• Title/Summary/Keyword: Cytochrome P-450 3A4

검색결과 278건 처리시간 0.029초

In vitro Ccovalent Binding of SC-42867, PGE2 Antagonist, to Rat Liver Microsomal Proteins

  • Lee, Kyung-Tae
    • Archives of Pharmacal Research
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    • 제18권6호
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    • pp.381-384
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    • 1995
  • Covalent binding of the reactive metabolites of SC_42867 to microsomal proteins has been examined. In the absence of inhibitor of cytochrome oxydase (.alpha.-naphtyl-isothiocyanate) or a radical scavenger (3-terthiobuty-4-hydroxyanisol), up to 4.0% of total redioactivity used in the assay could irreversibly bind to proteins. In the presence of an inhibitor, the highest percentage of covalent binding observed is 0.7% a significant decrease of the metabolism of SC42876 was observed. These results suggest in a cytochrome P-450 dependent generation of SC_42867 metabolites significantly take part in the covalent binding process.

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쇠무릎(Achyranthes japonica Nakai)으로부터 Ecdysteroid 생합성에 관련된 유전자의 분리 (Isolation of Genes Involved in Ecdysteroids Biosynthesis from Achyranthes japonica Nakai)

  • 부경환;김소미;진성범;채현병;이도승;김대운;조문제;류기중
    • Applied Biological Chemistry
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    • 제44권3호
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    • pp.153-161
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    • 2001
  • Ecdysteroid 생합성에 관련된 식물 유전자를 분리할 목적으로 ecdysteroid 생성능이 확인된 쇠무릎(Achyranthes japonica Nakai)으로부터 RNA를 분리하고, ecdysteroid 생합성에 관여한다고 알려진 cytochrome P45O family 유전자 4개와 곤충의 ecdysone 20-hydroxylase 유전자의 다중정렬 분석결과를 토대로 상동성이 높은 부위의 염기서열에 상응하는 degenerate primer를 사용하여 RT-PCR을 행하고, 고유한 염기서열을 가진 partical cDNA clone 14개를 선발했다. 기존에 ecdysteroid 생합성에 관여한다고 알려진 cytochrome P450 family 유전자들과의 염기 및 아미노산 서열의 상동성을 분석한 결과, 선발된 cDNA 중 6개가 cytochrome P45O fumily 유전자들과 상동성이 있는 것으로 나타났다. 이 중에서 4개의 clone은 곤충의 ecdysone 20-hydroxylase 유전자와도 상동성이 높아 ecdysteroid 생합성에 관련된 유전자로 추정되었다.

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Modulation of carcinogen-activating enzymes by synthetic trans-stilbene analogs

  • Lee, Sang-Kwang;Kim, Sang-Hee;Kim, Mie-Young;Chun, Young-Jin
    • 대한약학회:학술대회논문집
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    • 대한약학회 2002년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2
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    • pp.312.1-312.1
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    • 2002
  • Previous studies have demonstrated that 2,3',4,5' -tetramethoxystilbene (TMS) and 3,3',4',5,5'-pentamethoxystilbene (PMS) showed selective inhibition of human cytochrome P450 1 Bl and 1A1 in vitro., respectively, In the present study, the effects of synthetic stilbene analogs on the expression of cytochrome P450 1Al or lBl were investigated in human tumor cell lines such as HepG2, MCF-7 and MCF-l0A, TCDD caused a dramatic increase in the amount of P450 1A1 or 1B1 proteins and mRNA levels. (omitted)

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인체 간 microsome에서 pentoxifylline 대사체 M-1의 시험관내 대사 (In vitro Metabolism of Pentoxifylline Metabolite M-l in Human Liver Microsomes)

