Small molecule inhibitors of the VEGF and tyrosine kinase for the treatment of cervical cancer.

Cervical cancer accounts for most deaths due to cancer in women, majorly in developing nations. The culprit behind this disease is the human papillomavirus (HPV) which accounts for more than 90% of cervical cancer cases. The viral strains produce proteins that favor the knocking down of the apoptosis process and continuous growth of cells in the cervix leading to tumor growth. Proangiogenic growth factors, such as fibroblast growth factor (FGF), vascular endothelial growth factor (VEGF), angiopoietins, and other endothelial growth factors (EGF), are secreted by tumor cells and the surrounding microenvironment, which further advances the development of cancer. The extracellular domain of receptor tyrosine kinases is employed by ligands (like VEGF and EGF) to engage and activate them by inducing receptor dimerization, which facilitates the cascade impact of these factors. The tyrosine kinase domains of each receptor autophosphorylate each other, activating the receptor and initiating signaling cascades that promote angiogenesis, migration, proliferation, and survival of endothelial cells. Cancer cells benefit from its modified signaling pathways, which cause oncogenic activation. Upon early cervical cancer detection, the second-line therapy strategy involves blocking the signaling pathways with VEGF and small molecule tyrosine kinase inhibitors (TKIs). This review paper highlights the genesis of cervical cancer and combating it using VEGF and tyrosine kinase inhibitors by delving into the details of the currently available inhibitors. Further, we have discussed the inhibitor molecules that are currently in various phases of clinical trials. This paper will surely enhance the understanding of cervical cancer and its treatment approaches and what further interventions can be done to alleviate the disease currently serving as a major health burden in the developing world.

Bromelain: An Enzyme Expanding its Horizon from Food to Pharmaceutical Industry.

Bromelain is a protein digestive enzyme obtained from the extract of pineapple (steam, fruit, and leaves). It is a cocktail of several thiol endopeptidases and other components like peroxidase, cellulase, phosphatase, and several protease inhibitors. It is a glycoprotein with an oligosaccharide in its molecular structure that contains xylose, fucose, mannose, and N-acetyl glucosamine. Many approaches have been used in the extraction and purification of bromelain like filtration, membrane filtration, INT filtration, precipitation, aqueous two-phase system, ion-exchange chromatography, etc. This enzyme is widely used in the food industry for meat tenderization, baking, cheese processing, seafood processing, etc. However, this enzyme also expands its applicability in the food industry. It is reported to have the potential for the treatment of bronchitis, surgical trauma, sinusitis, etc. The in vitro and in vivo studies showed that it possesses fibrinolytic, antiinflammatory, antithrombotic, anti-edematous activity, etc. The human body absorbed bromelain without any side effects or reduction in its activity. However, in some cases, it shows side effects in those patients who are allergic to pineapple. To minimize such adverse effects bromelain is immobilized inside the nanoparticles. This paper gives an overview of the production, purification, and application of this industrially important enzyme in the food and pharmaceutical industry. It also discusses the various immobilization strategies used to enhance its efficiency.

Degradation product of curcumin restrain Salmonella typhimurium virulent protein L-asparaginase.

OBJECTIVES: Salmonella typhimurium is a pathogen responsible for causing a wide range of infectious diseases. The emergence of multi-drug resistance (MDR) in this microbe is a big challenge. L-asparaginase (less explored drug target) is selected as a drug target because it is actively involved in the virulence mechanism. To block this virulent enzyme, curcumin that is traditionally renowned for its medicinal properties was examined. However, its pharmacological behavior and targeting property is less understood because of its poor bioavailability. Therefore, the present work explores the antimicrobial effect of both curcumin and its degradation product against the MDR pathogen. METHODS: Molecular docking studies were carried out to evaluate the inhibitory effect of curcumin and its degradation product against the L-asparaginase enzyme using Schrodinger Maestro interface tools. The Absorption, Distribution, Metabolism, Excretion and Toxicity (ADMET) profile of all the test ligands was also performed. RESULTS: The docking score of curcumin was -5.465 kcal/mol while its degradation product curcumin glucuronide has the lowest i.e., -6.240 kcal/mol. All the test ligands showed better or comparable docking scores with respect to control (Ciprofloxacin). Arg 142 and Asn 84 amino acid residues of L-asparaginase were found to be interacting with test ligands inside the binding pocket of the target protein. ADME/toxicology study also indicated the potency of curcumin/curcumin degradation products as a potent inhibitor. CONCLUSIONS: It was found that both curcumin and its degradation products have the potential to inhibit Salmonella. This information could be valuable for futuristic drug candidate development against this pathogen and could be a potential lead for mitigation of MDR.

