William Riboulet | Université de Lorraine (original) (raw)

Papers by William Riboulet

Nature Medicine, 2014

Obesity and its comorbidities, including type 2 diabetes, represent a global health threat and a ... more Obesity and its comorbidities, including type 2 diabetes, represent a global health threat and a rapidly increasing burden to economic prosperity 1. Therapeutic intervention is urgently required because lifestyle modification has proven mostly ineffective. Despite this vast unmet need, potent and safe pharmacological options that effectively promote weight loss and improve metabolic health have largely remained elusive 2 , partly because many drug interventions historically directed at single molecular targets have exhibited insufficient efficacy or unacceptable safety when used chronically 3. However, new multimolecular therapies have shown enhanced clinical weight loss 4-6 , and singlemolecule peptides integrating the complementary actions of multiple endogenous metabolically-related hormones have emerged as one of the more promising clinical candidates for reversing obesity 7-13. We sought to explore the synergistic metabolic benefits of simultaneous modulation of glucagon, GLP-1 and GIP receptors through a single-molecule hybrid of the three hormones. Glucagon, GLP-1 and GIP are three distinct enteroinsular hormones with unique roles that complement, as well as oppose, each other in the regulation of energy and glucose homeostasis 14-16. Previously, we reported the ability to assemble balanced, high-potency coagonism for the GLP-1 and glucagon receptors (GLP-1R and GcgR, respectively) into a single peptide 8. This peptide exhibited a synergistic ability to lower body weight through coordinated thermogenic and anorectic actions, which can be attributed to the glucagon and GLP-1 components, respectively. Simultaneously, we discovered a high-potency, balanced coagonist for the GLP-1R and GIP receptors (GIPR) 9. This dual incretin coagonist displayed enhanced glycemic efficacy, diminished gastrointestinal toxicity and reduced body weight in preclinical studies, as well as the ability to lower hemoglobin A1C in humans with uncontrolled type 2 diabetes 9. Having established the unique efficacy of these two coagonists with glucagon and GIP independently complementing GLP-1 by different mechanisms, we hypothesized that, if chemically possible, simultaneous and aligned agonism at all three receptors through a single molecule would produce superior therapeutic outcomes. All three hormones are of comparable size and amino acid composition but sufficiently distinct to provide exquisite potency and specificity

In the context of metabolic syndrome, development of Type 2 Diabetes is associated with (and infl... more In the context of metabolic syndrome, development of Type 2 Diabetes is associated with (and influenced by) cardiac and renal comorbidities linked to micro- and macro-vasculature alterations. To assess the efficacy of new compounds on targeted organs in the context of metabolic syndrome, the Zucker fatty/Spontaneously hypertensive heart failure F1 hybrid (ZSF1) rat model could be suitable assuming cardiorenal alterations would develop in a short timeframe. Actually, the ZSF1 rat model recapitulates features of human metabolic syndrome, but develops relatively late (1year-time) and moderate cardiac and renal dysfunctions. The aim of our work was to exacerbate cardiorenal impairments in terms of onset and extent. Two options were explored. On one hand, unilateral nephrectomy was performed in ZSF1 rats, and cardiac and renal functions were longitudinally assessed. This surgical insult only significantly deteriorated renal function, which was prevented by standard of care, lisinopril an...

In the context of metabolic syndrome, development of Type 2 Diabetes is associated with (and infl... more In the context of metabolic syndrome, development of Type 2 Diabetes is associated with (and influenced by) cardiac and renal comorbidities linked to micro- and macro-vasculature alterations. To assess the efficacy of new compounds on targeted organs in the context of metabolic syndrome, the Zucker fatty/Spontaneously hypertensive heart failure F1 hybrid (ZSF1) rat model could be suitable assuming cardiorenal alterations would develop in a short timeframe. Actually, the ZSF1 rat model recapitulates features of human metabolic syndrome, but develops relatively late (1year-time) and moderate cardiac and renal dysfunctions. The aim of our work was to exacerbate cardiorenal impairments in terms of onset and extent. Two options were explored. On one hand, unilateral nephrectomy was performed in ZSF1 rats, and cardiac and renal functions were longitudinally assessed. This surgical insult only significantly deteriorated renal function, which was prevented by standard of care, lisinopril an...

