Isoenzymes

Linear Quantitative Structure-Activity Relationships for the Interaction of Small Molecules with Human Cytochrome P450 Isoenzymes 

2 Models based on alignment-independent molecular descriptors 69 

Drug-drug interactions have become an important issue in human health care [1- 

Many of the major pharmacokinetic interactions between drugs are due to hepatic cytochrome P450 (CYP) enzymes being inhibited by concomitant administration of other drugs. Additionally, genetic polymorphisms of important CYP 

Computational Chemistiy, Department of Lead Discovery, Boehringer Ingelheim Pharma GmbH Co. KG, 88397 Biberach, Germany. E-mail thomas.fox bc.boehringer-ingelheim.com Corresponding author. 

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Metabolism is still a barrier to be overcome. Some QSAR, pharmacophore, protein, and rule-based models are available to predict substrates and inhibitors of a specific cytochrome P450 isoenzyme [47-55]. 

Assay of Enzymes In body fluids, enzyme levels aie measured to help in diagnosis and for monitoiing treatment of disease. Some enzymes or isoenzymes are predominant only in a particular tissue. When such tissues are damaged because of a disease, these enzymes or isoenzymes are Hberated and there is an increase in the level of the enzyme in the semm. Enzyme levels are deterrnined by the kinetic methods described, ie, the assays are set up so that the enzyme concentration is rate-limiting. The continuous flow analyzers, introduced in the early 1960s, solved the problem of the high workload of clinical laboratories. In this method, reaction velocity is measured rapidly the change in absorbance may be very small, but within the capabiUty of advanced kinetic analyzers. 

Enzymes, measured in clinical laboratories, for which kits are available include y-glutamyl transferase (GGT), alanine transferase [9000-86-6] (ALT), aldolase, a-amylase [9000-90-2] aspartate aminotransferase [9000-97-9], creatine kinase and its isoenzymes, galactose-l-phosphate uridyl transferase, Hpase, malate dehydrogenase [9001 -64-3], 5 -nucleotidase, phosphohexose isomerase, and pymvate kinase [9001-59-6]. One example is the measurement of aspartate aminotransferase, where the reaction is followed by monitoring the loss of NADH ... 

Chronic use of these irreversible MAO inhibitors has been associated with life-threatening toxicity, ie, hepatotoxicity and hypertensive crisis. Interactions with tyramine contained in food and other drugs have severely limited use of irreversible MAO inhibitors. These MAO inhibitors are also nonselective, inhibiting both MAO-A and MAO-B isoenzymes. Furthermore, they interfere with the hepatic metabolism of many dmgs. 

Another interesting sperm specific antigen is lactic dehydrogenase-x (LDH-x or LDHC, an isoenzyme of LDH confined to male germ ceUs. 

Pyruvate kinase isoenzymes (from Salmonella typhimurium) [9001-59-6] Mr 64,000, [EC 2.7.1.40], amorphous. Purified by (NH4)2S04 fractionation and gel filtration, ion-exchange and affinity chromatography. [Garcia-Olalla and Garrido-Pertierra Sioc/iem 7 241573 1987.]... 

CDP840 is a selective inhibitor of the PDE-IV isoenzyme and interest in the compound arises from its potential application as an antiasthmatic agent. Chemists at Merck Co. used the asymmetric epoxidation reaction to set the stereochemistry of the carbon framework and subsequently removed the newly established C-O bonds." Epoxidation of the trisubstituted olefin 51 provided the desired epoxide in 89% ee and in 58% yield. Reduction of both C-O bonds was then accomplished to provide CDP840. 

Antithyroid drags have several side effects. The most frequent side effects are maculopapular rashes, pruritus, urticaria, fever, arthralgia and swelling of the joints. They occur in 1-5% of patients [1, 2]. Loss of scalp hair, gastrointestinal problems, elevations of bone isoenzyme of alkaline phosphatase and abnormalities of taste and smell are less common. The incidence of all these untoward reactions is similar with MMI and PTU. Side effects of MMI are dose-related, whereas those of PTU are less clearly related to dose [1]. PTU may cause slight transient increases of serum aminotransferase and y-glutamyl transpeptidase concentrations but also severe hq atotoxicity whereas methimazole or carbimazole can be associated with cholestasis. The side... 

The reactivity of the sodium pump isoforms toward ouabain can significantly differ between certain isoenzymes. The al-isoform from rat is reported to be 100-fold more resistant to ouabain than a2- or a3-isoform. In other species like rabbit, pig, dog or human these differences are not as distinct as in rat, due to a more sensitive al-isoform. At least two different types of ouabain binding sites are reported for the Na+/fC+-ATPase [1]. 

The development of the COXIBs has been based on the hypothesis COX-1 is the physiological COX and COX-2, the pathophysiological isoenzyme. Inhibition of the pathophysiological COX-2 only is assumed to result in fewer side effects as compared to non-selective inhibition of both COX isoenzymes (Fig. 2). Celecoxib, etoricoxib and lumiracoxib (in some countries also parecoxib) are the only COXIBs currently approved. 

