A vitamin B12 conjugate of exendin-4 improves glucose tolerance without associated nausea or hypophagia in rodents

Elizabeth G. Mietlicki-Baase; Claudia G. Liberini; Jayme L. Workinger; Ron L. Bonaccorso; Tito Borner; David J. Reiner; Kieran Koch-Laskowski; Lauren E. McGrath; Rinzin Lhamo; Lauren M. Stein; Bart C. De Jonghe; George G. Holz; Christian L. Roth; Robert P. Doyle; Matthew R. Hayes

doi:10.1111/dom.13222

A vitamin B12 conjugate of exendin-4 improves glucose tolerance without associated nausea or hypophagia in rodents

Elizabeth G. Mietlicki-Baase, Claudia G. Liberini, Jayme L. Workinger, Ron L. Bonaccorso, Tito Borner, David J. Reiner, Kieran Koch-Laskowski, Lauren E. McGrath, Rinzin Lhamo, Lauren M. Stein, Bart C. De Jonghe, George G. Holz, Christian L. Roth, Robert P. Doyle, Matthew R. Hayes

Research output: Contribution to journal › Article › peer-review

22 Scopus citations

Abstract

Aims: While pharmacological glucagon-like peptide-1 receptor (GLP-1R) agonists are FDA-approved for treating type 2 diabetes mellitus (T2DM) and obesity, a major side effect is nausea/malaise. We recently developed a conjugate of vitamin B12 (B12) bound to the GLP-1R agonist exendin-4 (Ex4), which displays enhanced proteolytic stability and retention of GLP-1R agonism. Here, we evaluate whether the conjugate (B12-Ex4) can improve glucose tolerance without producing anorexia and malaise. Materials and methods: We evaluated the effects of systemic B12-Ex4 and unconjugated Ex4 on food intake and body weight change, oral glucose tolerance and nausea/malaise in male rats, and on intraperitoneal glucose tolerance in mice. To evaluate whether differences in the profile of effects of B12-Ex4 vs unconjugated Ex4 are the result of altered CNS penetrance, rats received systemic injections of fluorescein-Ex4 (Flex), Cy5-B12 or Cy5-B12-Ex4 and brain penetrance was evaluated using confocal microscopy. Uptake of systemically administered Cy5-B12-Ex4 in insulin-containing pancreatic beta cells was also examined. Results: B12-Ex4 conjugate improves glucose tolerance, but does not elicit the malaise and anorexia produced by unconjugated Ex4. While Flex robustly penetrates into the brain (dorsal vagal complex, paraventricular hypothalamus), Cy5-B12 and Cy5-B12-Ex4 fluorescence were not observed centrally, supporting an absence of CNS penetrance, in line with observed reduction in CNS-associated Ex4 side effects. Cy5-B12-Ex4 colocalizes with insulin in the pancreas, suggesting direct pancreatic action as a potential mechanism underlying the hypoglycaemic effects of B12-Ex4. Conclusion: These novel findings highlight the potential clinical utility of B12-Ex4 conjugates as possible future T2DM therapeutics with reduced incidence of adverse effects.

Original language	English (US)
Pages (from-to)	1223-1234
Number of pages	12
Journal	Diabetes, Obesity and Metabolism
Volume	20
Issue number	5
DOIs	https://doi.org/10.1111/dom.13222
State	Published - May 2018

Keywords

antidiabetic drug
appetite control
drug development
exenatide

ASJC Scopus subject areas

Internal Medicine
Endocrinology, Diabetes and Metabolism
Endocrinology

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10.1111/dom.13222

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Mietlicki-Baase, E. G., Liberini, C. G., Workinger, J. L., Bonaccorso, R. L., Borner, T., Reiner, D. J., Koch-Laskowski, K., McGrath, L. E., Lhamo, R., Stein, L. M., De Jonghe, B. C., Holz, G. G., Roth, C. L., Doyle, R. P., & Hayes, M. R. (2018). A vitamin B12 conjugate of exendin-4 improves glucose tolerance without associated nausea or hypophagia in rodents. Diabetes, Obesity and Metabolism, 20(5), 1223-1234. https://doi.org/10.1111/dom.13222

