Day 1 - Thursday, November 6, 2007

7:00

Registration & Breakfast

7:55

Welcome and Opening Remarks: Chairman, Ming Wang, Amgen

[KEYNOTE PRESENTATION]

8:00

Adipokines, Myokines and Cardiovascular Disease

Kenneth Walsh, Ph.D., Professor of Medicine, Director, Whitaker Cardiovascular Institute, Boston University Medical Campus

It is recognized that obesity contributes to cardiovascular and metabolic disorders through alterations in the levels of adipocyte-derived cytokines (adipokines). Adiponectin is an adipokine that is down-regulated in obese individuals. We have found that adiponectin has beneficial actions on the cardiovascular system by directly acting on the heart and blood vessels, and acute administration of adiponectin can minimize tissue damage resulting from myocardial infarction. More recently, we have been attempting to identify novel adiponectin-like factors involved in metabolic and cardiovascular regulation. Activation of Akt, a protein kinase involved in cell signaling, has been implicated in the control of skeletal muscle hypertrophy. Using an experimental mouse model, we have demonstrated that substantial increases in muscle fiber hypertrophy, weight and strength occur upon induction of Akt signaling in skeletal muscle. In a mouse model of obesity, the increase in muscle mass caused by myogenic Akt induction results in diminished fat deposition and improvements in whole body metabolism. Based on these findings we further devised a protocol to identify novel muscle-secreted proteins (myokines) that confer the phenotypic changes brought on by myogenic Akt induction. One of these newly discovered factors, referred to a follistatin-like 1, is able to promote revascularization in ischemic limbs and protect the heart from ischemic stress.

Session I - Metabolic Syndrome

9:00

Vaspin, A Novel Adipokine with Insulin-Sensitizing Effects

Jun Wada, M.D., Assistant Professor, Okayama University Graduate School of Medicine

Differential screening of the genes up-regulated in visceral adipose tissues of obese OLETF rats and down-regulated in non-obese and diabetes-resistant LETO rats led to the identification of vaspin (visceral adipose tissue-derived serine protease inhibitor), which is a member of serine protease inhibitor (serpin) gene family. Vaspin mRNA is exclusively expressed in visceral adipose tissue in genetically obese rats and also expressed in white adipose tissues of human and its expression is correlated with BMI and insulin sensitivity. Recombinant vaspin-treated diet-induced obesity and db/db mice, and AP2-promotor driven vaspin Tg mice revealed improved insulin sensitivity. The body weight of Tg mice was ~8% less than wild type (WT) mice under HF diet. And serum leptin levels were significantly lower in Tg mice (2.7±2.9 ng/ml) compared with WT mice (10.9±6.4 ng/ml). Triglyceride accumulation in the liver is diminished in Tg mice rather than in WT mice. Collectively, vaspin improves insulin sensitivity by acting on insulin target organs and may be a new molecular target in the treatment of metabolic syndrome.

Conference delegates will learn that:
- Various adipokines differentially affect the insulin actions; TNF-a, resistin and RBP4 (retinol binding protein-4) cause insulin resistance, while leptin and adiponectin enhance the insulin sensitivity.
-  Vaspin is unique adipokine exclusively expressed in white adipose tissues.
- Vaspin improves the insulin sensitivity, obesity and fatty liver in metabolic syndrome.
- Vaspin is a new molecular target in the treatment of metabolic syndrome.

9:30

SCD1 as Therapeutic Target for Metabolic Syndrome

Jian Wang, Ph. D., Principle Research Scientist, Eli Lilly and Company

Stearoyl-CoA Desaturase (SCD) catalyzes the formation of monounsaturated fatty acids from saturated fatty acids. It plays a key role in lipid metabolism and energy expenditure in mammals. Studies in SCD1 null mice and with SCD1 antisense oligo suggest that targeting SCD1 could have potential benefit for dyslipidemia, insulin resistant and obesity. Four SCD isoforms, SCD1, SCD2, SCD3 and SCD4, have been identified in the mice. In contrast, only two SCD isoforms, SCD1 and SCD5 have identified in humans. Orthologue for the four SCD isoforms were also identified in rats and hamsters, suggesting a common evolution origin of these SCD isoforms in the rodents. A SCD inhibitor with therapeutic potential would have to selective to SCD but not inhibiting the other fatty acid desaturases. Animal models have been developed to demonstrate the in vivo target inhibition effect of these SCD inhibitors. SCD inhibition in animal models of obesity and dislipidemia will also be discussed.