Imidazole antimycotics, ketoconazole, clotrimazole, and miconazole are potent inhibitors of various cytochrome P450-isoenzymes that also affect the metabolism of retinoids. They were fust shown to inhibit the metabolism of RA in F9 embryonal carcinoma cells. When tested in vitm liarazole, a potent CYP-inhibitor, suppressed neoplastic transformation and upregulated gap junctional communication in murine and human fibroblasts, which appeared to be due to the presence of retinoids in the serum component of the cell culture medium. Furthermore, liarazole magnified the cancer chemopreventive activity of RA and (3-carotene in these experiments by inhibiting RA-catabolism as demonstrated by absence of a decrease in RA-levels in the culture medium in the presence of liarazole over 48 h, whereas without liarazole 99% of RA was catabolized. In vivo, treatment with liarazole and ketoconazole reduced the accelerated catabolism of retinoids and increased the mean plasma all-irans-RA-concentration in patients with acute promyelocytic leukemia and other cancels. 

Ethanol also inhibits ADH-catalyzed retinol oxidation in vitro, and ethanol treatment of mouse embtyos has been demonstrated to reduce endogenous RA levels. The inhibition of cytosolic RolDH activity and stimulation of microsomal RolDH activity could explain ethanol-mediated vitamin A depletion, separate from ADH isoenzymes. Although the exact mechanism of inhibition of retinoid metabolism by ethanol is unclear, these observations are consistent with the finding that patients with alcoholic liver disease have depletedhepatic vitamin A reserves [review see [2]. 

In resting muscle the high concentration of ADP does not decrease the proton gradient effectively and the high membrane potential slows electron transport. ADP, formed when ATP is hydrolyzed by myosin ATPase during contraction, may stimulate electron transport. However, the concentration of ATP (largely as its Mg salt) is buffered by its readily reversible formation from creatine phosphate catalyzed in the intermembrane space, and in other cell compartments, by the various isoenzymes of creatine kinase (reviewed by Walliman et al., 1992). 

Wallimann, T., Wyss. M., Brdicza, D., Nicolay, K.. Eppenberger, H.M. (1992). Intracellular compartmentation. structure and function of creatine kinase isoenzymes in tissues with high and fluctuating energy demands The phosphocreatine circuit for cellular energy homeostasis. Biochem. J. 281,21-40. 

The gene for myophosphoiylase has been assigned to chromosome 1 lql3. The enzyme is a dimer of two identical 97 IcDa subunits and is the sole isoform present in skeletal muscle. Heart and brain also contain this isoform in addition to a distinct brain isoenzyme and a hybrid muscle/brain isoform. Smooth muscle also contains a phosphorylase isoform distinct from the muscle isoenzyme. If regenerating muscle fibers are present they also contain phosphorylase activity due to the presence, in fetal and developing muscle, of an isoform said to be identical with brain phosphorylase. 

Chick J, Lehert P, Landron F, et al Does acamprosate improve reduction of drinking as well as aiding abstinence J Psychopharmacol 17 397-A02, 2003 Chrostek L, Jelski W, Szmitkowski M, et al Gender-related differences in hepatic activity of alcohol dehydrogenase isoenzymes and aldehyde dehydrogenase in humans. J Clin Lab Anal 17 93-96, 2003... 

Maksimov I. V. Gherepanova E. A. Surina O. B. (2010) Effect of chitooligosaccharides on peroxidase isoenzyme composition in wheat calli co cultured with bunt causal agent / / Rus. J. of Plant Physiol. V. 57. P. 131-138. 

Isoenzymes (isozymes) Enzymes that are very similar to one another in size and structure but with differences in catalytic ability. 

A non-allergic mechanism imderlying precipitation of asthmatic attacks by aspirin in hypersensitive patients was proposed over 30 years ago [4]. It was founded on pharmacological inhibition of COX of arachidonic acid and explained a cross-reactivity between different NSAIDs varying in chemical structure. This COX theory was confirmed by several studies [11] and was further refined following discovery of the second COX isoenzyme - COX-2. At least two COX isoenzymes, COX-1 and COX-2, are coded by separate genes. Their role in inflammation, asthma and anaphylaxis has been reviewed previously [12]. 

Succinyl-CoA is converted to succinate by the enzyme succinate thiokinase (succinyl-CoA synthetase). This is the only example in the citric acid cycle of substrate-level phosphorylation. Tissues in which glu-coneogenesis occurs (the hver and kidney) contain two isoenzymes of succinate thiokinase, one specific for GDP and the other for ADP. The GTP formed is used for the decarboxylation of oxaloacetate to phos-phoenolpymvate in gluconeogenesis and provides a regulatory hnk between citric acid cycle activity and the withdrawal of oxaloacetate for gluconeogenesis. Nongluconeogenic tissues have only the isoenzyme that uses ADP. 

The uptake of glucose into the liver is independent of insulin, but liver has an isoenzyme of hexokinase (glucokinase) with a high K, so that as the concentration of glucose entering the liver increases, so does the... 

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