Summary of <i>in vivo</i> studies performed in mice.

<p>Summary of <i>in vivo</i> studies performed in mice.</p

Efficacy of PTM in non-human primates (NHPs).

<p>A–C: Inhibition of <i>de novo</i> lipogenesis (DNL) by PTM in lean cynomolgus monkeys (A) (n = 5 for vehicle and 4 for PTM) and rhesus (B) (n = 4), and lean aged rhesus monkey (C) (n = 6). Animals were dosed with PTM (60 mpk BID p.o. in A and B, 20 and 60 mpk BID s.c. in C and blood samples were collected at 24 hrs post dosing. D: Effect of PTM on DNL of lean rhesus monkeys (60 mpk, p.o.). E–J: Effect of chronic treatment of PTM on body weight (E), fasting glucose levels for predose (F) and 2 hrs post dose at baseline, day 9 and day 22 (G) and insulin (H–I). PTM was dosed at 60 mpk mixed with yogurt for 28 days. J: Liver fat fraction was determined by MRS imaging at baseline and after 28 days of s.c. dosing of PTM (100 mpk, BID) in lean rhesus monkeys. Bars represent means ± SEM. Asterisks denote statistical significance of treatment group compared to vehicle or baseline. *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001.</p

Effect of Platensimycin on body weight gain and food intake in different mouse models.

<p>A–C: Subchronic treatment of PTM in eDIO mice (40, 125, and 400 μg/h, minipump for 10 days) (n = 8). D–E: Chronic treatment of PTM in <i>db/+</i> mice on high fructose diet (3, 10, 30, and 100 mpk in drinking water for 29 days) (n = 8). F–H: Subchronic treatment of PTM in <i>db/db</i> mice for 16 days (3, 10, and 30 mpk, BID, p.o. for 16 days). Bars represent means ± SEM. Asterisks denote statistical significance of treatment group compared to vehicle group. *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001.</p

Platensimycin inhibits enzymatic activity and expression of fatty acid synthase (FAS) and lipogenic genes.

<p>A: Effect of PTM on FAS enzymatic activity in the liver of eDIO mice treated with PTM (100 mpk, BID, p.o. for 4 days) determined by Malonyl-CoA dependent consumption of NADPH (n = 7–8). B–C: Protein level of FAS in the liver determined by western blot and quantitated by Li-COR. D: Relative RNA levels of FAS, sterol regulatory element-binding transcription factor 1 (SREBP-1c), liver X receptor alpha (LXRα), and carbohydrate-responsive element-binding protein (ChREBP) in the liver determined by quantitative RT-PCR. Bars represent means ± SEM. Asterisks denote statistical significance of treatment group compared to vehicle group. *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001.</p

Parallel comparison of PTM and CPT1 inhibitor on glucose and lipid metabolism in perfused liver of lean C57BL/6 mice.

<p>PTM and CPT1 inhibitor are used at 100 μM in the perfusion media (n = 3–5 per group). Bars represent means ± SEM. Asterisks denote statistical significance of treatment group compared to control. *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001.</p

Improved Stability of Proline-Derived Direct Thrombin Inhibitors through Hydroxyl to Heterocycle Replacement

Modification of the previously disclosed (<i>S</i>)-<i>N</i>-(2-(aminomethyl)-5-chlorobenzyl)-1-((<i>R</i>)-2-hydroxy-3,3-dimethylbutanoyl)­pyrrolidine-2-carboxamide <b>2</b> by optimization of the P3 group afforded novel, low molecular weight thrombin inhibitors. Heterocycle replacement of the hydroxyl functional group helped maintain thrombin<i> in vitro</i> potency while improving the chemical stability and pharmacokinetic profile. These modifications led to the identification of compound <b>10</b>, which showed excellent selectivity over related serine proteases as well as <i>in vivo</i> efficacy in the rat arteriovenous shunt. Compound <b>10</b> exhibited significantly improved chemical stability and pharmacokinetic properties over <b>2</b> and may be utilized as a structurally differentiated preclinical tool comparator to dabigatran etexilate (<b>Pro-1</b>) to interrogate the on- and off-target effects of oral direct thrombin inhibitors

Discovery of Chromane Propionic Acid Analogues as Selective Agonists of GPR120 with <i>in Vivo</i> Activity in Rodents

GPR120 (FFAR4) is a fatty acid sensing G protein coupled receptor (GPCR) that has been identified as a target for possible treatment of type 2 diabetes. A selective activator of GPR120 containing a chromane scaffold has been designed, synthesized, and evaluated <i>in vivo</i>. Results of these efforts suggest that chromane propionic acid <b>18</b> is a suitable tool molecule for further animal studies. Compound <b>18</b> is selective over the closely related target GPR40 (FFAR1), has a clean off-target profile, demonstrates suitable pharmacokinetic properties, and has been evaluated in wild-type/knockout GPR120 mouse oGTT studies

Discovery of MK-7725, A Potent, Selective Bombesin Receptor Subtype-3 Agonist for the Treatment of Obesity

Extensive structure–activity relationship studies of a series derived from atropisomer <b>1</b>, a previously described chiral benzodiazepine sulfonamide series, led to a potent, brain penetrant and selective compound with excellent preclinical pharmacokinetic across species. We also describe the utilization of a high throughput mouse pharmacodynamic assay which allowed for expedient assessment of pharmacokinetic and brain distribution

Microscale High-Throughput Experimentation as an Enabling Technology in Drug Discovery: Application in the Discovery of (Piperidinyl)pyridinyl‑1<i>H</i>‑benzimidazole Diacylglycerol Acyltransferase 1 Inhibitors

Miniaturization and parallel processing play an important role in the evolution of many technologies. We demonstrate the application of miniaturized high-throughput experimentation methods to resolve synthetic chemistry challenges on the frontlines of a lead optimization effort to develop diacylglycerol acyltransferase (DGAT1) inhibitors. Reactions were performed on ∼1 mg scale using glass microvials providing a miniaturized high-throughput experimentation capability that was used to study a challenging S_<i>N</i>Ar reaction. The availability of robust synthetic chemistry conditions discovered in these miniaturized investigations enabled the development of structure–activity relationships that ultimately led to the discovery of soluble, selective, and potent inhibitors of DGAT1

Discovery of a Potent and Selective DGAT1 Inhibitor with a Piperidinyl-oxy-cyclohexanecarboxylic Acid Moiety

We report the discovery of a novel series of DGAT1 inhibitors in the benzimidazole class with a piperdinyl-oxy-cyclohexanecarboxylic acid moiety. This novel series possesses significantly improved selectivity against the A_2A receptor, no ACAT1 off-target activity at 10 μM, and higher aqueous solubility and free fraction in plasma as compared to the previously reported pyridyl-oxy-cyclohexanecarboxylic acid series. In particular, <b>5B</b> was shown to possess an excellent selectivity profile by screening it against a panel of more than 100 biological targets. Compound <b>5B</b> significantly reduces lipid excursion in LTT in mouse and rat, demonstrates DGAT1 mediated reduction of food intake and body weight in mice, is negative in a 3-strain Ames test, and appears to distribute preferentially in the liver and the intestine in mice. We believe this lead series possesses significant potential to identify optimized compounds for clinical development