Attendees will learn about:
- SCD expression and regulation
- Role of SCD in lipid metabolism
- SCD as therapeutic target for metabolic disease
- SCD1 inhibitors

10:00

Networking & Refreshment Break

Session II: - Obesity: New Therapeutic Targets and Drug Development

10:30

Appetite and Weight Regulation by Peripheral TrkB Agonists

John Lin, Ph.D., Senior Director, Neuroscience, Pfizer

Loss of function mutations in the brain derived neurotrophic factor (BDNF) or tyrosine kinase receptor TrkB result in hyperphagia and obesity in human and rodents. Conversely, peripheral or central stimulation of TrkB by its natural ligands BDNF or NT4 reduced body weight and food intake in mice, supporting the idea that TrkB is a key anorexigenic signal. Here we show that in non-human primates TrkB agonists were anorexigenic when applied centrally, but unexpectedly orexigenic, leading to gain in appetite, body weight, fat deposits and serum leptin levels, when given peripherally. The orexigenic and pro-obesity effects of peripherally administered TrkB agonists appear to be dose dependent, not associated with fluid retention nor with evidence of receptor down regulation. Our findings indicate that TrkB signaling exerts dual control on energy homeostasis in the primates that could be targeted for the treatment of both wasting disorders and obesity.

11:00

Using the Transparent Zebrafish Animal Model for High Throughput Screening to Identify Non-toxic Molecules for Consideration in the Treatment of Clinical Obesity

W. Todd Penberthy, Ph. D., Assistant Professor, Genome Research Institute, University of Cincinnati

Amenable to placement into a 96 well plate, zebrafish larva has emerged as the highest throughput vertebrate animal model used for small molecule screens today! Inherent toxicity is quickly realized, thus eliminating toxic molecules early in the drug development pipeline. While mostly known as a co-factor, nicotinamide adenine dinucleotide (NAD) is in fact also used as a substrate in three categorical reactions: (1) ADP ribosyltransferase reactions, (2) deacetylation reactions, and (3) ADP cyclase reactions. The most studied representative for each of these classes is PARP-1, SIRT-1, and CD38 respectively, where SIRT-1 is the one of greatest focus for exploratory drug development in treatment of metabolic disease. Absence of CD38 however, leads to a situation where NAD levels are dramatically increased in the mouse leading to increased mitochondrial biogenesis through SIRT-1 activation. Amazingly, this mouse is completely resistant to high fat diet-induced obesity due to increased signaling: NAD-SIRT1-PGC1alpha-mitochondrial biogenesis. Thus, activators of SIRT1 and inhibitors of CD38 are being considered as potential drug targets for future development. Here we describe our work developing the zebrafish as a transparent animal model based assay for visualizing changes in total lipid content in response to treatment with NAD-centric pharmacotherapeutics.

Benefits:
(1) Visually introduces the advantages and utility of the zebrafish as a model organism for drug development.
(2) Describes the use of a novel animal model assay for identifying non-toxic molecules favorably altering lipid metabolism.
(3) Describes novel biochemical pathways for future drug development.
(4) Describes important biochemistry centered about NAD.

11:30

Dysregulation of the Endocannabinoid System with Obesity and Insulin Resistance; Evidence of a Role for CB-1 Receptor in Peripheral Metabolism

Tara D'Eon, Ph.D., Scientific Advisor, Metabolism Medical Team, Sanofi-Aventis

Growing evidence suggest the Endocannabinoid System (ECS) plays a significant role in energy balance, and that its involvement extends beyond regulation of food intake. Elevated central (hypothalamic) ECs induce food intake which can be prevented by Cannabinoid 1 receptor (CB1) antagonist, rimonabant. Obesity is associated with elevated EC levels; not only just in central tissues, but also in adipose tissue, liver, and muscle, suggesting further regulation of energy metabolism in metabolically active peripheral tissue. To date, there is little understanding of why ECs become elevated and/or the relative significance of ECs in central versus peripheral tissues.

