Academic literature on the topic 'Biochemistry|Pharmaceutical sciences'

Yalkin Halmatovich Turakulov is an internationally known medical researcher whose name is firmly engrained into the history of national endocrinology. He headed the Pharmaceutical Medical Institute, Tashkent Medical Institute, Regional Institute of Medicine, Institute of Nuclear Physics of Uzbekistan Academy of Sciences, Institute of Biochemistry and others institutions various times and was involved in the establishment of many of them. He was a teacher to many doctors and scientists. This article presents his biography and describes his impact on national and world science.

Advances in technological possibilities have made communication present in different media and spaces. By enabling interaction between different countries, by becoming a facilitator between knowledge and innovation in the globalized world, it has opened frontiers by providing innovations in various sectors of the knowledge society. In this sense, the objective in this article is to map the intersection of communication, innovation and knowledge in the globalized world. To that end, the methodology used in the research was the systematic search of literature that pointed out that the intersection is motivated by the use of innovative technologies in the process of knowledge sharing, and studies are still scarce in this area. It is possible to perceive, further, that this intersection is branched out, through Social Sciences, Business, Management and Accounting, Computer Science, Medicine, Engineering, Decision Sciences, Nursing, Arts and Humanities, Economics, Econometrics and Finance, Psychology, aligned Health Professions, Agricultural and Biological Sciences, Biochemistry, Genetics and Molecular Biology, Energy, Environmental Science, Mathematics, Materials Science, Multidisciplinary, Neuroscience, Pharmacology, Toxicology and Pharmaceutical and Veterinary.

Careful design of study plan is a key element of any successful educational program. Till 2018 Ministry of Health of Ukraine regulated the structure of Pharmacy study plans through the adoption of unified Ministerial study plan. Now the responsibility of educational programs and corresponding study plans design in Ukraine is fully transferred to universities. The purpose of this study is to compare the structure and content of pharmacy study plans in Visegrad Group countries with the most recent unified Pharmacy study plan in Ukraine. Methods. The official documents of Warsaw Medical University, Jagiellonian University in Krakow, Charles University, University of Veterinary and Pharmaceutical Sciences Brno, Comenius University, University of Veterinary Medicine and Pharmacy in Kosice, Semmelweis University and University of Debrecen were studied and data on required courses and corresponding ECTS credits extracted and compared with Ukrainian study plan. Results. Ukrainian unified study plan in Pharmacy pays much more attention to Humanity, Social and Economics section (9 ECTS credits plus 6 ECTS credits of Foreign Language), Computer and IT skills (8 ECTS credits), Hygiene and Ecology (3 ECTS credits), Life Safety, Labor Safety and Bioethics (6 ECTS credits in total), Extreme Medicine and Military Training (6 ECTS credits in total), Toxicological and Forensic Chemistry (4 ECTS credits), Organization and Economics of Pharmacy, Pharmaceutical Management and Marketing (12 ECTS credits in total) as compared to foreign universities. While natural science courses receive less ECTS credits in Ukraine, and some courses in rapidly evolving sciences like Molecular Biology, Immunology or Clinical Biochemistry are significantly underrepresented. Conclusions. The Pharmacy study plans of Visegrad Group universities show greater similarity with each other and tend to differ from the Ukrainian Ministerial study plan. The necessary steps to harmonize Pharmacy study plans of Ukrainian universities with V4 countries include the introduction of Molecular Biology, Immunology, Clinical Biochemistry courses, and strengthening the basic medical and chemical science courses like Human Anatomy and Physiology, Organic Chemistry, Analytical Chemistry, Pharmacology, Medicinal and Pharmaceutical Chemistry.

