Slow-onset inhibition of fumarylacetoacetate hydrolase by phosphinate mimics of the tetrahedral intermediate: kinetics, crystal structure and pharmacokinetics
FAH (fumarylacetoacetate hydrolase) catalyses the final step of tyrosine catabolism to produce fumarate and acetoacetate. HT1 (hereditary tyrosinaemia type 1) results from deficiency of this enzyme. Previously, we prepared a partial mimic of the putative tetrahedral intermediate in the reaction cata...
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Veröffentlicht in: | Biochemical journal 2007-03, Vol.402 (2), p.251-260 |
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description | FAH (fumarylacetoacetate hydrolase) catalyses the final step of tyrosine catabolism to produce fumarate and acetoacetate. HT1 (hereditary tyrosinaemia type 1) results from deficiency of this enzyme. Previously, we prepared a partial mimic of the putative tetrahedral intermediate in the reaction catalysed by FAH co-crystallized with the enzyme to reveal details of the mechanism [Bateman, Bhanumoorthy, Witte, McClard, Grompe and Timm (2001) J. Biol. Chem. 276, 15284-15291]. We have now successfully synthesized complete mimics CEHPOBA {4-[(2-carboxyethyl)-hydroxyphosphinyl]-3-oxobutyrate} and COPHPAA {3-[(3-carboxy-2-oxopropyl)hydroxyphosphinyl]acrylate}, which inhibit FAH in slow-onset tight-binding mode with K(i) values of 41 and 12 nM respectively. A high-resolution (1.35 A; 1 A=0.1 nm) crystal structure of the FAH.CEHPOBA complex was solved to reveal the affinity determinants for these compounds and to provide further insight into the mechanism of FAH catalysis. These compounds are active in vivo, and CEHPOBA demonstrated a notable dose-dependent increase in SA (succinylacetone; a metabolite seen in patients with HT1) in mouse serum after repeated injections, and, following a single injection (1 mumol/g; intraperitoneal), only a modest regain of FAH enzyme activity was detected in liver protein isolates after 24 h. These potent inhibitors provide a means to chemically phenocopy the metabolic defects of either HT1 or FAH knockout mice and promise future pharmacological utility for hepatocyte transplantation. |
doi_str_mv | 10.1042/BJ20060961 |
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HT1 (hereditary tyrosinaemia type 1) results from deficiency of this enzyme. Previously, we prepared a partial mimic of the putative tetrahedral intermediate in the reaction catalysed by FAH co-crystallized with the enzyme to reveal details of the mechanism [Bateman, Bhanumoorthy, Witte, McClard, Grompe and Timm (2001) J. Biol. Chem. 276, 15284-15291]. We have now successfully synthesized complete mimics CEHPOBA {4-[(2-carboxyethyl)-hydroxyphosphinyl]-3-oxobutyrate} and COPHPAA {3-[(3-carboxy-2-oxopropyl)hydroxyphosphinyl]acrylate}, which inhibit FAH in slow-onset tight-binding mode with K(i) values of 41 and 12 nM respectively. A high-resolution (1.35 A; 1 A=0.1 nm) crystal structure of the FAH.CEHPOBA complex was solved to reveal the affinity determinants for these compounds and to provide further insight into the mechanism of FAH catalysis. These compounds are active in vivo, and CEHPOBA demonstrated a notable dose-dependent increase in SA (succinylacetone; a metabolite seen in patients with HT1) in mouse serum after repeated injections, and, following a single injection (1 mumol/g; intraperitoneal), only a modest regain of FAH enzyme activity was detected in liver protein isolates after 24 h. These potent inhibitors provide a means to chemically phenocopy the metabolic defects of either HT1 or FAH knockout mice and promise future pharmacological utility for hepatocyte transplantation.