In my presentation, I will provide a topical overview of what we know of the ECS with regard to peripheral tissues and one of the mechanisms by which ECs become elevated with obesity/insulin resistance (D’Eon et al. Diabetes 2008). In addition, I will discuss clinical data in over 15,000 patients following treatment with CB1 receptor antagonist, rimonabant.

12:00

Lunch

1:30

Constitutive Activation of AMPK-alpha1 in Liver Prevents High-Fat Diet-Induced Obesity in Mice

Jian Yang, Ph.D., Assistant Professor, Department of Physiology, University of South Alabama College of Medicine

To assess the metabolic effects of chronic activation of AMP-activated protein kinase (AMPK) in liver, we generated a new transgenic (Tg) mouse model expressing constitutively active (CA)-AMPK-alpha1 in liver. In the short-term activation, the TgCA-AMPK-α1 mice exhibited minimal phenotype, but the Tg liver had elevated sterol regulatory element-binding protein (SREBP)-2 mRNA level and a parallel increase in transcripts of its target genes. UCP2 mRNA level was elevated. In the long-term activation, the TgCA-AMPK-α1 mice had markedly reduced white fat mass. The Tg liver had reduced mRNA expression of SREBP-1c and its target genes. Remarkably, the Tg mice were resistant to a high-fat diet-induced obesity. These data suggest that short-term chronic activation of AMPK-α1 in liver leads to compensatory increase in lipogenic gene expression due to increased SREBP-2 expression, and long-term chronic activation of AMPK-α1 decreases expression of SREBP-1c and its target genes, which results in reduced fat storage.

This talk will cover:
• AMPK
• Regulation of hepatic lipogenic gene expression
• Animal models for constitutive activation of AMPK in liver
• The consequence of chronic activation of AMPK in liver

2:00

Scaffold Hopping by Shape Matching - Experiences in the CB1 Antagonist Project

Jonas Boström, Ph.D., Assoc. Professor, AstraZeneca R&D

A scaffold hopping approach has been exploited to design a novel class of cannabinoid (CB1) receptor antagonists for the treatment of obesity. On the basis of shape-complementarity and synthetic feasibility the central fragment, a methylpyrazole, in Rimonabant was replaced by a pyrazine. The design, synthesis and CB1 antagonistic activities of a new series of 5,6-diaryl-pyrazine-2-amide derivatives will be described.

2:30

Antagonist of Nicotinic Receptors is a New Agent for Treatment of Obesity

Olga Taraschenko, M.D., Ph.D., Postdoctoral Fellow, Department of Neuropharmacology and Neuroscience, Albany Medical College

18-Methoxycoronaridine (18-MC), a selective antagonist of alpha3beta4 nicotinic receptors, has been previously shown to reduce the self-administration of several drugs of abuse and reduce operant responding for sucrose in rats. In the present experiments the acute administration of 18-MC (10-40 mg/kg i.p.) reduced ad libitum ingestion of sucrose and saccharin. In rats having unlimited access to sucrose (30 %), chronic treatment with 18-MC (20 mg/kg i.p.) prevented sucrose-induced increases in body weight, decreased fat deposition, and reduced consumption of sucrose while not altering food intake. In rats consuming a high fat diet, 18-MC (20 mg/kg i.p. for 14 days) reduced weight gain and fat deposition. The metabolic effects of such treatment are being investigated. These data suggest that α3β4 nicotinic receptors may participate in maintaining obesity, possibly being a new and important target for anti-obesity agents.