Optimized Production of Medically Significant Enzyme L-Asparaginase Under Submerged and Solid-State Fermentation From Agricultural Wastes.

L-asparaginase has prime medical significance due to its chemotherapeutic applications and action against a wide range of diseases. L-asparaginase obtained from commercial producer bacterial strains causes anaphylaxis, some sensitive reactions, and other side effects. To overcome these issues, eukaryotes like fungi can be used to produce L-asparaginase. Penicillium lilacinum was explored for its enhanced production through a statistically optimized fermentation approach. Firstly, the significant fermentation parameters (influencing the production) were screened through the Plackett-Burman approach. Thereafter, Response Surface Methodology (RSM) was employed for achieving higher production of L-asparaginase under specified fermentation conditions. The study explored both submerged (SmF) and solid-state (SSF) fermentation conditions to put forward a comparative analysis of the best suitable fermentation condition for the enhanced production of L-asparaginase. The Plackett-Burman optimization result suggested that pH, L-asparagine, ammonium nitrate, and temperature were significantly affecting enzyme production in SmF conditions, whereas wheat bran, arhar bran, kulthi bran, and temperature were identified as significant process parameters for SSF. The final enzyme activity obtained in SmF was 82.29 IU/mL and 190.439 U/gds in SSF after the implication of RSM. The statistical optimization tool fosters 2.47-fold and 6.36-fold enzyme activity in SmF and SSF, respectively. The study suggested that L-asparaginase can be efficiently produced using P. lilacinum at an optimized SSF condition.

Deciphering the blackbox of omics approaches and artificial intelligence in food waste transformation and mitigation.

It is necessary to stop the wastage of food during any stage of food chain to resolve the challenge of starvation, hunger and malnutrition in the world. Inception of modern techniques like omics (metagenomics, proteomics, transcriptomics, wasteomics, diseaseomics etc), enzymatic treatments, and artificial intelligence in food waste reduction and management can bring a sustainable solution for food loss management, starvation and environmental challenges. Acceptance of modern techniques while policies formulation by government bodies can substantially strengthen the idea of waste reduction, food security and can easily save the life of around 25,000 children and adults dying of starvation every day. Artificial Intelligence (AI) can bestead current agriculture and food supply chain system to overcome the challenges of nutrition demand, resource depletion, climate change, population growth, and pollution. This communication provides a thorough examination of the concept of food waste management with omics approaches linkages. In addition, the notion of artificial intelligence in food waste transformation and mitigation, as well as present challenges and future prospects have been covered. Overall, this communication would assist decision-makers in identifying economically and environmentally appropriate biorefinery solutions ahead of time.

Valorization of citrus peel waste for the sustainable production of value-added products.

Globally the generation and mismanagement of waste from fruit processing and post-harvest impose a severe burden on waste management strategies along with environmental pollution, health hazards. Citrus waste is one of such worrying fruit waste, which is rich in several value-added chemicals, including pectin. Pectin is a prebiotic polysaccharide possessing a multitude of health benefits. Citrus pectin has excellent gelling, thickening, water holding capacity, and encapsulating properties, which pave its functionality in versatile industrial fields including food processing and preservation, drug and therapeutic agents, cosmetics, and personal care products. The utilization of citrus wastes to derive valuable bioproducts can offer an effective approach towards sustainable waste management. With the ever-increasing demand, several strategies have been devised to increase the efficiency of pectin recovery from citrus waste. This review article discusses the sources, effect, and technology-mediated valorization of citrus waste, the functional and nutritive application of pectin along with its socio-economic and environmental perspective.

l-asparaginase: Need for an Expedition from an Enzymatic Molecule to Antimicrobial Drug.

Enzymes play a vital role in the biological system as a catalyst for biochemical reactions. They have a wide range of commercial applications in many areas like the pharmaceutical industry, food industry, etc. l-asparaginase is one of the commercial enzymes with prime importance in anticancer therapy. It is mainly used in chemotherapy; however, it has the potential to cure autoimmune disorders and infectious diseases also. Previous studies reported the antimicrobial potential of l-asparaginase. Therefore, we have discussed the possibility and challenges of the antimicrobial application of l-asparaginase in the treatment of infectious diseases. This is followed by a discussion on the effective delivery of this enzyme using biopolymeric nanocarriers that ensure safe and on target action. The present article gives a perspective on the l-asparaginase molecule that could be developed/established as an approved antimicrobial drug in the future.

Biological Degradation of Keratin by Microbial Keratinase for Effective Waste Management and Potent Industrial Applications.