Medical biochemistry (synonyms: clinical chemistry or clinical biochemistry) in the terms of professional and scientific discipline, stems from and/or has developed along with the natural sciences and its influences (mathematics, physics, chemistry and biochemistry) and medical sciences as well (physiology, genetics, cell biology). As a scientific discipline, medical biochemistry studies metabolic processes of physiological and pathological changes with humans and animals. Applying analytical chemistry's and biochemistry's techniques enables medical biochemists to gain plenty of information related to diagnosis and prognosis which serve physicians to asses the gravity of illness and prescribe healing therapy. Therefore medical biochemistry is an integral part of modern medicine. This discipline was dubbed various, often confusing names such as pathology, physiology, clinical biology, clinical pathology, chemical pathology, clinical biochemistry, medical biochemistry, clinical chemistry and laboratory medicine, all depending on place of origin. The official, internationally accepted name - clinical chemistry, was mentioned for the first time in 1912 by Johan Scherer, who described his laboratory as Clinical Chemistry Laboratory (Klinisch Chemische Laboratorium) in the hospital Julius in Wurzburg in Germany. After creating national societies of clinical chemists, Professor Earl J. King of Royal Postgraduate Medical School from London incited an initiative to unite national societies into the organization with worldwide character - it was the International Association of Clinical Biochemists, monitored by the International Union for Pure and Applied Chemistry (IUPAC). On 24 July 1952 in Paris, a Second International Congress of Biochemistry was held. A year later, in Stockholm, the name of a newly formed association was altered into International Federation of Clinical Chemistry, which was officially accepted in 1955 in Brussels. Today this federation-s name is International Federation for Clinical Chemistry and Laboratory Medicine (IFCC). Right after the World War II our medical biochemists began to gather within their expert societies. Even before 1950 Pharmaceutical Society of Serbia hosted laboratory experts among whom the most active were Prof. Dr. Aleksandar Damanski for bromatology, Prof. Dr. Momcilo Mokranjac for toxicology and Docent Dr. Pavle Trpinac for biochemistry. When the Managing Board of the Pharmaceutical Society of National Republic of Serbia held its session on 22 December 1950, an issue was raised with reference to creation of a Section that would gather together the laboratory experts. Section for Sanitary Chemistry, combining all three profiles of laboratory staff, i.e. medical biochemists, sanitary chemists and toxicologists, was founded on 1st of January 1951. On 15 May 1955, during the sixth plenum of the Society of Pharmaceutical Societies of Yugoslavia (SFRY) held in Split, the decision was passed to set up a Section for Medical Biochemistry in SFDJ. The Section for Medical Biochemistry in SFDJ was renamed into Society for Medical Biochemistry of SFDJ based on the decision passed during the 16th plenum of SFDJ, held on 15 May 1965 in Banja Luka. Pursuant to the decision passed by SMBY on 6 April 1995 and based on the historic data, 15 May was declared as being the official Day of the Society of Medical Biochemists of Yugoslavia. The purpose of YuSMB (currently SMBSCG) is to gather medical biochemists who would develop and enhance all the branches of medical biochemistry in health industry. Its tasks are as following: to standardize operations in clinical-biochemical laboratories, education of young biochemists on all levels, encouraging scientific research, setting up of working norms and implementation, execution and abiding by the ethics codices with health workers. SMBSCG is to promote the systemized standards in the field of medical biochemistry with the relevant federal and republican institutions. SMBSCG is to enable exchange of experiences of its members with the members of affiliate associations in the country and abroad. .

Monoclonal antibodies (MAbs) are an old immunological tool with applications in the fields of immunology, biotechnology, biochemistry, and applied biology. Production of monoclonal antibodies using hybridoma technology was discovered in 1975 by Georges Kohler of West Germany and Cesar Milstein of Argentina. Modern-day research on MAbs from laboratories worldwide is revealing additional applications in diverse branches of sciences. Recently, MAbs have been widely applied in the field of clinical medicine. Currently, MAbs account for one-third of all the new therapeutic treatments for breast cancer, leukemia, arthritis, transplant rejection, asthma, and psoriasis, with many more late-stage clinical trials being conducted. In this review, we outline the (i) production of MAbs, (ii) application of MAbs, (iii) antibody engineering, and (iv) pharmaceutical application of MAbs. The future prospect of this review lies in the applicability of monoclonal antibodies as a molecule for understanding and monitoring the biology of disease and its role in research, clinical, diagnostic, analytical, and pharmaceutical applications.

