Synthesis, Characterization, and Pharmacokinetic Studies of PEGylated Glucagon-like Peptide-1
Glucagon-like peptide-1-(7−36) (GLP-1) is a hormone derived from the proglucagon molecule, which is considered a highly desirable antidiabetic agent mainly due to its unique glucose-dependent stimulation of insulin secretion profiles. However, the development of a GLP-1-based pharmaceutical agent ha...
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| Veröffentlicht in: | Bioconjugate chemistry 2005-03, Vol.16 (2), p.377-382 |
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| creator | Lee, Sang-Heon Lee, Seulki Youn, Yu Seok Na, Dong Hee Chae, Su Young Byun, Youngro Lee, Kang Choon |
| description | Glucagon-like peptide-1-(7−36) (GLP-1) is a hormone derived from the proglucagon molecule, which is considered a highly desirable antidiabetic agent mainly due to its unique glucose-dependent stimulation of insulin secretion profiles. However, the development of a GLP-1-based pharmaceutical agent has a severe limitation due to its very short half-life in plasma, being primarily degraded by dipeptidyl peptidase IV (DPP-IV) enzyme. To overcome this limitation, in this article we propose a novel and potent DPP-IV-resistant form of a poly(ethylene glycol)-conjugated GLP-1 preparation and its pharmacokinetic evaluation in rats. Two series of mono-PEGylated GLP-1, (i) N-terminally modified PEG2k-Nter-GLP-1 and (ii) isomers of Lys26, Lys34 modified PEG2k-Lys-GLP-1, were prepared by using mPEG-aldehyde and mPEG-succinimidyl propionate, respectively. To determine the optimized condition for PEGylation, the reactions were monitored at different pH buffer and time intervals by RP-HPLC and MALDI-TOF-MS. The in vitro insulinotropic effect of PEG2k-Lys-GLP-1 showed comparable biological activity with native GLP-1 (P = 0.11) in stimulating insulin secretion in isolated rat pancreatic islet and was significantly more potent than the PEG2k-Nter-GLP-1 (P < 0.05) that showed a marked reduced potency. Furthermore, PEG2k-Lys-GLP-1 was clearly resistant to purified DPP-IV in buffer with 50-fold increased half-life compared to unmodified GLP-1. When PEG2k-Lys-GLP-1 was administered intravenously and subcutaneously into rats, PEGylation improved the half-life, which resulted in substantial improvement of the mean plasma residence time as a 16-fold increase for iv and a 3.2-fold increase for sc. These preliminary results suggest a site specifically mono-PEGylated GLP-1 greatly improved the pharmacological profiles; thus, we anticipated that it could serve as potential candidate as an antidiabetic agent for the treatment of non-insulin-dependent diabetes patients. |
| doi_str_mv | 10.1021/bc049735+ |
| format | Article |
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However, the development of a GLP-1-based pharmaceutical agent has a severe limitation due to its very short half-life in plasma, being primarily degraded by dipeptidyl peptidase IV (DPP-IV) enzyme. To overcome this limitation, in this article we propose a novel and potent DPP-IV-resistant form of a poly(ethylene glycol)-conjugated GLP-1 preparation and its pharmacokinetic evaluation in rats. Two series of mono-PEGylated GLP-1, (i) N-terminally modified PEG2k-Nter-GLP-1 and (ii) isomers of Lys26, Lys34 modified PEG2k-Lys-GLP-1, were prepared by using mPEG-aldehyde and mPEG-succinimidyl propionate, respectively. To determine the optimized condition for PEGylation, the reactions were monitored at different pH buffer and time intervals by RP-HPLC and