Current Pharmaceutical Biotechnology - Volume 21, Issue 15, 2020

Triple negative breast cancer is the highly aggressive form of breast cancer with high reoccurrence rate and is short of effective treatment strategy. The prognostic markers of it are also not well understood. miRNAs are the global regulators of various cancers on the virtue of its ability to post transcriptional regulation of genes involved in various pathways involved in complicating TNBC. In this review we studied the expression of miRNAs at different stages of TNBC and the role of miRNAs as a tumor suppressor to inhibit cell proliferation, angiogenesis, invasion and metastasis and to induce apoptosis and thereby proposing these miRNAs as an effective treatment strategy against TNBC. miRNA also acts as chemosenstizer in enhancing chemosensitivity of conventional drugs against resistant TNBC cells. The present review emphasizes the importance of miRNAs as prognostic markers to determine the overall survival, disease free survival and distant metastasis free survival rate in TNBC patients. We speculate that miRNA can present themselves as an effective treatment strategy and prognostic marker against TNBC.

In recent decades, functional foods with ingredients comprising probiotics, prebiotics and postbiotics have been gaining a lot of attention from scientists. Probiotics and postbiotics are usually applied in pharmaceutical formulations and/or commercial food-based products. These bioactive agents can be associated with host eukaryotic cells and have a key role in maintaining and restoring host health. The review describes the concept of postbiotics, their quality control and potential applications in pharmaceutical formulations and commercial food-based products for health promotion, prevention of disease and complementary treatment. Despite the effectiveness of probiotic products, researchers have introduced the concept of postbiotic to optimize their beneficial effects as well as to meet the needs of consumers to provide a safe product. The finding of recent studies suggests that postbiotics might be appropriate alternative agents for live probiotic cells and can be applied in medical, veterinary and food practice to prevent and to treat some diseases, promote animal health status and develop functional foods. Presently scientific literature confirms that postbiotics, as potential alternative agents, may have superiority in terms of safety relative to their parent live cells, and due to their unique characteristics in terms of clinical, technological and economical aspects, can be applied as promising tools in the drug and food industry for developing health benefits, and therapeutic aims.

With the emergence and evolution of bacterial strains, it is now becoming difficult for pharmaceutical industries to provide remedies required for the betterment of mankind. Continuous exposure with available medication leads to the development of new strains with a significant amount of resistance, due to which pharmaceutical industries are facing various challenges. Antimicrobial resistance is the phenomenon causing a challenge in new drug development through conventional methods. Therefore, the requirement of alternative medicine is in high demand. Recently, allopathic medicines have seen a disinterest and people are preferring natural solutions due to their fewer side effects comparable to conventional medicine. Worldwide plants are utilized for various disease treatments such as bacterial infection, skin disorders, cancer, asthma, respiratory problems, etc. The presence of a wide range of phytocompounds in different plants provides an alternative to the pharmaceutical industries to counter the problem of bacterial infections. Different plants contain various phytochemicals that possess numerous therapeutic activities and provide a remedy to suppress various bacterial strains. Therefore, in this review, an overview of various plants and their phytocompounds which are responsible for antibacterial activity has been discussed.

In recent decades, there has been a very rapid increase in the prevalence of diabetes globally, with serious health and economic implications. Although today there are several therapeutic treatments for this disease, these do not address the causes of the disease and have serious side effects, so it is necessary to seek new treatments to replace or complement the existing ones. Among these complementary treatments, a strong link between the intestinal microbiota and diabetes has been demonstrated, which has focused attention on the use of biotherapy to regulate the function of the intestinal microbiota and, thus, treat diabetes. In this way, the main objective of this work is to provide a review of the latest scientific evidence on diabetes, gathering information about new trends in its management, and especially, the influence of the intestinal microbiota and microbiome on this pathology. It is possible to conclude that the relationship between the intestinal microbiota and diabetes is carried out through alterations in energy metabolism, the immune system, changes in intestinal permeability, and a state of low-intensity systemic inflammation. Although, currently, most of the experimental work, using probiotics for diabetes management, has been done on experimental animals, the results obtained are promising. Thus, the modification of the microbiota through biotherapy has shown to improve the symptoms and severity of diabetes through various mechanisms related to these alterations.

Adherence to Omega-3 fatty acids (O3FAs) as Nutraceuticals for medicinal applications provides health improvement. The prevention and treatment of diseases with O3FAs hold promise in clinical therapy and significantly reduces the risk of chronic disorders. Polyunsaturated fatty acids (PUFA) O3FAs have beneficial effects in the treatment of cardiovascular disorders, diabetic disease, foetal development, Alzheimer’s disease, retinal problem, growth and brain development of infants and antitumor effects. Association to current analysis promotes the application of algal biomass for production of O3FAs, mode of action, fate, weight management, immune functions, pharmaceutical and therapeutic applications serving potent sources in healthcare management. A search of the literature was conducted in the databases of WHO website, Sci.org, PubMed, academics and Google. The authors performed search strategies and current scenario of O3FAs in health associated disorders. Promising outcomes and future strategies towards O3FAs may play a pivotal role in Nutraceutical industries in the cure of human health in the future.