Enzymes are the biocatalysts synthesized by living organisms having high specificity, catalytic activity, and a broad range of applicability. One such biotechnologically relevant enzyme is keratinase with various industrial applications that capture a significant place in the enzyme market. It belongs to the proteolytic enzyme group that cleaves the highly stable and fibrous protein, keratin through hydrolysis. Keratins are hard-degrading fibrous proteins insoluble in natural solvents and water. It is frequently aggregated in nature and expressively present in the plumes, hair, nail, horn, skins, feet, etc. The broad range of microorganisms, such as bacteria, fungi, and actinomycetes, have been accounted for producing keratinases with significant biotechnological applications. Successful application of this group of enzymes has been seen in various industries such as farming, laundry detergent, cosmetics, animal feed, pharmaceutical, leather, and textile. Moreover, they have found remarkable usability in environment friendly waste management also. This paper focuses on the structure, sources, and various applications of this industrially important enzyme.

Target shortage and less explored multiple targeting: hurdles in the development of novel antifungals but overcome/addressed effectively through structural bioinformatics.

Billions of people are affected by fungal infection worldwide, which is a major cause of morbidity and mortality in humans. Regardless of development in the field of antifungal therapeutics over the last three decades, multidrug resistance and limited efficacy of available antifungal drugs are very prominent and still a great hurdle in the patient treatment. The current antifungal pipeline is dry, which is needed to be strengthened. Although several strategies have been implemented over time to discover novel promising antifungal leads, but very little emphasis has been given to address the gap of fungal target identification. Undeniably, the need for identifying novel cellular fungal targets is as vital as discovering novel antifungal leads and a structural bioinformatics approach could be an effective strategy in this regard. To address the issue, we have performed in silico screening to identify a few potent multiple targeting ligands and their respective antifungal targets. Thus, we offer a perspective on the phenomena of 'target shortage' and least explored 'multiple targeting' being the most underrated challenges in antifungal drug discovery. 'Structural bioinformatics' could be an effective approach in the recognition of new/innovative antifungal target and identification/development of novel antifungal lead molecule aiming multiple molecular targets of the fungal pathogen.

An Improved Synthesis of Key Intermediate to the Formation of Selected Indolin-2-Ones Derivatives Incorporating Ultrasound and Deep Eutectic Solvent (DES) Blend of Techniques, for Some Biological Activities and Molecular Docking Studies.

We have developed a new idea to synthesize a key intermediate molecule by utilizing deep eutectic solvent (DES) and ultrasound in a multistep reaction to ensure process cost-effectiveness. To confirm the stability of reagents with DES, electronic energies were calculated at the B3LYP/6-31+G(d,p) level of theory. DES stabilized the reagents mainly due to strong intermolecular hydrogen bonding. Key intermediate (3) and final compounds (4a-n) were synthesized in a higher yield of 95% and 80%-88%, respectively. Further, final compounds (4a-n) were assessed for their anti-inflammatory, analgesic, ulcerogenic, and lipid peroxidation. The compounds 4f, 4g, 4j, 4l, and 4m showed good anti-inflammatory activity, while 4f, 4i, and 4n exhibited very good analgesic activity as compared to the standard drug. The ulcerogenicity of selected compounds was far less than the indomethacin. The ligands had also shown a good docking score (4f = -6.859 kcal/mol and 4n = -7.077 kcal/mol) as compared to control indomethacin (-6.109 kcal/mol) against the target protein COX-2. These derivatives have the potential to block this enzyme and can be used as NSAID. The state-of-art DFT theory was used to validate the lipid peroxidation mechanism of the active compounds which was in good agreement with the variations of BDEs and IP of the tested compounds.

l-Asparaginase: a feasible therapeutic molecule for multiple diseases.

This note highlights our understanding and thinking about the feasibility of l-asparaginase as therapeutics for multiple diseases. l-asparaginase enzyme (l-asparagine amidohydrolase, EC 3.5.1.1) is prominently known for its chemotherapeutic application. It is primarily used in the treatment of acute lymphoblastic leukemia in children. It is also used in the treatment of other forms of cancer Hodgkin disease, lymphosarcoma, acute myelomonocytic leukemia, acute myelogenous leukemia, chronic lymphocytic leukemia, reticulosarcoma and melanosarcoma (Lopes et al. Crit Rev Biotechnol 23:1-18, 2015). It deaminates l-asparagine present in the plasma pool causing the demise of tumor cell due to nutritional starvation. The anti-tumorigenic property of this enzyme has been exploited for over four decades and evidenced as a boon for the cancer patients. Presently, the medical application of l-asparaginase is limited only in curing various forms of cancer.