MALDI-TOF-MS. The in vitro insulinotropic effect of PEG2k-Lys-GLP-1 showed comparable biological activity with native GLP-1 (P = 0.11) in stimulating insulin secretion in isolated rat pancreatic islet and was significantly more potent than the PEG2k-Nter-GLP-1 (P < 0.05) that showed a marked reduced potency. Furthermore, PEG2k-Lys-GLP-1 was clearly resistant to purified DPP-IV in buffer with 50-fold increased half-life compared to unmodified GLP-1. When PEG2k-Lys-GLP-1 was administered intravenously and subcutaneously into rats, PEGylation improved the half-life, which resulted in substantial improvement of the mean plasma residence time as a 16-fold increase for iv and a 3.2-fold increase for sc. These preliminary results suggest a site specifically mono-PEGylated GLP-1 greatly improved the pharmacological profiles; thus, we anticipated that it could serve as potential candidate as an antidiabetic agent for the treatment of non-insulin-dependent diabetes patients.</description><identifier>ISSN: 1043-1802</identifier><identifier>EISSN: 1520-4812</identifier><identifier>DOI: 10.1021/bc049735+</identifier><identifier>PMID: 15769092</identifier><language>eng</language><publisher>United States: American Chemical Society</publisher><subject>Animals ; Chemical synthesis ; Diabetes ; Dipeptidyl Peptidase 4 - metabolism ; Drug Stability ; Enzymes ; Glucagon - administration & dosage ; Glucagon - chemistry ; Glucagon - pharmacokinetics ; Glucagon-Like Peptide 1 ; Glucagon-Like Peptides ; Half-Life ; Hypoglycemic Agents - chemical synthesis ; Hypoglycemic Agents - pharmacokinetics ; Insulin - metabolism ; Insulin Secretion ; Islets of Langerhans - drug effects ; Islets of Langerhans - metabolism ; Male ; Peptide Fragments - administration & dosage ; Peptide Fragments - chemistry ; Peptide Fragments - pharmacokinetics ; Peptides ; Pharmacokinetics ; Pharmacology ; Polyethylene Glycols - chemistry ; Polyethylene Glycols - pharmacology ; Protein Precursors - administration & dosage ; Protein Precursors - chemistry ; Protein Precursors - pharmacokinetics ; Rats ; Rats, Sprague-Dawley ; Structure-Activity Relationship</subject><ispartof>Bioconjugate chemistry, 2005-03, Vol.16 (2), p.377-382</ispartof><rights>Copyright © 2005 American Chemical Society</rights><rights>Copyright American Chemical Society Mar/Apr 2005</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-a441t-1f765e21d85b24cf7cf72e0ebb7706add5b5309905d4b276484d25006386f9ed3</citedby><cites>FETCH-LOGICAL-a441t-1f765e21d85b24cf7cf72e0ebb7706add5b5309905d4b276484d25006386f9ed3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://pubs.acs.org/doi/pdf/10.1021/bc049735+$$EPDF$$P50$$Gacs$$H</linktopdf><linktohtml>$$Uhttps://pubs.acs.org/doi/10.1021/bc049735+$$EHTML$$P50$$Gacs$$H</linktohtml><link.rule.ids>314,780,784,2765,27076,27924,27925,56738,56788</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/15769092$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Lee, Sang-Heon</creatorcontrib><creatorcontrib>Lee, Seulki</creatorcontrib><creatorcontrib>Youn, Yu Seok</creatorcontrib><creatorcontrib>Na, Dong Hee</creatorcontrib><creatorcontrib>Chae, Su Young</creatorcontrib><creatorcontrib>Byun, Youngro</creatorcontrib><creatorcontrib>Lee, Kang Choon</creatorcontrib><title>Synthesis, Characterization, and Pharmacokinetic Studies of PEGylated Glucagon-like Peptide-1</title><title>Bioconjugate chemistry</title><addtitle>Bioconjugate Chem</addtitle><description>Glucagon-like peptide-1-(7−36) (GLP-1) is a hormone derived from the proglucagon molecule, which