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Selective Substrates (selective + substrate)

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Medical Sciences	87%

Selected Abstracts

Prediction of polymorphic N -acetylation of new drug candidates by correlation with human NAT1 and NAT2

DRUG DEVELOPMENT RESEARCH, Issue 1 2002
Katalin Jemnitz
Abstract Due to interindividual variation in N -acetyltransferase 2 (NAT2) activity, pharmaceutical companies face the problem of polymorphic metabolism in drugs that are metabolized mainly or exclusively by this enzyme. An in vitro method has been developed to predict in vivo polymorphic N -acetylation at an early stage of drug development. Two new type 5H-2,3-benzodiazepine derivatives, Nerisopam (NER) with anxiolytic activity and GYKI47261 with antiepileptic activity, are metabolized mainly by N -acetylation in the rat and human. The selectivity of human N -acetyltransferases (NAT1,2) to form the acetylated metabolites has been investigated by correlation analysis. Twelve human liver samples were characterized for NAT1 and NAT2 phenotype based on their enzyme activity toward two selective NAT1 (p -aminobenzoic acid, PABA; p -aminosalicylic acid, PAS) and two selective NAT2 (sulfamethazine, SMZ; procainamide, PROC) substrates. Significant correlation was found between enzyme activities NAT1PABA/NAT1PAS and NAT2SMZ/NAT2PROC, respectively, and no correlation was observed comparing enzyme activities toward NAT1PABA/NAT2PROC. Enzyme activities using NER and GYKI 47261 as substrates were compared to activities obtained with NAT1 and NAT2 selective substrates, and the correlation coefficients were calculated. Good correlation was established between the rates of acetylation of the two drugs and that of the NAT2 selective substrate (NER/NAT2SMZ, r2=0.91, GYKI 47261/NAT2SMZ, r2=0.91). In contrast, no correlation was found between the rate of conjugation of the drugs and that of NAT1 selective substrate (NER/NAT1PABA, r2=0.022, GYKI 47261/NAT1PABA, r2=0.0004), suggesting polymorphic in vivo metabolism, since both drugs are acetylated preferably by NAT2. According to our results, correlation analysis based on in vitro acetylation activity may be used to predict in vivo polymorphic metabolism. Drug Dev. Res. 56:17,22, 2002. © 2002 Wiley-Liss, Inc. [source]

Cultured mammary epithelial monolayers (BME-UV) express functional organic anion and cation transporters

JOURNAL OF VETERINARY PHARMACOLOGY & THERAPEUTICS, Issue 5 2009
M. M. AL-BATAINEH
There is ongoing concern about the potential adverse effects of xenobiotic residues in cows' milk to the human consumer. Although drugs that are intentionally administered to lactating dairy cattle are rigorously regulated to prevent harmful residues, there are numerous other potential sources of exposure that are not as easily controlled. For example, cattle may be exposed to mycotoxins, pesticides and/or persistent organic pollutants through feed, water and inhalation of polluted air. Accurate estimates of the rate and extent of excretion of these compounds into milk is important to assess the risk of exposure through cows' milk. In the present study, the expression of carrier mediated transport processes in cultured monolayers of an immortalized bovine mammary epithelial cell line (BME-UV) was determined using a flow-through diffusion cell system, selective substrates and inhibitors of organic cation transporters (OCT) and organic anion transporters (OAT). The basal-to-apical (BL-to-Ap) flux of tetraethylammonium and estrone sulfate significantly exceeded their flux in the opposite direction. The addition of selective inhibitors to the donor compartment significantly decreased the BL-to-Ap flux of either selective substrate. These results suggest that both OCT and OAT are functionally expressed by BME-UV cells. [source]

Dynamic Chemical Devices: Photoinduced Electron Transfer and Its Ion-Triggered Switching in Nanomechanical Butterfly-Type Bis(porphyrin)terpyridines,

