NNC2215 is an insulin analog with glucose-sensitive properties on activation of the insulin receptor. In NNC2215, a glucoside and a macrocycle with strong affinity to glucose were conjugated to insulin. This introduced a switch that is open (activated) or closed (inactivated) at high or low glucose, respectively. The current study investigated the interaction between NNC2215 and paracetamol glucuronide (PaG), a major human metabolite of paracetamol. Interactions between PaG or d-glucose and NNC2215 were investigated using native mass spectrometry and insulin receptor binding studies in vitro. Interactions between PaG or l-glucose and NNC2215 were studied during glucose clamp conditions in domestic pigs. NNC2215 bound >10-fold stronger to PaG than to d-glucose in vitro. At a clinically relevant PaG concentration (70 μmol/L), the binding affinity of NNC2215 to the insulin receptor was increased, particularly at low glucose concentrations. In the clamp experiment, intravenous administration of 2.0 g PaG versus 0.3 g/kg l-glucose led to a similar glucodynamic effect. These data demonstrate a relevant interaction between PaG and NNC2215, potentially introducing hypoglycemia risk with NNC2215 versus nonglucose-sensitive insulin after paracetamol administration. When engineering a glucose-sensitive insulin, it should be considered whether molecules other than glucose can alter the insulin activity.

The risk of inducing hypoglycaemia (low blood glucose) constitutes the main challenge associated with insulin therapy for diabetes1,2. Insulin doses must be adjusted to ensure that blood glucose values are within the normal range, but matching insulin doses to fluctuating glucose levels is difficult because even a slightly higher insulin dose than needed can lead to a hypoglycaemic incidence, which can be anything from uncomfortable to life-threatening. It has therefore been a long-standing goal to engineer a glucose-sensitive insulin that can auto-adjust its bioactivity in a reversible manner according to ambient glucose levels to ultimately achieve better glycaemic control while lowering the risk of hypoglycaemia3. Here we report the design and properties of NNC2215, an insulin conjugate with bioactivity that is reversibly responsive to a glucose range relevant for diabetes, as demonstrated in vitro and in vivo. NNC2215 was engineered by conjugating a glucose-binding macrocycle4 and a glucoside to insulin, thereby introducing a switch that can open and close in response to glucose and thereby equilibrate insulin between active and less-active conformations. The insulin receptor affinity for NNC2215 increased 3.2-fold when the glucose concentration was increased from 3 to 20 mM. In animal studies, the glucose-sensitive bioactivity of NNC2215 was demonstrated to lead to protection against hypoglycaemia while partially covering glucose excursions. NNC2215 is an insulin conjugate that can reversibly adjust its bioactivity in response to a diabetes-relevant glucose range in vivo.

During recent years combining GLP-1 and glucagon receptor agonism with the purpose of achieving superior weight loss and metabolic control compared to GLP-1 alone has received much attention. The superior efficacy has been shown by several in preclinical models but has been difficult to reproduce in humans. In this paper, we present the pre-clinical evaluation of NN1177, a long-acting GLP-1/glucagon receptor co-agonist previously tested in clinical trials. To further investigate the contribution from the respective receptors, two other co-agonists (NN1151, NN1359) with different GLP-1-to-glucagon receptor ratios were evaluated in parallel. In the process of characterizing NN1177, species differences and pitfalls in traditional pre-clinical evaluation methods were identified, highlighting the translational challenges in predicting the optimal receptor balance in humans. In diet-induced obese (DIO) mice, NN1177 induced a dose-dependent body weight loss, primarily due to loss of fat mass, and improvement in glucose tolerance. In DIO rats, NN1177 induced a comparable total body weight reduction, which was in contrast mainly caused by loss of lean mass, and glucose tolerance was impaired. Furthermore, despite long half-lives of the three co-agonists, glucose control during steady state was seen to depend on compound exposure at time of evaluation. When evaluated at higher compound exposure, glucose tolerance was similarly improved for all three co-agonists, independent of receptor balance. However, at lower compound exposure, glucose tolerance was gradually impaired with higher glucagon receptor preference. In addition, glucose tolerance was found to depend on study duration where the effect of glucagon on glucose control became more evident with time. To conclude, the pharmacodynamic effects at a given GLP-1-to-glucagon ratio differs between species, depends on compound exposure and study length, complicating the identification of an optimally balanced clinical candidate. The present findings could partly explain the low number of clinical successes for this dual agonism.