Attendees will benefit from:
1. Introduction of 18-methoxycoronaridine (18-MC) - an iboga alkaloid congener
2. 18-MC's pharmacological properties
3. 18-MC's mechanism(s) of action
4. Assessment of 18-MC's efficacies to decrease consumption of sweet substances and to reduce weight gain in animal models of obesity

3:00

Networking & Refreshment Break

Session III - Diabetes: New Therapeutic Targets - Part I

3:30

Antidiabetic Drugs: What Has Worked and What’s in the Making?

Ming Wang, Ph.D., Scientific Director, Metabolic Disorders Research, Amgen, Inc.

Currently there are multiple treatment options for type 2 diabetes and related cardiovascular complications. However, none of the existing therapies or any combination therapy is capable of completely mitigating insulin resistance and hyperglycemia. Further improvement of insulin sensitivity and glycemic control remains a challenge. This presentation will examine the attributes of the existing antidiabetic therapies and identify unmet medical needs. New targets and strategies to meet these needs will be discussed and examples of drug discovery programs will be presented in the context of future challenges.

4:00

Targeting JNK for Diabetes

Ruojing Yang, Ph.D., Senior research biologist, Metabolic disorders, Merck & Co. Inc.

Type 2 diabetes develops from insulin resistance and has become a worldwide epidemic. Clinical evidence indicates a strong correlation between insulin resistance and abnormal inflammatory responses. The c-Jun N-terminal kinases (JNK) can be activated by inflammatory cytokines leading to insulin resistance. Genetic disruption of c-Jun N-terminal kinase-1 (JNK1) gene prevents the development of insulin resistance in obese and diabetic mice. Inhibition of c-Jun N-terminal kinases by a small cell-permeable peptide improves insulin sensitivity in mice. Using adenovirus-mediated delivery of short-hairpin RNA (shRNA), we specifically and efficiently knock down hepatic JNK1 gene expression and observe significant reduction of blood glucose and insulin levels and enhanced hepatic insulin signaling in diet-induced obese mice. Unexpectedly, plasma triglyceride levels were robustly elevated upon hepatic JNK1 knockdown in concomitant with increased expression of transcription co-activator PGC-1beta. Several lines of evidence suggest that hepatic JNK pathway plays an important role in lipid and lipoprotein homeostasis in mice. Furthermore, knockdown of JNK1 significantly increases hepatic phosphorylation of eukaryotic translation initiation factor 2, subunit 1 alpha, (eIF2alpha) and ER chaperone protein GRP78 expression. Significant progress has been achieved in understanding the function of the JNK pathway in metabolic control as well as identifying selective JNK small molecule inhibitors. This helps us to better understand the potential of JNK pathway as a target for diabetes.

[Oral Presentation from Outstanding Abstracts]

4:30

Metformin Inhibits Hepatic Lactate Gluconeogenesis in Zucker Diabetic Fatty ZDF Rats: A Cellular Metabolomic Approach Using Carbon 13 NMR Applicable to Drug Development

Gabriel Baverel, Metabolomics and Metabolic Diseases, INSERM Unit # 820 and Metabolys Inc.