Eugenol derivatives prospectively inhibit l-asparaginase: A heady target protein of Salmonella typhimurium.

Salmonella typhimurium is the causative agent of severe human infections and mortality throughout the world. Pacing advent of new resistance mechanisms in this microorganism exists, rendering treatment of infectious disease difficult. Ciprofloxacin is no longer considered the first choice of antimicrobial agent due to the emergence of resistance. Therefore, the need for scenario is to find out novel drug target and its potential inhibitor to fight against this pathogen. The present study was undertaken to find out a novel drug target and its inhibitor for improving the current therapeutic methods for treating Salmonella infections. It is found that l-asparaginase is exploited by the pathogen for its survival benefit. Therefore, it could be targeted to fight against lethality caused by Salmonella infections. In the present in silico study, the 3-D structure of the enzyme l-asparaginase was modelled by using homology modeling technique. Thereafter, molecular docking studies and ADMET prediction to assess pharmacokinetic profiles of test ligands (eugenol and its derivative) was performed. The results show that eugenol and its derivative are capable of inhibiting the Salmonella virulent protein l-asparaginase. There were 18 ligands including ciprofloxacin (used as reference) were docked. The lowest binding energy was observed with eugenol derivative 8 i.e -5.836 kcal/mol while for ciprofloxacin was -4.661 kcal/mol. The docking of the eugenol derivative 8 with l-asparaginase revealed a strong interaction between them with two hydrogen bonds. Thr 35 and Asp 116 residues are actively participating in this interaction. The result of ADMET profiling suggests the potency of eugenol and its derivatives against Salmonellal-asparaginase-II as a compelling drug candidate. These findings provide useful information on the biological role, structure-based drug design and potent inhibitor of l-asparaginase for the development of effective therapeutic molecule against Salmonella infection.

Biotechnological production and practical application of L-asparaginase enzyme.

L-asparaginase is a vital enzyme of medical importance, and renowned as a chemotherapeutic agent. The relevance of this enzyme is not only limited as an anti-cancer agent, it also possesses a wide range of medical application. The application includes the antimicrobial property, treatment of infectious diseases, autoimmune diseases, canine and feline cancer. Apart from the health care industry, its significance is also established in the food sector as a food processing agent to reduce the acrylamide concentration. L-asparaginase is known to be produced from various bacterial, fungal and plant sources. However, there is a huge market demand due to its wide range of application. Therefore, the industry is still in the search of better-producing source in terms of high yield and low immunogenicity. It can be produced by both submerged and solid state fermentation, and each fermentation process has its own merits and demerits. This review paper focuses on its improved production strategy by adopting statistical experimental optimization techniques, development of recombinant strains, through mutagenesis and nanoparticle immobilization, adopting advanced and cost-effective purification techniques. Available research literature proves the competence and therapeutic potential of this enzyme. Therefore, research orientation toward the exploration of this clinical significant enzyme has to be accelerated. The objectives of this review are to discuss the high yielding sources, current production strategies, improvement of production, effective downstream processing and therapeutic application of L-asparaginase.

Eucalyptol, sabinene and cinnamaldehyde: potent inhibitors of salmonella target protein L-asparaginase.

Salmonella typhimurium is a severe threat to human life. The treatment became more difficult with the emergence of multidrug resistance. In the present in silico study, a novel drug target L-asparaginase was tested against three ligands eucalyptol, sabinene, and cinnamaldehyde, major components of cardamom, nutmeg, and cinnamon, respectively. The lowest docking score was obtained for sabinene followed by eucalyptol and cinnamaldehyde i.e. -5.648, -3.939 and -3.469. The docking score of sabinene is also better than the standard drug, Ciprofloxacin (-4.661) and natural substrate L-asparagine (-5.497). The amino acid residues involved in interactions inside the binding pocket are threonine 115 and threonine 35. The ADMET profile studied, also suggests the potency of the test ligands as a drug candidate. The results suggest they could be safe alternatives of chemical compounds to treat infections and combat multidrug-resistant bacteria.