A series of compounds based on the mGluR5-selective ligand 2-methyl-6-(phenylethynyl)pyridine (MPEP) were designed and synthesized. The compounds were found to be either structural analogues of MPEP, substituted monomers, or dimeric analogues. All compounds retained mGluR5 selectivity with only weak or no activity at other mGluRs or iGluRs. The substituted analogue, 1,3-bis(pyridin-2-ylethynyl)benzene (19), is a potent negative modulator at mGluR5, whereas all other compounds lost potency relative to MPEP and showed that activity is highly dependent on the position of the nitrogen atom in the pyridine moieties. A homology modeling and ligand docking study was used to understand the binding mode and the observed selectivity of compound 19.

BACKGROUND AND PURPOSE The peroxisome proliferator‐activated receptor (PPAR)δ has been considered a therapeutic target for diabetes and obesity through enhancement of fatty acid oxidation. The present study aimed to characterize the effects of PPARδ agonists during insulin resistance of the whole body, muscle and liver.EXPERIMENTAL APPROACH Wistar rats and C57BL/J6 mice were fed a high fat diet (HF) and then treated with PPARδ agonists NNC61‐5920 and GW501516. The effects on insulin resistance were evaluated by hyperinsulinaemic clamp or glucose tolerance tests combined with glucose tracers.KEY RESULTS In HF rats, 3 weeks of treatment with NNC61‐5920 reduced the glucose infusion rate (by 14%, P < 0.05) and glucose disposal into muscle (by 20–30%, P < 0.01) during hyperinsulinaemic clamp. Despite increased mRNA expression of carnitine palmitoyltransferase‐1, pyruvate dehydrogenase kinase 4 and uncoupling protein 3 in muscle, plasma and muscle triglyceride levels were raised (P < 0.01). Similar metabolic effects were observed after extended treatment with NNC61‐5920 and GW501516 to 6 weeks. However, HF mice treated with NNC61‐5920 improved their plasma lipid profile, glucose tolerance and insulin action in muscle. In both HF rats and mice, NNC61‐5920 treatment attenuated hepatic insulin resistance and decreased expression of stearoyl‐CoA desaturase 1, fatty acid translocase protein CD36 and lipoprotein lipase in liver.CONCLUSIONS AND IMPLICATIONS PPARδ agonists exacerbated insulin resistance in HF rats in contrast to their beneficial effects on metabolic syndrome in HF mice. These opposing metabolic consequences result from their different effects on lipid metabolism and insulin sensitivity in skeletal muscle of these two species.

L'invention concerne un derive d'un analogue de GPL-1, ledit analogue comprenant un premier residu K a une position correspondant a la position 18 de GLP-1 (7-37) (SEQ ID NO : 1), un second residu K a une autre position, et un maximum de douze modifications d'acides amines en comparaison a GLP-1 (7-37) ; le derive comprenant deux fractions etendues et fixees auxdits premier et second residus K, respectivement, par l'intermediaire d'un liant, la fraction etendue etant choisie parmi Chem. 1, Chem. 2 et Chem.3 : Chem. : HOOC-(CH 2 ) x -CO-* Chem. 2: HOOC-C 6 H 4 -0-(CH 2 ) y -CO-* Chem. 3: R 2 -C 6 H 4 -(CH 2 ) z -CO-*, ou x est un entier se situant dans la plage de 6-18, y est un entier se situant dans la plage de 3-17, z est un entier se situant dans la plage de 1-5, et R 2 est un groupe ayant une masse molaire non superieure a 150 Da ; et le liant comprenant Chem. 4: *-NH-(CH 2 ) 2 -(0-(CH 2 ) 2 ) k -0-(CH 2 ) n -CO-*, ou k est un entier se situant dans la plage de 1-5, et n est un entier se situant dans la plage de 1-5 ; ou un sel, un amide ou un ester pharmaceutiquement acceptable de celui-ci. L'invention concerne egalement son utilisation pharmaceutique, par exemple dans le traitement et/ou la prevention de toutes formes de diabete et de maladie associees, ainsi que des nouveaux peptides et intermediaires de chaine laterale correspondants. Les derives sont adaptes a une administration par voie orale.