Metformin, one of the most widely used antidiabetic drugs, has been previously shown to decrease hyperglycemia in diabetic subjects and in diabetic animal models. This effect has been shown to result in part from the inhibition of hepatic gluconeogenesis but the mechanism of this effect remains poorly understood. Since lactate is the main gluconeogenic precursor, the objective of this study was to investigate in vitro the effect of metformin on hepatic lactate gluconeogenesis, which contributes to hyperglycemia in diabetes, by using a cellular metabolomic approach. Precision-cut liver slices from adult male ZDF rats were incubated in oxygenated Krebs-Henseleit medium with variously 13C-labeled lactates 5 mM or with 5 mM unlabeled lactate plus 25 mM 13C-bicarbonate. Substrate removal lactate and glycogen and product formation were measured by enzymatic and 13C-NMR spectroscopy methods. In slices from fasted lean ZDF rats, metformin dose-dependently inhibited 13C-glucose synthesis from [2-13C]-lactate IC50 2.3 mM. The labeling patterns of the glucoses synthesized from [1-13C]-, [2-13C]- and [3-13C]-lactate as well as from lactate plus 13C-bicarbonate indicated that lactate gluconeogenesis involved not only the passage of carbons through pyruvate carboxylase and the reversible equilibration of oxaloacetate with fumarate but also pyruvate dehydrogenase and the entire tricarboxylic acid cycle. In slices from fed diabetic ZDF rats, 2 mM metformin inhibited lactate consumption by 97%, glucose production from glycogen plus lactate by 18% and the synthesis of all the 13C-labeled glucose carbons from all 13C-labeled substrates by 70, 70, 71 and 63%, respectively but metformin did not consistently change glycogen degradation. By contrast, metformin increased lactate production by 96% and ketogenesis by 78%. Metformin also caused a reduction of cellular ATP levels and of 13CO2 production from 13C-lactates but increased the beta-hydroxybutyrate/acetoacetate ratio which reflects the mitochondrial redox state. These results show that metformin produced a strong inhibition of hepatic lactate gluconeogenesis that could be partially explained by a more reduced mitochondrial redox state and by a fall of cellular ATP levels.

4:45

3,5-Diiodothyronine T2 Treatment Drastically Prevents Fat Accumulation in High-Fat-Diet-Treated Rats Multi-Organ Involvement of the AMP-Activated Protein Kinase AMPK

Pieter de Lange, Seconda Università degli Studi di Napoli

3,5-diiodo-L-thyronine T2 prevents body weight-gain and fat accumulation when administered to rats receiving a high -fat diet HFD. It induces an increase in hepatic fatty acid FA oxidation concomitant to their less efficient utilization. We investigated whether the effects of T2 in tissues such as liver and white adipose tissue WAT involved the AMP-activated protein kinase AMPK. Three groups of rats were used, either receiving a 6 hour, 1 day, 1 -week or 4-week high fat diet with group HFD-T2, or without group HFD, a daily i.p. injection of T2 25 microg/100 g BW, or a standard diet for the same periods group N. Within 6 hours, T2 increased liver FA oxidation without induction of AMPK phosphorylation. As FA oxidation proceeded, liver AMPK was phosphorylated and as a consequence the expression of genes involved in gluconeogenesis was inhibited, resulting in a higher glucose tolerance. In WAT, between 1 day and 1 week, T2 induced an decrease in AMPK phosphorylation compared to the HFD rats, concomitant with an increase in lipolysis and serum fatty acid levels, which ceases after 4 weeks, possibly because WAT cell volume reached a minimum. At this time-point hepatic lipids were barely present and serum lipids were not increased compared to N controls. In conclusion, treatment with T2 increases hepatic FA utilization which does not depend on AMPK phosphorylation in this tissue, but decreases AMPK phosphorylation in WAT, resulting in increased lipolysis.

5:00

Panel Discussion - Partnering and Licensing

Moderator:

Mike McCully, Deloitte Recap LLC

Panelists:

Vince Wacher, Ph.D., CEO, Verva Pharmaceuticals Ltd.

Robert Knowles, Ph.D., Senior Director, Worldwide Business Development, Pfizer Inc.

David Glass, Ph.D., Director, Technology Transfer, Joslin Diabetes Center

5:45

Poster Session & Networking Reception

Day 2 - Friday, November 7 2007

7:30

Breakfast

[Keynote Presentation]

8:00

How Can We Improve the Treatment of Diabetes and Is There Hope For a Cure?

Allan Ertmann Karlsen, Ph.D., Vice President, Diabetes Biology, Novo Nordisk A/S

Diabetes is a continuously growing health care problem and challenge in the world. In the US alone 24 million people (8% of the population) have diabetes today. Even with the presently available treatments aiming at improving metabolic and blood glucose control, a large number of complications are developed as a result of the disease and a reduced lifespan is still a consequence. Therefore a major focus of diabetes research is still the identification of means to improve the treatment or even intervene or prevent disease development.

This presentation will answer the following questions:
- What are the key challenges in diabetes treatment?
- How can research help?
- GLP-1 as a good example?
- Can we regain the lost beta-cells and cure diabetes?