is considered a highly desirable antidiabetic agent mainly due to its unique glucose-dependent stimulation of insulin secretion profiles. However, the development of a GLP-1-based pharmaceutical agent has a severe limitation due to its very short half-life in plasma, being primarily degraded by dipeptidyl peptidase IV (DPP-IV) enzyme. To overcome this limitation, in this article we propose a novel and potent DPP-IV-resistant form of a poly(ethylene glycol)-conjugated GLP-1 preparation and its pharmacokinetic evaluation in rats. Two series of mono-PEGylated GLP-1, (i) N-terminally modified PEG2k-Nter-GLP-1 and (ii) isomers of Lys26, Lys34 modified PEG2k-Lys-GLP-1, were prepared by using mPEG-aldehyde and mPEG-succinimidyl propionate, respectively. To determine the optimized condition for PEGylation, the reactions were monitored at different pH buffer and time intervals by RP-HPLC and MALDI-TOF-MS. The in vitro insulinotropic effect of PEG2k-Lys-GLP-1 showed comparable biological activity with native GLP-1 (P = 0.11) in stimulating insulin secretion in isolated rat pancreatic islet and was significantly more potent than the PEG2k-Nter-GLP-1 (P < 0.05) that showed a marked reduced potency. Furthermore, PEG2k-Lys-GLP-1 was clearly resistant to purified DPP-IV in buffer with 50-fold increased half-life compared to unmodified GLP-1. When PEG2k-Lys-GLP-1 was administered intravenously and subcutaneously into rats, PEGylation improved the half-life, which resulted in substantial improvement of the mean plasma residence time as a 16-fold increase for iv and a 3.2-fold increase for sc. These preliminary results suggest a site specifically mono-PEGylated GLP-1 greatly improved the pharmacological profiles; thus, we anticipated that it could serve as potential candidate as an antidiabetic agent for the treatment of non-insulin-dependent diabetes patients.</description><subject>Animals</subject><subject>Chemical synthesis</subject><subject>Diabetes</subject><subject>Dipeptidyl Peptidase 4 - metabolism</subject><subject>Drug Stability</subject><subject>Enzymes</subject><subject>Glucagon - administration & dosage</subject><subject>Glucagon - chemistry</subject><subject>Glucagon - pharmacokinetics</subject><subject>Glucagon-Like Peptide 1</subject><subject>Glucagon-Like Peptides</subject><subject>Half-Life</subject><subject>Hypoglycemic Agents - chemical synthesis</subject><subject>Hypoglycemic Agents - pharmacokinetics</subject><subject>Insulin - metabolism</subject><subject>Insulin Secretion</subject><subject>Islets of Langerhans - drug effects</subject><subject>Islets of Langerhans - metabolism</subject><subject>Male</subject><subject>Peptide Fragments - administration & dosage</subject><subject>Peptide Fragments - chemistry</subject><subject>Peptide Fragments - pharmacokinetics</subject><subject>Peptides</subject><subject>Pharmacokinetics</subject><subject>Pharmacology</subject><subject>Polyethylene Glycols - chemistry</subject><subject>Polyethylene Glycols - pharmacology</subject><subject>Protein Precursors - administration & dosage</subject><subject>Protein Precursors - chemistry</subject><subject>Protein Precursors - pharmacokinetics</subject><subject>Rats</subject><subject>Rats, Sprague-Dawley</subject><subject>Structure-Activity