Journal of Experimental and Applied Animal Sciences, Nov 10, 2013

Journal of medicinal chemistry, Jan 2, 2015

Inappropriately high levels of aldosterone are associated with many serious medical conditions, i... more Inappropriately high levels of aldosterone are associated with many serious medical conditions, including renal and cardiac failure. A focused screen hit has been optimized into a potent and selective aldosterone synthase (CYP11B2) inhibitor with in vitro activity against rat, mouse, human, and cynomolgus monkey enzymes, showing a selectivity factor of 160 against cytochrome CYP11B1 in the last species. The novel tetrahydroisoquinoline compound (+)-(R)-6 selectively reduced aldosterone plasma levels in vivo in a dose-dependent manner in db/db mice and cynomolgus monkeys. The selectivity against CYP11B1 as predicted by cellular inhibition data and free plasma fraction translated well to Synacthen challenged cynomolgus monkeys up to a dose of 0.1 mg kg(-1). This compound, displaying good in vivo potency and selectivity in mice and monkeys, is ideally suited to perform mechanistic studies in relevant rodent models and to provide the information necessary for translation to non-human pr...

Nature Medicine, 2014

We report the discovery of a new monomeric peptide that reduces body weight and diabetic complica... more We report the discovery of a new monomeric peptide that reduces body weight and diabetic complications in rodent models of obesity by acting as an agonist at three key metabolically-related peptide hormone receptors: glucagon-like peptide-1 (GLP-1), glucose-dependent insulinotropic polypeptide (GIP) and glucagon receptors. This triple agonist demonstrates supraphysiological potency and equally aligned constituent activities at each receptor, all without cross-reactivity at other related receptors. Such balanced unimolecular triple agonism proved superior to any existing dual coagonists and best-in-class monoagonists to reduce body weight, enhance glycemic control and reverse hepatic steatosis in relevant rodent models. Various loss-of-function models, including genetic knockout, pharmacological blockade and selective chemical knockout, confirmed contributions of each constituent activity in vivo. We demonstrate that these individual constituent activities harmonize to govern the overall metabolic efficacy, which predominantly results from synergistic glucagon action to increase energy expenditure, GLP-1 action to reduce caloric intake and improve glucose control, and GIP action to potentiate the incretin effect and buffer against the diabetogenic effect of inherent glucagon activity. These preclinical studies suggest that, so far, this unimolecular, polypharmaceutical strategy has potential to be the most effective pharmacological approach to reversing obesity and related metabolic disorders.

Type 2 Diabetes (T2D) is recognized by the World Health Organization as a global epidemic affecti... more Type 2 Diabetes (T2D) is recognized by the World Health Organization as a global epidemic affecting 284-347 million people in the world depending on the inclusion criteria used (Danaei et al. 2011; American Diabetes Association 2013). Co-morbidities associated with T2D are hypertension, cardiovascular diseases (CVD) (Castelli 1984; Wilson et al. 2005) and diabetic nephropathy (DN) (Shah et al. 2009; Shao et al. 2013). There is an unmet need for therapies curing or preventing the progression of T2D and combined pathologies (Grundy 2008). In the metabolic diseases field, transgenic or chemically-induced models (mainly rodents) were characterized that focused on one disease or disturbance (i.e. dyslipidemia, T2D, obesity) or pathway abnormalities (insulin resistance, glucose intolerance). The main limitation for the use of those models is the lack of translatability to human T2D and comorbidities. Metabolic syndrome (MS, MetS, Syndrome X) was described years ago as a combination of at ...