The health professionals require scientific knowledge to advise their patients on complementary and alternative medicines (CAM). Previously, several studies were conducted regarding the CAM perception, attitude and use on health care professionals only; in contrast, our study encompasses both students of health care professionals and other disciplines. The aim of this study was to compare the attitudes and perception about CAM practices between students of biological sciences (Biochemistry and Molecular Biology, Microbiology and Botany) and health care professionals (Pharmacy and medical students) in public and private universities located in the southeastern region in Bangladesh. The questionnaire-based study conducted on 332 systematically sampled students (four private and one public universities) located in the southeastern region in Bangladesh. The cross-sectional study was conducted from March to July, 2018. All students showed a positive attitude towards CAM use. Herbal medicine, homeopathy, nutritional supplements followed by hypnosis, massage, spiritual healing, and meditation were the most commonly known and used CAM modalities. Most of the students (40.7%) believed that the integration of CAM and conventional medicine should be essential in health care setting. The major obstacles for CAM use are patient interest (48.2%) and lack of physician interest (43.4%). In addition, 36.7% students believed that CAM practices should be included in their school’s curriculum. Moreover, 39.5% students assumed that CAM knowledge is important to their daily life. Biological sciences and healthcare professional students of Bangladesh showed positive perception on CAM uses. Bangladesh Pharmaceutical Journal 24(2): 159-167, 2021

Pharmacogenomics combines traditional pharmaceutical sciences such as biochemistry with annotated knowledge of genetics, protein chemistry, and DNA polymorphisms. The difference of therapeutic efficacy of the same drug in different individual can be best explained by the study of genetic polymorphisms that underlie individual differences in drug response. The small change of the genome in one individual may make a drug inefficacious as oppose to the other patients. Such variability will bring the individualization of the therapy to obtain the best effects of the drug, as an example autologous dendrimer got a tremendous success in the cancer therapy of the individual. Markers of exposure can determine whether the desired target tissues of a subject have been exposed to a drug at physiological concentrations. Gene expression profiling is a tool that can be used to characterize chemically induced toxicity in cells and/or animal models, in order to provide plausible explanations for observed toxicity in preclinical testing. Pharmacogenomics holds the promise that drugs might one day in future be tailor-made for individuals and adapted to each person's own genetic makeup by the use of microarray technology to express the gene which in turn helps to develop receptor protein. The stability of the drug and its toxicity can be predicted prior to administration of the drug to the subject through the knowledge of computer assisted structural simulation and DNA reactivity technology respectively. This article provides some critical aspects of drug developments, clinical trial design and ethical issues which are centered with pharmacogenomics.

The two strands of treatment available for coronary artery disease and associated pathologies are pharmaceutical and physical. However, these treatments are typically only available too late to help tackle the early underlying processes. Better understanding of the underlying processes is key to the development of preventative solutions. One of the key processes understanding coronary problems is atherosclerosis, which is when arteries lose elasticity. Associate Professor Seisuke Mimori, Department of Clinical Medicine, Chiba Institute of Science, Japan, is examining the underlying biochemistry of atherosclerosis. Professor Tetsuto Kanzaki has succeeded in cloning a protein called LTBP-1 and, under his direction, Seisuke has created a mutant of the protein and is analysing it. He intends to produce variants of LTBP-1 in order to investigate the function of various domains of the protein. This will involve firstly producing and isolating the protein and its variants in sufficient quantities. Then, he will test cell migration rates through an assay that he and the team have designed. This will enable the researchers to clarify exactly how LTBP-1 functions. In the future, Seisuke and the team will investigate the exact mechanism of action of the domains involved and explore the impact of LTBP-1 on the relevant organs and cells.

