved in the management of diabetes mellitus. From the available literature and data presented, it can be concluded that these compounds hold tremendous potential for use as monotherapies or in combination with current treatments, which can result in the development of better efficacy and safety profiles

Determination of Caffeine Content in Commercial Energy Beverages Available in Saudi Arabian Market by Gas Chromatography-Mass Spectrometric Analysis

The popularity of energy beverages among young adult population is high. These drinks are claimed to boost energy and performance and contain high concentration of caffeine as one of the several ingredients. Discrepancies have been encountered by some of the previous studies between the actual quantity of caffeine present in the product and the amount mentioned on the label, making the determination of caffeine content in these drinks very important. Thus, in this study, we analyzed the caffeine concentration in most popular energy drinks available in Saudi Arabia. The energy drink samples (n = 9) were procured from retail outlets. Sample solutions were prepared in methanol and analyzed for caffeine content by GC-MS. Chromatographic parameters were optimized to achieve optimum resolution and various validation parameters were evaluated. The method was successfully applied for the quantification of caffeine in energy drinks by directly injecting the multifold diluted samples in methanol. The method was linear (r2 = 0.999) over a concentration range of 5–25 µg/mL, specific, precise (%RSD of peak area = 0.56–0.78), and accurate (%recovery = 99.3–101.2%). The amounts of caffeine determined were found in the range of 20.82–33.72 mg/100 mL (52.05–84.3 mg/pack). Results revealed that the amount of caffeine actually present in the tested drinks varied within ±10% range from the amount specified on the product labels. The amounts of caffeine detected in tested beverages were within the USFDA safe upper limit of per-day caffeine consumption, which has indicated that the consumption of one serving of energy drink is unlikely to produce any adverse health effect