Science Translational Medicine, 2013

We report the discovery and translational therapeutic efficacy of a peptide with potent, balanced... more We report the discovery and translational therapeutic efficacy of a peptide with potent, balanced co-agonism at both of the receptors for the incretin hormones glucagon-like peptide-1 (GLP-1) and glucose-dependent insulinotropic polypeptide (GIP). This unimolecular dual incretin is derived from an intermixed sequence of GLP-1 and GIP, and demonstrated enhanced antihyperglycemic and insulinotropic efficacy relative to selective GLP-1 agonists. Notably, this superior efficacy translated across rodent models of obesity and diabetes, including db/db mice and ZDF rats, to primates (cynomolgus monkeys and humans). Furthermore, this co-agonist exhibited synergism in reducing fat mass in obese rodents, whereas a selective GIP agonist demonstrated negligible weight-lowering efficacy. The unimolecular dual incretins corrected two causal mechanisms of diabesity, adiposity-induced insulin resistance and pancreatic insulin deficiency, more effectively than did selective mono-agonists. The duration of action of the unimolecular dual incretins was refined through site-specific lipidation or PEGylation to support less frequent administration. These peptides provide comparable pharmacology to the native peptides and enhanced efficacy relative to similarly modified selective GLP-1 agonists. The pharmacokinetic enhancement lessened peak drug exposure and, in combination with less dependence on GLP-1-mediated pharmacology, avoided the adverse gastrointestinal effects that typify selective GLP-1-based agonists. This discovery and validation of a balanced and high-potency dual incretin agonist enables a more physiological approach to management of diseases associated with impaired glucose tolerance.

Bioconjugate Chemistry, 2013

Recombinant human serum albumin (HSA) conjugates of a 15-amino-acid truncated peptide YY (PYY) an... more Recombinant human serum albumin (HSA) conjugates of a 15-amino-acid truncated peptide YY (PYY) analogue were prepared using three heterobifunctional linkers [succinimidyl 4-[N-maleimidomethyl]cyclohexane-1-carboxylate (SMCC), 6-maleimidohexanoic acid N-hydroxysuccinimide ester (MHS), and N-[γ-maleimidobutyryloxy]sulfosuccinimide ester (GMBS)] in 2 synthetic steps involving (1) reaction of succinimidyl ester on linker with ε-amine of Lys2 on the peptide and (2) reaction of maleimide on peptide linker with free thiol of Cysteine 34 (Cys34) on albumin. In-process controls using ESI LC-MS were used to follow reactions and identify reaction products. Proteolytic digests of the conjugate revealed that peptide conjugation occurs at Cys34 on HSA. Conjugates were assayed in cell-based assays to determine potency at the human Y2-receptor, and selectivity at the human Y1-, Y4-, and Y5-receptors using a calcium flux assay. All three conjugates assayed were selective agonists of the Y2-receptor, and displayed nanomolar potencies. MCC and MH conjugates were selected for acute PK/PD studies in DIO mice. Significant reduction in food intake was observed with the MH conjugate, which lasted for 24 h at the 10 mg (or 4 μmol)/kg dose. While the MCC conjugate exhibited greater potency in vitro, it was slightly less effective than the MH conjugate in vivo with respect to reduction in food intake. Both conjugates were significantly less active than the peptide coupled to a 30 kDa PEG. The observed T1/2 (8-9 h) for both conjugates was significantly lower than that observed for the PEGylated peptide (∼25 h). These results suggest that, as compared with the unmodified and PEGylated peptide, the extended circulation half-life of albumin conjugates is mediated through uptake and recirculation by FcRn, and allometric scaling methods are necessary to account for interspecies variation in pharmacokinetic properties.

Nature Medicine, 2014

We report the discovery of a new monomeric peptide that reduces body weight and diabetic complica... more We report the discovery of a new monomeric peptide that reduces body weight and diabetic complications in rodent models of obesity by acting as an agonist at three key metabolically-related peptide hormone receptors: glucagon-like peptide-1 (GLP-1), glucose-dependent insulinotropic polypeptide (GIP) and glucagon receptors. This triple agonist demonstrates supraphysiological potency and equally aligned constituent activities at each receptor, all without cross-reactivity at other related receptors. Such balanced unimolecular triple agonism proved superior to any existing dual coagonists and best-in-class monoagonists to reduce body weight, enhance glycemic control and reverse hepatic steatosis in relevant rodent models. Various loss-of-function models, including genetic knockout, pharmacological blockade and selective chemical knockout, confirmed contributions of each constituent activity in vivo. We demonstrate that these individual constituent activities harmonize to govern the overall metabolic efficacy, which predominantly results from synergistic glucagon action to increase energy expenditure, GLP-1 action to reduce caloric intake and improve glucose control, and GIP action to potentiate the incretin effect and buffer against the diabetogenic effect of inherent glucagon activity. These preclinical studies suggest that, so far, this unimolecular, polypharmaceutical strategy has potential to be the most effective pharmacological approach to reversing obesity and related metabolic disorders.