Phenylketonuria (PKU) is an inborn error of metabolism characterized by a loss of phenylalanine hydroxylase activity; an enzyme that metabolizes phenylalanine to tyrosine. Phenylalanine ammonia lyase (PAL) is currently being evaluated as a possible therapy for the management of PKU. PAL catalyzes the conversion of phenylalanine to transcinnamic acid (TCA). Our proposed therapy involves encapsulation of PAL enzyme in ethyl cellulose microcapsules using the spray drying method. PAL activity is markedly reduced due to product inhibition. We hypothesized that the addition of albumin to the PAL reaction mixture would bind the TCA and prevent it from inhibiting PAL. In the first phase of our research, we developed an HPLC assay to quantitate phenylalanine and TCA in the presence of albumin. In the second phase, we determined that albumin completely alleviated product inhibition and enhanced PAL activity. Subsequent ultrafiltration studies showed that albumin acted by extensively binding to and sequestering TCA. PAL microcapsules will be taken orally. In the final phase, we studied the activity of encapsulated PAL in simulated GIT conditions to evaluate the ability of microcapsules to protect PAL enzyme against pH and protease mediated degradation. The activity of encapsulated PAL was lower than an equivalent amount of free PAL possibly due to diffusional limitations to the entry of phenylalanine into the microcapsules. Encapsulation of PAL in ethyl cellulose microcapsules did not protect against acidic pH mediated reduction of PAL activity or pepsin mediated proteolytic degradation.

Human epidermal growth factor receptor 2 (HER2) is a well-studied therapeutic target as well as a biomarker of breast cancer. HER2-targeting affibody (Z _HER2:342) is a novel small scaffold protein with an extreme high affinity against HER2 screened by phage display. However, the small molecular weight of Z_HER2:342 has limited its pharmaceutical application. Human serum albumin (HSA), as the main protein in plasma, has been commonly used to extend the small peptides serum half-life. Its high solubility, stability and excellent ability to carry multiple ligands in blood stream make it a good candidate for drug delivery.

Two HSA and Z_HER2:342 fusion proteins, one with a single Z _HER2:342 domain fused to the C terminus of HSA (rHSA-ZHER2) and the other with two tandem copies of Z_HER2:342 (rHSA-(ZHER2)₂), have been constructed, expressed, and purified. Both fusion proteins possessed the HER2 and fatty acid (FA) binding abilities demonstrated by in vitro assays. Interestingly, rHSA-(ZHER2)₂, not rHSA-ZHER2, was able to inhibit the proliferation of SK-BR-3 cells at a relatively low concentration, and the increase of HER2 and ERK1/2 phosphorylation followed by rHSA-(ZHER2) ₂ treatment has been observed. However, the inhibition effect on HER2-overexpressing cells is cell linedependent. Fusion protein rHSA-(ZHER2)₂ showed preferred accumulation in tumor tissues in xenograft model.

HSA fusion proteins are easy and economical to express, purify, and formulate. Two formulation strategies have been explored, one is to complex the fusion protein with FA modified chemo drugs, and the other is to make them into nanoparticles. As expected, HSA fusion proteins and fusion protein-bound fatty acid-modified fluorescein isothiocyanate (FITC) could be efficiently taken up by cells. FA-Taxol/albumin formulation showed its advantages over Taxol/albumin treatments on in vitro cell growth inhibition. Nanoparticles containing rHSA-ZHER2 produced by desolvation method displayed optimal size distribution, satisfactory stability and preferred binding/uptake on HER2-overexpressing cells. These results proved the feasibility of using HSA fusion proteins as therapeutic agents as well as carriers for targeted drug delivery.

Inappropriate expression of nicotinic acetylcholine receptors in the central nervous system is associated with nicotine addiction, Alzheimer’s disease, Parkinson’s disease and other disorders. Modulators (drugs) have the potential to restore circuit properties that arise from inappropriate expression of nicotinic receptor’s. Compounds that interact with allosteric sites have a distinct advantage over agonists and partial agonists, in that, they retain normal activation patterns by allowing binding of the endogenous ligand. Positive allosteric modulators boost the receptors ability to respond to agonist. Studies of these modulators constitute a first step toward the identification and development of better compounds that minimize signaling errors at cholinergic synapses. We have used single molecule methods to investigate the action of a novel positive allosteric modulator, desformylflustrabromine (dFBr), on nicotinic receptors. Our studies were focused on the α4β2 subtype of nicotinic receptors in the brain. These receptors exist in two forms with low sensitivity (α4₂β2₃) or, alternatively, high sensitivity (α4₂β2₃) to agonist. Our experiments allowed us to develop detailed gating models for high and low sensitivity receptors, as well as gain new insights regarding the mechanisms that underlie potentiation by allosteric modulators. We found that dFBr potentiates low sensitivity receptors by destabilizing desensitized states of the receptor. In contrast, potentiation of high sensitivity receptors arises from a synchronization of openings following an application of agonist due to an increase in the opening rate. Based on our results we now have a better understanding of the advantages of dFBr on high and low sensitivity receptors.

