Cytochromes P450 reconstituted with NADPH: P450 reductase mimic the activating and detoxicating metabolism of the anticancer drug ellipticine in microsomes
Ellipticine is a potent antineoplastic agent exhibiting multiple action mechanisms. Recently, we found that after cytochrome P450 (CYP)-mediated oxidation ellipticine forms covalent DNA adducts. Ellipticine oxidation by isolated CYP and its binding to DNA is the target of this study. High performanc...
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| Veröffentlicht in: | Neuro-endocrinology letters 2006-12, Vol.27 Suppl 2, p.18 |
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| container_title | Neuro-endocrinology letters |
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| creator | Kotrbová, Vera Aimová, Dagmar Brezinová, Anna Janouchová, Katerina Poljaková, Jitka Frei, Eva Stiborová, Marie |
| description | Ellipticine is a potent antineoplastic agent exhibiting multiple action mechanisms. Recently, we found that after cytochrome P450 (CYP)-mediated oxidation ellipticine forms covalent DNA adducts. Ellipticine oxidation by isolated CYP and its binding to DNA is the target of this study.
High performance liquid chromatography (HPLC) was employed for separation and characterization of ellipticine metabolites generated by CYPs. The (32)P-postlabeling technique was utilized to determine ellipticine-DNA adducts.
Purified CYP enzymes reconstituted with NADPH:CYP reductase oxidized ellipticine to up to five metabolites, 7-hydroxy-, 9-hydroxy-, 12-hydroxy-, 13-hydroxyellipticine and ellipticine N(2)-oxide. However, only CYP1A1 was capable to form all metabolites. Using the reconstituted enzymatic system, we demonstrated that the detoxication ellipticine metabolites, 7-hydroxyellipticine and 9-hydroxyellipticine, are mainly generated by CYP1A1 and 1A2, while those responsible for DNA binding, 13-hydroxy-, 12-hydroxyellipticine and ellipticine N(2)-oxide, by CYP3A1 and 2C3. Likewise, the most efficient CYPs forming DNA adducts from ellipticine were CYP3A1 and 2C3.
The results showed that the system of purified CYPs reconstituted with NADPH: CYP reductase proved for ellipticine oxidation provide a true reflection of the situation in the microsomal membrane. |
| format | Article |
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High performance liquid chromatography (HPLC) was employed for separation and characterization of ellipticine metabolites generated by CYPs. The (32)P-postlabeling technique was utilized to determine ellipticine-DNA adducts.
Purified CYP enzymes reconstituted with NADPH:CYP reductase oxidized ellipticine to up to five metabolites, 7-hydroxy-, 9-hydroxy-, 12-hydroxy-, 13-hydroxyellipticine and ellipticine N(2)-oxide. However, only CYP1A1 was capable to form all metabolites. Using the reconstituted enzymatic system, we demonstrated that the detoxication ellipticine metabolites, 7-hydroxyellipticine and 9-hydroxyellipticine, are mainly generated by CYP1A1 and 1A2, while those responsible for DNA binding, 13-hydroxy-, 12-hydroxyellipticine and ellipticine N(2)-oxide, by CYP3A1 and 2C3. Likewise, the most efficient CYPs forming DNA adducts from ellipticine were CYP3A1 and 2C3.
The results showed that the system of purified CYPs reconstituted with NADPH: CYP reductase proved for ellipticine oxidation provide a true reflection of the situation in the microsomal membrane.</description><identifier>ISSN: 0172-780X</identifier><identifier>PMID: 17159771</identifier><language>eng</language><publisher>Sweden</publisher><subject>Animals ; Antineoplastic Agents - pharmacokinetics ; Cytochrome P-450 Enzyme System - chemistry ; Cytochrome P-450 Enzyme System - isolation & purification ; Cytochrome P-450 Enzyme System - metabolism ; DNA Adducts - metabolism ; Ellipticines - pharmacokinetics ; Metabolic Detoxication, Phase I ; Microsomes, Liver - chemistry ; Microsomes, Liver - metabolism ; Models, Biological ; NADP - chemistry ; NADP - metabolism ; NADPH-Ferrihemoprotein Reductase - isolation & purification ; NADPH-Ferrihemoprotein Reductase - metabolism ; Rabbits ; Rats</subject><ispartof>Neuro-endocrinology letters, 2006-12, Vol.27 Suppl 2, p.18</ispartof><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,777,781</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/17159771$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Kotrbová, Vera</creatorcontrib><creatorcontrib>Aimová, Dagmar</creatorcontrib><creatorcontrib>Brezinová, Anna</creatorcontrib><creatorcontrib>Janouchová, Katerina</creatorcontrib><creatorcontrib>Poljaková, Jitka</creatorcontrib><creatorcontrib>Frei, Eva</creatorcontrib><creatorcontrib>Stiborová, Marie</creatorcontrib><title>Cytochromes P450 reconstituted with NADPH: P450 reductase mimic the activating and detoxicating metabolism of the anticancer drug ellipticine in microsomes</title><title>Neuro-endocrinology letters</title><addtitle>Neuro Endocrinol Lett</addtitle><description>Ellipticine is a potent antineoplastic agent exhibiting multiple action mechanisms. Recently, we found that after cytochrome P450 (CYP)-mediated oxidation ellipticine forms covalent DNA adducts. Ellipticine oxidation by isolated CYP and its binding to DNA is the target of this study.