CHEMISTRY - A EUROPEAN JOURNAL, Issue 7 2006
Myriam Linke-Schaetzel Dr.
Abstract A series of butterfly-type molecular constructs has been prepared in good yield by using a double Stille coupling synthetic protocol. They are composed of a terpyridine (terpy) scaffold and two wings composed of appended porphyrins that are capable of switching from an extended W geometry to a compact U geometry upon cation coordination of the terpy unit. The porphyrin moieties exist in the constructs either as free bases or they can be sequentially metallated, thus giving rise to wings of different "colours". Stationary and time-resolved emission studies of the HZn, ZnAu and Zn2Au constructs show that the electronic properties are strongly dependent on the geometry. In the extended W conformation an energy-transfer process is seen from the free base to the Zn-metallated porphyrin. In the U conformation in Zn2Au the donor luminescence resulting from the singlet excited state of the Zn wing is strongly, quenched not only due to the heavy atom effect but also due to a fast electron-transfer process to the ground state of the Au wing. Furthermore, the binding of (,,,)-diamine substrates to the ZnII,porphyrin sites can also influence the conformation of the system. For the Zn2Zn construct, single-crystal diffraction experiments with synchrotron radiation allowed the structure to be solved by direct methods and fully refined; it shows the expected U conformation. The central Zn atom is six-coordinate, whereby the zinc atom is coordinated by the ,3 -terpy ligand as well by monodentate and semi-chelating acetate anions. The structure is made rigid by hydrogen bonds involving the aqua ligands on the outer Zn centres and acetate oxygen atoms. The present system thus represents a double-trigger-modulated optomechanical switching device with selective substrate binding for either metal atoms or tailored ligands. Both energy- and electron-transfer processes can be controlled opening a means of improving the on/off ratio in future constructs. Une série d'architectures moléculaires, présentant une forme de papillon, a été préparée avec de bons rendements en utilisant une méthodologie synthétique comprenant comme étape clé un double couplage de Stille. Ces architectures sont composées d'un squelette terpyridine (terpy), qui compose le torse du papillon, sur lequel deux ailes porphyriniques ont été greffées; la géométrie peut varier entre une conformation étendue en forme de W, et une conformation compacte en forme de U, dû à la complexation du torse terpy par un cation de taille et coordination adéquate. Les ailes porphyriniques se présentent soit comme bases libres ou, peuvent être métallées successivement avec différentes métaux, pouvant ainsi avoir différentes "couleurs". Des études de fluorescence non seulement stationnaire mais aussi résolues dans le temps sur les architectures HZn, ZnAu et Zn2Au, montrent que les propriétés électroniques sont très dépendantes de la géométrie adoptée. Dans la conformation W étendue, nous avons mis en évidence un processus de transfert d'énergie de la porphyrine base libre vers la porphyrine metalée avec du Zn. Dans la conformation U de Zn2Au la luminescence de l'aile donneuse, à cause de l'état excité singulet, est fortement éteinte, non seulement à cause de l'effet d'atome lourd, mais aussi à cause d'un processus de transfert d'électron vers l'état fondamental de l'aile metalée avec de l'or. De plus, par compléxation avec des (,,,)-diamines des atomes de ZnIIdans les tetrapyrroles porphyrinques, la conformation induite du système peut être profondément influencée. Pour le composé Zn2Zn, que nous avons pu obtenir en monocristaux, des expériences de diffraction en utilisant comme source lumineuse un synchrotron, ont fourni des données qui ont permis de résoudre la structure par des méthodes directes et de la raffiner pour montrer la conformation en U attendue. L'atome central de Zn a une coordination de six dont trois provenant du ligand ,3 -terpy, et les autres de deux anions acetates, un monodenté et l'autre semi-chelatizé. La structure est ligotée par des multiples liaisons hydrogène entre des ligands aqueux sur les atomes de Zn externes et les atomes d'oxygène des anions acetates. Le système présenté est un double effecteur qui montre une commutation opto-mechanique dirigée soit par des ions métalliques, soit par d'autres ligands de taille adaptée. En même temps, les processus de transfert d'énergie et d'électrons peuvent être commuté en laissant de la place pour améliorer le rapport "on/off" dans de futures architectures. Abstract in Romanian:O serie de arhitecturi moleculare având o form, de fluture au fost preparate cu randamente bune utilizând o metodologie sintetic, cuprinzând ca etap, cheie un dublu cuplaj Stille. Aceste architecturi sunt compuse dintr-un schelet terpiridinic (terpy) alc,tuind trupul fluturelui, la care au fost implementate dou, aripi porfirinice care sunt capabile sa varieze între o conforma,ie extins, în forma de liter, W ,i o conformai,ie compact, în form, de litera U, datorit, complexarii trupului terpy printr-un cation de talie ,i coordina,ie adecvat,. Aripile porfirinice se prezintã fie ca base libere sau, datorita unei metalari succesive cu diferite metale, pot avea diferite "culori". Studii de fluorescen,a atât sta,ionar, cât ,i rezolvat, în timp, asupra arhitecturilor HZn, ZnAu ,i Zn2Au, arat, c, proprit,,ile electronice sunt foarte dependente de geometria adoptat,. In conforma,ia W extins,, am putut pune în eviden,, un proces de transfer de energie dinspre baza liber, c,tre porfirina metalat, cu Zn. In conforma,ia U a Zn2Au luminescen,a aripei donoare, datorate st,rii singulet excitate, este puternic stins, nu numai datorit, efectului de atom greu, cât ,i a unui proces de transfer de electron c,tre starea fundamental, a aripei metalat, cu aur. Pe deasupra, complexând (,,,)-diamine c,tre atomii de ZnIIîntre tetrapirolii porfirinici, sunt induse profunde influen,e asupra conforma,iei sistemului. Pentru compusul Zn2Zn, care a putut fi crescut monocristalin, experimente de difrac,ie folosind ca surs, luminoas, un sincrotron au condus la un set de date care a permis ca structura sa fie elucidat, prin metode directe ,i rafinat,, ar,tând conforma,ia în U a,teptat,. Atomul central de Zn are o coordina,ie de sase unde pe lang, lignadul ,3 -terpy, atomul de Zn este coordinat de c,tre doi anioni acetat, unul monodentat, iar cel,lalt semi-chelatizant. Structura este br,zdat, de leg,turi de hidrogen care implic, liganzii apo,i situa,i pe centrii de Zn exteriori, ,i atomii de oxigen din ionii acetat. Sistemul prezentat este un dutblu efector, ar,tând comutare opto-mecanic,, datorat, fie lig,rii prin ioni metalici, fie prin al,i liganzi perfect croitori,i. In acel,i timp, atât procese de transfer de energie, cât ,i de electroni, pot fi perfect comutate, l,sând loc pentru imbun,t,,irea raportului "on/off" în arhitecturi viitoare. [source]

