Dynamic combinatorial mass spectrometry for 2-oxoglutarate oxygenase inhibition
<p>In the last decade, dynamic combinatorial mass spectrometry (DCMS) with protein targets has emerged as a promising method for the identification of enzyme-inhibitors. 2-Oxoglutarate (2OG) oxygenases are involved in important biological processes related to many diseases; several human 2OG o...
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| Format: | Thesis |
| Language: | English |
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2013
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| _version_ | 1826314615482482688 |
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| author | Demetriades, M |
| author2 | Schofield, C |
| author_facet | Schofield, C Demetriades, M |
| author_sort | Demetriades, M |
| collection | OXFORD |
| description | <p>In the last decade, dynamic combinatorial mass spectrometry (DCMS) with protein targets has emerged as a promising method for the identification of enzyme-inhibitors. 2-Oxoglutarate (2OG) oxygenases are involved in important biological processes related to many diseases; several human 2OG oxygenases are targeted for pharmaceutical intervention. This thesis describes inhibition studies on three 2OG oxygenases using DCMS and structure activity relation (SAR) studies.</p> <p>Disulphide based DCMS was used for the identification of N-oxalyl based lead inhibitors for the 2OG oxygenase AlkB from <em>Escherichia coli</em>. Crystallographic analyses of AlkB with a lead inhibitor assisted in the design of a second generation of inhibitors using N-oxalyl, pyridyl and quinolinyl scaffolds. Crystallographic and kinetic data of three potent and selective AlkB inhibitors validates the DCMS approach for the development of 2OG oxygenase inhibitors.</p> <p>The hypoxia inducible factor hydroxylase, prolyl hydroxylase domain 2 (PHD2), was then used as the model enzyme for the development of a novel DCMS approach employing the reversible reaction of boronic acids with diols to form boronate esters. The ‘boronate’ DCMS method was used to identify pyridyl- substituted lead compounds. Further modification of the pyridine scaffold, based on structural analyses, led to the development of highly potent and selective PHD2 inhibitors.</p> <p>To identify inhibitors for the fat mass and obesity associated protein (FTO), another 2OG oxygenase, an inhibition assay was developed. The inhibition assay was used in conjunction with a differential scanning fluorimetry (DSF) binding assay to identify isoquinolinyl and pyridyl inhibitor scaffolds, related to those used in the DCMS studies. FTO complexed structures of these compounds, and with a natural product anthraquinone, enabled the design and synthesis of new inhibitors that are both co-substrate and substrate competitors of FTO. One such compound proved to be a potent FTO inhibitor with improved selectivity over other 2OG oxygenases.</p> <p>Overall, the work validates the use of DCMS methods for the development of potent and selective inhibitors for 2OG oxygenases, and by implication of other enzyme families.</p> |
| first_indexed | 2024-03-07T01:08:42Z |
| format | Thesis |
| id | oxford-uuid:8c3a3a33-bc1a-407e-8b86-70c5eca58f38 |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-12-09T03:09:52Z |
| publishDate | 2013 |
| record_format | dspace |
| spelling | oxford-uuid:8c3a3a33-bc1a-407e-8b86-70c5eca58f382024-09-27T11:30:00ZDynamic combinatorial mass spectrometry for 2-oxoglutarate oxygenase inhibition Thesishttp://purl.org/coar/resource_type/c_db06uuid:8c3a3a33-bc1a-407e-8b86-70c5eca58f38Chemical biologyMass spectrometryOrganic chemistryEnglishOxford University Research Archive - Valet2013Demetriades, MSchofield, C<p>In the last decade, dynamic combinatorial mass spectrometry (DCMS) with protein targets has emerged as a promising method for the identification of enzyme-inhibitors. 2-Oxoglutarate (2OG) oxygenases are involved in important biological processes related to many diseases; several human 2OG oxygenases are targeted for pharmaceutical intervention. This thesis describes inhibition studies on three 2OG oxygenases using DCMS and structure activity relation (SAR) studies.</p> <p>Disulphide based DCMS was used for the identification of N-oxalyl based lead inhibitors for the 2OG oxygenase AlkB from <em>Escherichia coli</em>. Crystallographic analyses of AlkB with a lead inhibitor assisted in the design of a second generation of inhibitors using N-oxalyl, pyridyl and quinolinyl scaffolds. Crystallographic and kinetic data of three potent and selective AlkB inhibitors validates the DCMS approach for the development of 2OG oxygenase inhibitors.</p> <p>The hypoxia inducible factor hydroxylase, prolyl hydroxylase domain 2 (PHD2), was then used as the model enzyme for the development of a novel DCMS approach employing the reversible reaction of boronic acids with diols to form boronate esters. The ‘boronate’ DCMS method was used to identify pyridyl- substituted lead compounds. Further modification of the pyridine scaffold, based on structural analyses, led to the development of highly potent and selective PHD2 inhibitors.</p> <p>To identify inhibitors for the fat mass and obesity associated protein (FTO), another 2OG oxygenase, an inhibition assay was developed. The inhibition assay was used in conjunction with a differential scanning fluorimetry (DSF) binding assay to identify isoquinolinyl and pyridyl inhibitor scaffolds, related to those used in the DCMS studies. FTO complexed structures of these compounds, and with a natural product anthraquinone, enabled the design and synthesis of new inhibitors that are both co-substrate and substrate competitors of FTO. One such compound proved to be a potent FTO inhibitor with improved selectivity over other 2OG oxygenases.</p> <p>Overall, the work validates the use of DCMS methods for the development of potent and selective inhibitors for 2OG oxygenases, and by implication of other enzyme families.</p> |
| spellingShingle | Chemical biology Mass spectrometry Organic chemistry Demetriades, M Dynamic combinatorial mass spectrometry for 2-oxoglutarate oxygenase inhibition |
| title | Dynamic combinatorial mass spectrometry for 2-oxoglutarate oxygenase inhibition |
| title_full | Dynamic combinatorial mass spectrometry for 2-oxoglutarate oxygenase inhibition |
| title_fullStr | Dynamic combinatorial mass spectrometry for 2-oxoglutarate oxygenase inhibition |
| title_full_unstemmed | Dynamic combinatorial mass spectrometry for 2-oxoglutarate oxygenase inhibition |
| title_short | Dynamic combinatorial mass spectrometry for 2-oxoglutarate oxygenase inhibition |
| title_sort | dynamic combinatorial mass spectrometry for 2 oxoglutarate oxygenase inhibition |
| topic | Chemical biology Mass spectrometry Organic chemistry |
| work_keys_str_mv | AT demetriadesm dynamiccombinatorialmassspectrometryfor2oxoglutarateoxygenaseinhibition |