The Mode of SN38 Derivatives Interacting with Nicked DNA Mimics Biological Targeting of Topo I Poisons
The compounds 7-ethyl-9-(<i>N</i>-methylamino)methyl-10-hydroxycamptothecin (<b>2</b>) and 7-ethyl-9-(<i>N</i>-morpholino)methyl-10-hydroxycamptothecin (<b>3</b>) are potential topoisomerase I poisons. Moreover, they were shown to have favorable anti-n...
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| Language: | English |
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MDPI AG
2021-07-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/22/14/7471 |
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| author | Wojciech Bocian Beata Naumczuk Magdalena Urbanowicz Jerzy Sitkowski Anna Bierczyńska-Krzysik Elżbieta Bednarek Katarzyna Wiktorska Małgorzata Milczarek Lech Kozerski |
| author_facet | Wojciech Bocian Beata Naumczuk Magdalena Urbanowicz Jerzy Sitkowski Anna Bierczyńska-Krzysik Elżbieta Bednarek Katarzyna Wiktorska Małgorzata Milczarek Lech Kozerski |
| author_sort | Wojciech Bocian |
| collection | DOAJ |
| description | The compounds 7-ethyl-9-(<i>N</i>-methylamino)methyl-10-hydroxycamptothecin (<b>2</b>) and 7-ethyl-9-(<i>N</i>-morpholino)methyl-10-hydroxycamptothecin (<b>3</b>) are potential topoisomerase I poisons. Moreover, they were shown to have favorable anti-neoplastic effects on several tumor cell lines. Due to these properties, the compounds are being considered for advancement to the preclinical development stage. To gain better insights into the molecular mechanism with the biological target, here, we conducted an investigation into their interactions with model nicked DNA (<b>1</b>) using different techniques. In this work, we observed the complexity of the mechanism of action of the compounds <b>2</b> and <b>3</b>, in addition to their decomposition products: compound <b>4</b> and SN38. Using DOSY experiments, evidence of the formation of strongly bonded molecular complexes of SN38 derivatives with DNA duplexes was provided. The molecular modeling based on cross-peaks from the NOESY spectrum also allowed us to assign the geometry of a molecular complex of DNA with compound <b>2</b>. Confirmation of the alkylation reaction of both compounds was obtained using MALDI–MS. Additionally, in the case of <b>3</b>, alkylation was confirmed in the recording of cross-peaks in the <sup>1</sup>H/<sup>13</sup>C HSQC spectrum of <sup>13</sup>C-enriched compound <b>3.</b> In this work, we showed that the studied compounds—parent compounds <b>2</b> and <b>3</b>, and their potential metabolite <b>4</b> and SN38—interact inside the nick of <b>1</b>, either forming the molecular complex or alkylating the DNA nitrogen bases. In order to confirm the influence of the studied compounds on the topoisomerase I relaxation activity of supercoiled DNA, the test was performed based upon the measurement of the fluorescence of DNA stain which can differentiate between supercoiled and relaxed DNA. The presented results confirmed that studied SN38 derivatives effectively block DNA relaxation mediated by Topo I, which means that they stop the machinery of Topo I activity. |
| first_indexed | 2024-03-10T09:37:05Z |
| format | Article |
| id | doaj.art-333cfa843c6943e8844782d6f38eeb5b |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-10T09:37:05Z |
| publishDate | 2021-07-01 |
| publisher | MDPI AG |
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| series | International Journal of Molecular Sciences |