The aim of this thesis is to evaluate the impact of somatostatin receptor subtype-4 (SSTR4) actions on microglia cell viability, towards the understanding and advance of pharmacological treatments for Alzheimer’s disease (AD). As of March 2016, there were 5 million people living in the United States with AD. Current drugs for AD have highly variable effects from patient to patient and are palliative at best. This thesis project focuses on the study of the BV2 cell line and the compound NNC 26-9100 (NNC). BV2 cells are immortalized microglial cells that maintain most of their morphology. The data collected suggests that BV2 cells can phagocytize amyloid-? peptides (A?), respond to lipopolysaccharide (LPS), and have the somatostatin receptor subtype-4 (SSTR4). NNC is a selective agonist of the SSTR4 and we have found that it causes BV2 cells to increase in number. The effects of NNC were able to be reduced with a somatostatin receptor pan-antagonist, cyclosomatostatin, and the adenyl cyclase activator forskolin. NNC can alter BV2 numbers by binding to the SSTR4, creating an intracellular cascade that results in the inhibition of adenyl cyclase and an increase in cell count. Collectively, a potential therapeutic mechanism for AD is increasing the number of microglial cells to increase both amyloid beta (A?) phagocytosis and degradation of A? by neprilysin.

Recent strategies for anticancer drug design have been focused on utilizing antibody as a drug or targeted moiety for targeted drug delivery. Antibody−drug conjugates (ADCs) have become a promising new class of targeted therapeutic agents for treatment of cancer. ADCs are designed to preferentially direct a cytotoxic drug to a cell-surface antigen recognized by an antibody. However, there are some challenges in developing ADCs, such as limited solid tumor penetration, high manufacturing costs and antibody-drug stoichiometry. Smaller molecules such as peptides have been shown to specifically bind to cancer related targets. These peptides can be used to form peptide-drug conjugates (PDCs) to overcome above-mentioned drawbacks presented by ADCs. In this study, it was hypothesized that novel synthesized PDCs can be a strategy for breast cancer therapy. HER2 specific binding peptides, MARAKE and MARSGL, were modified by addition of a cysteine at C-terminus. The modified peptides were coupled with monomethylauristatin E (MMAE) by using maleimidocaproyl (MC) as a non-cleavable linker to form peptide-drug conjugates (YW1, YW2) and maleimidocaproyl-valine-citrulline (MC-VC) as a cleavable linker to form peptide-drug conjugates (YW3 and YW4). The peptides, peptide-drug conjugates and MC-MMAE, MC-VC-MMAE were characterized using ESI-MS and purified by using high-performance liquid chromatography (HPLC). Cellular uptake study was performed to determine binding specificity and internalization of two HER2 specific peptides and cysteine-modified peptides (MARAKEC, MARSGLC). In vitro cell viability assay was conducted to assess the cytotoxicity and determine the targeting specificity as well as the potency of the peptide-drug conjugates. The purity of each compound was greater than 90%. Internalization of both HER2 specific binding peptides and cysteine-modified peptides were significantly higher than random peptides in HER2 over-expressed cell lines, MDA-MB361 and ZR75, while negligible uptake in HER2 negative cell line, HEK293. MC linked PDCs showed similar cytotoxicity as peptide in all cell lines; while MC-VC linked PDCs have higher cytotoxicity than MMAE in HER2 positive cell line and significant lower cytotoxicity than MMAE in normal cell line HEK293. However, PDCs with MC link do not show significant difference in cytotoxicity compared to the peptide in all cell lines. In conclusion, specificity of HER2 binding for both peptides was preserved after modification with cysteine. The derivation of MMAE to link drug and peptide played a crucial role in the anticancer activity. Peptide-MMAE conjugates with cleavable linker showed a promising targeting capability for delivery of MMAE to HER2 overexpressed cancer cells.

