CCK-8S increases the firing frequency of CCK-positive neurons and facilitates excitatory synaptic transmission in primary rat hippocampal neurons.

The neuropeptide cholecystokinin octapeptide (CCK) is involved in a variety of brain functions. In the hippocampus, most CCK is released from CCK-positive (CCK+) neurons, but the effects of CCK on CCK+ neurons are poorly understood. We employed primary hippocampal cultures to explore the modulatory effect of CCK on CCK+ neurons. CCK-8S (0.2 µM) was added to the culture medium from day in vitro 2 (DIV-2) to DIV-11. An adenovirus integrated with the CCK promoter was used to label CCK+ neurons. Whole-cell patch clamp recording was carried on to record the electrophysiology properties. The results show that: (1) CCK-8S significantly decreased membrane capacity but increased the membrane resistance (Rm) of CCK+ neurons, (2) CCK-8S increased action potential (AP) firing frequency of CCK+ neurons but did not affect the firing pattern, (3) CCK-8S facilitated CCK+ neuron excitatory synaptic transmission but attenuated inhibitory synaptic transmission, and (4) the expression of postsynaptic density-95 (PSD-95) in cultured hippocampal neurons was elevated by CCK-8S treatment. Our results demonstrate that CCK-8S significantly alters the membrane electrophysiological characteristics and synaptic activity of cultured hippocampal CCK+ neurons. These findings may enhance our understanding of the modulatory effect of CCK in the brain.

CCK-8S increased the filopodia and spines density in cultured hippocampal neurons of APP/PS1 and wild-type mice.

Cholecystokinin (CCK), a neuropeptide, is widely distributed in the brain. The function of CCK is involved in many brain functions including learning and memory, but the cellular mechanism is poorly understood. In the present study, we investigated the effect of CCK on dendritic filopodia and spines of cultured hippocampal neurons from wild-type and APP/PS1 mice. The cultured hippocampal neurons were infected with CMV-GFP (CMV promoter with green fluorescent protein) adenovirus 24h before image acquisition to display the subtle structure of dendrites. Cholecystokinin octapeptide sulfated (CCK-8S, 0.2µM) was added into the cultured solution from divided in vitro day 2 (DIV 2). A decrease of filopodia and spines density was observed in APP/PS1 mice compared with that of wild type mice. CCK-8S increased the density of filopodia and spines at DIV 7, DIV 14 and DIV 21 in hippocampal neurons of both wild-type and APP/PS1 mice. In addition, this effect was inhibited by CI988, an antagonist of CCK-2 receptor. Those results indicate that CCK-8S can influence the dendritic development and spine genesis of cultured hippocampal neurons derived from both wild-type and APP/PS1 mice. These data suggest that CCK may play an important role in learning and memory.

Divergent C-H functionalizations directed by sulfonamide pharmacophores: late-stage diversification as a tool for drug discovery.

Modern drug discovery is contingent on identifying lead compounds and rapidly synthesizing analogues. The use of a common pharmacophore to direct multiple and divergent C-H functionalizations of lead compounds is a particularly attractive approach. Herein, we demonstrate the viability of late-stage diversification through the divergent C-H functionalization of sulfonamides, an important class of pharmacophores found in nearly 200 drugs currently on the market, including the non-steroidal anti-inflammatory blockbuster drug celecoxib. We developed a set of six categorically different sulfonamide C-H functionalization reactions (olefination, arylation, alkylation, halogenation, carboxylation, and carbonylation), each representing a distinct handle for further diversification to reach a large number of analogues. We then performed late-stage, site-selective diversification of a sulfonamide drug candidate containing multiple potentially reactive C-H bonds to synthesize directly novel celecoxib analogues as potential cyclooxygenase-II (COX-2)-specific inhibitors. Together with other recently developed practical directing groups, such as CONHOMe and CONHC(6)F(5), sulfonamide directing groups demonstrate that the auxiliary approach established in asymmetric catalysis can be equally effective in developing broadly useful C-H activation reactions.

Pd(II)-catalyzed enantioselective C-H olefination of diphenylacetic acids.

Pd(II)-catalyzed enantioselective C-H olefination of diphenylacetic acid substrates has been achieved through the use of monoprotected chiral amino acid ligands. The absolute configuration of the resulting olefinated products is consistent with that of a proposed C-H insertion intermediate.

Pd(II)-catalyzed hydroxylation of arenes with 1 atm of O(2) or air.

Pd(II)-catalyzed ortho-hydroxylation of variously substituted benzoic acids under 1 atm of O(2) or air is achieved under nonacidic conditions. Extensive labeling studies support a direct oxygenation of aryl C-H bonds with molecular oxygen.

Pd(II)-catalyzed olefination of electron-deficient arenes using 2,6-dialkylpyridine ligands.

Pd(II)-catalyzed meta-olefination of highly electron-deficient arenes is achieved through the use of a rationally designed mutually repulsive ligand. The combination of directed and nondirected C-H functionalization of arenes provides a versatile route for the synthesis of highly sought after 1,2,4-trisubstituted arenes.