Discovery of a clinical candidate from the structurally unique dioxa-bicyclo[3.2.1]octane class of sodium-dependent glucose cotransporter 2 inhibitors.

Compound 4 (PF-04971729) belongs to a new class of potent and selective sodium-dependent glucose cotransporter 2 inhibitors incorporating a unique dioxa-bicyclo[3.2.1]octane (bridged ketal) ring system. In this paper we present the design, synthesis, preclinical evaluation, and human dose predictions related to 4. This compound demonstrated robust urinary glucose excretion in rats and an excellent preclinical safety profile. It is currently in phase 2 clinical trials and is being evaluated for the treatment of type 2 diabetes.

C-Aryl glycoside inhibitors of SGLT2: Exploration of sugar modifications including C-5 spirocyclization.

Modifications to the sugar portion of C-aryl glycoside sodium glucose transporter 2 (SGLT2) inhibitors were explored, including systematic deletion and modification of each of the glycoside hydroxyl groups. Based on results showing activity to be quite tolerant of structural change at the C-5 position, a series of novel C-5 spiro analogues was prepared. Some of these analogues exhibit low nanomolar potency versus SGLT2 and promote urinary glucose excretion (UGE) in rats. However, due to sub-optimal pharmacokinetic parameters (in particular half-life), predicted human doses did not meet criteria for further advancement.

Characterization of blackberry extract and its antiproliferative and anti-inflammatory properties.

Blackberries are rich in polyphenols, including anthocyanins. Polyphenols are hypothesized to have biological activities that may impact positively on human health. In these studies, an anthocyanin-rich extract from Hull blackberries grown in Kentucky was obtained and fully characterized in terms of total anthocyanin and phenolic content, polymeric color, anthocyanin composition, and total antioxidant capacity. In vitro cell culture studies showed that the blackberry extract inhibited HT-29 colon tumor cell growth in a concentration-dependent manner with 49.2 microg of total anthocyanins/mL inhibiting HT-29 cell growth up to 66% at 72 hours. Likewise, in a concentration-dependent manner, total anthocyanin concentrations in the range of 0-40 microg/mL suppressed both high-dose (10 microg/mL) and low-dose (0.1 microg/mL) lipid A-induced interleukin-12 release from mouse bone marrow-derived dendritic cells. These results suggest that Hull blackberry extract (HBE) has potent antioxidant, antiproliferative, and anti-inflammatory activities and that HBE-formulated products may have the potential for the treatment and/or prevention of cancer and/or other inflammatory diseases.

Preparation and characterization of nickel nanoparticles for binding to his-tag proteins and antigens.

PURPOSE: The purpose of these studies was to prepare nanoparticles (NPs) with a small amount of surface-chelated nickel for obtaining enhanced binding of histidine-tagged (his-tag) proteins compared to non-histidine-tagged protein binding to charged nanoparticles. MATERIALS AND METHODS: NPs were prepared from oil-in-water microemulsion precursors using emulsifying wax, 3 mM Brij 78 and 0.1 mM DOGS-NTA-Ni lipid (referred to as Ni-NPs). The amount of lipid entrapped in the NPs was quantitated by atomic emission spectroscopy (AES). The Ni-NPs were investigated for binding to two his-tag proteins, green fluorescent protein (GFP) and his-tag HIV-1 Gag p24. In vivo studies in mice were carried out to evaluate the immune responses obtained to his-tag Gag p24 bound to Ni-NPs. RESULTS: AES studies demonstrated that approximately 5% of the DOGS-NTA-Ni lipid used was entrapped in the NPs. The optimal binding ratio his-tag GFP and his-tag Gag p24 to Ni-NPs was found to be 1:33.7 and 1:35.4 w/w, respectively. This interaction was stable at 37 degrees C in PBS, pH 7.4 over 4 h and the interaction of his-tag GFP with the Ni-NPs was enhanced compared to control NPs prepared with no Ni on the surface (NTA-NPs). The in vivo studies demonstrated enhanced serum IgG and IgG2a responses to his-tag Gag p24 bound to Ni-NPs compared to protein adjuvanted with Alum or adsorbed on the surface of control NTA-NPs. CONCLUSIONS: Ni-NPs can be used to bind strongly to his-tag proteins. This system was demonstrated to have potential applications in vaccine delivery for enhancing immune responses to protein-based vaccines.