Relationship</subject><issn>1043-1802</issn><issn>1520-4812</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2005</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpl0F2L1DAUBuAgiruuXvgHpIiI4FZP0ny0lzKsHXHBgRkvJaTJqZudTjsmKTj-erPM6IJCICF5ePNyCHlO4R0FRt93FnijKvH2ATmngkHJa8oe5jPwqqQ1sDPyJMZbAGhozR6TMyqUbKBh5-Tb-jCmG4w-XhaLGxOMTRj8L5P8NF4WZnTFKt_ujJ22fsTkbbFOs_MYi6kvVlftYTAJXdEOszXfp7Ec_BaLFe6Td1jSp-RRb4aIz077Bfn68WqzWJbXX9pPiw_XpeGcppL2Sgpk1NWiY9z2Ki-GgF2nFEjjnOhEBU0DwvGOKclr7pgAkFUt-wZddUFeH3P3YfoxY0x656PFYTAjTnPUUgkmKacZvvwH3k5zGHM3zaikdU68Q2-OyIYpxoC93ge_M-GgKei7ges_A8_0xSlv7nbo7uFpwBmUR-Bjwp9_303Y5lKVEnqzWutl-1mKdrnRi-xfHb2x8b7bf__-BvjHk8w</recordid><startdate>20050301</startdate><enddate>20050301</enddate><creator>Lee, Sang-Heon</creator><creator>Lee, Seulki</creator><creator>Youn, Yu Seok</creator><creator>Na, Dong Hee</creator><creator>Chae, Su Young</creator><creator>Byun, Youngro</creator><creator>Lee, Kang Choon</creator><general>American Chemical Society</general><scope>BSCLL</scope><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>7QO</scope><scope>7TM</scope><scope>8FD</scope><scope>FR3</scope><scope>P64</scope><scope>7X8</scope></search><sort><creationdate>20050301</creationdate><title>Synthesis, Characterization, and Pharmacokinetic Studies of PEGylated Glucagon-like Peptide-1</title><author>Lee, Sang-Heon ; Lee, Seulki ; Youn, Yu Seok ; Na, Dong Hee ; Chae, Su Young ; Byun, Youngro ; Lee, Kang Choon</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-a441t-1f765e21d85b24cf7cf72e0ebb7706add5b5309905d4b276484d25006386f9ed3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2005</creationdate><topic>Animals</topic><topic>Chemical synthesis</topic><topic>Diabetes</topic><topic>Dipeptidyl Peptidase 4 - metabolism</topic><topic>Drug Stability</topic><topic>Enzymes</topic><topic>Glucagon - administration & dosage</topic><topic>Glucagon - chemistry</topic><topic>Glucagon - pharmacokinetics</topic><topic>Glucagon-Like Peptide 1</topic><topic>Glucagon-Like Peptides</topic><topic>Half-Life</topic><topic>Hypoglycemic Agents - chemical synthesis</topic><topic>Hypoglycemic Agents - pharmacokinetics</topic><topic>Insulin - metabolism</topic><topic>Insulin Secretion</topic><topic>Islets of Langerhans - drug effects</topic><topic>Islets of Langerhans - metabolism</topic><topic>Male</topic><topic>Peptide Fragments - administration & dosage</topic><topic>Peptide Fragments - chemistry</topic><topic>Peptide Fragments - pharmacokinetics</topic><topic>Peptides</topic><topic>Pharmacokinetics</topic><topic>Pharmacology</topic><topic>Polyethylene Glycols - chemistry</topic><topic>Polyethylene Glycols - pharmacology</topic><topic>Protein Precursors - administration & dosage</topic><topic>Protein Precursors - chemistry</topic><topic>Protein Precursors - pharmacokinetics</topic><topic>Rats</topic><topic>Rats, Sprague-Dawley</topic><topic>Structure-Activity Relationship</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Lee, Sang-Heon</creatorcontrib><creatorcontrib>Lee, Seulki</creatorcontrib><creatorcontrib>Youn, Yu Seok</creatorcontrib><creatorcontrib>Na, Dong Hee</creatorcontrib><creatorcontrib>Chae, Su Young</creatorcontrib><creatorcontrib>Byun, Youngro</creatorcontrib><creatorcontrib>Lee, Kang Choon</creatorcontrib><collection>Istex</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Biotechnology Research Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Bioconjugate chemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Lee, Sang-Heon</au><au>Lee, Seulki</au><au>Youn, Yu Seok</au><au>Na, Dong Hee</au><au>Chae, Su Young</au><au>Byun, Youngro</au><au>Lee, Kang Choon</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Synthesis, Characterization, and Pharmacokinetic Studies of PEGylated Glucagon-like Peptide-1</atitle><jtitle>Bioconjugate