Session IV - Diabetes: New Therapeutic Targets - Part II

9:00

Small Molecule Ghrelin Receptor Antagonists for the Treatment of Type II Diabetes

Brad Geddes, Ph.D., Senior Director of Drug Discovery, Elixir Pharmaceuticals Inc.

FIt is well established that loss of ghrelin signaling is protective against the negative metabolic effects of chronic high-fat diet (HFD) feeding. For example, GhrR KO mice fed a HFD resist body weight gain and are insulin sensitive relative to WT littermates. We have developed orally bioavailable, small molecule antagonists of the ghrelin receptor (GhrR) for the treatment of type II diabetes. We present data demonstrating that treatment of HFD-fed C57BL6 mice with our GhrR antagonists is able to recapitulate the HFD-fed GhrR KO phenotype. Mice subjected to a long-term (56 day) dose schedule with a GhrR antagonist (60 mg/kg, bid) experienced a significant, 10.3% decrease in body weight. White adipose tissue mass was lower in compound treated mice, with no change in lean tissue mass. In an oral glucose tolerance test, these mice displayed significantly improved insulin sensitivity characterized by a striking reduction in the amount of insulin needed for glucose disposal. The insulin sensitivity of both the GhrR KO mice and GhrR antagonist-treated mice has been confirmed in glycemic clamp assays. It is hoped that this novel, insulin-sparing mechanism of action may also protect the pancreas. In summary, we believe that antagonism of the GhrR is an attractive therapeutic modality for the treatment of metabolic disease in humans.

Attendees will learn about:
1. Ghrelin as a target for type II diabetes
2. The effects of blocking GhrR signaling on insulin sensitivity
3. A novel approach to conducting glycemic clamp assays in pharmacology studies
4. Preclinical efficacy data for a late stage small molecule GhrR antagonist program

9:30

FGF21 and its Metabolic Actions

Alex Kharontonnekov, Research Advisor, BioTechnology Discovery Research, Eli Lilly

Fibroblast growth factor21 (FGF21) is a unique member of FGF family and a pleiotropic metabolic regulator able to rectify multiple abnormal characteristics of metabolic diseases. When administered systemically to animals, FGF21 causes an effective and sustained lowering of plasma glucose and triglycerides, preservation of β-cell function and mass, beneficial changes in LDLc/HDLc and cardiovascular risk factors, amelioration of hepatosteatosis and correction of obesity even though it induces a hyperphagic response in rodents. Of equal importance, the effects of FGF21 in animal models are not associated with side effects such as hypoglycemia, edema, or increased adiposity, highlighting the therapeutic advantage of FGF21 over therapies currently available for diabetic patients and suggesting FGF21 as an effective and safe drug candidate in the context of various metabolic diseases.
While FGF21 was discovered relatively recently, the understanding of its biology as a major metabolic regulator and its therapeutic utility is rapidly evolving. The specifics of FGF21 activities both in cell culture and in vivo, its potential as a therapeutic agent to treat diabetes mellitus and other diseases as well as up-to-date insights into the molecular mechanisms of FGF21 metabolic actions, will be discussed at this talk.

10:00

Networking and Refreshment Break

10:30

Biochemical and Pharmacological Characterizations of Fibroblast Growth Factor 21

Yang Li, Ph.D., Principal Scientist, Amgen, Inc.

Fibroblast growth factor 19 (FGF19) subfamily proteins (FGF19, FGF21, and FGF23) are unique members of fibroblast growth factors (FGFs) that regulate energy, bile acid, glucose, lipid, phosphate, and vitamin D homeostasis in an endocrine fashion. Here, we will discuss our studies both in vivo and in vitro to better understand and characterize the effects of the subfamily members, in particular FGF21, on metabolism as well as their receptor requirements for signaling. We will report our pharmacological studies using recombinant murine FGF21 (mFGF21) in high fat diet-induced obese (DIO) mice. Results show that mFGF21 treatment improved peripheral tissue insulin sensitivity and increased energy metabolism. In addition, we and other have shown that one of the unique features of this subfamily is in their requirement for the presence of either ? or βKlotho as co-receptors for FGFR signaling. We will also present results describing the different specificity of the subfamily members toward ? or βKlotho proteins, as well as the domains/regions on the FGF molecules that are responsible for Klotho co-receptor interactions.