Background: Chiral safety, especially chiral drug inversion in vivo, is the top priority of current scientific research. Medicine researchers and pharmacists often ignore that one enantiomer will be converted or partially converted to another enantiomer when it is ingested in vivo. So that, in the context that more than 50% of the listed drugs are chiral drugs, it is necessary and important to pay attention to the inversion of chiral drugs. Methods: The metabolic and stereoselective pharmacokinetic characteristics of seven chiral drugs with one chiral center in the hydroxy group were reviewed in vivo and in vitro including the possible chiral inversion of each drug enantiomer. These seven drugs include (S)-Mandelic acid, RS-8359, Tramadol, Venlafaxine, Carvedilol, Fluoxetine and Metoprolol. Results: The differences in stereoselective pharmacokinetics could be found for all the seven chiral drugs, since R and S isomers often exhibit different PK and PD properties. However, not every drug has shown the properties of one direction or two direction chiral inversion. For chiral hydroxyl group drugs, the redox enzyme system may be one of the key factors for chiral inversion in vivo. Conclusion: In vitro and in vivo chiral inversion is a very complex problem and may occur during every process of ADME. Nowadays, research on chiral metabolism in the liver has the most attention, while neglecting the chiral transformation of other processes. Our review may provide the basis for the drug R&D and the safety of drugs in clinical therapy.

Background: Icariin has been shown to enhance bone formation. Objective: The present study aimed to investigate whether icariin also promotes bone fracture healing and its mechanisms. Methods: First, we isolated and cultured rat bone marrow stromal cells (rBMSCs) with icariincontaining serum at various concentrations (0%, 2.5%, 5% and 10%) and then measured alkaline phosphatase (ALP) activity and the expression of Core-binding factor, alpha 1 (Cbfα1), bone morphogenetic protein-2 (BMP-2) and bone morphogenetic protein-4 (BMP-4) in the rBMSCs. Second, we established a model of fracture healing in rats and performed gavage treatment for 20 days. Then, we detected bone biochemical markers (ELISA kits) in the serum, fracture healing (digital radiography, DR), and osteocalcin expression (immunohistochemistry). Results: Icariin treatment increased ALP activity and induced the expression of Cbfα1, BMP-2 and BMP-4 in rBMSCs in a dose-dependent manner. In addition, Icariin increased the serum levels of osteocalcin (OC), bone-specific alkaline phosphatase (BAP), N-terminal telopeptides of type I collagen (NTX-1), C-terminal telopeptide of type I collagen (CTX-1) and tartrate-resistant acid phosphatase 5b (TRACP-5b); promoted osteocalcin secretion at the fracture site; and accelerated fracture healing. Conclusion: Icariin can promote the levels of bone-formation markers, accelerate fracture healing, and activate the WNT1/β-catenin osteogenic signaling pathway.

Background: The re-emerging of targeting Dihydroorotate Dehydrogenase (DHODH) in cancer treatment particularly Acute Myelogenous Leukemia (AML) has corroborated the substantial role of DHODH in cancer and received the attention of many pharmaceutical industries. Objective: The effects of Brequinar Sodium (BQR) and 4SC-101 on lymphoblastoid cell lines were investigated. Methods: DHODH expression and cell proliferation inhibition of lymphoblastoid and lymphoma cell lines were analyzed using Western blot analysis and XTT assay, respectively. JC-1 probe and ATP biochemiluminescence kit were used to evaluate the mitochondrial membrane potential and ATP generation in these cell lines. Furthermore, we explored the cell cycle progression using Muse™ Cell Cycle Kit. Results: Ramos, SUDHL-1 and RPMI-1788 cells are fast-growing cells with equal expression of DHODH enzyme and sensitivity to DHODH inhibitors that showed that the inhibition of DHODH was not cancer-specific. In ATP depletion assay, the non-cancerous RPMI-1788 cells showed only a minor ATP reduction compared to Ramos and SUDHL-1 (cancer) cells. In the mechanistic impact of DHODH inhibitors on non-cancerous vs cancerous cells, the mitochondrial membrane potential assay revealed that significant depolarization and cytochrome c release occurred with DHODH inhibitors treatment in Ramos but not in the RPMI-1788 cells, indicating a different mechanism of proliferation inhibition in normal cells. Conclusion: The findings of this study provide evidence that DHODH inhibitors perturb the proliferation of non-cancerous cells via a distinct mechanism compared to cancerous cells. These results may lead to strategies for overcoming the impact on non-cancerous cells during treatment with DHODH inhibitors, leading to a better therapeutic window in patients.