chemistry</jtitle><addtitle>Bioconjugate Chem</addtitle><date>2005-03-01</date><risdate>2005</risdate><volume>16</volume><issue>2</issue><spage>377</spage><epage>382</epage><pages>377-382</pages><issn>1043-1802</issn><eissn>1520-4812</eissn><abstract>Glucagon-like peptide-1-(7−36) (GLP-1) is a hormone derived from the proglucagon molecule, which is considered a highly desirable antidiabetic agent mainly due to its unique glucose-dependent stimulation of insulin secretion profiles. However, the development of a GLP-1-based pharmaceutical agent has a severe limitation due to its very short half-life in plasma, being primarily degraded by dipeptidyl peptidase IV (DPP-IV) enzyme. To overcome this limitation, in this article we propose a novel and potent DPP-IV-resistant form of a poly(ethylene glycol)-conjugated GLP-1 preparation and its pharmacokinetic evaluation in rats. Two series of mono-PEGylated GLP-1, (i) N-terminally modified PEG2k-Nter-GLP-1 and (ii) isomers of Lys26, Lys34 modified PEG2k-Lys-GLP-1, were prepared by using mPEG-aldehyde and mPEG-succinimidyl propionate, respectively. To determine the optimized condition for PEGylation, the reactions were monitored at different pH buffer and time intervals by RP-HPLC and MALDI-TOF-MS. The in vitro insulinotropic effect of PEG2k-Lys-GLP-1 showed comparable biological activity with native GLP-1 (P = 0.11) in stimulating insulin secretion in isolated rat pancreatic islet and was significantly more potent than the PEG2k-Nter-GLP-1 (P < 0.05) that showed a marked reduced potency. Furthermore, PEG2k-Lys-GLP-1 was clearly resistant to purified DPP-IV in buffer with 50-fold increased half-life compared to unmodified GLP-1. When PEG2k-Lys-GLP-1 was administered intravenously and subcutaneously into rats, PEGylation improved the half-life, which resulted in substantial improvement of the mean plasma residence time as a 16-fold increase for iv and a 3.2-fold increase for sc. These preliminary results suggest a site specifically mono-PEGylated GLP-1 greatly improved the pharmacological profiles; thus, we anticipated that it could serve as potential candidate as an antidiabetic agent for the treatment of non-insulin-dependent diabetes patients.</abstract><cop>United States</cop><pub>American Chemical Society</pub><pmid>15769092</pmid><doi>10.1021/bc049735+</doi><tpages>6</tpages></addata></record> |
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| ispartof | Bioconjugate chemistry, 2005-03, Vol.16 (2), p.377-382 |
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| subjects | Animals Chemical synthesis Diabetes Dipeptidyl Peptidase 4 - metabolism Drug Stability Enzymes Glucagon - administration & dosage Glucagon - chemistry Glucagon - pharmacokinetics Glucagon-Like Peptide 1 Glucagon-Like Peptides Half-Life Hypoglycemic Agents - chemical synthesis Hypoglycemic Agents - pharmacokinetics Insulin - metabolism Insulin Secretion Islets of Langerhans - drug effects Islets of Langerhans - metabolism Male Peptide Fragments - administration & dosage Peptide Fragments - chemistry Peptide Fragments - pharmacokinetics Peptides Pharmacokinetics Pharmacology Polyethylene Glycols - chemistry Polyethylene Glycols - pharmacology Protein Precursors - administration & dosage Protein Precursors - chemistry Protein Precursors - pharmacokinetics Rats Rats, Sprague-Dawley Structure-Activity Relationship |
| title | Synthesis, Characterization, and Pharmacokinetic Studies of PEGylated Glucagon-like Peptide-1 |
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