William H. Martin

Isozyme-nonselective N-Substituted Bipiperidylcarboxamide Acetyl-CoA Carboxylase Inhibitors Reduce Tissue Malonyl-CoA Concentrations, Inhibit Fatty Acid Synthesis, and Increase Fatty Acid Oxidation in Cultured Cells and in Experimental Animals

Inhibition of acetyl-CoA carboxylase (ACC), with its resultant inhibition of fatty acid synthesis and stimulation of fatty acid oxidation, has the potential to favorably affect the multitude of cardiovascular risk factors associated with the metabolic syndrome. To achieve maximal effectiveness, an ACC inhibitor should inhibit both the lipogenic tissue isozyme (ACC1) and the oxidative tissue isozyme (ACC2). Herein, we describe the biochemical and acute physiological properties of CP-610431, an isozyme-nonselective ACC inhibitor identified through high throughput inhibition screening, and CP-640186, an analog with improved metabolic stability. CP-610431 inhibited ACC1 and ACC2 with IC50s of ∼50 nm. Inhibition was reversible, uncompetitive with respect to ATP, and non-competitive with respect to bicarbonate, acetyl-CoA, and citrate, indicating interaction with the enzymatic carboxyl transfer reaction. CP-610431 also inhibited fatty acid synthesis, triglyceride (TG) synthesis, TG secretion, and apolipoprotein B secretion in HepG2 cells (ACC1) with EC50s of 1.6, 1.8, 3.0, and 5.7 μm, without affecting either cholesterol synthesis or apolipoprotein CIII secretion. CP-640186, also inhibited both isozymes with IC50sof ∼55 nm but was 2–3 times more potent than CP-610431 in inhibiting HepG2 cell fatty acid and TG synthesis. CP-640186 also stimulated fatty acid oxidation in C2C12 cells (ACC2) and in rat epitrochlearis muscle strips with EC50s of 57 nm and 1.3 μm. In rats, CP-640186 lowered hepatic, soleus muscle, quadriceps muscle, and cardiac muscle malonyl-CoA with ED50s of 55, 6, 15, and 8 mg/kg. Consequently, CP-640186 inhibited fatty acid synthesis in rats, CD1 mice, and ob/ob mice with ED50s of 13, 11, and 4 mg/kg, and stimulated rat whole body fatty acid oxidation with an ED50 of ∼30 mg/kg. Taken together, These observations indicate that isozyme-nonselective ACC inhibition has the potential to favorably affect risk factors associated with the metabolic syndrome.

CP-481,715, a Potent and Selective CCR1 Antagonist with Potential Therapeutic Implications for Inflammatory Diseases

The chemokines CCL3 and CCL5, as well as their shared receptor CCR1, are believed to play a role in the pathogenesis of several inflammatory diseases including rheumatoid arthritis, multiple sclerosis, and transplant rejection. In this study we describe the pharmacological properties of a novel small molecular weight CCR1 antagonist, CP-481,715 (quinoxaline-2-carboxylic acid [4(R)-carbamoyl-1(S)-(3-fluorobenzyl)-2(S),7-dihydroxy-7-methyloctyl]amide). Radiolabeled binding studies indicate that CP-481,715 binds to human CCR1 with a Kd of 9.2 nm and displaces 125I-labeled CCL3 from CCR1-transfected cells with an IC50 of 74 nm. CP-481,715 lacks intrinsic agonist activity but fully blocks the ability of CCL3 and CCL5 to stimulate receptor signaling (guanosine 5′-O-(thiotriphosphate) incorporation; IC50 = 210 nm), calcium mobilization (IC50 = 71 nm), monocyte chemotaxis (IC50 = 55 nm), and matrix metalloproteinase 9 release (IC50 = 54 nm). CP-481,715 retains activity in human whole blood, inhibiting CCL3-induced CD11b up-regulation and actin polymerization (IC50 = 165 and 57 nm, respectively) on monocytes. Furthermore, it behaves as a competitive and reversible antagonist. CP-481,715 is >100-fold selective for CCR1 as compared with a panel of G-protein-coupled receptors including related chemokine receptors. Evidence for its potential use in human disease is suggested by its ability to inhibit 90% of the monocyte chemotactic activity present in 11/15 rheumatoid arthritis synovial fluid samples. These data illustrate that CP-481,715 is a potent and selective antagonist for CCR1 with therapeutic potential for rheumatoid arthritis and other inflammatory diseases.