Background: The prevalence of inflammatory bowel disease is rapidly increasing around the world. Quercetin is a flavonoid commonly found in vegetables and fruits and has been reported to exert numerous pharmacological activities such as enhancing antioxidant capacity or suppressing inflammation. Objective: We aimed to explore whether quercetin was effective for IBD and the underlying mechanism of quercetin for the ameliorative effects on the DSS-induced colitis in mice. Methods: Thirty-six mice were randomly assigned to three treatments, including the control group (Ctr), DSS-induced colitis group (DSS) and DSS-induced colitis supplemented with 500 ppm quercetin (DQ500). Colitis was induced by DSS intake, and body weight was recorded every day. After six days administration of DSS, intestinal permeability was measured, and the liver was taken for antioxidant enzyme tests. Colonic tissue was taken for the histopathlogical score and RNA-sequencing analysis. Results: In this experiment, dietary quercetin for 500ppm alleviated the DSS-induced colitis, possibly by strengthening intestinal integrity, liver antioxidant capacity. Based on the results of the transcriptome of colon tissue, several key genes were modulated by quercetin. ERK1/2-FKBP pathway and RXR-STAT3 pathway were involved in the development of IBD, furthermore, in the down-regulation of S100a8/9, FBN2 contributed to lowering the risk of colongenesis. Conclusion: We demonstrated that dietary quercetin alleviated the DSS-induced colitis in mice. This is most likely due to its beneficial effects on intestinal integrity and modulation of several key pathways. Based on our research, quercetin was a promising candidate for IBD and its pharmaceutical effects on both IBD and colongenesis need further research.

Background: Metal Nanoparticles (NPs) have been widely used for various applications in biomedical sciences, including in drug delivery, and as therapeutic agents, but limited owing to their toxicity towards the healthy tissue. This warrants an alternative method, which can achieve the desired activity with much reduced or no toxicity. Being a biological product, Withania somnifera (W. somnifera) is environment friendly, besides being less toxic as compared to metal-based NPs. However, the exact mechanism of action of W. somnifera for its antibacterial activities has not been studied so far. Objective: To develop “silver nanoparticles with root extract of W. somnifera (AgNPs-REWS)” for antimicrobial and anticancer activities. Furthermore, the analysis of their mechanism of action will be studied. Methods: Using the in-silico approach, the molecular docking study was performed to evaluate the possible antibacterial mechanism of W. somnifera phytochemicals such as Anaferine, Somniferine, Stigmasterol, Withaferin A, Withanolide- A, G, M, and Withanone by the inhibition of Penicillin- Binding Protein 4 (PBP4). Next, we utilized a bottom-up approach for the green synthesis of AgNPs- REWS, performed an in-detail phytochemical analysis, confirmed the AgNPs-REWS by SEM, UVvisible spectroscopy, XRD, FT-IR, and HPLC. Eventually, we examined their antibacterial activity. Results: The result of molecular docking suggests that WS phytochemicals (Somniferine, Withaferin A, Withanolide A, Withanolide G, Withanolide M, and Withanone) possess the higher binding affinity toward the active site of PBP4 as compared to the Ampicillin (-6.39 kcal/mol) reference molecule. These phytochemicals predicted as potent inhibitors of PBP4. Next, as a proof-of-concept, AgNPs- REWS showed significant antibacterial effect as compared to crude, and control; against Xanthomonas and Ralstonia species. Conclusion: The in-silico and molecular docking analysis showed that active constituents of W. somnifera such as Somniferine, Withaferin A, Withanolide A, Withanolide G, Withanolide M, and Withanone possess inhibition potential for PBP4 and are responsible for the anti-bacterial property of W. somnifera extract. This study also establishes that AgNPs via the green synthesis with REWS showed enhanced antibacterial activity towards pathogenic bacteria.