Mietlicki-Baase, EG, Liberini, CG, Workinger, JL, Bonaccorso, RL, Borner, T, Reiner, DJ, Koch-Laskowski, K, McGrath, LE, Lhamo, R, Stein, LM, De Jonghe, BC, Holz, GG, Roth, CL, Doyle, RP & Hayes, MR 2018, 'A vitamin B12 conjugate of exendin-4 improves glucose tolerance without associated nausea or hypophagia in rodents', Diabetes, Obesity and Metabolism, vol. 20, no. 5, pp. 1223-1234. https://doi.org/10.1111/dom.13222

@article{bc8905ff6f8249559b9e298485107b90,

title = "A vitamin B12 conjugate of exendin-4 improves glucose tolerance without associated nausea or hypophagia in rodents",

abstract = "Aims: While pharmacological glucagon-like peptide-1 receptor (GLP-1R) agonists are FDA-approved for treating type 2 diabetes mellitus (T2DM) and obesity, a major side effect is nausea/malaise. We recently developed a conjugate of vitamin B12 (B12) bound to the GLP-1R agonist exendin-4 (Ex4), which displays enhanced proteolytic stability and retention of GLP-1R agonism. Here, we evaluate whether the conjugate (B12-Ex4) can improve glucose tolerance without producing anorexia and malaise. Materials and methods: We evaluated the effects of systemic B12-Ex4 and unconjugated Ex4 on food intake and body weight change, oral glucose tolerance and nausea/malaise in male rats, and on intraperitoneal glucose tolerance in mice. To evaluate whether differences in the profile of effects of B12-Ex4 vs unconjugated Ex4 are the result of altered CNS penetrance, rats received systemic injections of fluorescein-Ex4 (Flex), Cy5-B12 or Cy5-B12-Ex4 and brain penetrance was evaluated using confocal microscopy. Uptake of systemically administered Cy5-B12-Ex4 in insulin-containing pancreatic beta cells was also examined. Results: B12-Ex4 conjugate improves glucose tolerance, but does not elicit the malaise and anorexia produced by unconjugated Ex4. While Flex robustly penetrates into the brain (dorsal vagal complex, paraventricular hypothalamus), Cy5-B12 and Cy5-B12-Ex4 fluorescence were not observed centrally, supporting an absence of CNS penetrance, in line with observed reduction in CNS-associated Ex4 side effects. Cy5-B12-Ex4 colocalizes with insulin in the pancreas, suggesting direct pancreatic action as a potential mechanism underlying the hypoglycaemic effects of B12-Ex4. Conclusion: These novel findings highlight the potential clinical utility of B12-Ex4 conjugates as possible future T2DM therapeutics with reduced incidence of adverse effects.",

keywords = "antidiabetic drug, appetite control, drug development, exenatide",

author = "Mietlicki-Baase, {Elizabeth G.} and Liberini, {Claudia G.} and Workinger, {Jayme L.} and Bonaccorso, {Ron L.} and Tito Borner and Reiner, {David J.} and Kieran Koch-Laskowski and McGrath, {Lauren E.} and Rinzin Lhamo and Stein, {Lauren M.} and {De Jonghe}, {Bart C.} and Holz, {George G.} and Roth, {Christian L.} and Doyle, {Robert P.} and Hayes, {Matthew R.}",

