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P Su

Publications and source records attributed to P Su.

31 records · Page 2Linked to original sources
PubMed1994

Involvement of CYP2D6, CYP3A4, and other cytochrome P-450 isozymes in N-dealkylation reactions.

Metabolic N-dealkylation is a commonly observed biotransformation with tertiary and secondary amine drugs and related N-alkylated amides, but surprisingly little is known about the cytochrome P-450 isozymes involved in these dealkylation reactions. In this review, evidence is provided that supports the involvement of various P-450 isozymes, but especially CYP3A4 and other isozymes of the CYP3A subfamily. Although CYP2D6 is generally not considered to be capable of catalyzing the N-dealkylation of basic drugs, some examples of the involvement of this important isozyme in N-dealkylation reactions are identified. Procedures used to identify individual P-450 isozymes involved in N-dealkylation reactions are discussed.

Amitriptyline↗
PubMed1993

Metabolism of imipramine in vitro by isozyme CYP2D6 expressed in a human cell line, and observations on metabolite stability.

A metabolism study of imipramine (IMI) has been conducted in vitro with commercially available human CYP2D6 isozyme expressed in a human AHH-1 TK +/- cell line. This enzyme system catalyzed the anticipated ring-oxidative biotransformation of IMI to 2-hydroxyimipramine (2-OH-IMI). In addition, however, the human CYP2D6 isozyme preparation was found to be unequivocally involved in the N-dealkylation of IMI to desmethylimipramine (DMI). 2-Hydroxydesipramine was also identified as a trace metabolite of IMI, but no 10-hydroxyimipramine was detected. The 2-OH-IMI metabolite was unstable and disappeared from metabolic solutions on standing. A procedure involving the O-acetylation of 2-OH-IMI was developed to minimize this decomposition. When an authentic sample of 10-OH-IMI was subjected to the same acetylation procedure, it was partially dehydrated to 10,11-dehydroimipramine, but also underwent unexpected degradations to two other products in which the dimethylaminopropyl side-chain was deaminated. Plausible structures for these two decomposition products are suggested from their gas chromatographic-mass spectrometric behaviour.

Acetylation↗
PubMed1993

Analysis of imipramine and three metabolites produced by isozyme CYP2D6 expressed in a human cell line.

1. A commercially-available human cytochrome P450 isozyme (CYP2D6) preparation was used in imipramine metabolism studies. This isozyme catalysed both aromatic C-oxidation and N-demethylation. 2-Hydroxyimipramine was the major metabolite; desipramine was isolated in a significant amount and 2-hydroxydesipramine was a trace metabolite. 2. To prevent decomposition of metabolites during the analytical procedure, the metabolism mixture was derivatized with acetic anhydride prior to extraction, and the derivatized metabolites were separated and quantified by g.l.c. with N/P detection. The analytical procedure had excellent sensitivity and was capable of routinely quantifying imipramine and its metabolites down to the 0.36 nmol level. 3. In excess of 90% of drug and metabolites was consistently recovered when metabolism was conducted over a 5-60-min duration. 4. The formation of the secondary metabolite, 2-hydroxydesimipramine, from imipramine proceeds by two pathways, via desipramine and via 2-hydroxyimipramine; the former is the preferred pathway. 5. CYP2D6 catalyses C-hydroxylation of imipramine to 2-hydroxyimipramine more efficiently than its N-demethylation to desipramine. Also, the C-hydroxylation of imipramine to 2-hydroxyimipramine proceeds more efficiently than the conversion of desipramine to 2-hydroxydesipramine.

Catalysis↗
PubMed1990

A cDNA encoding a rat mitochondrial cytochrome P450 catalyzing both the 26-hydroxylation of cholesterol and 25-hydroxylation of vitamin D3: gonadotropic regulation of the cognate mRNA in ovaries.

A cDNA expression library prepared from rat liver RNA was screened with a polyclonal antibody specific for mitochondrial vitamin D3 25-hydroxylase and a cDNA for rabbit liver mitochondrial cytochrome P450c26 (CYP 26), yielding cDNA clones with identical sequences. The deduced amino acid sequence derived from a 1.9-kb full-length cDNA was 73% identical to that of rabbit cytochrome P450c26. A monoclonal antibody was used to demonstrate that the product of the 1.9-kb cDNA clone was targeted to the mitochondrial compartment when expressed in COS cells. Mitochondrial membranes containing the expressed protein showed both vitamin D3 25-hydroxylase and cholesterol 26-hydroxylase activities when reconstituted with ferredoxin reductase and ferredoxin, demonstrating that the same P450, designated as P450c26/25, can catalyze both reactions. Northern blot analysis revealed that the P450c26/25 cDNA hybridizes with a 2.4-kb RNA from rat liver and unstimulated ovaries. Treatment of rats with pregnant mare's serum gonadotropin resulted in a fivefold increase in the 2.4-kb mRNA as well as the appearance of a 2.1-kb mRNA species in the ovaries. Our findings document the presence of a regulated bifunctional mitochondrial cytochrome P450 capable of catalyzing the 25-hydroxylation of vitamin D3 and the 26-hydroxylation of cholesterol.