</description><identifier>ISSN: 0264-6021</identifier><identifier>EISSN: 1470-8728</identifier><identifier>DOI: 10.1042/BJ20060961</identifier><identifier>PMID: 17064256</identifier><language>eng</language><publisher>England: Portland Press Ltd</publisher><subject>Animals ; Binding Sites ; Biomimetic Materials - chemical synthesis ; Biomimetic Materials - chemistry ; Biomimetic Materials - pharmacokinetics ; Butyrates - blood ; Butyrates - chemical synthesis ; Butyrates - chemistry ; Butyrates - pharmacokinetics ; Crystallography, X-Ray ; Humans ; Hydrolases - antagonists & inhibitors ; Hydrolases - chemistry ; Hydrolases - metabolism ; Kinetics ; Male ; Mice ; Mice, Inbred C57BL ; Models, Molecular ; Molecular Structure ; Organophosphorus Compounds - blood ; Organophosphorus Compounds - chemical synthesis ; Organophosphorus Compounds - chemistry ; Organophosphorus Compounds - pharmacokinetics ; Phosphinic Acids - blood ; Phosphinic Acids - chemical synthesis ; Phosphinic Acids - chemistry ; Phosphinic Acids - pharmacokinetics ; Protease Inhibitors - blood ; Protease Inhibitors - chemical synthesis ; Protease Inhibitors - chemistry ; Protease Inhibitors - pharmacokinetics ; Protein Structure, Tertiary ; Time Factors</subject><ispartof>Biochemical journal, 2007-03, Vol.402 (2), p.251-260</ispartof><rights>The Biochemical Society, London 2007</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c376t-2e546d377577fc7e9a3e0d910c6189ed04c05fa77ecb7b6a2a2d1adf15f1242f3</citedby><cites>FETCH-LOGICAL-c376t-2e546d377577fc7e9a3e0d910c6189ed04c05fa77ecb7b6a2a2d1adf15f1242f3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC1798426/pdf/$$EPDF$$P50$$Gpubmedcentral$$H</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC1798426/$$EHTML$$P50$$Gpubmedcentral$$H</linktohtml><link.rule.ids>230,315,729,782,786,887,27931,27932,53798,53800</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/17064256$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Bateman, Raynard L</creatorcontrib><creatorcontrib>Ashworth, Justin</creatorcontrib><creatorcontrib>Witte, John F</creatorcontrib><creatorcontrib>Baker, L-J</creatorcontrib><creatorcontrib>Bhanumoorthy, Pullooru</creatorcontrib><creatorcontrib>Timm, David E</creatorcontrib><creatorcontrib>Hurley, Thomas D</creatorcontrib><creatorcontrib>Grompe, Markus</creatorcontrib><creatorcontrib>McClard, Ronald W</creatorcontrib><title>Slow-onset inhibition of fumarylacetoacetate hydrolase by phosphinate mimics of the tetrahedral intermediate: kinetics, crystal structure and pharmacokinetics</title><title>Biochemical journal</title><addtitle>Biochem J</addtitle><description>FAH (fumarylacetoacetate hydrolase) catalyses the final step of tyrosine catabolism to produce fumarate and acetoacetate. HT1 (hereditary tyrosinaemia type 1) results from deficiency of this enzyme. Previously, we prepared a partial mimic of the putative tetrahedral intermediate in the reaction catalysed by FAH co-crystallized with the enzyme to reveal details of the mechanism [Bateman, Bhanumoorthy, Witte, McClard, Grompe and Timm (2001) J. Biol. Chem. 276, 15284-15291]. We have now successfully synthesized complete mimics CEHPOBA {4-[(2-carboxyethyl)-hydroxyphosphinyl]-3-oxobutyrate} and COPHPAA {3-[(3-carboxy-2-oxopropyl)hydroxyphosphinyl]acrylate}, which inhibit FAH in slow-onset tight-binding mode with K(i) values of 41 and 12 nM respectively. A high-resolution (1.35 A; 1 A=0.1 nm) crystal structure of the FAH.CEHPOBA complex was solved to reveal the affinity determinants for these compounds and to provide further insight into the mechanism of FAH catalysis. These compounds are active in vivo, and CEHPOBA demonstrated a notable dose-dependent increase in SA (succinylacetone; a metabolite seen in patients with HT1) in mouse serum after repeated injections, and, following a single injection (1 mumol/g; intraperitoneal), only a modest regain of FAH enzyme activity was detected in liver protein isolates after 24 h. These potent inhibitors provide a means to chemically phenocopy the metabolic defects of either HT1 or FAH knockout mice and promise future pharmacological utility for hepatocyte transplantation.