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The use of positional scanning synthetic combinatorial libraries (PS-SCLs)1 enables the most active amino acids at each position of a peptide or non-peptide to be determined directly from the initial screening data.

This information can then be used to synthesize highly active individual compounds.

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Screening of the same combinatorial library in separate assays selective for each of the three receptors provides not only new ligands for these receptors but yields insights into the ability of combinatorial libraries to discriminate between closely related receptors.

A combinatorial library of 6,250,000 tetrapeptides, made using 50 different amino acids, was prepared in the positional scanning format (6, 7).

It should be noted that each of the four positional sublibraries are made up of the same 6,250,000 tetrapeptides.

Screening the four sets of mixtures in the three separate opiate specific assays yielded information about the most important amino acids of each position in the tetrapeptide and led to the identification of three different series of active individual tetrapeptides selective for the μ, δ, and κ receptors.

This study demonstrates the power of mixture-based combinatorial libraries to identify distinctly different ligands for closely related receptors.

Synthetic combinatorial libraries have gained widespread acceptance for the rapid identification of new drug leads.Concurrently, a second combinatorial library containing 52 million non-acetylated hexapeptides was subsequently used to identify ligands for the opioid receptors, which were closely related to the natural ligands methionine- and leucine-enkephalin (YGGFM, YGGFL) (4).In later studies, six distinctly hexapeptide sequences that specifically bound to the μ opioid receptor were identified from the acetylated and nonacetylated libraries (5).A combinatorial library of 6,250,000 tetrapeptides in the mixture based positional scanning format was screened in binding assays for the three opioid receptors, μ, δ, and κ. Individual peptides were synthesized representing all possible combinations of the active amino acids identified from the screening data.New, highly active peptides selective for each of the three receptors were chosen.Complete combinatorial libraries contain all possible arrangements of building blocks used in their synthesis.

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