In vitro screening and in silico validation revealed key microbes for higher production of significant therapeutic enzyme l-asparaginase.

l-asparaginase is an enzyme of medical prominence and reputable as a chemotherapeutic agent. It also has immense potential to cure autoimmune and infectious diseases. The vast application of this enzyme in healthcare sector increases its market demand. However, presently the huge market demand is not achieved completely. This serves the basis to explore better producer microbial strains to bridge the gap between huge demand and supply of this therapeutic enzyme. The present study deals with the successful screening of potent microorganisms producing l-asparaginase. 47 microorganisms were screened including bacteria, fungi, and yeasts. Among all, Penicillium lilacinum showed the highest enzyme activity i.e., 39.67 IU/ml. Shigella flexneri has 23.21 IU/ml of enzyme activity (highest among all the bacterial strain tested). Further, the 3-D structure of l-asparaginase from higher producer strains was developed and validated in silico for its activity. l-asparagine (substrate for l-asparaginase) was docked inside the binding pocket of P. lilacinum and S. flexneri. Docking score for the most common substrate l-asparagine is -6.188 (P. lilacinum), -5.576 (S. flexneri) which is quite good. Moreover, the chemical property of the binding pocket revealed that amino acid residues Phe 243, Gln 260, Gly 365, Asp 386 in P. lilacinum and residues Asp 181, Thr 318, Asn 320 in S. flexneri have an important role in H-bonding. The in silico results supports and strengthen the wet lab results. The outcome obtained motivates to take the present study result from lab to industry for the economic/massive production of this enzyme for the diverse therapeutic application.

The morpheein model of allosterism: a remedial step for targeting virulent l-asparaginase.

Allosterism has emerged as an innovative and significant mode of drug discovery. It facilitates the targeting of an allosteric site that is unique and more specific. A relatively new approach to allosteric regulation is the morpheein model, a concerted dissociative model that describes the equilibrium of alternate quaternary structure assemblies, whose architectures are dictated by alternate conformations in the dissociated state. It is involved in various biological phenomena, including enzyme regulation. One such enzyme is l-asparaginase, which is exploited by pathogenic microbes to cause infectious disease in humans. Thus, the morpheein model can be applied as a novel approach to the discovery of allosteric modulators that regulate l-asparaginase function to control virulence and serves as the basis for novel drug discovery research.

Why Chitosan? From properties to perspective of mucosal drug delivery.

Non-parenteral drug delivery routes primarily remove the local pain at the injection site. The drugs administered through the oral route encounter the process of hepatic first pass metabolism. Among the alternative delivery routes, mucosal route is being investigated as the most preferred route. Different mucosal routes include the gastrointestinal tract (oral), vagina, buccal cavity and nasal cavity. Novel formulations are being developed using natural and synthetic polymers that could increase the residence time of the drug at mucosal surface in order to facilitate permeation and reduce (or bypass) the first pass metabolism. For recombinant drugs, the formulations are accompanied by enzyme inhibitors and penetration enhancers. Buccal cavity (buccal and sublingual mucosa) has smaller surface area than the gastrointestinal tract but the drugs can easily escape the first pass metabolism. Chitosan is the most applied natural polymer while synthetic polymers include Carbopol and Eudragit. Chitosan has inherent properties of mucoadhesion and penetration enhancement apart from biodegradability and efflux pump inhibition. This review hoards the important research purview of chitosan as a compatible drug carrier macromolecule for mucosal delivery on single platform.

Docking and ADMET prediction of few GSK-3 inhibitors divulges 6-bromoindirubin-3-oxime as a potential inhibitor.

GSK-3 is a member of cellular kinases with diversified functions such as cellular differentiation, metabolic signaling, neuronal functions and apoptosis. It has been validated as an important therapeutic target in Alzheimer's disease and type 2 diabetes. Few molecules targeting GSK-3 are currently in clinical trials. In this study, we have compared certain docking and computational ADME (Absorption, Distribution, Metabolism, Excretion) parameters of a few GSK-3 targeted ligands (Indirubin, Hymenialdisine, Meridianins, 6-bromoindirubin-3-oxime) against two control molecules (Tideglusib and LY-2090314) to derive and analyze the basic drug-like properties of the test compounds. Docking between the GSK-3 and various ligands was done using AutoDock while ADME parameters were derived from ADMET server PreADMET and admetSAR. Various docked images were retrieved from docking, indicating the docking sites in the target protein. Out of four compounds tested, 6-bromoindirubin-3-oxime (6-BIO) was found as the best docking and ADME parameters, followed by Hymenialdisine (HMD). The LigPlot interaction results show two residues Leu (188) and Thr (138) to be common at the interaction site. The LD50 of 6-BIO is better than one of the control ligands while very similar to the other. Some of the parameters were very similar to the control ligands, thus, making it a suitable candidate among the test ligands. From this in-silico study, we concluded that 6-BIO is a potent drug candidate which could be further tested in vitro and in vivo to establish a drug molecule. Since, 6-BIO is a chemically modified form of the basic molecule Indirubin, we can hypothesize that certain other modified indirubins could be tested as GSK-3 targeted ligands.