</description><subject>Animals</subject><subject>Binding Sites</subject><subject>Biomimetic Materials - chemical synthesis</subject><subject>Biomimetic Materials - chemistry</subject><subject>Biomimetic Materials - pharmacokinetics</subject><subject>Butyrates - blood</subject><subject>Butyrates - chemical synthesis</subject><subject>Butyrates - chemistry</subject><subject>Butyrates - pharmacokinetics</subject><subject>Crystallography, X-Ray</subject><subject>Humans</subject><subject>Hydrolases - antagonists & inhibitors</subject><subject>Hydrolases - chemistry</subject><subject>Hydrolases - metabolism</subject><subject>Kinetics</subject><subject>Male</subject><subject>Mice</subject><subject>Mice, Inbred C57BL</subject><subject>Models, Molecular</subject><subject>Molecular Structure</subject><subject>Organophosphorus Compounds - blood</subject><subject>Organophosphorus Compounds - chemical synthesis</subject><subject>Organophosphorus Compounds - chemistry</subject><subject>Organophosphorus Compounds - pharmacokinetics</subject><subject>Phosphinic Acids - blood</subject><subject>Phosphinic Acids - chemical synthesis</subject><subject>Phosphinic Acids - chemistry</subject><subject>Phosphinic Acids - pharmacokinetics</subject><subject>Protease Inhibitors - blood</subject><subject>Protease Inhibitors - chemical synthesis</subject><subject>Protease Inhibitors - chemistry</subject><subject>Protease Inhibitors - pharmacokinetics</subject><subject>Protein Structure, Tertiary</subject><subject>Time Factors</subject><issn>0264-6021</issn><issn>1470-8728</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2007</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpVkU2P1SAUhonRONfRjT_AsHJhrB4ohVsXJjrxM5O4UNeEwqlFW7gC1dw_42-VZq6ObmDxPjwceAm5z-AJA8GfvnzPAST0kt0gOyYUNHvF9zfJDrgUjQTOzsidnL8CMAECbpMzpkAK3skd-fVxjj-bGDIW6sPkB198DDSOdFwXk46zsVjitpiCdDq6FGeTkQ5HephiPkw-bMHiF2_zdqxMSAuWZCZ0ycxVWjAt6HzFntFvPmCp5GNq0zGXmueSVlvWhNQEV50mLcbGP9xdcms0c8Z7p_2cfH796tPF2-byw5t3Fy8uG9sqWRqOnZCuVapTarQKe9MiuJ6BlWzfowNhoRuNUmgHNUjDDXfMuJF1I-OCj-05eX7lPaxDHdZiqC-Y9SH57RN0NF7_nwQ_6S_xh2aq3wsuq-DhSZDi9xVz0YvPFufZBIxr1rIHaCVvK_joCrQp5pxw_HsJA73Vqa_rrPCDf8e6Rk_9tb8BFAWhBg</recordid><startdate>20070301</startdate><enddate>20070301</enddate><creator>Bateman, Raynard L</creator><creator>Ashworth, Justin</creator><creator>Witte, John F</creator><creator>Baker, L-J</creator><creator>Bhanumoorthy, Pullooru</creator><creator>Timm, David E</creator><creator>Hurley, Thomas D</creator><creator>Grompe, Markus</creator><creator>McClard, Ronald W</creator><general>Portland Press Ltd</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>20070301</creationdate><title>Slow-onset inhibition of fumarylacetoacetate hydrolase by phosphinate mimics of the tetrahedral intermediate: kinetics, crystal structure and pharmacokinetics</title><author>Bateman, Raynard L ; Ashworth, Justin ; Witte, John F ; Baker, L-J ; Bhanumoorthy, Pullooru ; Timm, David E ; Hurley, Thomas D ; Grompe, Markus ; McClard, Ronald W</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c376t-2e546d377577fc7e9a3e0d910c6189ed04c05fa77ecb7b6a2a2d1adf15f1242f3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2007</creationdate><topic>Animals</topic><topic>Binding Sites</topic><topic>Biomimetic Materials - chemical synthesis</topic><topic>Biomimetic Materials - chemistry</topic><topic>Biomimetic Materials - pharmacokinetics</topic><topic>Butyrates - blood</topic><topic>Butyrates - chemical synthesis</topic><topic>Butyrates - chemistry</topic><topic>Butyrates - pharmacokinetics</topic><topic>Crystallography, X-Ray</topic><topic>Humans</topic><topic>Hydrolases - antagonists & inhibitors</topic><topic>Hydrolases - chemistry</topic><topic>Hydrolases - metabolism</topic><topic>Kinetics</topic><topic>Male</topic><topic>Mice</topic><topic>Mice, Inbred C57BL</topic><topic>Models, Molecular</topic><topic>Molecular Structure</topic><topic>Organophosphorus Compounds - blood</topic><topic>Organophosphorus Compounds - chemical synthesis</topic><topic>Organophosphorus Compounds - chemistry</topic><topic>Organophosphorus Compounds - pharmacokinetics</topic><topic>Phosphinic Acids - blood</topic><topic>Phosphinic Acids - chemical synthesis</topic><topic>Phosphinic Acids - chemistry</topic><topic>Phosphinic Acids - pharmacokinetics</topic><topic>Protease Inhibitors - blood</topic><topic>Protease Inhibitors - chemical synthesis</topic><topic>Protease Inhibitors - chemistry</topic><topic>Protease Inhibitors - pharmacokinetics</topic><topic>Protein Structure, Tertiary</topic><topic>Time