Prediction of polymorphic N -acetylation of new drug candidates by correlation with human NAT1 and NAT2

Cultured mammary epithelial monolayers (BME-UV) express functional organic anion and cation transporters

Inhibitory Effects of Silibinin on Cytochrome P-450 Enzymes in Human Liver Microsomes

BASIC AND CLINICAL PHARMACOLOGY & TOXICOLOGY, Issue 6 2000
Svane Beckmann-Knopp
Silibinin, the main constituent of silymarin, a flavonoid drug from silybum marianum used in liver disease, was tested for inhibition of human cytochrome P-450 enzymes. Metabolic activities were determined in liver microsomes from two donors using selective substrates. With each substrate, incubations were carried out with and without silibinin (concentrations 3.7,300 ,M) at 37° in 0.1 M KH2PO4 buffer containing up to 3% DMSO. Metabolite concentrations were determined by HPLC or direct spectroscopy. First, silibinin IC50 values were determined for each substrate at respective KM concentrations. Silibinin had little effect (IC50>200 ,M) on the metabolism of erythromycin (CYP3A4), chlorzoxazone (CYP2E1), S(+)-mephenytoin (CYP2C19), caffeine (CYP1A2) or coumarin (CYP2A6). A moderate effect was observed for high affinity dextromethorphan metabolism (CYP2D6) in one of the microsomes samples tested only (IC50=173 ,M). Clear inhibition was found for denitronifedipine oxidation (CYP3A4; IC50=29 ,M and 46 ,M) and S(,)-warfarin 7-hydroxylation (CYP2C9; IC50=43 ,M and 45 ,M). When additional substrate concentrations were tested to assess enzyme kinetics, silibinin was a potent competitive inhibitor of dextromethorphan metabolism at the low affinity site, which is not CYP2D6 (Ki,c=2.3 ,M and 2.4 ,M). Inhibition was competitive for S(,)-warfarin 7-hydroxylation (Ki,c=18 ,M and 19 ,M) and mainly non-competitive for denitronifedipine oxidation (Ki,n=9 ,M and 12 ,M). With therapeutic silibinin peak plasma concentrations of 0.6 ,M and biliary concentrations up to 200 ,M, metabolic interactions with xenobiotics metabolised by CYP3A4 or CYP2C9 cannot be excluded. [source]

Pharmacokinetic assessment of a five-probe cocktail for CYPs 1A2, 2C9, 2C19, 2D6 and 3A

