Inhibition Of Different Isoforms Research Articles

Identification, crystallization, and first X-ray structure analyses of phenyl boronic acid-based inhibitors of human carbonic anhydrase-II

Human carbonic anhydrases (hCAs) play a central role in various physiological processes in the human body. HCAs catalyze the reversible hydration of CO2 into HCO3−, and hence maintains the fluid and pH balance. Overexpression of CA II is associated with diseases, such as glaucoma, and epilepsy. Therefore, CAs are important clinical targets and inhibition of different isoforms, especially hCA II is used in treatment of glaucoma, altitude sickness, and epilepsy. Therapeutically used CA inhibitors (CAI) are sulfonamide-based, such as acetazolamide, dichlorphenamide, methazolamide, ethoxzolamide, etc. However, they exhibit several undesirable effects such as numbness, tingling of extremities, malaise, metallic taste, fatigue, renal calculi, and metabolic acidosis. Therefore, there is an urgent need to identify safe and effective inhibitors of the hCAs. In this study, different phenyl boronic acids 1–5 were evaluated against bovine (bCA II) and hCA II. Among all, compound 1 (4-acetylphenyl boronic acid) was found to be active against bCAII and hCA II with IC50 values of 246 ± 0.48 and 281.40 ± 2.8 μM, respectively, while the remaining compounds were found in-active. Compound 1 was identified as competitive inhibitor of hCA II enzyme (Ki = 283.7 ± 0.002 μM). Additionally, compound 1 was found to be non-toxic against BJ Human fibroblast cell line. The X-ray crystal structure for hCA II in-complex with compound 1 was evaluated to a resolution of 2.6 Å. In fact, this the first structural analysis of a phenyl boron-based inhibitor bound to hCA II, allowing an additional structure-activity analysis of the compounds. Compound 1 was found to be directly bound in the active site of hCA II by interacting with His94, His119, and Thr199 residues. In addition, a bond of 3.11 Å between the zinc ion and coordinated boron atom of the boronic acid moiety of compound 1 was also observed, contributing to binding affinity of compound 1 for hCA II. PDB ID8IGF.

Overview of the New Bioactive Heterocycles as Targeting Topoisomerase Inhibitors Useful Against Colon Cancer.

Colorectal cancer (CRC) is the third most common cancer globally, with high mortality. Metastatic CRC is incurable in most cases, and multiple drug therapy can increase patients' life expectancy by 2 to 3 years. Efforts are being made to understand the relationship between topoisomerase enzymes and colorectal cancer. Some studies have shown that higher expression of these enzymes is correlated to a poor prognosis for this type of cancer. One of the primary drugs used in the treatment of CRC is Irinotecan, which can be used in monotherapy or, more commonly, in therapeutic schemes such as FOLFIRI (Fluorouracil, Leucovorin, and Irinotecan) and CAPIRI (Capecitabine and Irinotecan). Like Camptothecin, Irinotecan and other compounds have a mechanism of action based on the formation of a ternary complex with topoisomerase I and DNA providing damage to it, therefore leading to cell death. Thus, this review focused on the principal works published in the last ten years that demonstrate a correlation between the inhibition of different isoforms of topoisomerase and in vitro cytotoxic activity against CRC by natural products, semisynthetic and synthetic compounds of pyridine, quinoline, acridine, imidazoles, indoles, and metal complexes. The results revealed that natural compounds, semisynthetic and synthetic derivatives showed potential in vitro cytotoxicity against several colon cancer cell lines, and this activity was often accompanied by the ability to inhibit both isoforms of topoisomerase (I and II), highlighting that these enzymes can be promising targets for the development of new chemotherapy against CRC. Pyridine analogs were considered the most promising for this study, while the evaluation of the real potential of natural products was limited by the lack of information in their work. Moreover, the complexes, although promising, presented as the main limitation the lack of selectivity.

Selective Inhibition of Different Isoforms of Connexin Hemichannels by New Aminoglycosides

Selective Inhibition of Different Isoforms of Connexin Hemichannels by New Aminoglycosides

New pharmacologic interventions to increase cardiac contractility: challenges and opportunities.

The most extensively studied inotropic agents in patients with heart failure are phosphodiesterase (PDE) 3 inhibitors, which increase contractility by raising intracellular cyclic adenosine monophosphate content. In clinical trials, the inotropic benefits of these agents have been outweighed by an increase in sudden cardiac death. Here, I review recent findings that help explain what are likely to be distinct mechanisms involved in the beneficial and adverse effects of PDE3 inhibition. The proapoptotic consequences of PDE3 inhibition are becoming more apparent. Moreover, it has also become clear that individual PDE3 isoforms in cardiac myocytes are selectively regulated to interact with different proteins in different intracellular compartments. The beneficial and adverse effects of PDE3 inhibition may thus be attributable to the inhibition of different isoforms in different intracellular domains. In particular, PDE3A1 has been shown to interact directly with sarcoplasmic/endoplasmic reticulum Ca ATPase (SERCA2) in the sarcoplasmic reticulum through a phosphorylation of a site in its unique N-terminal domain, making it possible that this isoform can be selectively targeted to increase intracellular Ca cycling. Conventional PDE3 inhibitors target several functionally distinct isoforms of these enzymes. Isoform-selective and/or compartment-selective targeting of PDE3, through its protein-protein interactions, may produce the inotropic benefits of PDE3 inhibition without the adverse consequences.

A novel fluorescence-based biosynthetic trafficking method provides pharmacologic evidence that PI4-kinase IIIα is important for protein trafficking from the endoplasmic reticulum to the plasma membrane.

