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Novel Zn2+ Modulated GPR39 Receptor Agonists Do Not Drive Acute Insulin Secretion in Rodents.

Fjellström O, Larsson N, Yasuda S, Tsuchida T, Oguma T, Marley A, Wennberg-Huldt C, Hovdal D, Fukuda H, Yoneyama Y, Sasaki K, Johansson A, Lundqvist S, Brengdahl J, Isaacs RJ, Brown D, Geschwindner S, Benthem L, Priest C, Turnbull A - PLoS ONE (2015)

Bottom Line: A high throughput screen, followed by a medicinal chemistry program, identified three novel potent Zn2+ modulated GPR39 agonists.The results showed a lack of glucose lowering and insulinotropic effects not only in lean mice, but also in diet-induced obese (DIO) mice and Zucker fatty rats.It is concluded that Zn2+ modulated GPR39 agonists do not acutely stimulate insulin release in rodents.

View Article: PubMed Central - PubMed

Affiliation: Medicinal Chemistry CVMD iMed, AstraZeneca R&D Gothenburg, Mölndal, Sweden.

ABSTRACT
Type 2 diabetes (T2D) occurs when there is insufficient insulin release to control blood glucose, due to insulin resistance and impaired β-cell function. The GPR39 receptor is expressed in metabolic tissues including pancreatic β-cells and has been proposed as a T2D target. Specifically, GPR39 agonists might improve β-cell function leading to more adequate and sustained insulin release and glucose control. The present study aimed to test the hypothesis that GPR39 agonism would improve glucose stimulated insulin secretion in vivo. A high throughput screen, followed by a medicinal chemistry program, identified three novel potent Zn2+ modulated GPR39 agonists. These agonists were evaluated in acute rodent glucose tolerance tests. The results showed a lack of glucose lowering and insulinotropic effects not only in lean mice, but also in diet-induced obese (DIO) mice and Zucker fatty rats. It is concluded that Zn2+ modulated GPR39 agonists do not acutely stimulate insulin release in rodents.

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AZ7914 (A), AZ4237 (B) and AZ1395 (C) ten point concentration response curves for in vitro screens.Concentration responses of compounds were measured with DMR, IP1 and cAMP assays in the absence (-) or presence of 5 μM Zn2+ (+) as indicated in the figure. Lines represent fits to Eq 1. Cells employed were HEK293s-hGPR39 (hGPR39), untransfected HEK293s (HEK), and NIT-1 cells endogenously expressing GPR39. Responses were normalized to AZ1395 in each assay and presented as % effect of control. For HEK293s and HEK293s-hGPR39 cells, individual DMR assays were run in singlicates and IP1 and cAMP assays were run in triplicates. Values shown are means ± SEM of typically three (two to five) independent experiments. For NIT-1 cells, IP1 assays were typically run in duplicates (one to four experiments), and values are shown as means with error bars representing range. EC50 values calculated from the data in Fig 2 are summarized in Table 1.
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pone.0145849.g002: AZ7914 (A), AZ4237 (B) and AZ1395 (C) ten point concentration response curves for in vitro screens.Concentration responses of compounds were measured with DMR, IP1 and cAMP assays in the absence (-) or presence of 5 μM Zn2+ (+) as indicated in the figure. Lines represent fits to Eq 1. Cells employed were HEK293s-hGPR39 (hGPR39), untransfected HEK293s (HEK), and NIT-1 cells endogenously expressing GPR39. Responses were normalized to AZ1395 in each assay and presented as % effect of control. For HEK293s and HEK293s-hGPR39 cells, individual DMR assays were run in singlicates and IP1 and cAMP assays were run in triplicates. Values shown are means ± SEM of typically three (two to five) independent experiments. For NIT-1 cells, IP1 assays were typically run in duplicates (one to four experiments), and values are shown as means with error bars representing range. EC50 values calculated from the data in Fig 2 are summarized in Table 1.

Mentions: EC50 values (μM) and pEC50 ± SD (shown within brackets) in the absence (w/o) or presence of 5 μM Zn2+ in HEK293s cells transfected with human (h), rat (r) and mouse (m) GPR39, and in NIT-1 cells endogenously expressing GPR39. Data is typically from three (two to five) independent experiments. For HEK293s-hGPR39 and NIT-1, calculations were performed on the data shown in Fig 2. Compounds with sufficient potency to reach a plateau in assays employing transfected cells displayed similar efficacy as the AZ1395 100% reference. For compounds not reaching a plateau at highest tested concentration, potencies are shown as >33 μM. For binding affinity measured by BSI, KD values (μM) ± SE and pKD (within brackets) are shown. Data are from three or four independent experiments and calculation performed on the data shown in Fig 3. AZ7914, AZ4237 and AZ1395 were fitted to a one site binding model and Zn2+ to a two site binding model. KD1 and KD2 correspond to high and low affinity sites, respectively.