11:00

RBP4 Inhibitors and Insulin Sensitivity in Rodent Models of Diabetes

Ganesh Bhat, Group Leader Pharmacology, Genomics Institute of the Novartis Research Foundation

Insulin resistance is a key factor in the etiology of type 2 diabetes. Serum levels of retinol-binding protein 4 (RBP4), a protein secreted by adipocytes, have been shown to be elevated not only in several mouse models of obesity but also in human subjects who are obese and insulin-resistant or who have type 2 diabetes. The magnitude of this elevation correlates highly with the degree of insulin resistance and is reduced upon intervention. Genetic manipulation of RBP4 in mice also showed strong correlation of circulating RBP4 levels to insulin resistance. These evidences suggest a rationale for antidiabetic therapies aimed at lowering serum RBP4 levels (Nature. 2006 436[2006] 337, NEJM 354 [2006] 2552). Therefore, the goal of this work is to develop RBP4 inhibitors for the treatment of insulin resistance. Precedent for this concept has been established with the synthetic retinoid, fenretinide. Fenretinide disrupts the RBP4-TTR dimmer, which results in RBP4 being excreted in urine. Treatment of diet-induced obese mice (DIO) with fenretinide reduced the serum level of RBP4, resulting in improved insulin sensitivity.
We developed a HTS assay with RBP4/retinol displacement HTRF and a secondary RBP4/TTR dissociation assay to screen >1.5 million compounds. Through HTS and medicinal chemistry efforts, we identified several inhibitors of RBP4 from several chemical series and characterized three compounds, representing two different chemical scaffolds, in mouse PD and insulin resistance models. Fenretinide and GNF RBP4 inhibitor compounds effectively and similarly decreased serum RBP4 levels in DIO-C57Bl and ob/ob mice while rosiglitazone treatment caused a significant increase in RBP4 levels. Both fenretinide and rosiglitazone improved glucose challenge in ob/ob mice. Lowering serum RBP4 levels by GNF compounds did not result in improved glucose tolerance. Our data suggest that pharmacological RBP4 lowering may not be a viable approach to treat insulin resistance and/or diabetes.

Benefits to participants:
-First ever report of RBP4 inhibitors
-Pharmacology of RBP4 inhibitors and relevant animal models
-Target invalidation
-Biomarker vs biomarkers as targets.

11:30

Targeting Inflammation in Type 2 Diabetes: Rationale and Results from the TINSAL Trial

Steven Shoelson, M.D., Ph.D., Professor of Medicine, Head of the Section on Cellular and Molecular Physiology, Harvard Medical School

Obesity is accompanied by a chronic, subacute inflammatory state thought to play roles in the pathogenesis of insulin resistance, type 2 diabetes and cardiovascular disease. Many of our preclinical studies have focused on identifying specific cell types in adipose tissue, liver and the circulation, interactions between different cells types, and relevant intracellular pathways including NF-kB. We have also conducted smaller, 2-4 week clinical trials to determine the effects of targeting inflammation using the salicylate, salsalate, as an NF-kB inhibitor, in patients with diabetes or impaired glucose tolerance. Results were sufficiently promising that we initiated a larger, multicenter clinical trial, referred to as TINSAL-T2D (Targeting Inflammation with Salsalate in Type 2 Diabetes). The first stage of TINSAL-T2D, conducted for 14 weeks using 3 salsalate doses, has been completed and the results, which will be presented, provide proof-of-principle for targeting inflammation to treat diabetes.