Gamma-Hydroxybutyrate (GHB) is an endogenous short-chain fatty acid formed from Gamma-aminobutyric acid (GABA). Clinically, GHB is marketed in the United States as Xyrem to treat narcolepsy with cataplexy and in Europe for the treatment of alcohol withdrawal and narcolepsy. However, the illicit use and abuse of GHB occurs due to its sedative/hypnotic and euphoric effects. Monocarboxylate transporters (MCTs and SMCTs) are integral membrane proteins that control the bidirectional transport of endogenous substrates including lactate, acetate and pyruvate. They have also been found to transport and mediate the clearance and distribution of GHB. MCTs demonstrate a wide tissue distribution, including brain, kidney, liver, and intestine, all of which play an important role in determining the disposition of GHB. Sex differences in drug elimination pathways contribute to the wide range of inter-individual variability observed between sexes with respect to drug disposition and effect. Sex differences in MCT expression have been observed in the brain, muscle, liver and kidney with variations potentially driven by sex hormones; however, there is an absence of information on how these expression differences translate into sex differences in GHB toxicokinetics. The objective of this study was to evaluate sex differences and the influence of the estrus cycle on GHB toxicokinetics after IV administration. Our hypothesis is that renal clearance and toxicokinetics will vary over the estrus cycle. Estrus cycle stage in female rats was determined by vaginal lavage prior to GHB administration. Ovariectomized (OVX) females were included in the study to evaluate GHB toxicokinetics in the absence of female sex hormones. Our results demonstrated that sex and the estrus cycle influence GHB toxicokinetics. Total and renal clearance varies over the estrus cycle with the highest renal clearance observed in proestrus females. In contrast, males and OVX females demonstrated significantly lower renal clearance. These results suggest that GHB toxicity and risk of overdose varies over the estrus cycle due to expression changes in renal MCTs and SMCTs. Future studies will evaluate higher GHB doses to determine the role of sex hormones in GHB overdose and fatality. In addition, hormone replacement studies will be conducted to confirm the role of individual sex hormones on GHB toxicokinetics.

The novel knob-socket (KS) model provides a construct to interpret and analyze the direct contributions of amino acid residues to the stability in α-helical protein structures. Based on residue preferences derived from a set of protein structures, the KS construct characterizes intra- and inter-helical packing into regular patterns of simple motifs. The KS model was used in the de novo design of an α-helical homodimer, KSα1.1. Using site-directed mutagenesis, KSα1.1 point mutants were designed to selectively increase and decrease stability by relating KS propensities with changes to α-helical structure. This study suggests that the sockets from the KS Model can be used as a measure of α-helical structure and stability. The KS model was also used to investigate coiled-coil specificity in bZIP proteins. Identifying and characterizing the interactions that determine the dimerization specificity between bZIP proteins is a crucial factor in better understanding disease formation and proliferation, as well as developing drugs or therapeutics to combat these diseases. Knob-Socket mapping methods identified Asn residues at a positions within the helices, and were determined to be crucial factors in coiled-coil specificity. Site-directed mutagenesis was conducted to investigate the role of the Asn residues, as well as the role played by the neighboring residues at the g and b positions. The results indicate that the Asn at the a position defines coiled-coil specificity, and that the Knob-Socket model can be used to determine bZIP protein quaternary interactions.

Hepatitis C virus (HCV) is the etiologic agent responsible for the majority of cases of non-A, non-B hepatitis. It is estimated that 2% of the world's population is infected with hepatitis C with roughly 75% of infections progressing into a chronic state. Current interferon therapy is largely ineffective against chronic Hepatitis C due to the induction of interferon-resistance by HCV. The underlying mechanisms for resistance to interferon therapy have been widely studied and documented and as is the case in interferon resistant malignancies, there is some evidence that HCV infected cells are more susceptible to infection by interferon sensitive viruses. Most current research into novel HCV therapeutics has focused on the discovery of inhibitors of viral replication. However, the high mutation rate of the HCV genome makes it a virtual certainty that viral variants resistant to protease and/or polymerase inhibitors will arise. As an alternative approach, we have designed a therapeutic strategy that exploits both the expression of a viral protease in infected cells as well as the defects in the interferon pathway to build selectively active therapeutics that target and kill HCV infected cells. Initial experiments showed some promise using this approach and indicated areas for further development. Conditionally replicating viruses failed to rescue in HCV expressing cell lines, thus requiring further modification. A separate approach involving conditional stabilization of the suicide gene HSV-1 TK was successful with a two-fold increase in the expression levels of TK in cells co-expressing HCV protease.