Purpose: The work was performed to investigate the feasibility of preparing ocular inserts loaded with Poly-ε-Caprolactone (PCL) nanoparticles as a sustained ocular delivery system. Methods: First, Atorvastatin Calcium-Poly-ε-Caprolactone (ATC-PCL) nanoparticles were prepared and characterized. Then, the optimized nanoparticles were loaded within inserts formulated with Methylcellulose (MC) and Polyvinyl Alcohol (PVA) by a solvent casting technique and evaluated physically, for in-vitro drug release profile. Finally, an in-vivo study was performed on the selected formulation to prove non-irritability and sustained ocular anti-inflammatory efficacy compared with free drug-loaded ocuserts. Results: The results revealed (ATC-PCL) nanoparticles prepared with 0.5% pluronic F127 were optimized with 181.72±3.6 nm particle size, 0.12±0.02 (PDI) analysis, -27.4± 0.69 mV zeta potential and 62.41%±4.7% entrapment efficiency. Nanoparticles loaded ocuserts manifested compatibility between drug and formulation polymers. Moreover, formulations complied with average weight 0.055±0.002 to 0.143±0.023 mg, and accepted pH. ATC-PCL nanoparticles loaded inserts prepared by 5% MC showed more sustained, prolonged in-vitro release over 24h. In-vivo study emphasized non-irritability, ocular anti-inflammatory effectiveness represented by smaller lid closure scores, and statistically significant lowering in PMN count after 3h. Conclusion: These findings proposed a possibly simple, new and affordable price technique to prepare promising (ATC-PCL) nanoparticles loaded inserts to achieve sustained release with prolonged antiinflammatory efficacy.

Background: Oral inflammatory ulcers are one of the common complaints of patients attending out-patient clinics. Previous in vivo studies had shown that an Aqueous M. citrifolia Fruit Extract (AMFE) possessed anti-inflammatory and ulcer healing activities. Therefore, a standardized topical bioadhesive gel containing AMFE-phytosome was developed and determined for its oral ulcer healing efficacy in a rabbit model. Methods: The AMFE phytosome (AMFE-P) was prepared by a complexation method with the required amount of AMFE: Phosphatidylcholine: Tween 80 to weigh ratio of 2:1:0.2. Poloxamer 407 was used as a gelling agent. The oral ulcer was induced in male New Zealand white rabbits by topical application of acetic acid. Each test compound was applied to the ulcer for 10 days beginning on the second day after the ulcer induction. Complete ulcer healing on the specimen obtained on day 12 was observed histologically using the histological scoring protocol. Results: The optimized gel containing AMFE-P equivalent to AMFE 10%w/w (10%AMFE-P gel) showed the best bioadhesive gel quality, a smooth and homogeneous texture with an optimum viscosity and pH range used in human oral cavity, a good physical and chemical stability and the highest percentage cumulative release of total phenolic and scopoletin content. It was found that a daily application of 10% AMFE-P gel exerted a superior ulcer healing efficacy and a significantly rapid ulcer healing process than a twice daily application of topical gel containing AMFE 10%w/w or chlorhexidine 0.2%. Conclusion: These findings demonstrated that 10% AMFE-P gel has potential as a safe and effective alternative therapeutic agent for oral ulcers.

Background: Artemisia absinthium L is an ornamental plant widespread in Saudi Arabia. Traditionally, the plant has been used in the Arabic medicine. But the scientific evidence of the bioactive compounds and their medicinal value was not yet explored widely. Objective: The study was designed to analyse the bioactive principles and medicinal properties of Artemisia absinthium L, a traditional herb grown in southern part of Saudi Arabia. Methods: The bioactive compounds present in Hot Methanolic Extract of the Leaves (HMEL) of Artemisia absinthium L. was explored by GC-MS analysis. The cytotoxicity effect of HMEL was determined against MCF-7 breast cancer cells ATCC and human colon cancer cells HCT 116 ATCC by performing MTT assay. Morphological changes of HMEL treated MCF-7 were observed under a phasecontrast microscope by staining the cells with neutral red. A Reaction Mixture (RM) of HMEL was prepared in Milli-Q water and antibacterial susceptibility was performed against both Gram-positive and Gram-negative bacteria. Furthermore, in vivo wound healing properties of the RM was screened in male rats and their efficacy was compared with standard povidone iodine cream. Biomarkers such as IL-1β, IL- 6, TNF-α, caspase-9 and caspase-3 levels were determined to qualify the wound healing property. Results: Epiyangambin, flavone, octadecanoic acid, 2,3-dihydroxypropyl ester, palmitic acid β - monoglyceride, -mannofuranoside, camphor, and terpineol were identified as possible compounds through GC-MS analysis. The HMEL of Artemisia absinthium L was actively inhibiting the proliferation of breast cancer cells MCF-7 ATCC at the concentration of 80.96 ± 3.94 μg/ml as IC50 value but failed to inhibit the proliferation against the treated human colon cancer cells HCT 116 cells ATCC. HMEL of Artemisia absinthium L was showing a moderate spectrum of antibacterial effect against the screened bacteria. RM showed better wound healing property than standard povidone iodine cream that modulates cytokine networks and apoptosis markers levels indicated the healing of wound. Conclusion: The study suggested that novel anticancer, antibacterial and immune modulatory molecules can be developed from the leaves of Artemisia absinthium L.