BRITISH JOURNAL OF CLINICAL PHARMACOLOGY, Issue 6 2009
Sandrine Turpault
WHAT IS ALREADY KNOWN ABOUT THIS SUBJECT , Numerous cocktails using concurrent administration of several cytochrome P450 (CYP) isoform-selective probe drugs have been reported to investigate drug,drug interactions in vivo. , This approach has several advantages: characterize the inhibitory or induction potential of compounds in development toward the CYP enzymes identified in vitro in an in vivo situation, assess several enzymes in the same trial, and have complete in vivo information about potential CYP-based drug interactions. WHAT THIS STUDY ADDS , This study describes a new cocktail containing five probe drugs that has never been published. , This cocktail can be used to test the effects of a new chemical entity on multiple CYP isoforms in a single clinical study: CYP1A2 (caffeine), CYP2C9 (warfarin), CYP2C19 (omeprazole), CYP2D6 (metoprolol), and CYP3A (midazolam) and was designed to overcome potential liabilities of other reported cocktails. AIMS To assess the pharmacokinetics (PK) of selective substrates of CYP1A2 (caffeine), CYP2C9 (S-warfarin), CYP2C19 (omeprazole), CYP2D6 (metoprolol) and CYP3A (midazolam) when administered orally and concurrently as a cocktail relative to the drugs administered alone. METHODS This was an open-label, single-dose, randomized, six-treatment six-period six-sequence William's design study with a wash-out of 7 or 14 days. Thirty healthy male subjects received 100 mg caffeine, 100 mg metoprolol, 0.03 mg kg,1 midazolam, 20 mg omeprazole and 10 mg warfarin individually and in combination (cocktail). Poor metabolizers of CYP2C9, 2C19 and 2D6 were excluded. Plasma samples were obtained up to 48 h for caffeine, metoprolol and omeprazole, 12 h for midazolam, 312 h for warfarin and the cocktail. Three different validated liquid chromatography tandem mass spectrometry methods were used. Noncompartmental PK parameters were calculated. Log-transformed Cmax, AUClast and AUC for each analyte were analysed with a linear mixed effects model with fixed term for treatment, sequence and period, and random term for subject within sequence. Point estimates (90% CI) for treatment ratios (individual/cocktail) were computed for each analyte Cmax, AUClast and AUC. RESULTS There was no PK interaction between the probe drugs when administered in combination as a cocktail, relative to the probes administered alone, as the 90% CI of the PK parameters was within the prespecified bioequivalence limits of 0.80, 1.25. CONCLUSION The lack of interaction between probes indicates that this cocktail could be used to evaluate the potential for multiple drug,drug interactions in vivo. [source]

Relationship of glutathione S-transferase genotypes with side-effects of pulsed cyclophosphamide therapy in patients with systemic lupus erythematosus

BRITISH JOURNAL OF CLINICAL PHARMACOLOGY, Issue 4 2006
Shilong Zhong
Aims Cyclophosphamide (CTX) is an established treatment of severe systemic lupus erythematosus (SLE). Cytotoxic CTX metabolites are mainly detoxified by multiple glutathione S-transferases (GSTs). However, data are lacking on the relationship between the short-term side-effects of CTX therapy and GST genotypes. In the present study, the effects of common GSTM1, GSTT1, and GSTP1 genetic mutations on the severity of myelosuppression, gastrointestinal (GI) toxicity, and infection incidences induced by pulsed CTX therapy were evaluated in patients SLE. Methods DNA was extracted from peripheral leucocytes in patients with confirmed SLE diagnosis (n = 102). GSTM1 and GSTT1 null mutations were analyzed by a polymerase chain reaction (PCR)-multiplex procedure, whereas the GSTP1 codon 105 polymorphism (Ile,Val) was analyzed by a PCR-restriction fragment length polymorphism (RFLP) assay. Results Our study demonstrated that SLE patients carrying the genotypes with GSTP1 codon 105 mutation [GSTP1*-105I/V (heterozygote) and GSTP1*-105 V/V (homozygote)] had an increased risk of myelotoxicity when treated with pulsed high-dose CTX therapy (Odds ratio (OR) 5.00, 95% confidence interval (CI) 1.96, 12.76); especially in patients younger than 30 years (OR 7.50, 95% CI 2.14, 26.24), or in patients treated with a total CTX dose greater than 1.0 g (OR 12.88, 95% CI 3.16, 52.57). Similarly, patients with these genotypes (GSTP1*I/V and GSTP1*V/V) also had an increased risk of GI toxicity when treated with an initial pulsed high-dose CTX regimen (OR 3.33, 95% CI 1.03, 10.79). However, GSTM1 and GSTT1 null mutations did not significantly alter the risks of these short-term side-effects of pulsed high-dose CTX therapy in SLE patients. Conclusions The GSTP1 codon 105 polymorphism, but not GSTM1 or GSTT1 null mutations, significantly increased the risks of short-term side-effects of pulsed high-dose CTX therapy in SLE patients. Because of the lack of selective substrates for a GST enzyme phenotyping study, timely detection of this mutation on codon 105 may assist in optimizing pulsed high-dose CTX therapy in SLE patients. [source]