| spelling | doaj.art-333cfa843c6943e8844782d6f38eeb5b2023-11-22T03:59:06ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672021-07-012214747110.3390/ijms22147471The Mode of SN38 Derivatives Interacting with Nicked DNA Mimics Biological Targeting of Topo I PoisonsWojciech Bocian0Beata Naumczuk1Magdalena Urbanowicz2Jerzy Sitkowski3Anna Bierczyńska-Krzysik4Elżbieta Bednarek5Katarzyna Wiktorska6Małgorzata Milczarek7Lech Kozerski8National Medicines Institute, 00-725 Warsaw, PolandNational Medicines Institute, 00-725 Warsaw, PolandNational Medicines Institute, 00-725 Warsaw, PolandNational Medicines Institute, 00-725 Warsaw, PolandŁukasiewicz Industrial Chemistry Institute, 01-793 Warsaw, PolandNational Medicines Institute, 00-725 Warsaw, PolandNational Medicines Institute, 00-725 Warsaw, PolandNational Medicines Institute, 00-725 Warsaw, PolandNational Medicines Institute, 00-725 Warsaw, PolandThe compounds 7-ethyl-9-(<i>N</i>-methylamino)methyl-10-hydroxycamptothecin (<b>2</b>) and 7-ethyl-9-(<i>N</i>-morpholino)methyl-10-hydroxycamptothecin (<b>3</b>) are potential topoisomerase I poisons. Moreover, they were shown to have favorable anti-neoplastic effects on several tumor cell lines. Due to these properties, the compounds are being considered for advancement to the preclinical development stage. To gain better insights into the molecular mechanism with the biological target, here, we conducted an investigation into their interactions with model nicked DNA (<b>1</b>) using different techniques. In this work, we observed the complexity of the mechanism of action of the compounds <b>2</b> and <b>3</b>, in addition to their decomposition products: compound <b>4</b> and SN38. Using DOSY experiments, evidence of the formation of strongly bonded molecular complexes of SN38 derivatives with DNA duplexes was provided. The molecular modeling based on cross-peaks from the NOESY spectrum also allowed us to assign the geometry of a molecular complex of DNA with compound <b>2</b>. Confirmation of the alkylation reaction of both compounds was obtained using MALDI–MS. Additionally, in the case of <b>3</b>, alkylation was confirmed in the recording of cross-peaks in the <sup>1</sup>H/<sup>13</sup>C HSQC spectrum of <sup>13</sup>C-enriched compound <b>3.</b> In this work, we showed that the studied compounds—parent compounds <b>2</b> and <b>3</b>, and their potential metabolite <b>4</b> and SN38—interact inside the nick of <b>1</b>, either forming the molecular complex or alkylating the DNA nitrogen bases. In order to confirm the influence of the studied compounds on the topoisomerase I relaxation activity of supercoiled DNA, the test was performed based upon the measurement of the fluorescence of DNA stain which can differentiate between supercoiled and relaxed DNA. The presented results confirmed that studied SN38 derivatives effectively block DNA relaxation mediated by Topo I, which means that they stop the machinery of Topo I activity.https://www.mdpi.com/1422-0067/22/14/7471bioconjugatecamptothecinDNA complexesmolecular modeling<sup>1</sup>H/<sup>13</sup>C NMRDOSY |
| spellingShingle | Wojciech Bocian Beata Naumczuk Magdalena Urbanowicz Jerzy Sitkowski Anna Bierczyńska-Krzysik Elżbieta Bednarek Katarzyna Wiktorska Małgorzata Milczarek Lech Kozerski The Mode of SN38 Derivatives Interacting with Nicked DNA Mimics Biological Targeting of Topo I Poisons International Journal of Molecular Sciences bioconjugate camptothecin DNA complexes molecular modeling <sup>1</sup>H/<sup>13</sup>C NMR DOSY |
| title | The Mode of SN38 Derivatives Interacting with Nicked DNA Mimics Biological Targeting of Topo I Poisons |
| title_full | The Mode of SN38 Derivatives Interacting with Nicked DNA Mimics Biological Targeting of Topo I Poisons |
| title_fullStr | The Mode of SN38 Derivatives Interacting with Nicked DNA Mimics Biological Targeting of Topo I Poisons |
| title_full_unstemmed | The Mode of SN38 Derivatives Interacting with Nicked DNA Mimics Biological Targeting of Topo I Poisons |
| title_short | The Mode of SN38 Derivatives Interacting with Nicked DNA Mimics Biological Targeting of Topo I Poisons |
| title_sort | mode of sn38 derivatives interacting with nicked dna mimics biological targeting of topo i poisons |
| topic | bioconjugate camptothecin DNA complexes molecular modeling <sup>1</sup>H/<sup>13</sup>C NMR DOSY |
| url | https://www.mdpi.com/1422-0067/22/14/7471 |
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