Developing a new drug from original idea to the launch of a finished product is a complex process which can take 12–15 years and cost in excess of $1 billion. The idea for a target can come from a variety of sources including academic and clinical research and from the commercial sector. It may take many years to build up a body of supporting evidence before selecting a target for a costly drug discovery programme. Once a target has been chosen, the pharmaceutical industry and more recently some academic centres have streamlined a number of early processes to identify molecules which possess suitable characteristics to make acceptable drugs. This review will look at key preclinical stages of the drug discovery process, from initial target identification and validation, through assay development, high throughput screening, hit identification, lead optimization and finally the selection of a candidate molecule for clinical development.

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Summary Changes in plasma 15‐keto‐13, 14‐dihydro‐prostaglandin F2α were monitored at frequent intervals before, during and after spontaneous deliveries (three mares) and foalings induced by oxytocin (eight mares). No evidence of increased concentrations of the prostaglandin metabolite was observed in the final 10 days of gestation. In spontaneously delivering mares, there was a marked increase from 3 ng/ml at −125 mins to 18 ng/ml at −65 mins to the highest observed value of 182 ng/ml at 20 mins pre‐partum. Following delivery, concentrations declined rapidly to around 0.2 ng/ml. Further release of prostaglandins was seen on Days 1 and 3 post partum. In oxytocin induced mares, maximal concentrations of about 100 ng/ml were observed to occur very close to the time of delivery. Large increases were observed as early as 2 mins following oxytocin injection. The significance of these findings is discussed in relation to parturient and post parturient events and changes in levels of the hormone relaxin during the same period.

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Endogenous, cysteine-rich antimicrobial peptides known as defensins are prominent components of human, rabbit, and rat neutrophils, yet little is known about their occurrence in other mammalian species. Although we did not detect mature (i.e., processed) defensins in equine neutrophil granules, we found that these granules contained small amounts of other cysteine-rich peptides with antimicrobial activity. One of these, eNAP-1, was purified by a combination of gel permeation and reversed-phase high-performance liquid chromatography from acid extracts prepared from the cytoplasmic granules of equine neutrophils. The molecular mass of eNAP-1 was approximately 7.2 kDa, as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis. Amino acid analysis revealed that eNAP-1 had an unusually high cysteine content and that it was relatively enriched in alanine, glycine, lysine, and proline residues. The partial (N-terminal) amino acid sequence of eNAP-1 was DVQCGEGHFCHDXQTCCRASQGGXACCPYSQGVCCADQRHCCPVGF. Thirtysix of these residues (78.3%) were identical to those of a recently cloned human neutrophil peptide of unknown function and belonging to the granulin family. Homologous peptides have also been noted in rat bone marrow cells and rat kidney epithelins. We tested the ability of eNAP-1 to kill several equine uterine pathogens. Streptococcus zooepidemicus was killed most effectively, sustaining a >99.8% decrease in CFU per milliliter after a 2-h exposure to 100 ,g of eNAP-1 per ml (-15 FM). Escherichia coli and Pseudomonas aeruginosa were somewhat less susceptible, manifesting 87.0 and 87.1% mean decreases in CFU per milliliter, respectively, after incubation for 2 h with 200 ,ug of eNAP-1 per ml. KlebsieUla pneumoniae numbers were not significantly reduced after exposure to eNAP-1. These antimicrobial properties suggest that eNAP-1 may contribute to phagocyte-mediated host defense against equine infections.