Amino Acid Sequence↗
PubMed1990

Preparative slab electrofocusing of methane monooxygenase from a type I methanotroph Methylomonas GYJ3.

The isolation and purification of methane monooxygenase from a type I methanotroph Methylomonas GYJ3 to near homogeneity is reported. The isoelectric focusing in flat-bed granulated gels has resolved the methane monooxygenase into three protein components. The specific activity of the enzyme is 198.4 n mol per min per mg protein, degree of purification 4.13-fold. Recovery of the focused proteins is high and elution simple. Several purification steps may be omitted from the previously published scheme by other techniques.

Isoelectric Focusing↗
PubMed1987

[Preliminary application of carbohydrate antigen CA-50 radioimmunoinhibition test in diagnosing and monitoring cancers].

Serum CA-50 antigen in the normal individuals and cancer patients was assayed by radioimmunoassay. CA-50, one type of carbohydrate antigens, consisting mainly of sialylated glycolipid (a ganglioside) and sialylated glycoprotein, can be isolated from the primary or metastatic tumors. Using the latest CA-50 antigen radioimmunoinhibition test kits prepared by Steana Diagnostics AB Company, Sweden, the serum CA-50 content was measured in 62 normal individuals, 9 primary hepatocellular carcinomas, 9 liver benign tumors, 23 ovarian or uterine carcinomas, 22 other malignancies and 21 benign tumors. The normal value was 6.05 +/- 4.93 U/ml. The test values in cancer patients were significantly higher than those in normals or benign tumor patients. Taking the normal mean value plus two times of standard deviation as positive, the positive rate was 77.7% for liver cancer, 65.2% for ovary and uterus carcinomas, 50% for the other cancers. There was only one positive in 62 normals with a false positive rate of 1.6%. Furthermore, 19 patients, being in the remission period or cured by effective treatment, gave normal CA-50 value. It is shown that this method can be used in cancer diagnosis, differential diagnosis and the monitor, prediction of prognosis.

Antigens, Neoplasm↗
PubMed1987

Effects of zinc deficiency on thyroid function.

Interactive combinations of altered zinc and thyroid states were studied in rats to assess pathophysiologic effects. Clinical signs of zinc deficiency or thyroid alteration were limited to effects on growth rate. Changes in organ and glandular weights and serum thyrotropin levels reflected changes in serum thyroid hormone concentrations. Significantly (probability less than .001), zinc-deficient rats had enhanced hepatic thyroxine-5'-monodeiodinase activity. In addition, the zinc-deficient state was found to be protective against thiouracil-induced suppression of the microsomal-monooxygenase and thyroxine-5'-monodeiodinase enzyme complex. This protective effect was evident by greater thyroxine-5'-monodeiodinase and reduced nicotinamide-adenine dinucleotide phosphate cytochrome c reductase activities, as well as cytochrome P-450 content, in zinc-deficient/thiouracil-treated animals. Thus, the enzyme complex had increased triiodothyronine-generating capacity in conditions of zinc deficiency, which may be important because of the greater biological reactivity of triiodothyronine. Primary zinc deficiency conditions of the magnitude seen in this study and in this-age rat did not appear to alter serum thyroid hormone levels or organ/glandular function. However, concurrent zinc deficiency and altered thyroid status did change thyroid hormone response and disposition, which may be important to populations at risk because of thyroid dysfunctional states.

Animals↗
PubMedDate not supplied

Metabolism of methoxyphenamine in vitro by a CYP2D6 microsomal preparation.

Metabolism of methoxyphenamine (MP) was conducted in vitro using commercially available microsomes prepared from human AHH-1 TK+/-cells in which CYP2D6 had been expressed. This study has confirmed the involvement of CYP2D6 in the metabolism of MP to O-desmethylmethoxyphenamine (ODMP) and 5-hydroxymethoxyphenamine (5HMP), but not to N-desmethylmethoxyphenamine. It has also revealed that CYP2D6 catalyzes the formation of another, hitherto unknown, ring-hydroxylated metabolite of MP, isomeric with 5HMP. The analytical procedure used to identify and quantify MP metabolites involved an acetylation procedure that had distinct advantages. MP and its basic and amphoteric metabolites were all converted to neutral products that were efficiently extracted into organic solvent. The acetylated products also had good chromatographic properties and provided mass spectra that were readily interpretable. MP metabolism studies were also conducted with CYP2D6 microsomes in the presence of quinidine and quinine. The former was the more potent inhibitor of CYP2D6-catalyzed oxidations of MP. Its inclusion resulted in complete inhibition of metabolism of MP to ODMP, 5HMP, and its novel isomer. This study shows that the in vitro use of human cytochrome P450 isozyme preparations in drug metabolism studies can aid in the identification of possible in vivo metabolites of these drugs in humans and can provide information on putative drug-drug interactions.

Acetylation↗