12:00

Luncheon

Session V: Drug Development for Diabetes

1:00

Discovery and Development of Liraglutide, The First Once-Daily GLP-1 Analog

Lotte Bjerre Knudsen, Ph.D., Senior Principal Scientist, Diabetes Research Unit, Novo Nordisk A/S

Liraglutide is the first once-daily GLP-1 (Glucagon-Like Peptide-1) analog for the treatment of type 2 diabetes and potentially also obesity. The LEAD (Liraglutide Effect and Action in Diabetes) phase 3 studies have been finalized and an NDA has been submitted in both US, EU and Japan. The presentation will describe the discovery and development of liraglutide.

• Discovery of GLP-1 and the incretin effect
• Lack of incretin effect in type 2 diabetes
• Multiple effects of GLP-1 in the control of blood glucose and body weight
• Importance of pharmacokinetics for GLP-1
• Design of liraglutide
• Preclinical and clinical effects of liraglutide.

1:30

Clinical Pharmacokinetics and Pharmacodynamics of Vildagliptin

Yan-Ling He, Ph.D., Translational Medicine Expert, Novartis Inc.

Vildagliptin (V) is an orally active, potent and selective dipeptidyl-peptidase IV (DPP-4) inhibitor, shown to be effective and well tolerated in patients with type 2 diabetes (T2D) as monotherapy and in combination with other anti-diabetic agents. Following oral dose, V is rapidly and almost completely absorbed (85%). Food had no impact on the pharmacokinetics (PK) of V, suggesting that V can be taken without regard to food. Vildagliptin is minimally bound to plasma proteins (<10%) and distributes extensively into extravascular spaces (Vss=71 L). The primary elimination pathway of V is hydrolysis, while P450 CYP enzymes showed little contribution. Approximately 33% of the total body clearance (41 L/hr) after intravenous dosing can be attributed to renal clearance (13 L/hr). Vildagliptin has a low potential for drug interaction based on its PK characteristics, which was confirmed the by lack of drug interaction with several drugs frequently co-administered to patients with T2D (metformin, glyburide, pioglitazone, amlodipine, valsartan, ramipril, simvastatin, digoxin and warfarin). Exposure to vildagliptin increased by approximately 30% in the elderly and population pharmacokinetic analysis based on phase III data indicates that age has no significant effect on the exposure once the renal function is incorporated in the model. Renal impairment led to approximately two-fold increase in subjects with various degrees of renal impairment, while the increase in exposure to V did not correlated with the severity of renal impairment. Hepatic impairment showed minimal impact on the PK including severe hepatic impairment (<30% change in exposure). Gender, BMI and ethnicity do not have influence on the PK of vildagliptin, indicating that it can be used in a diverse patient population without dose adjustment. Oral administration of vildagliptin to patients with type 2 diabetes resulted in significant inhibition of DPP-4 activity at doses of 10-400 mg. The onset of DPP-4 inhibition was rapid and the duration of DPP-4 inhibition is dose-dependent. Vildagliptin is a potent DPP-4 inhibitor with an EC50 of 4.5 nM in patients with T2D. Vildagliptin significantly increased the intact GLP-1 by approximately 2-fold and GIP levels by approximately 5-fold, and significantly suppressed the postprandial glucagon levels in response to a meal and during an OGTT in patients with T2D. Fasting and postprandial glucose levels were significantly reduced over the dose range of 50-100 mg daily. Although insulin levels in response to a meal were not altered by vildagliptin treatment, insulin secretion was significantly improved when the reduced glucose levels were included in the pharmacodynamic modeling, indicating improvement of beta-cell function. Increased insulin levels were observed during OGTT, indicating stimulation of insulin secretion after V treatment is glucose-dependent. Mechanistically, reduction of endogenous glucose production appears to contribute to the glucose lowering effects. Vildagliptin did not affect the gastric emptying. Long-term clinical trials (Twenty four to 2 year) have demonstrated that vildagliptin is effective and well tolerated in patients with type 2 diabetes as monotherapy or in combination with metformin, pioglitazone or insulin at 50-100 mg daily dose. Vildagliptin and its fixed dose combination with metformin have been approved in >50 countries around the world for treatment of T2D.

2:00

Conference Concludes