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50 values represent the mean of at least three experiments. Levels of partial agonism are from one to four experiments.
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Experimental protocols involving animals in this study were reviewed by the Animal Care and Use Committee of Digital Biotech Co., according to the NIH guidelines (NIH publication number 85-23, revised 1985) of 'Principles of Laboratory Animal Care'.
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Since 1 and 12 are structurally related, we have studied whether 1 may be generated after the systemic administration of 12. Thus, 5 mg/kg of 12 was administered intravenously to male Sprague-Dawley rats (250-280 g) and 0.25 mL of blood was collected at various times up to 120 min. Plasma was collected and an aliquot (0.15 mL) was deproteinated by the addition of 0.3 mL acetonitrile. An aliquot of the supernatant (0.35 mL) was evaporated to dryness under a stream of nitrogen and the residue was reconstituted by the addition of 0.1 mL of mobile phase (water/methanol/acetonitrile=20:40:40). An aliquot of the mixture was injected directly onto the LC-MS system. Although both 1 and 12 could be readily quantified by the LC-MS system, the concentration of 12, but not 1, was detected in the sample. Since we could not have detected any conversion of 12 into 1, we conclude that the action of 12 cannot be explained by its conversion to 1 in vivo.
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