New quinoline NK3 receptor antagonists with CNS activity.

Lead optimisation starting from the previously reported selective quinoline NK(3) receptor antagonists talnetant 2 (SB-223412) and 3 (SB-222200) led to the identification of 3-aminoquinoline NK(3) antagonist 10 (GSK172981) with excellent CNS penetration. Investigation of a structurally related series of sulfonamides with reduced lipophilicity led to the discovery of 20 (GSK256471). Both 10 and 20 are high affinity, potent NK(3) receptor antagonists which despite having different degrees of CNS penetration produced excellent NK(3) receptor occupancy in an ex vivo binding study in gerbil cortex.

NanoStore: A Concept for Logistical Improvements of Compound Handling in High-Throughput Screening

Small molecule screening, the systematic encounter of biology space with chemical space, has provoked the emergence of a whole industry that recreates itself by constant iterative improvements to this process. The authors describe an approach to tackle the problem for one of the most time-consuming steps in the execution of a screening campaign, namely, the reformatting of high-throughput screening test compounds from master plates to daughter assay plates used in the execution of the screen. Through an engineered storage procedure, they prepare plates ahead of the screening process with the respective compounds in a ready-to-use format. They show the biological inertness of the method and how it facilitates efficient recovery of compound activity. This uncoupling of normally interconnected processes provides time and compound savings, avoids repeated freeze-thaw cycles of compound solutions, and removes the problems associated with the DMSO sensitivity of certain assays types.

Rational design, synthesis, and X-ray structure of renin inhibitors with extended P1 sidechains

Abstract Structural information from a complex of a tetrapeptide renin inhibitor, CP-85, 339, with human renin, led to the design of inhibitors with extended P1 groups that span the S1-S3 active site pocket. A m-biphenyl sidechain at P1 and glycine at P3 led to a 160X increase in potency over the analogous phenyl compound. A crystal structure of this compound in complex with recombinant human renin (rHR) was solved.

A phase I, double-blind, placebo-controlled, randomized, multiple dose study to evaluate the safety, tolerability, and pharmacokinetics of PF 04447943 in mild-to-moderate Alzheimer's disease subjects on stable donepezil therapy

ing established diets found a combination of nutrients may be more powerful than any single class of nutrients. Brain health related evidence was used to design a whole foods Memory Preservation Nutrition (MPN) program emphasizing synergistic contributions of increasing foods with various properties including anti-oxidant, anti-inflammatory attenuate insulin resistance and/or improve lipid balance. Poly-nutrient interventions for clinical research on AD and MCI were derived from the MPN, including the Memory Preservation Nutrition Supplement Program (MPNSP), and the Nutritional Supplement Combination Therapy (NSCT). The MPNSP consists of a phyto-nutrient powder comprised of 100% organic freeze-dried fruits and vegetables, spices, grains and probiotics; an amalgam of herbs and spices chosen for their reported anti-inflammatory properties; and cod liver oil. The NCST added to MPNSP a DHA-enriched fish oil and 2000 IU of vitamin D. Dr. Jon Valla’s Arizona-based team administered the MPNSP incorporated into standard mouse chow, to triply-transgenic mice known to develop features of AD with age and achieved promising results, especially with regard to improving mitochondrial function. Methods: The NSCT study is an open label Phase 1 clinical trial of a combination of over-the-counter nutritional supplements in 25 cognitively normal healthy older adults over an 8 month period including two months for titration to full dosage of the NCST, which comprises 19 pills. Blood and urine specimens, blood pressure, BMI measurements and FFQs will be taken at baseline, 5 and 8 months. Outcome biomarkers include: inflammation (C-RP, IL-6), blood glucose (HbA1c), lipids, oxidative stress (8OH2dG in urine), homocysteine, CoQ10, creatine and creatinine, neurotransmitters tryptophan and tyrosine, and metabolomic profiles in plasma. For adherence and bioavailability: vit Bs, Vit D, Vit E, Vit K, fatty acids & beta-carotene. Results: Recruitment started Spring 2010. Will report whether subjects were able to ingest 19 pills. Conclusions: Combination nutritional interventions for brain and body health may yield significant knowledge.