Exopeptidase combination enhances the degradation of isotopically labelled gluten immunogenic peptides in humans
IntroductionCeliac disease is a common autoimmune-like enteropathy caused by an aberrant response to incompletely digested dietary gluten. Gluten immunogenic peptides including the immunodominant 33-mer are thought to be resistant to proteolytic digestion by human gastrointestinal peptidases. We dev...
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Frontiers Media S.A.
2024-10-01
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| Series: | Frontiers in Immunology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fimmu.2024.1425982/full |
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| author | Sulayman Mourabit Sarah Römer Erin R. Bonner Fabian Winter Julian Tschollar Mladen V. Tzvetkov Werner Weitschies Stefan Engeli Werner Tschollar |
| author_facet | Sulayman Mourabit Sarah Römer Erin R. Bonner Fabian Winter Julian Tschollar Mladen V. Tzvetkov Werner Weitschies Stefan Engeli Werner Tschollar |
| author_sort | Sulayman Mourabit |
| collection | DOAJ |
| description | IntroductionCeliac disease is a common autoimmune-like enteropathy caused by an aberrant response to incompletely digested dietary gluten. Gluten immunogenic peptides including the immunodominant 33-mer are thought to be resistant to proteolytic digestion by human gastrointestinal peptidases. We developed a novel enzyme therapy approach to support gluten peptide digestion using a combination of two tandem-acting exopeptidases, AMYNOPEP, that complement the intrinsic enzymatic activity of intestinal brush border enterocytes.MethodsWe evaluated the effects of AMYNOPEP supplementation on 33-mer degradation in vitro and in vivo. In a cross-over clinical study, healthy volunteers with no gastrointestinal disorders were given stable isotope (SI) labelled 33-mer peptides in the presence of varying peptide substrates and caloric loads, with and without AMYNOPEP. 33-mer degradation products (SI-labelled single amino acids) were measured in the blood plasma using LC-MS/MS.ResultsAMYNOPEP achieved rapid, complete amino-to-carboxyl terminal degradation of the 33-mer in vitro, generating single amino acids and dipeptides. In healthy volunteers, AMYNOPEP supplementation significantly increased 33-mer degradation and absorption of SI-labelled amino acids even in the presence of competing substrates. Specifically, we observed a 2.8-fold increase in the Cmax of stable isotope-labelled amino acids in the presence of wheat gluten. The absorption kinetics of labelled amino acids derived from 33-mer digestion with AMYNOPEP closely resembled that of SI-labelled X-Proline dipeptides administered without enzyme supplementation, highlighting the rapid hydrolytic activity of AMYNOPEP on polypeptides.ConclusionsAMYNOPEP achieved complete degradation of the 33-mer into single amino acids and dipeptides in vitro and significantly improved 33-mer degradation kinetics in healthy volunteers, as measured by labelled amino acid detection, warranting further investigation into the potential therapeutic benefits of exopeptidase combinations for patients with gluten-related health disorders including celiac disease. |
| format | Article |
| id | doaj-art-bde4e3f06cc24660b833ea9a15c69414 |
| institution | OA Journals |
| issn | 1664-3224 |
| language | English |
| publishDate | 2024-10-01 |
| publisher | Frontiers Media S.A. |
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| series | Frontiers in Immunology |
| spelling | doaj-art-bde4e3f06cc24660b833ea9a15c694142025-08-20T01:47:37ZengFrontiers Media S.A.Frontiers in Immunology1664-32242024-10-011510.3389/fimmu.2024.14259821425982Exopeptidase combination enhances the degradation of isotopically labelled gluten immunogenic peptides in humansSulayman Mourabit0Sarah Römer1Erin R. Bonner2Fabian Winter3Julian Tschollar4Mladen V. Tzvetkov5Werner Weitschies6Stefan Engeli7Werner Tschollar8AMYRA Biotech AG, Basel, SwitzerlandDepartment of General Pharmacology, Institute of Pharmacology Center of Drug Absorption and Transport (C_DAT), University Medicine Greifswald, Greifswald, GermanyAMYRA Biotech AG, Basel, SwitzerlandDepartment of Pharmaceutical Technology and Biopharmaceutics, Institute of Pharmacy, Center of Drug