  • 신혜순
    • 약학회지
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    • 제43권6호
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    • pp.834-842
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    • 1999
  • The metabolism and pharmacokinetics of M-l, which is metabolite of pentoxifylline, have been studied in human liver microsomes. Biphasic kinetics was observed from the Eadie-Hofstee plot for the formation of both metabolites of M-l. For the kinetics of pentoxifylline, mean values of $V_{max1}{\;}and{\;}V_{max2}$ were 1,648 and 5,622 nmol/min/mg protein, and the estimated values of $K_{ml}{\;}and{\;}K_{m2}$ were 0.180 and 4.829 mM, respectively. For M-3, mean values of $V_{max1}{\;}and{\;}V_{max2}$ were 0.062 and 0.491 nmol/min/mg protein, and estimated values of $K_{ml}{\;}and{\;}K_{m2}$ were 0.025 and 1.216 mM. The formations of pentoxifylline and M-3 from M-1 were indentified by using several selective inhibitors of cytochrome P450 isoformes at 0.05-5 mM concentration of M-1 in human liver microsomes. For the analysis of low (0.05 mM) concentration of M-1, where the affinity was expected as low, indicated that CYPlA2 and CYP3A4 were major P450 isoforms responsible for pentoxifylline and M-3 formation. CYP3A4 and CYP2A6 appeared to be P450 isoforms responsible for M-3 formation at high (5 mM) concentration of M-1.

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In Vitro Enhancement of Microsomal Cytochrome P450-Dependent Monooxygenases by Organic Solvents in Rat Liver

  • Lee, Dong-Wook;Lim, Heung-Bin;Moon, Ja-Young;Park, Ki-Hyun
    • BMB Reports
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    • 제31권4호
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    • pp.391-398
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    • 1998
  • In vitro effects of acetone, methanol, and dimethylsulfoxide (DMSO) on liver microsomal cytochrome P450 (P450) content, and P450-dependent arylhydrocarbon hydroxylase (AHH) and 7-ethoxycoumarin O-deethylase (ECOD) activities were studied in rats. Acetone at 1% (v/v) enhanced the content ofP450, assayed spectrally in 3-methylcholanethrene (MC)- and ${\beta}-naphthoflavone$ (BNF)-inducible microsomes by 18 and 7%, respectively. Methanol, up to 5% (v/v) applied, also showed enhancement effects on P450 content in liver microsomes from rats treated with phenobarbital (PB), MC, and BNF, as well as uninduced microsomes with similar but low strength. DMSO, however, did not show such enhancing effects at the ranges of the concentrations applied. AHH and ECOD activities in MC-inducible microsomes were also enhanced by acetone at 1%, which was in proportion to the increase in P450 content by the same concentration. However, the P450 content, and AHH and ECOD activities, were decreased by increasing the concentration of acetone. Methanol at the same concentration with acetone also enhanced ECOD activity but not AHH activity in MCinducible microsomes. The enhancing effect of acetone on the enzymes was negligible when the microsomes were pretreated with a specific monoclonal antibody of MC-inducible isozyme. The difference in the effects of these solvents on P450 system might be due to their different properties that cause the P450 active site to be exposed in milieu.

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N-(2-Ethylhexyl)-8, 9, 10-trinorborn-5-ene-2, 3-dicorhoxymide가 Rat의 Cytochrome P-450 및 생화학적 혈액상에 미치는 독성작용 (The Toxicity of N-(2-Ethylhexyl)-8, 9, 10-trinorborn-5-ene-2, 3-dicorhoxymide on Cytochrome P-450 and Biochemical Parameter of Serum in Rats)