Glycosaminoglycans (GAGs) are complex linear chain carbohydrate molecules found on virtually all animal cell surfaces. Owing to their negatively charged nature, GAGs interact with a number of different proteins. Thus, although they have great potential as therapeutic agents, their apparent promiscuous interactions increase their side effect risk. GAG mimetics, including GAG oligosaccharides and non-saccharide GAG mimetics (NSGMs) are viable approaches to address this. This work discusses sulfated benzofuran thrombin inhibitors with submaximal protease inhibition, sulfated diflavonoid inhibitors of plasmin and GAG oligosaccharides with selectivity for human neutrophil elastase (HNE). Anticoagulants are very important for the treatment of thrombotic diseases. The adverse effects associated with current clinically used anticoagulants warrant the continuous search for new agents. Thrombin, being the central player in the coagulation cascade, remains a very important target for anticoagulant therapy, however drugs inhibiting its activity carry the risk of prolonged bleeding. Based on a previously identified sulfated benzofuran thrombin inhibitor, we have developed analogs with submaximal inhibition of the protease. These agents inhibit thrombin with efficacies approaching 50%, for both chromogenic and macromolecular substrates, ensuring a basal level of thrombin activity even at saturating inhibitor concentrations. The most potent of these compounds had a potency of 1.8 µM, 2-3 fold better than the lead. Additionally, these compounds utilize an allosteric mechanism for thrombin inhibition. Further, studies have revealed structural features responsible for submaximal thrombin inhibition. Fibrinolysis is an important part of hemostasis and plasmin is the most important fibrinolytic enzyme. Anti-plasmin agents are thus important for conditions such as hemophilia; however, there are no clinically used direct plasmin inhibitors. By structural modifications of a previously identified sulfated diflavonoid plasmin inhibitor, we have achieved a compound with 12-fold better potency (IC50 = 6.3 ± 0.4 µM), and a selectivity index of at least 22 over closely related serine proteases. We have shown that this compound inhibits plasmin mediated clot lysis, and further demonstrated that its activity is reversible using protamine sulfate, indicating its potential as a lead for the development of clinical anti-plasmin agents. HNE, a serine protease associated with inflammatory diseases is known to be inhibited by GAGs. However, the interactions at the molecular level have remained elusive. Using biochemical methods, and by studying the inhibitory potency of different GAGs and GAG oligosaccharides, we have shown that an octasaccharide may be the ideal GAG length for the achievement of potent HNE inhibition. Under our assay conditions, the inhibition of HNE by an octasaccharide species was only 5-fold less than that of unfractionated heparin, whereas the hexasaccharide species was 30-fold less active. The data also suggests that the inhibition of HNE by GAGs is via an allosteric mechanism and using molecular modeling, we have identified putative GAG binding sites on HNE and further identified GAG species with potential selectivity for anti-HNE activity

Aryl hydrocarbon receptor (AHR), commonly known as an environmental sensor involved in the metabolism and elimination of xenobiotic substances, is also an important modulator in the development and functioning of the immune system. AHR expression is varied in the T cell subsets with the highest expression in T-helper 17 and T regulatory cells. Work from many researchers has suggested that AHR can act as a tumor promoter or a tumor suppressor depending on the tumor type. Our goal is to understand the role played by AHR in MC38 syngeneic colon carcinoma tumor model. In the absence of AHR, MC38 tumor progresses by an increase in tumor associated macrophages (TAMs), M2 macrophages and a decrease in CD8a positive cytotoxic lymphocytes. Analysis of the intratumoral cytokines reveals a pro-inflammatory phenotype. This has been assessed by pharmacologic blocking of the receptor using CH223191 and in AHR deficient (AHR-/-) mice. Therefore AHR acts as a tumor suppressor gene in colon carcinoma tumor model and silencing it may lead to colon cancer progression.