Nature Medicine, 2014

Obesity and its comorbidities, including type 2 diabetes, represent a global health threat and a ... more Obesity and its comorbidities, including type 2 diabetes, represent a global health threat and a rapidly increasing burden to economic prosperity 1. Therapeutic intervention is urgently required because lifestyle modification has proven mostly ineffective. Despite this vast unmet need, potent and safe pharmacological options that effectively promote weight loss and improve metabolic health have largely remained elusive 2 , partly because many drug interventions historically directed at single molecular targets have exhibited insufficient efficacy or unacceptable safety when used chronically 3. However, new multimolecular therapies have shown enhanced clinical weight loss 4-6 , and singlemolecule peptides integrating the complementary actions of multiple endogenous metabolically-related hormones have emerged as one of the more promising clinical candidates for reversing obesity 7-13. We sought to explore the synergistic metabolic benefits of simultaneous modulation of glucagon, GLP-1 and GIP receptors through a single-molecule hybrid of the three hormones. Glucagon, GLP-1 and GIP are three distinct enteroinsular hormones with unique roles that complement, as well as oppose, each other in the regulation of energy and glucose homeostasis 14-16. Previously, we reported the ability to assemble balanced, high-potency coagonism for the GLP-1 and glucagon receptors (GLP-1R and GcgR, respectively) into a single peptide 8. This peptide exhibited a synergistic ability to lower body weight through coordinated thermogenic and anorectic actions, which can be attributed to the glucagon and GLP-1 components, respectively. Simultaneously, we discovered a high-potency, balanced coagonist for the GLP-1R and GIP receptors (GIPR) 9. This dual incretin coagonist displayed enhanced glycemic efficacy, diminished gastrointestinal toxicity and reduced body weight in preclinical studies, as well as the ability to lower hemoglobin A1C in humans with uncontrolled type 2 diabetes 9. Having established the unique efficacy of these two coagonists with glucagon and GIP independently complementing GLP-1 by different mechanisms, we hypothesized that, if chemically possible, simultaneous and aligned agonism at all three receptors through a single molecule would produce superior therapeutic outcomes. All three hormones are of comparable size and amino acid composition but sufficiently distinct to provide exquisite potency and specificity

In the context of metabolic syndrome, development of Type 2 Diabetes is associated with (and infl... more In the context of metabolic syndrome, development of Type 2 Diabetes is associated with (and influenced by) cardiac and renal comorbidities linked to micro- and macro-vasculature alterations. To assess the efficacy of new compounds on targeted organs in the context of metabolic syndrome, the Zucker fatty/Spontaneously hypertensive heart failure F1 hybrid (ZSF1) rat model could be suitable assuming cardiorenal alterations would develop in a short timeframe. Actually, the ZSF1 rat model recapitulates features of human metabolic syndrome, but develops relatively late (1year-time) and moderate cardiac and renal dysfunctions. The aim of our work was to exacerbate cardiorenal impairments in terms of onset and extent. Two options were explored. On one hand, unilateral nephrectomy was performed in ZSF1 rats, and cardiac and renal functions were longitudinally assessed. This surgical insult only significantly deteriorated renal function, which was prevented by standard of care, lisinopril an...

In the context of metabolic syndrome, development of Type 2 Diabetes is associated with (and infl... more In the context of metabolic syndrome, development of Type 2 Diabetes is associated with (and influenced by) cardiac and renal comorbidities linked to micro- and macro-vasculature alterations. To assess the efficacy of new compounds on targeted organs in the context of metabolic syndrome, the Zucker fatty/Spontaneously hypertensive heart failure F1 hybrid (ZSF1) rat model could be suitable assuming cardiorenal alterations would develop in a short timeframe. Actually, the ZSF1 rat model recapitulates features of human metabolic syndrome, but develops relatively late (1year-time) and moderate cardiac and renal dysfunctions. The aim of our work was to exacerbate cardiorenal impairments in terms of onset and extent. Two options were explored. On one hand, unilateral nephrectomy was performed in ZSF1 rats, and cardiac and renal functions were longitudinally assessed. This surgical insult only significantly deteriorated renal function, which was prevented by standard of care, lisinopril an...