note = "Funding Information: A portion of this work was presented in abstract form at the 2017 meeting of the Society for the Study of Ingestive Behavior, Montreal, Quebec, Canada. M. R. H. and E. G. M-B. receive funding from Zealand Pharma that was not used in support of these studies. M. R. H. receives funding from Novo Nordisk that was not used in support of these studies. R. P. D. is a scientific advisory board member and receives funding from Xeragenx LLc, St. Louis, Missouri that was used, in part, to support these studies. R. P. D. is a scientific advisory board member of Ichor Therapeutics, Lafayette, New York and of Balchem, New Hampton, New York and receives funds from both that were not used in support of these studies. R. P. D. is the named author of a patent pursuant to this work that is owned by Syracuse University. The authors declare no other competing financial interests or conflicts of interest. R. P. D. and M. R. H. conceived of the project. R. P. D. and M. R. H. conceptualized the hypotheses and designed the experiments. J. L. W., R. L. B. and R. P. D. designed and synthesized the compounds. E. G. M-B., C. G. L., J. L. W., R. L. B., T. B., D. J. R., K. K-L., L. E. M., R. L. and L. M. S. conducted experiments and collected data. E. G. M.-B, C. G. L., T. B., D. J. R., L. M. S., B. C. D. J., G. G. H., R. P. D., and M. R. H. analyzed data. E. G. M-B., C. G. L., T. B., B. C. D. J., R. P. D. and M. R. H. wrote the manuscript. All authors edited the manuscript. R. P. D. and M. R. H. are guarantors for this work. Publisher Copyright: {\textcopyright} 2018 John Wiley & Sons Ltd",

year = "2018",

month = may,

doi = "10.1111/dom.13222",

language = "English (US)",

volume = "20",

pages = "1223--1234",

journal = "Diabetes, Obesity and Metabolism",

issn = "1462-8902",

publisher = "Wiley-Blackwell",

number = "5",

}

TY - JOUR

T1 - A vitamin B12 conjugate of exendin-4 improves glucose tolerance without associated nausea or hypophagia in rodents

AU - Mietlicki-Baase, Elizabeth G.

AU - Liberini, Claudia G.

AU - Workinger, Jayme L.

AU - Bonaccorso, Ron L.

AU - Borner, Tito

AU - Reiner, David J.

AU - Koch-Laskowski, Kieran

AU - McGrath, Lauren E.

AU - Lhamo, Rinzin

AU - Stein, Lauren M.

AU - De Jonghe, Bart C.

AU - Holz, George G.

AU - Roth, Christian L.

AU - Doyle, Robert P.

AU - Hayes, Matthew R.

N1 - Funding Information: A portion of this work was presented in abstract form at the 2017 meeting of the Society for the Study of Ingestive Behavior, Montreal, Quebec, Canada. M. R. H. and E. G. M-B. receive funding from Zealand Pharma that was not used in support of these studies. M. R. H. receives funding from Novo Nordisk that was not used in support of these studies. R. P. D. is a scientific advisory board member and receives funding from Xeragenx LLc, St. Louis, Missouri that was used, in part, to support these studies. R. P. D. is a scientific advisory board member of Ichor Therapeutics, Lafayette, New York and of Balchem, New Hampton, New York and receives funds from both that were not used in support of these studies. R. P. D. is the named author of a patent pursuant to this work that is owned by Syracuse University. The authors declare no other competing financial interests or conflicts of interest. R. P. D. and M. R. H. conceived of the project. R. P. D. and M. R. H. conceptualized the hypotheses and designed the experiments. J. L. W., R. L. B. and R. P. D. designed and synthesized the compounds. E. G. M-B., C. G. L., J. L. W., R. L. B., T. B., D. J. R., K. K-L., L. E. M., R. L. and L. M. S. conducted experiments and collected data. E. G. M.-B, C. G. L., T. B., D. J. R., L. M. S., B. C. D. J., G. G. H., R. P. D., and M. R. H. analyzed data. E. G. M-B., C. G. L., T. B., B. C. D. J., R. P. D. and M. R. H. wrote the manuscript. All authors edited the manuscript. R. P. D. and M. R. H. are guarantors for this work. Publisher Copyright: © 2018 John Wiley & Sons Ltd