  • 홍사욱;장준식
    • Environmental Analysis Health and Toxicology
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    • 제7권1_2호
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    • pp.1-16
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    • 1992
  • Biologically, MGK-264 (N-octyl bicycloheptene dicarboximide) acts as a synergists for insecticides mainly pyrethrins and pyrethroids. It's used extensively in combination with pyrethrin and piperonyl butoxide and also with personal insect repellent and cockroach repellents. But the toxic effect of MGK-264 in mamalians was a relatively little known therefore in this studies it was initiated to examine the toxic effect of MGK -264 in rats. For 5 weeks it administrated daily in each 250 mg and 500 mg of MGK-264 per kg of body weight in rats. 1) The body weight gain and the LYMPH (%) value in blood were observed a slight tendency to reduce in accordance with amount of dose and number treatment time. 2) The content of cytochrome P-450 and activity of NADPH-cytochrome c reductase were decreased in liver and those were observed some tendency in the kidney as liver but not significant. 3) The liver cholinesterase activity in the both 250 mg/kg and 500 mg/kg per kg of body weight with treated groups and the liver aniline hydroxylase in 500 mg/kg treated group were gradually decreased from 4 weeks after treated groups. In consequence it would sugested that the toxic effect of MGK-264 was low but in could offer hazard effect in liver and nervers system of rats if it was administrated move dose of MGK-264 and agumented in number of treated time.

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Inhibition of Carcinogen-Activating Cytochrome P450 Enzymes by Xenobiotic Chemicals in Relation to Antimutagenicity and Anticarcinogenicity

  • Shimada, Tsutomu
    • Toxicological Research
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    • 제33권2호
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    • pp.79-96
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    • 2017
  • A variety of xenobiotic chemicals, such as polycyclic aromatic hydrocarbons (PAHs), aryl- and heterocyclic amines and tobacco related nitrosamines, are ubiquitous environmental carcinogens and are required to be activated to chemically reactive metabolites by xenobiotic-metabolizing enzymes, including cytochrome P450 (P450 or CYP), in order to initiate cell transformation. Of various human P450 enzymes determined to date, CYP1A1, 1A2, 1B1, 2A13, 2A6, 2E1, and 3A4 are reported to play critical roles in the bioactivation of these carcinogenic chemicals. In vivo studies have shown that disruption of Cyp1b1 and Cyp2a5 genes in mice resulted in suppression of tumor formation caused by 7,12-dimethylbenz[a]anthracene and 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone, respectively. In addition, specific inhibitors for CYP1 and 2A enzymes are able to suppress tumor formation caused by several carcinogens in experimental animals in vivo, when these inhibitors are applied before or just after the administration of carcinogens. In this review, we describe recent progress, including our own studies done during past decade, on the nature of inhibitors of human CYP1 and CYP2A enzymes that have been shown to activate carcinogenic PAHs and tobacco-related nitrosamines, respectively, in humans. The inhibitors considered here include a variety of carcinogenic and/or non-carcinogenic PAHs and acethylenic PAHs, many flavonoid derivatives, derivatives of naphthalene, phenanthrene, biphenyl, and pyrene and chemopreventive organoselenium compounds, such as benzyl selenocyanate and benzyl selenocyanate; o-XSC, 1,2-, 1,3-, and 1,4-phenylenebis(methylene)selenocyanate.

체외배양 생쥐정소세포에서 합성에스트로겐이 P450 등위효소의 발현에 미치는 영향 (Effects of Xenoestrogens on Gene Expression of Cytochrome P450 Genes in in vitro Cultured Mice Spermatogenic Cells)