Journal of Experimental and Applied Animal Sciences, Nov 10, 2013

Journal of medicinal chemistry, Jan 2, 2015

Inappropriately high levels of aldosterone are associated with many serious medical conditions, i... more Inappropriately high levels of aldosterone are associated with many serious medical conditions, including renal and cardiac failure. A focused screen hit has been optimized into a potent and selective aldosterone synthase (CYP11B2) inhibitor with in vitro activity against rat, mouse, human, and cynomolgus monkey enzymes, showing a selectivity factor of 160 against cytochrome CYP11B1 in the last species. The novel tetrahydroisoquinoline compound (+)-(R)-6 selectively reduced aldosterone plasma levels in vivo in a dose-dependent manner in db/db mice and cynomolgus monkeys. The selectivity against CYP11B1 as predicted by cellular inhibition data and free plasma fraction translated well to Synacthen challenged cynomolgus monkeys up to a dose of 0.1 mg kg(-1). This compound, displaying good in vivo potency and selectivity in mice and monkeys, is ideally suited to perform mechanistic studies in relevant rodent models and to provide the information necessary for translation to non-human pr...

Nature Medicine, 2014

We report the discovery of a new monomeric peptide that reduces body weight and diabetic complica... more We report the discovery of a new monomeric peptide that reduces body weight and diabetic complications in rodent models of obesity by acting as an agonist at three key metabolically-related peptide hormone receptors: glucagon-like peptide-1 (GLP-1), glucose-dependent insulinotropic polypeptide (GIP) and glucagon receptors. This triple agonist demonstrates supraphysiological potency and equally aligned constituent activities at each receptor, all without cross-reactivity at other related receptors. Such balanced unimolecular triple agonism proved superior to any existing dual coagonists and best-in-class monoagonists to reduce body weight, enhance glycemic control and reverse hepatic steatosis in relevant rodent models. Various loss-of-function models, including genetic knockout, pharmacological blockade and selective chemical knockout, confirmed contributions of each constituent activity in vivo. We demonstrate that these individual constituent activities harmonize to govern the overall metabolic efficacy, which predominantly results from synergistic glucagon action to increase energy expenditure, GLP-1 action to reduce caloric intake and improve glucose control, and GIP action to potentiate the incretin effect and buffer against the diabetogenic effect of inherent glucagon activity. These preclinical studies suggest that, so far, this unimolecular, polypharmaceutical strategy has potential to be the most effective pharmacological approach to reversing obesity and related metabolic disorders.

Type 2 Diabetes (T2D) is recognized by the World Health Organization as a global epidemic affecti... more Type 2 Diabetes (T2D) is recognized by the World Health Organization as a global epidemic affecting 284-347 million people in the world depending on the inclusion criteria used (Danaei et al. 2011; American Diabetes Association 2013). Co-morbidities associated with T2D are hypertension, cardiovascular diseases (CVD) (Castelli 1984; Wilson et al. 2005) and diabetic nephropathy (DN) (Shah et al. 2009; Shao et al. 2013). There is an unmet need for therapies curing or preventing the progression of T2D and combined pathologies (Grundy 2008). In the metabolic diseases field, transgenic or chemically-induced models (mainly rodents) were characterized that focused on one disease or disturbance (i.e. dyslipidemia, T2D, obesity) or pathway abnormalities (insulin resistance, glucose intolerance). The main limitation for the use of those models is the lack of translatability to human T2D and comorbidities. Metabolic syndrome (MS, MetS, Syndrome X) was described years ago as a combination of at ...