Absorption and Transport (C_DAT), University of Greifswald, Greifswald, GermanyAMYRA Biotech AG, Basel, SwitzerlandDepartment of General Pharmacology, Institute of Pharmacology Center of Drug Absorption and Transport (C_DAT), University Medicine Greifswald, Greifswald, GermanyDepartment of Pharmaceutical Technology and Biopharmaceutics, Institute of Pharmacy, Center of Drug Absorption and Transport (C_DAT), University of Greifswald, Greifswald, GermanyDepartment of Clinical Pharmacology, Institute of Pharmacology, Center of Drug Absorption and Transport (C_DAT), University Medicine Greifswald, Greifswald, GermanyAMYRA Biotech AG, Basel, SwitzerlandIntroductionCeliac disease is a common autoimmune-like enteropathy caused by an aberrant response to incompletely digested dietary gluten. Gluten immunogenic peptides including the immunodominant 33-mer are thought to be resistant to proteolytic digestion by human gastrointestinal peptidases. We developed a novel enzyme therapy approach to support gluten peptide digestion using a combination of two tandem-acting exopeptidases, AMYNOPEP, that complement the intrinsic enzymatic activity of intestinal brush border enterocytes.MethodsWe evaluated the effects of AMYNOPEP supplementation on 33-mer degradation in vitro and in vivo. In a cross-over clinical study, healthy volunteers with no gastrointestinal disorders were given stable isotope (SI) labelled 33-mer peptides in the presence of varying peptide substrates and caloric loads, with and without AMYNOPEP. 33-mer degradation products (SI-labelled single amino acids) were measured in the blood plasma using LC-MS/MS.ResultsAMYNOPEP achieved rapid, complete amino-to-carboxyl terminal degradation of the 33-mer in vitro, generating single amino acids and dipeptides. In healthy volunteers, AMYNOPEP supplementation significantly increased 33-mer degradation and absorption of SI-labelled amino acids even in the presence of competing substrates. Specifically, we observed a 2.8-fold increase in the Cmax of stable isotope-labelled amino acids in the presence of wheat gluten. The absorption kinetics of labelled amino acids derived from 33-mer digestion with AMYNOPEP closely resembled that of SI-labelled X-Proline dipeptides administered without enzyme supplementation, highlighting the rapid hydrolytic activity of AMYNOPEP on polypeptides.ConclusionsAMYNOPEP achieved complete degradation of the 33-mer into single amino acids and dipeptides in vitro and significantly improved 33-mer degradation kinetics in healthy volunteers, as measured by labelled amino acid detection, warranting further investigation into the potential therapeutic benefits of exopeptidase combinations for patients with gluten-related health disorders including celiac disease.https://www.frontiersin.org/articles/10.3389/fimmu.2024.1425982/fullglutengluten immunogenic peptideceliac diseaseexopeptidaseglutenase33-mer |
| spellingShingle | Sulayman Mourabit Sarah Römer Erin R. Bonner Fabian Winter Julian Tschollar Mladen V. Tzvetkov Werner Weitschies Stefan Engeli Werner Tschollar Exopeptidase combination enhances the degradation of isotopically labelled gluten immunogenic peptides in humans Frontiers in Immunology gluten gluten immunogenic peptide celiac disease exopeptidase glutenase 33-mer |
| title | Exopeptidase combination enhances the degradation of isotopically labelled gluten immunogenic peptides in humans |
| title_full | Exopeptidase combination enhances the degradation of isotopically labelled gluten immunogenic peptides in humans |
| title_fullStr | Exopeptidase combination enhances the degradation of isotopically labelled gluten immunogenic peptides in humans |
| title_full_unstemmed | Exopeptidase combination enhances the degradation of isotopically labelled gluten immunogenic peptides in humans |
| title_short | Exopeptidase combination enhances the degradation of isotopically labelled gluten immunogenic peptides in humans |
| title_sort | exopeptidase combination enhances the degradation of isotopically labelled gluten immunogenic peptides in humans |
| topic | gluten gluten immunogenic peptide celiac disease exopeptidase glutenase 33-mer |
| url | https://www.frontiersin.org/articles/10.3389/fimmu.2024.1425982/full |
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