  • 이호준;김묘경;고덕성;김길수;강희규;김동훈
    • Clinical and Experimental Reproductive Medicine
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    • 제28권2호
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    • pp.131-140
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    • 2001
  • Objective: To know the effects of xenoestrogen on spermatogenesis, we investigated the expression of cytochrome P450s enzymes (CYPscc, $CYP_{17{\alpha}}$, CYP19) and $3{\beta}$-HSD genes involved in steroidogenesis. Methods: Mouse testicular cells were prepared from 15-day-old ICR mice which had only pre-meiotic germ cells by enzyme digestion using collagenase and trypsin. Testicular cells were cultured in DMEM supplemented with FSH (0.1 IU/ml) and 10% FBS or medium with estrogen ($E_2$), bisphenol-A (BPA), octylphenol (OP; $10^{-9},\;10^{-7},\;10^{-6},\;10^{-5},\;10^{-4}M$, respectively) and aroclor 1254 (A1254) known as PCBs for 48 hours. The gene expression of cytochrome P450 enzymes were examined by semi-quantitive RT-PCR. The production of estrogen and testosterone was examined by RIA. Results: As results, expression of CYPscc mRNA was not significantly decreased, but $3{\beta}$-HSD and $CYP_{17{\alpha}}$. mRNA were significantly dose-dependent decreased. And production of testosterone and estrogen were not different except BPA and OP group ($10^{-5}M$). Conclusion: BPA, OP and A1254 might inhibit steroidogenesis by decreasing CYPscc, $3{\beta}$-HSD and $CYP_{17{\alpha}}$. mRNA expression in the mouse testis. These results suggest that BPA, OP and PCBs like as an endocrine disruptors inhibit the productions of steroidogenic enzymes and decrease the production of T and E by negative feedback mechanism. Therefore, these might disrupt steroidogenesis in Leydig cells of testis and would disturb testicular function and subsequently impair spermatogenesis.

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향신료의 약물대사효소 CYP3A4 저해효과 (Inhibitory Effect of a Drug Metabolizing Enzyme CYP3A4 on Spices)

  • 차배천
    • 생약학회지
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    • 제34권1호통권132호
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    • pp.86-90
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    • 2003
  • For the determination of inhibiting cytochrome P450(CYP)3A4 activity, an improvement HPLC method was established by using a new internal standard and solvent system. Moreover, CYP3A4 amount for a optimum reaction of enzyme was determined by a comparative study with a variety concentration of enzyme. Using a established method, inhibitory effect of CYP3A4 that is drug metabolizing enzyme Investigated on EtOAc extracts of 5-class spices. As a result of experiment, EtOAc extract of white pepper (Piper nigrum L.) showed strong inhibitory activity. On a continuous experiment, the fraction 2, 4 and 5 of while pepper extract showed remarkable inhibitory activity. Pipeline, a main constituent of pepper was not included in these fraction. It is suggested that major compounds for the inhibitory activity of white pepper may be other ingredient that is not piperine.

Purification and Characterization of a Cytochrome P-450 from Pravastatin-Producing Streptomyces sp. Y-110.

  • Park, Joo-Woong;Lee, Joo-Kyung;Kwon, Tae-Jong;Yi, Dong-Hee;Park, Yong-Il;Kang, Sang-Mo
    • Journal of Microbiology and Biotechnology
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    • 제11권6호
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    • pp.1011-1017
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    • 2001
  • Streptomyces sp. Y-110 cytochrome P-450, induced by the addition of compactin -Na into the culture medium, was purified from the cell extract to apparent homogeniety, mainly by DEAE-Sepharose, hydroxyapatite, and Mono Q column chromatyography. The sepcific activity of purified enzyme on its substrate, compactin-Na, was determined to be 15 nmol of pravastatin per mg protein. The molecular mass of this enzyme on SDS-PAGE was $37{\pm}0.5$ kDa, pI was 4.5, and its CO difference spectrum showed maximum absorption peaks at 452 and 550nm, respectively. The N-terminal amino acid sequence was determined to be Met>Thr>Cys>Thr>Pro>Val>Thr>Val>The>Gly>Ala>Ala>Gly>Gln>Ile>Gly>Tyr>Ala>Leu. Its apparent $K_m$ on compactin-Na was $1.294{\mu}M{\cdot}min^-1,\;and\;V_{max}\;was\;1.028{\mu}M{\cdot}min^-1$. The maximum substrate concentration ($K_s$) for reaction was $270 {\mu}M$and thus $1/[K_s]$ was $3.7{\mu}M$. These physicochemical characteristics and kinetic behavior of this enzyme were compared and shown to be different from those of Streptomyces cytochrome P-450 enzymes reported, suggesting that this enzyme may be an additional member of the Streptomyces cytochrome P-450 family.

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