PY - 2018/5

Y1 - 2018/5

N2 - Aims: While pharmacological glucagon-like peptide-1 receptor (GLP-1R) agonists are FDA-approved for treating type 2 diabetes mellitus (T2DM) and obesity, a major side effect is nausea/malaise. We recently developed a conjugate of vitamin B12 (B12) bound to the GLP-1R agonist exendin-4 (Ex4), which displays enhanced proteolytic stability and retention of GLP-1R agonism. Here, we evaluate whether the conjugate (B12-Ex4) can improve glucose tolerance without producing anorexia and malaise. Materials and methods: We evaluated the effects of systemic B12-Ex4 and unconjugated Ex4 on food intake and body weight change, oral glucose tolerance and nausea/malaise in male rats, and on intraperitoneal glucose tolerance in mice. To evaluate whether differences in the profile of effects of B12-Ex4 vs unconjugated Ex4 are the result of altered CNS penetrance, rats received systemic injections of fluorescein-Ex4 (Flex), Cy5-B12 or Cy5-B12-Ex4 and brain penetrance was evaluated using confocal microscopy. Uptake of systemically administered Cy5-B12-Ex4 in insulin-containing pancreatic beta cells was also examined. Results: B12-Ex4 conjugate improves glucose tolerance, but does not elicit the malaise and anorexia produced by unconjugated Ex4. While Flex robustly penetrates into the brain (dorsal vagal complex, paraventricular hypothalamus), Cy5-B12 and Cy5-B12-Ex4 fluorescence were not observed centrally, supporting an absence of CNS penetrance, in line with observed reduction in CNS-associated Ex4 side effects. Cy5-B12-Ex4 colocalizes with insulin in the pancreas, suggesting direct pancreatic action as a potential mechanism underlying the hypoglycaemic effects of B12-Ex4. Conclusion: These novel findings highlight the potential clinical utility of B12-Ex4 conjugates as possible future T2DM therapeutics with reduced incidence of adverse effects.

AB - Aims: While pharmacological glucagon-like peptide-1 receptor (GLP-1R) agonists are FDA-approved for treating type 2 diabetes mellitus (T2DM) and obesity, a major side effect is nausea/malaise. We recently developed a conjugate of vitamin B12 (B12) bound to the GLP-1R agonist exendin-4 (Ex4), which displays enhanced proteolytic stability and retention of GLP-1R agonism. Here, we evaluate whether the conjugate (B12-Ex4) can improve glucose tolerance without producing anorexia and malaise. Materials and methods: We evaluated the effects of systemic B12-Ex4 and unconjugated Ex4 on food intake and body weight change, oral glucose tolerance and nausea/malaise in male rats, and on intraperitoneal glucose tolerance in mice. To evaluate whether differences in the profile of effects of B12-Ex4 vs unconjugated Ex4 are the result of altered CNS penetrance, rats received systemic injections of fluorescein-Ex4 (Flex), Cy5-B12 or Cy5-B12-Ex4 and brain penetrance was evaluated using confocal microscopy. Uptake of systemically administered Cy5-B12-Ex4 in insulin-containing pancreatic beta cells was also examined. Results: B12-Ex4 conjugate improves glucose tolerance, but does not elicit the malaise and anorexia produced by unconjugated Ex4. While Flex robustly penetrates into the brain (dorsal vagal complex, paraventricular hypothalamus), Cy5-B12 and Cy5-B12-Ex4 fluorescence were not observed centrally, supporting an absence of CNS penetrance, in line with observed reduction in CNS-associated Ex4 side effects. Cy5-B12-Ex4 colocalizes with insulin in the pancreas, suggesting direct pancreatic action as a potential mechanism underlying the hypoglycaemic effects of B12-Ex4. Conclusion: These novel findings highlight the potential clinical utility of B12-Ex4 conjugates as possible future T2DM therapeutics with reduced incidence of adverse effects.

KW - antidiabetic drug

KW - appetite control

KW - drug development

KW - exenatide

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A vitamin B12 conjugate of exendin-4 improves glucose tolerance without associated nausea or hypophagia in rodents

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