The peroxisome proliferator-activated receptors (PPARs) are transcription factors belonging to the nuclear receptor superfamily. Several reports have shown that PPARδ is involved in lipid metabolism, increasing fat oxidation and depleting lipid accumulation. Whether PPARδ is involved in the regulation of glucose metabolism is not completely understood. In this study, we examined effects of long-term PPARδ activation on glycemic control, islet function and insulin sensitivity in diabetic db/db mice. Male db/db mice were administered orally once daily with a selective and partial PPARδ agonist (NNC 61-5920, 30 mg/kg) for eight weeks; control mice received vehicle. Fasting and non-fasting plasma glucose were reduced, reflected in reduced hemoglobinA1c (3.6 ± 1.6% vs. 5.4 ± 1.8 in db/db controls, P < 0.05) and furthermore, the AUCglucose after oral glucose (3 g/kg) was reduced by 67% (P < 0.05) after long-term PPARδ activation. Following intravenous glucose (1 g/kg), glucose tolerance was improved after PPARδ activation (KG 1.3 ± 0.6 vs. − 0.05 ± 0.7 %/min, P = 0.048). Insulin sensitivity, measured as the glucose clearance after intravenous injection of glucose (1 g/kg) and insulin (0.75 or 1.0 U/kg), during inhibition of endogenous insulin secretion by diazoxide (25 mg/kg), was improved (KG 2.9 ± 0.6 vs. 1.3 ± 0.3 %/min in controls, P < 0.05) despite lower insulin levels. Furthermore, islets isolated from PPARδ agonist treated mice demonstrated improved glucose responsiveness as well as improved cellular topography. In conclusion, PPARδ agonism alleviates insulin resistance and improves islet function and topography, resulting in improved glycemia in diabetic db/db mice. This suggests that activation of PPARδ improves glucose metabolism and may therefore potentially be target for treatment of type 2 diabetes.

Balaglitazone is a novel thiazolidinedione in clinical development for the treatment of type 2 diabetes. Common side effects associated with PPARgamma receptor agonists are weight gain, oedema and adipogenesis. Balaglitazone is a selective partial PPARgamma agonist and it has been speculated that such compounds have a more favourable safety margin than full agonists. We have compared impact of equi-efficacious antihyperglycaemic doses of balaglitazone with full PPARgamma agonist rosiglitazone on body fluid accumulation, cardiac enlargement, and adipogenesis. Equi-efficacious antihyperglycaemic doses (ED(90)) of balaglitazone (3 mg/kg/day) and rosiglitazone (6 mg/kg/day) were determined in male diabetic db/db mice. In adult male rats treated for up to 42 days, feeding, drinking, anthropometry, and plasma volumes were measured. Total plasma volume was measured with dye dilution technique. Compared to vehicle, rosiglitazone consistently increased food intake throughout the 42 day treatment period. In contrast, balaglitazone increased food intake in the last week of the experiment. However, both rosiglitazone and balaglitazone increased water intake. After 42 days, rosiglitazone treated rats displayed significantly elevated adiposity. Rosiglitazone increased total blood and plasma volumes throughout the treatment. Twenty-one days of balaglitazone treatment had no significant impact on blood or plasma volumes, whilst 42 days of balaglitazone increased plasma volume but to a significantly lesser extent than seen for rosiglitazone (vehicle: 46.1+/-1.5; balaglitazone: 50.8+/-1.21; rosiglitazone: 54.6+/-1.6 ml/kg). Heart weight was significantly elevated only in rosiglitazone treated animals. At doses inducing comparable antihyperglycaemic control, the full PPARgamma agonist, rosiglitazone, induces more pronounced body fluid retention and heart enlargement than seen for the partial PPARgamma agonist, balaglitazone. Thus, partial agonists may pose safer alternative to current anti-diabetic therapy with full PPARgamma agonist.

Y-shaped molecules bearing alkynylallylic moieties were found to be potent and selective PPAR delta activators. The alkynylallylic moiety was synthesized from alkyn-l-ols by hydroalumination followed by a cross-coupling reaction. Series of active compounds 6 were obtained by stepwise changing the structure of the known PPARpan agonist 5 into Y-shaped compounds. The most active and selective compound, 6f, had a PPAR delta potency of 0.13 mu M, which is 50-fold more potent than compound 5. (c) 2007 Elsevier Ltd. All rights reserved.