BackgroundBiosynthetic trafficking of receptors and other membrane-associated proteins from the endoplasmic reticulum (ER) to the plasma membrane (PM) underlies the capacity of these proteins to participate in crucial cellular roles. Phosphoinositides have been shown to mediate distinct biological functions in cells, and phosphatidylinositol 4-phosphate (PI4P), in particular, has emerged as a key regulator of biosynthetic trafficking.ResultsTo investigate the source of PI4P that orchestrates trafficking events, we developed a novel flow cytometry based method to monitor biosynthetic trafficking of transiently transfected proteins. We demonstrated that our method can be used to assess the trafficking of both type-1 transmembrane and GPI-linked proteins, and that it can accurately monitor the pharmacological disruption of biosynthetic trafficking with brefeldin A, a well-documented inhibitor of early biosynthetic trafficking. Furthermore, utilizing our newly developed method, we applied pharmacological inhibition of different isoforms of PI 4-kinase to reveal a role for a distinct pool of PI4P, synthesized by PI4KIIIα, in ER-to-PM trafficking.ConclusionsTaken together, these findings provide evidence that a specific pool of PI4P plays a role in biosynthetic trafficking of two different classes of proteins from the ER to the Golgi complex. Furthermore, our simple, flow cytometry-based biosynthetic trafficking assay can be widely applied to the study of multiple classes of proteins and varied pharmacological and genetic perturbations.

Studies on the antioxidant effect and interaction of diphenyl diselenide and dicholesteroyl diselenide with hepatic δ-aminolevulinic acid dehydratase and isoforms of lactate dehydrogenase

Studies on the interaction of dicholesteroyl diselenide (DCDS) and diphenyl diselenide (DPDS) with hepatic delta-aminolevulinic acid dehydratase (ALA-D) and different isoforms of lactate dehydrogenase (LDH) from different tissues were investigated. In addition, their antioxidant effects were tested in vitro by measuring the ability of the compounds to inhibit the formation of hepatic thiobarbituric acid reactive species (TBARS) induced by both iron (II) and sodium nitroprusside (SNP). The results show that while DPDS markedly inhibited the formation of TBARS induced by both iron (II) and SNP, DCDS did not. Also, the activities of hepatic delta-aminolevulinic acid dehydratase (ALA-D) and different isoforms of lactate dehydrogenase (LDH) were significantly inhibited by both DPDS and DCDS. Moreover, we further observed that the in vitro inhibition of different isoforms of lactate dehydrogenase by DCDS and DPDS likely involves the modification of the groups at the NAD+ binding site of the enzyme. Since organoselenides interacts with thiol groups on proteins, we conclude that the inhibition of different isoforms of lactate dehydrogenase by DPDS and DCDS possibly involves the modification of the thiol groups at the NAD+ binding site of the enzyme.

Differing effects of N(G)-monomethyl L-arginine and 7-nitroindazole on detrusor activity.

Previous studies reported that nitric oxide (NO) synthase (NOS) inhibition decreases micturition volume threshold (MVT), the volume required to produce a centrally mediated micturition contraction, and that NO can be released from urothelium by means of certain stimuli. With elucidation of multiple isoforms of NOS, studies were performed to determine whether inhibition of specific isoforms of NOS altered MVT in different ways. In naive, anesthetized cats, the urinary bladder was exposed by means of a midline abdominal incision and cannulated through a slit in the internal urethra approximately 4-5 cm distal to the neck of the bladder. The left renal artery and left radial vein were cannulated for the intra-arterial and intravenous administration of drugs, respectively. All nerves were left intact. A control MVT was determined by slowly infusing saline into the bladder at a rate of 0.018 mL/kg per minute. Varying doses of L-NMMA (N(G)-monomethyl-L-arginine) or 7-NI (7-nitro indazole) were administered and the MVT was again determined. Inhibition of endothelial NOS (eNOS), by L-NMMA, or neuronal NOS (nNOS), by 7-NI, produces varying effects on certain detrusor activities and that inhibition of different isoforms of NOS produces qualitatively different effects. L-NMMA significantly decreases MVT (up to 60% decrease), whereas 7-NI significantly increases MVT (over 300% increase). L-NMMA increases frequency and onset of small bladder contractions, whereas 7-NI produces opposite effects. The results suggest that detrusor relaxation and contractility may be modulated by NO levels and that NO released from the urothelium may be a mediator of detrusor relaxation during the storage phase of micturition.

PDE3 cyclic nucleotide phosphodiesterases and the compartmentation of cyclic nucleotide-mediated signalling in cardiac myocytes.

PDE3 cyclic nucleotide phosphodiesterase inhibitors raise cAMP and cGMP content in cardiac and vascular myocytes. Their administration to patients with dilated cardiomyopathy leads to improvements in hemodynamic parameters in the short term but reduces survival with chronic administration. The reasons for this 'biphasic' response have not been elucidated, but it is likely that beneficial and harmful effects of PDE3 inhibition reflect the phosphorylation of different substrates of cAMP- and cGMP-dependent protein kinases (PK-A and PK-G). It is now apparent that cardiac and vascular myocytes contain several isoforms of PDE3 that differ in their intracellular distribution and thus regulate cAMP and cGMP levels in different subcellular compartments. These isoforms also differ in their regulation by extracellular signals that may be important in the pathophysiology of dilated cardiomyopathy. An intriguing possibility is that the beneficial and harmful effects of PDE3 inhibition may be attributable to the inhibition of different isoforms of these enzymes.

P-444 - 5HT causes either activation or inhibition of different isoforms of adenylyl cyclases (ACs) and phospholipase C (PLC) through G protein βγ subunits in oocytes expressing 5HT1A receptors (5HT1ARS).

P-444 - 5HT causes either activation or inhibition of different isoforms of adenylyl cyclases (ACs) and phospholipase C (PLC) through G protein βγ subunits in oocytes expressing 5HT1A receptors (5HT1ARS).