Science Translational Medicine, 2013

We report the discovery and translational therapeutic efficacy of a peptide with potent, balanced... more We report the discovery and translational therapeutic efficacy of a peptide with potent, balanced co-agonism at both of the receptors for the incretin hormones glucagon-like peptide-1 (GLP-1) and glucose-dependent insulinotropic polypeptide (GIP). This unimolecular dual incretin is derived from an intermixed sequence of GLP-1 and GIP, and demonstrated enhanced antihyperglycemic and insulinotropic efficacy relative to selective GLP-1 agonists. Notably, this superior efficacy translated across rodent models of obesity and diabetes, including db/db mice and ZDF rats, to primates (cynomolgus monkeys and humans). Furthermore, this co-agonist exhibited synergism in reducing fat mass in obese rodents, whereas a selective GIP agonist demonstrated negligible weight-lowering efficacy. The unimolecular dual incretins corrected two causal mechanisms of diabesity, adiposity-induced insulin resistance and pancreatic insulin deficiency, more effectively than did selective mono-agonists. The duration of action of the unimolecular dual incretins was refined through site-specific lipidation or PEGylation to support less frequent administration. These peptides provide comparable pharmacology to the native peptides and enhanced efficacy relative to similarly modified selective GLP-1 agonists. The pharmacokinetic enhancement lessened peak drug exposure and, in combination with less dependence on GLP-1-mediated pharmacology, avoided the adverse gastrointestinal effects that typify selective GLP-1-based agonists. This discovery and validation of a balanced and high-potency dual incretin agonist enables a more physiological approach to management of diseases associated with impaired glucose tolerance.

Bioconjugate Chemistry, 2013

Recombinant human serum albumin (HSA) conjugates of a 15-amino-acid truncated peptide YY (PYY) an... more Recombinant human serum albumin (HSA) conjugates of a 15-amino-acid truncated peptide YY (PYY) analogue were prepared using three heterobifunctional linkers [succinimidyl 4-[N-maleimidomethyl]cyclohexane-1-carboxylate (SMCC), 6-maleimidohexanoic acid N-hydroxysuccinimide ester (MHS), and N-[γ-maleimidobutyryloxy]sulfosuccinimide ester (GMBS)] in 2 synthetic steps involving (1) reaction of succinimidyl ester on linker with ε-amine of Lys2 on the peptide and (2) reaction of maleimide on peptide linker with free thiol of Cysteine 34 (Cys34) on albumin. In-process controls using ESI LC-MS were used to follow reactions and identify reaction products. Proteolytic digests of the conjugate revealed that peptide conjugation occurs at Cys34 on HSA. Conjugates were assayed in cell-based assays to determine potency at the human Y2-receptor, and selectivity at the human Y1-, Y4-, and Y5-receptors using a calcium flux assay. All three conjugates assayed were selective agonists of the Y2-receptor, and displayed nanomolar potencies. MCC and MH conjugates were selected for acute PK/PD studies in DIO mice. Significant reduction in food intake was observed with the MH conjugate, which lasted for 24 h at the 10 mg (or 4 μmol)/kg dose. While the MCC conjugate exhibited greater potency in vitro, it was slightly less effective than the MH conjugate in vivo with respect to reduction in food intake. Both conjugates were significantly less active than the peptide coupled to a 30 kDa PEG. The observed T1/2 (8-9 h) for both conjugates was significantly lower than that observed for the PEGylated peptide (∼25 h). These results suggest that, as compared with the unmodified and PEGylated peptide, the extended circulation half-life of albumin conjugates is mediated through uptake and recirculation by FcRn, and allometric scaling methods are necessary to account for interspecies variation in pharmacokinetic properties.

Nature Medicine, 2014

We report the discovery of a new monomeric peptide that reduces body weight and diabetic complica... more We report the discovery of a new monomeric peptide that reduces body weight and diabetic complications in rodent models of obesity by acting as an agonist at three key metabolically-related peptide hormone receptors: glucagon-like peptide-1 (GLP-1), glucose-dependent insulinotropic polypeptide (GIP) and glucagon receptors. This triple agonist demonstrates supraphysiological potency and equally aligned constituent activities at each receptor, all without cross-reactivity at other related receptors. Such balanced unimolecular triple agonism proved superior to any existing dual coagonists and best-in-class monoagonists to reduce body weight, enhance glycemic control and reverse hepatic steatosis in relevant rodent models. Various loss-of-function models, including genetic knockout, pharmacological blockade and selective chemical knockout, confirmed contributions of each constituent activity in vivo. We demonstrate that these individual constituent activities harmonize to govern the overall metabolic efficacy, which predominantly results from synergistic glucagon action to increase energy expenditure, GLP-1 action to reduce caloric intake and improve glucose control, and GIP action to potentiate the incretin effect and buffer against the diabetogenic effect of inherent glucagon activity. These preclinical studies suggest that, so far, this unimolecular, polypharmaceutical strategy has potential to be the most effective pharmacological approach to reversing obesity and related metabolic disorders.