Factors</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Bateman, Raynard L</creatorcontrib><creatorcontrib>Ashworth, Justin</creatorcontrib><creatorcontrib>Witte, John F</creatorcontrib><creatorcontrib>Baker, L-J</creatorcontrib><creatorcontrib>Bhanumoorthy, Pullooru</creatorcontrib><creatorcontrib>Timm, David E</creatorcontrib><creatorcontrib>Hurley, Thomas D</creatorcontrib><creatorcontrib>Grompe, Markus</creatorcontrib><creatorcontrib>McClard, Ronald W</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Biochemical journal</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Bateman, Raynard L</au><au>Ashworth, Justin</au><au>Witte, John F</au><au>Baker, L-J</au><au>Bhanumoorthy, Pullooru</au><au>Timm, David E</au><au>Hurley, Thomas D</au><au>Grompe, Markus</au><au>McClard, Ronald W</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Slow-onset inhibition of fumarylacetoacetate hydrolase by phosphinate mimics of the tetrahedral intermediate: kinetics, crystal structure and pharmacokinetics</atitle><jtitle>Biochemical journal</jtitle><addtitle>Biochem J</addtitle><date>2007-03-01</date><risdate>2007</risdate><volume>402</volume><issue>2</issue><spage>251</spage><epage>260</epage><pages>251-260</pages><issn>0264-6021</issn><eissn>1470-8728</eissn><abstract>FAH (fumarylacetoacetate hydrolase) catalyses the final step of tyrosine catabolism to produce fumarate and acetoacetate. HT1 (hereditary tyrosinaemia type 1) results from deficiency of this enzyme. Previously, we prepared a partial mimic of the putative tetrahedral intermediate in the reaction catalysed by FAH co-crystallized with the enzyme to reveal details of the mechanism [Bateman, Bhanumoorthy, Witte, McClard, Grompe and Timm (2001) J. Biol. Chem. 276, 15284-15291]. We have now successfully synthesized complete mimics CEHPOBA {4-[(2-carboxyethyl)-hydroxyphosphinyl]-3-oxobutyrate} and COPHPAA {3-[(3-carboxy-2-oxopropyl)hydroxyphosphinyl]acrylate}, which inhibit FAH in slow-onset tight-binding mode with K(i) values of 41 and 12 nM respectively. A high-resolution (1.35 A; 1 A=0.1 nm) crystal structure of the FAH.CEHPOBA complex was solved to reveal the affinity determinants for these compounds and to provide further insight into the mechanism of FAH catalysis. These compounds are active in vivo, and CEHPOBA demonstrated a notable dose-dependent increase in SA (succinylacetone; a metabolite seen in patients with HT1) in mouse serum after repeated injections, and, following a single injection (1 mumol/g; intraperitoneal), only a modest regain of FAH enzyme activity was detected in liver protein isolates after 24 h. These potent inhibitors provide a means to chemically phenocopy the metabolic defects of either HT1 or FAH knockout mice and promise future pharmacological utility for hepatocyte transplantation.</abstract><cop>England</cop><pub>Portland Press Ltd</pub><pmid>17064256</pmid><doi>10.1042/BJ20060961</doi><tpages>10</tpages><oa>free_for_read</oa></addata></record> |
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subjects | Animals Binding Sites Biomimetic Materials - chemical synthesis Biomimetic Materials - chemistry Biomimetic Materials - pharmacokinetics Butyrates - blood Butyrates - chemical synthesis Butyrates - chemistry Butyrates - pharmacokinetics Crystallography, X-Ray Humans Hydrolases - antagonists & inhibitors Hydrolases - chemistry Hydrolases - metabolism Kinetics Male Mice Mice, Inbred C57BL Models, Molecular Molecular Structure Organophosphorus Compounds - blood Organophosphorus Compounds - chemical synthesis Organophosphorus Compounds - chemistry Organophosphorus Compounds - pharmacokinetics Phosphinic Acids - blood Phosphinic Acids - chemical synthesis Phosphinic Acids - chemistry Phosphinic Acids - pharmacokinetics Protease Inhibitors - blood Protease Inhibitors - chemical synthesis Protease Inhibitors - chemistry Protease Inhibitors - pharmacokinetics Protein Structure, Tertiary Time Factors |
title | Slow-onset inhibition of fumarylacetoacetate hydrolase by phosphinate mimics of the tetrahedral intermediate: kinetics, crystal structure and pharmacokinetics |
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