Pharma's Developing Interest in Stem Cells

Human stem cell biology is driving the promise of novel regenerative therapies into clinical trials. Although the pharmaceutical industry has embraced stem cells as tools in drug discovery, few companies have taken the risk to deliver stem cell-based medicines. Here, we evaluate the various cell-based opportunities and corporate strategies

Effect of subunit on allosteric modulation of ion channel function in stably expressed human recombinant -aminobutyric acidA receptors determined using 36Cl ion flux.

ABSTRACT Inhibitory ␥-aminobutyric acid (GABA) A receptors are subject to modulation at a variety of allosteric sites, with pharmacology dependent on receptor subunit combination. The influence of different ␣ subunits in combination with ␤3␥2s was examined in stably expressed human recombinant GABA A receptors by measuring 36 Cl influx through the ion channel pore. Muscimol and GABA exhibited similar maximal efficacy at each receptor subtype, although muscimol was more potent, with responses blocked by picrotoxin and bicuculline. Receptors containing the ␣3 subunit exhibited slightly lower potency. The comparative pharmacology of a range of benzodiazepine site ligands was examined, revealing a range of intrinsic efficacies at different receptor subtypes. Of the diazepam-sensitive GABA A receptors (␣1, ␣2, ␣3, ␣5), ␣5 showed the most divergence, being discriminated by zolpidem in terms of very low affinity, and CL218,872 and CGS9895 with different efficacies. Benzodiazepine potentiation at ␣3␤3␥2s with nonselective agonist chlordiazepoxide was greater than at ␣1, ␣2, or ␣5 (P Ͻ 0.001). The presence of an ␣4 subunit conferred a unique pharmacological profile. The partial agonist bretazenil was the most efficacious benzodiazepine, despite lower ␣4 affinity, and FG8205 displayed similar efficacy. Most striking were the lack of affinity/ efficacy for classical benzodiazepines and the relatively high efficacy of Ro15-1788 (53 Ϯ 12%), CGS8216 (56 Ϯ 6%), CGS9895 (65 Ϯ 6%), and the weak partial inverse agonist Ro15-4513 (87 Ϯ 5%). Each receptor subtype was modulated by pentobarbital, loreclezole, and 5␣-pregnan-3␣-ol-20-one, but the type of ␣ subunit influenced the level of potentiation. The maximal pentobarbital response was significantly greater at ␣4␤3␥2s (226 Ϯ 10% increase in the EC 20 response to GABA) than any other modulator. The rank order of potentiation for pregnanolone was ␣5 Ͼ ␣2 Ͼ ␣3 ϭ ␣4 Ͼ ␣1, for loreclezole ␣1 ϭ ␣2 ϭ ␣3 Ͼ ␣5 Ͼ ␣4, and for pentobarbital ␣4 ϭ ␣5 ϭ ␣2 Ͼ ␣1 ϭ ␣3