Discovery of a novel fibroblast activation protein (FAP) inhibitor, BR103354, with anti-diabetic and anti-steatotic effects.

Fibroblast growth factor (FGF) 21 is a class of hepatokines that plays a protective role against obesity, insulin resistance, and liver damage. Despite this, protective effects of FGF21 in human appear to be minimal, possibly due to its proteolytic cleavage by the fibroblast activation protein (FAP). Here, we presented a novel FAP inhibitor, BR103354, and described its pharmacological activities as a potential therapeutic agent for the treatment of metabolic disorders. BR103354 inhibited FAP with an IC50 value of 14 nM, showing high selectivity against dipeptidyl peptidase (DPP)-related enzymes and prolyl oligopeptidase (PREP). In differentiated 3T3/L1 adipocytes, the addition of FAP diminished hFGF21-induced Glut1 and phosphorylated levels of ERK, which were restored by BR103354. BR103354 exhibited good pharmacokinetic properties as evidenced by oral bioavailability of 48.4% and minimal hERG inhibition. Single co-administration of BR103354 with hFGF21 reduced nonfasting blood glucose concentrations, in association with increased intact form of hFGF21 in ob/ob mice. Additionally, chronic treatment of BR103354 for 4 weeks reduced nonfasting blood glucose concentrations with improved glucose tolerance and with reduced triglyceride (TG) content in liver of ob/ob mice. Consistently, BR103354 improved hepatic steatosis and fibrosis in a choline-deficient, L-amino acid-defined, high-fat diet (CDAHFD)-induced non-alcoholic steatohepatitis (NASH) mouse model. FAP inhibitory effects of BR103354 were confirmed in normal cynomolgus monkeys. Together, BR103354 acts as an effective FAP inhibitor in vitro and in vivo, thereby demonstrating its potential application as an anti-diabetic and anti-NASH agent.

Role of hypothalamic autophagy in the control of whole body energy balance.

Autophagy is a catabolic process involving the rearrangement of subcellular membranes to sequester cytoplasm and organelles for delivery to lysosomes, where the sequestered material is degraded and recycled. Autophagy is important for maintenance of intracellular energy homeostasis and the quality control of organelles such as the endoplasmic reticulum (ER) and mitochondria, which suggests that dysregulated autophagy might play a role in the pathogenesis of metabolic disorders and diabetes. In an attempt to elucidate the role of autophagy in metabolic disorders, diverse in vivo and in vitro models have been employed. Site-specific autophagy knockout models that are autophagy-deficient specifically in pancreatic ß-cells, skeletal muscle, adipose tissues or liver have been produced. These models have generated valuable information regarding the role of autophagy in body metabolism. The role of autophagy in the hypothalamus, which controls whole body energy balance, appetite and energy expenditure, has also been investigated. Thus, mice with autophagy deficiency in the hypothalamus have shown diverse phenotypes (lean vs. obese) depending on the site of autophagy deficiency or the method of autophagy abrogation.

Role of hypothalamic proopiomelanocortin neuron autophagy in the control of appetite and leptin response.

Autophagy is a catabolic cellular process involving the degradation of the cell's own components. Although the role of autophagy of diverse tissues in body metabolism has been investigated, the importance of autophagy in hypothalamic proopiomelanocortin (POMC) neurons, key regulators of energy balance, has not been addressed. The role of autophagy in leptin sensitivity that is critical for the control of body weight and appetite has also not been investigated. We produced mice with specific deletion of autophagy-related 7 (Atg7), an essential autophagy gene, in hypothalamic POMC neurons (Atg7(ΔPOMC) mice). Atg7 expression was deficient in the arcuate nucleus of the hypothalamus of Atg7(ΔPOMC) mice. p62, a specific substrate of autophagy, accumulated in the hypothalamus of Atg7(ΔPOMC) mice, which colocalized with ubiquitin. Atg7(ΔPOMC) mice had increased body weight due to increased food intake and decreased energy expenditure. Atg7(ΔPOMC) mice were not more prone to diet-induced obesity compared with control mice but more susceptible to hyperglycemia after high-fat diet. The ability of leptin to suppress fasting-elicited hyperphagia and weight gain during refeeding was attenuated in Atg7(ΔPOMC) mice. Deficient autophagy did not significantly affect POMC neuron number but impaired leptin-induced signal transducer and activation of transcription 3 activation. Our findings indicate a critical role for autophagy of POMC neurons in the control of energy homeostasis and leptin signaling.

Anti-allergic prenylated flavonoids from the roots of Sophora flavescens.

The bioassay-guided fractionation of the MeOH extract from the root of Sophora flavescens led to the isolation of eight known prenylated flavonoids ( 1 - 8) responsible for the IN VITRO anti-allergic activity. Among them, kushenol N ( 3), sophoraflavanone G ( 6), and leachianone A ( 7) demonstrated significant inhibition of the release of beta-hexosaminidase from cultured RBL-2H3 cells with IC (50) values ranging from 15 to 30 muM.