High performance liquid chromatography (HPLC) was employed for separation and characterization of ellipticine metabolites generated by CYPs. The (32)P-postlabeling technique was utilized to determine ellipticine-DNA adducts.
Purified CYP enzymes reconstituted with NADPH:CYP reductase oxidized ellipticine to up to five metabolites, 7-hydroxy-, 9-hydroxy-, 12-hydroxy-, 13-hydroxyellipticine and ellipticine N(2)-oxide. However, only CYP1A1 was capable to form all metabolites. Using the reconstituted enzymatic system, we demonstrated that the detoxication ellipticine metabolites, 7-hydroxyellipticine and 9-hydroxyellipticine, are mainly generated by CYP1A1 and 1A2, while those responsible for DNA binding, 13-hydroxy-, 12-hydroxyellipticine and ellipticine N(2)-oxide, by CYP3A1 and 2C3. Likewise, the most efficient CYPs forming DNA adducts from ellipticine were CYP3A1 and 2C3.
The results showed that the system of purified CYPs reconstituted with NADPH: CYP reductase proved for ellipticine oxidation provide a true reflection of the situation in the microsomal membrane.</description><subject>Animals</subject><subject>Antineoplastic Agents - pharmacokinetics</subject><subject>Cytochrome P-450 Enzyme System - chemistry</subject><subject>Cytochrome P-450 Enzyme System - isolation & purification</subject><subject>Cytochrome P-450 Enzyme System - metabolism</subject><subject>DNA Adducts - metabolism</subject><subject>Ellipticines - pharmacokinetics</subject><subject>Metabolic Detoxication, Phase I</subject><subject>Microsomes, Liver - chemistry</subject><subject>Microsomes, Liver - metabolism</subject><subject>Models, Biological</subject><subject>NADP - chemistry</subject><subject>NADP - metabolism</subject><subject>NADPH-Ferrihemoprotein Reductase - isolation & purification</subject><subject>NADPH-Ferrihemoprotein Reductase - metabolism</subject><subject>Rabbits</subject><subject>Rats</subject><issn>0172-780X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2006</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNo1kEtOwzAYhLMA0VK4AvKKXSQ7ieOYXVUeRULQBUjsIj_-tkaxHWwH6Fm4LEFtVyPNfBqN5iSbYsKKnDX4fZKdx_iBccFpUZ5lE8II5YyRafa72CWvtsFbiGhVUYwCKO9iMmlIoNG3SVv0PL9dLW-OsR5UEhGQNdYolLaAhErmSyTjNkg4jTQk_2PU3rCQhPSdiRb59Z52aQydgoB0GDYIus70o2UcIOPGWhV8_N9zkZ2uRRfh8qCz7O3-7nWxzJ9eHh4X86e8JyVPudKqEZiWRIIqa1E2WjBWEcVIU9dCFkwoKglQygkmFdFKVLTWAjiTXErg5Sy73vf2wX8OEFNrTVTjLOHAD7EtSEFwxfEIXh3AQVrQbR-MFWHXHu8s_wCC_3NO</recordid><startdate>20061201</startdate><enddate>20061201</enddate><creator>Kotrbová, Vera</creator><creator>Aimová, Dagmar</creator><creator>Brezinová, Anna</creator><creator>Janouchová, Katerina</creator><creator>Poljaková, Jitka</creator><creator>Frei, Eva</creator><creator>Stiborová, Marie</creator><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>7QO</scope><scope>7TK</scope><scope>8FD</scope><scope>FR3</scope><scope>P64</scope></search><sort><creationdate>20061201</creationdate><title>Cytochromes P450 reconstituted with NADPH: P450 reductase mimic the activating and detoxicating metabolism of the anticancer drug ellipticine in microsomes</title><author>Kotrbová, Vera ; Aimová, Dagmar ; Brezinová, Anna ; Janouchová, Katerina ; Poljaková, Jitka ; Frei, Eva ; Stiborová, Marie</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-p139t-cdc8a0531bec36a38da7741c71866ab27ac5b1e55910141dca456dae97b9bbe93</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2006</creationdate><topic>Animals</topic><topic>Antineoplastic Agents - pharmacokinetics</topic><topic>Cytochrome P-450 Enzyme System - chemistry</topic><topic>Cytochrome P-450 Enzyme System - isolation & purification</topic><topic>Cytochrome P-450 Enzyme System - metabolism</topic><topic>DNA Adducts - metabolism</topic><topic>Ellipticines - pharmacokinetics</topic><topic>Metabolic Detoxication, Phase I</topic><topic>Microsomes, Liver - chemistry</topic><topic>Microsomes, Liver - metabolism</topic><topic>Models, Biological</topic><topic>NADP - chemistry</topic><topic>NADP - metabolism</topic><topic>NADPH-Ferrihemoprotein Reductase - isolation & purification</topic><topic>NADPH-Ferrihemoprotein Reductase - metabolism</topic><topic>Rabbits</topic><topic>Rats</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Kotrbová, Vera</creatorcontrib><creatorcontrib>Aimová, Dagmar</creatorcontrib><creatorcontrib>Brezinová, Anna</creatorcontrib><creatorcontrib>Janouchová, Katerina</creatorcontrib><creatorcontrib>Poljaková, Jitka</creatorcontrib><creatorcontrib>Frei, Eva</creatorcontrib><creatorcontrib>Stiborová, Marie</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>Biotechnology Research Abstracts</collection><collection>Neurosciences Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><jtitle>Neuro-endocrinology letters</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Kotrbová, Vera</au><au>Aimová, Dagmar</au><au>Brezinová, Anna</au><au>Janouchová, Katerina</au><au>Poljaková, Jitka</au><au>Frei, Eva</au><au>Stiborová, Marie</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Cytochromes P450 reconstituted with NADPH: P450 reductase mimic the activating and detoxicating metabolism of the anticancer drug ellipticine in microsomes</atitle><jtitle>Neuro-endocrinology letters</jtitle><addtitle>Neuro Endocrinol Lett</addtitle><date>2006-12-01</date><risdate>2006</risdate><volume>27 Suppl 2</volume><spage>18</spage><pages>18-</pages><issn>0172-780X</issn><abstract>Ellipticine is a potent antineoplastic agent exhibiting multiple action mechanisms. Recently, we found that after cytochrome P450 (CYP)-mediated oxidation ellipticine forms covalent DNA adducts. Ellipticine oxidation by isolated CYP and its binding to DNA is the target of this study.
High performance liquid chromatography (HPLC) was employed for separation and characterization of ellipticine metabolites generated by CYPs. The (32)P-postlabeling technique was utilized to determine ellipticine-DNA adducts.
Purified CYP enzymes reconstituted with NADPH:CYP reductase oxidized ellipticine to up to five metabolites, 7-hydroxy-, 9-hydroxy-, 12-hydroxy-, 13-hydroxyellipticine and ellipticine N(2)-oxide. However, only CYP1A1 was capable to form all metabolites. Using the reconstituted enzymatic system, we demonstrated that the detoxication ellipticine metabolites, 7-hydroxyellipticine and 9-hydroxyellipticine, are mainly generated by CYP1A1 and 1A2, while those responsible for DNA binding, 13-hydroxy-, 12-hydroxyellipticine and ellipticine N(2)-oxide, by CYP3A1 and 2C3. Likewise, the most efficient CYPs forming DNA adducts from ellipticine were CYP3A1 and 2C3.
The results showed that the system of purified CYPs reconstituted with NADPH: CYP reductase proved for ellipticine oxidation provide a true reflection of the situation in the microsomal membrane.</abstract><cop>Sweden</cop><pmid>17159771</pmid></addata></record> |
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| ispartof | Neuro-endocrinology letters, 2006-12, Vol.27 Suppl 2, p.18 |
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| source | MEDLINE; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals |
| subjects | Animals Antineoplastic Agents - pharmacokinetics Cytochrome P-450 Enzyme System - chemistry Cytochrome P-450 Enzyme System - isolation & purification Cytochrome P-450 Enzyme System - metabolism DNA Adducts - metabolism Ellipticines - pharmacokinetics Metabolic Detoxication, Phase I Microsomes, Liver - chemistry Microsomes, Liver - metabolism Models, Biological NADP - chemistry NADP - metabolism NADPH-Ferrihemoprotein Reductase - isolation & purification NADPH-Ferrihemoprotein Reductase - metabolism Rabbits Rats |
| title | Cytochromes P450 reconstituted with NADPH: P450 reductase mimic the activating and detoxicating metabolism of the anticancer drug ellipticine in microsomes |
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