Chemical,Pharmacological, and in vitro Metabolic Stability Studies on Enantiomerically Pure RC‐33 Compounds: Promising Neuroprotective Agents Acting as σ1 Receptor Agonists

Our recent research efforts identified racemic RC‐33 as a potent and metabolically stable σ₁ receptor agonist. Herein we describe the isolation of pure RC‐33 enantiomers by chiral chromatography, assignment of their absolute configuration, and in vitro biological studies in order to address the role of chirality in the biological activity of these compounds and their metabolic processing. The binding of enantiopure RC‐33 to the σ₁ receptor was also investigated in silico by molecular dynamics simulations. Both RC‐33 enantiomers showed similar affinities for the σ₁ receptor and appeared to be almost equally effective as σ₁ receptor agonists. However, the R‐configured enantiomer showed higher in vitro hepatic metabolic stability in the presence of NADPH than the S enantiomer. Overall, the results presented herein led us to select (R)‐RC‐33 as the optimal candidate for further in vivo studies in an animal model of amyotrophic lateral sclerosis.     

Skeletal Muscle Mitochondrial Energetic Efficiency and Aging

Aging is associated with a progressive loss of maximal cell functionality, and mitochondria are considered a key factor in aging process, since they determine the ATP availability in the cells. Mitochondrial performance during aging in skeletal muscle is reported to be either decreased or unchanged. This heterogeneity of results could partly be due to the method used to assess mitochondrial performance. In addition, in skeletal muscle the mitochondrial population is heterogeneous, composed of subsarcolemmal and intermyofibrillar mitochondria. Therefore, the purpose of the present review is to summarize the results obtained on the functionality of the above mitochondrial populations during aging, taking into account that the mitochondrial performance depends on organelle number, organelle activity, and energetic efficiency of the mitochondrial machinery in synthesizing ATP from the oxidation of fuels.     

Selective Aryl α‐Diimine/Palladium‐Catalyzed Bis‐Alkoxy‐ carbonylation of Olefins for the Synthesis of Substituted Succinic Diesters

Aryl α‐diimine derivatives have been used, for the first time, as efficient new ligands for the palladium‐catalyzed oxidative bis‐alkoxycarbonylation reaction of olefins. The most active catalyst was formed in situ from bis(9‐anthryl)‐2,3‐dimethyl‐1,4‐diazabutadiene and palladium(II) trifluoroacetate [Pd(TFA)₂]. This catalytic system was able to selectively convert olefins into succinic diesters in good yields (up to 97%) and low catalyst loading (up to 0.5 mol%) under mild reaction conditions [4 bar of carbon monoxide (CO) at 20 °C in the presence of p‐toluenesulphonic acid as additive and p‐benzoquinone as oxidant]. The optimized conditions could be successfully applied to both aromatic and aliphatic olefins, by using methanol, benzyl alcohol or isopropyl alcohol as nucleophiles.

     

Chiral Resolution and Pharmacological Characterization of the Enantiomers of the Hsp90 Inhibitor 2‐Amino‐7‐[4‐fluoro‐2‐(3‐pyridyl)phenyl]‐4‐methyl‐7,8‐dihydro‐6H‐quinazolin‐5‐one Oxime

Heat‐shock protein 90 (Hsp90) is a molecular chaperone involved in the stabilization of key oncogenic signaling proteins, and therefore, inhibition of Hsp90 represents a new strategy in cancer therapy. 2‐Amino‐7‐[4‐fluoro‐2‐(3‐pyridyl)phenyl]‐4‐methyl‐7,8‐dihydro‐6H‐quinazolin‐5‐one oxime is a racemic Hsp90 inhibitor that targets the N‐terminal adenosine triphosphatase site. We developed a method to resolve the enantiomers and evaluated their inhibitory activity on Hsp90 and the consequent antitumor effects. The (S) stereoisomer emerged as a potent Hsp90 inhibitor in biochemical and cellular assays. In addition, this enantiomer exhibited high oral bioavailability in mice and excellent antitumor activity in two different human cancer xenograft models.     

Multicomponent Cascade Reactions: A Novel and Expedient Approach to Functionalized Indoles by an Unprecedented Nucleophilic Addition‐Heterocyclization‐Oxidative Alkoxycarbonylation Sequence

A novel multicomponent cascade process is reported, based on the sequential combination between an initial nucleophilic attack step to an imine moiety and a palladium‐catalyzed oxidative heterocyclization‐alkoxycarbonylation process. By this new process, five simple molecules [2‐alkynylaniline imines, alcohol (ROH), carbon monoxide (CO), alcohol (ROH), and oxygen (O₂)] are sequentially activated, selectively leading to high value‐added functionalized indole derivatives in a single operation.     