Novel Zn2+ Modulated GPR39 Receptor Agonists Do Not Drive Acute Insulin Secretion in Rodents.

Fjellström O, Larsson N, Yasuda S, Tsuchida T, Oguma T, Marley A, Wennberg-Huldt C, Hovdal D, Fukuda H, Yoneyama Y, Sasaki K, Johansson A, Lundqvist S, Brengdahl J, Isaacs RJ, Brown D, Geschwindner S, Benthem L, Priest C, Turnbull A - PLoS ONE (2015)

AZ7914 (A), AZ4237 (B) and AZ1395 (C) ten point concentration response curves for in vitro screens.Concentration responses of compounds were measured with DMR, IP1 and cAMP assays in the absence (-) or presence of 5 μM Zn2+ (+) as indicated in the figure. Lines represent fits to Eq 1. Cells employed were HEK293s-hGPR39 (hGPR39), untransfected HEK293s (HEK), and NIT-1 cells endogenously expressing GPR39. Responses were normalized to AZ1395 in each assay and presented as % effect of control. For HEK293s and HEK293s-hGPR39 cells, individual DMR assays were run in singlicates and IP1 and cAMP assays were run in triplicates. Values shown are means ± SEM of typically three (two to five) independent experiments. For NIT-1 cells, IP1 assays were typically run in duplicates (one to four experiments), and values are shown as means with error bars representing range. EC50 values calculated from the data in Fig 2 are summarized in Table 1.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4697807&req=5

pone.0145849.g002: AZ7914 (A), AZ4237 (B) and AZ1395 (C) ten point concentration response curves for in vitro screens.Concentration responses of compounds were measured with DMR, IP1 and cAMP assays in the absence (-) or presence of 5 μM Zn2+ (+) as indicated in the figure. Lines represent fits to Eq 1. Cells employed were HEK293s-hGPR39 (hGPR39), untransfected HEK293s (HEK), and NIT-1 cells endogenously expressing GPR39. Responses were normalized to AZ1395 in each assay and presented as % effect of control. For HEK293s and HEK293s-hGPR39 cells, individual DMR assays were run in singlicates and IP1 and cAMP assays were run in triplicates. Values shown are means ± SEM of typically three (two to five) independent experiments. For NIT-1 cells, IP1 assays were typically run in duplicates (one to four experiments), and values are shown as means with error bars representing range. EC50 values calculated from the data in Fig 2 are summarized in Table 1.
Mentions: EC50 values (μM) and pEC50 ± SD (shown within brackets) in the absence (w/o) or presence of 5 μM Zn2+ in HEK293s cells transfected with human (h), rat (r) and mouse (m) GPR39, and in NIT-1 cells endogenously expressing GPR39. Data is typically from three (two to five) independent experiments. For HEK293s-hGPR39 and NIT-1, calculations were performed on the data shown in Fig 2. Compounds with sufficient potency to reach a plateau in assays employing transfected cells displayed similar efficacy as the AZ1395 100% reference. For compounds not reaching a plateau at highest tested concentration, potencies are shown as >33 μM. For binding affinity measured by BSI, KD values (μM) ± SE and pKD (within brackets) are shown. Data are from three or four independent experiments and calculation performed on the data shown in Fig 3. AZ7914, AZ4237 and AZ1395 were fitted to a one site binding model and Zn2+ to a two site binding model. KD1 and KD2 correspond to high and low affinity sites, respectively.

Bottom Line: A high throughput screen, followed by a medicinal chemistry program, identified three novel potent Zn2+ modulated GPR39 agonists.The results showed a lack of glucose lowering and insulinotropic effects not only in lean mice, but also in diet-induced obese (DIO) mice and Zucker fatty rats.It is concluded that Zn2+ modulated GPR39 agonists do not acutely stimulate insulin release in rodents.

View Article: PubMed Central - PubMed

Affiliation: Medicinal Chemistry CVMD iMed, AstraZeneca R&D Gothenburg, Mölndal, Sweden.

ABSTRACT
Type 2 diabetes (T2D) occurs when there is insufficient insulin release to control blood glucose, due to insulin resistance and impaired β-cell function. The GPR39 receptor is expressed in metabolic tissues including pancreatic β-cells and has been proposed as a T2D target. Specifically, GPR39 agonists might improve β-cell function leading to more adequate and sustained insulin release and glucose control. The present study aimed to test the hypothesis that GPR39 agonism would improve glucose stimulated insulin secretion in vivo. A high throughput screen, followed by a medicinal chemistry program, identified three novel potent Zn2+ modulated GPR39 agonists. These agonists were evaluated in acute rodent glucose tolerance tests. The results showed a lack of glucose lowering and insulinotropic effects not only in lean mice, but also in diet-induced obese (DIO) mice and Zucker fatty rats. It is concluded that Zn2+ modulated GPR39 agonists do not acutely stimulate insulin release in rodents.

Show MeSH
Related in: MedlinePlus