Conformationally constrained CCK8 analogues obtained from a rationally designed peptide library as ligands for cholecystokinin type B receptor

A library of 14 cyclic peptide analogues derived from the octapeptide C-terminal sequence of the human cholecystokinin hormone (CCK(26-33), or CCK8) was designed, synthesized, and characterized. The 14 peptide analogues were rationally designed to specifically interact with the CCK type B receptor (CCK(B)-R) on the basis of the structure of the bimolecular complex between CCK8 and the third extracellular loop of CCK(B)-R, namely CCK(B)-R(352-379). The rational design of new ligands for CCK(B)-R has relied on stabilization by cyclic constraints of the structural motifs that bring the key residues of the ligand (especially Trp 30, Met 31, and Phe 33) in the proper spatial orientation for optimal interaction with the receptor. The binding affinity of the new ligands for CCK(B)-R was assessed by displacement experiments of (111)In-radiolabeled CCK8 in cells that overexpress the CCK(B) receptor. The new ligands generally showed binding affinities lower than that of parent CCK8, with the best compounds having IC50 values around 10 microM. Structure-activity relationship data show that preservation of the Trp 30-Met 31 motif is essential and that the Phe 33 side chain must be present. NMR conformational studies of the compound with maximal binding affinity (cyclo-B11, IC50=11 microM) in DPC micelles shows that this compound presents a turn-like conformation centered at the Trp 30-Met 31 segment, as planned by rational design. Such a conformation is stabilized by its interaction with the micelle rather than by the cyclic constraint.     

Effect of atmosphere composition on the quality of ready‐to‐use broccoli raab (Brassica rapa L.)

BACKGROUND: Many leafy brassicas are widely used for ready‐to‐use salads. Broccoli raab (Brassica rapa L.), also called turnip top, or rapini, is extensively cultivated in southern Italy. The edible portion is made up of the green, immature inflorescences and the stem with its most tender leaves. Recently, interest in this crop has increased among European consumers; moreover, a substantial increase in consumption could come from the ready‐to‐use product. RESULTS: The effects of four different atmosphere compositions (air, 3% O₂ in nitrogen, 3% O₂ + 10% CO₂ in air, and 10% CO₂ in air) on quality attributes of ready‐to‐use broccoli raab were studied. Controlled atmosphere affected appearance, composition, respiration rate, weight loss, and presence of off‐odours. Storage of broccoli raab florets under low oxygen conditions delayed post‐cutting deterioration during storage at 5 °C and preserved appearance and typical odour, up to 17 days. Moreover, respiration rate as well as loss of green colour, chlorophyll and vitamin C were also slowed down in this condition. Finally, in this study no effect of controlled atmosphere storage was found on total phenols content and antioxidant activity. CONCLUSION: Results showed that cold storage in 3% O₂ can be beneficial in order to maintain quality of ready‐to‐use, broccoli raab for up to 17 days. Copyright © 2010 Society of Chemical Industry     

Effects of free fatty acids on the oxidative processes in purified olive oil

Several studies have investigated the interaction of free fatty acids with the oxidative phenomena involving oils during processing and storage. Nevertheless, the available information is insufficient and, in some cases, contradictory.The aim of this work was to throw light upon this matter, by evaluating – by means of different analytical approaches – the effect of adding increasing amounts of free fatty acids (0.25%, 0.5%, 1%, 3%) on the oxidative processes occurring in purified olive oil during oxidation at 60 °C.The results obtained showed that oxidized forms of triglycerides and polar oligopolymers of triglycerides increased during oxidation. Low amounts of added FFA caused a further increase of the levels of oxidized triglycerides and triglyceride oligopolymers – pointing out a pro-oxidant activity – while higher doses of added free fatty acids lead to lower amounts of oxidized forms of triglycerides respect to the purified oil. This could be due to an increase in peroxide decomposition exerted by free fatty acids when present in higher amounts.     

Shear and longitudinal ultrasonic measurements of solid fat dispersions

Dispersions of coating fat in corn oil (2.5–12.5 wt%) were prepared following two different protocols: Type A dispersions had an average crystal size of 30–36 μm, whereas type B dispersions were less than 1 μm. In both dispersions the fat crystals were aggregated into larger structures (up to 80 μm). The longitudinal ultrasonic properties (i.e., velocity, attenuation, and reflectance) were linearly related to the solid fat content, but only attenuation was sensitive to the different microstructures. The velocity and reflectance measurements were modeled using the Urick equation. Shear ultrasonic reflectance and oscillatory viscometry were used to measure the dynamic viscosity of all dispersions. According to both methods, type B samples were always more viscous than type A at a similar solids content. The correlation between the two techniques was good (r ²>0.99), but the numeric agreement was different for both systems.     

Structure-based drug design to the discovery of new 2-aminothiazole CDK2 inhibitors

N-(5-Bromo-1,3-thiazol-2-yl)butanamide (compound 1) was found active (IC50=808 nM) in a high throughput screening (HTS) for CDK2 inhibitors. By exploiting crystal structures of several complexes between CDK2 and inhibitors and applying structure-based drug design (SBDD), we rapidly discovered a very potent and selective CDK2 inhibitor 4-[(5-isopropyl-1,3-thiazol-2-yl)amino] benzenesulfonamide (compound 4, IC50=20 nM). The syntheses, structure-based analog design, kinases inhibition data and X-ray crystallographic structures of CDK2/inhibitor complexes are reported.