Insufficient oral bioavailability is considered as a key limitation for the widespread development of peptides as therapeutics. While the oral bioavailability of small organic compounds is often estimated from simple rules, similar rules do not apply to peptides, and even the high oral bioavailability that is described for a small number of peptides is not well understood. Here we present two highly Caco-2 permeable template structures based on a library of 54 cyclo(-d-Ala-Ala5-) peptides with different N-methylation patterns. The first (all-trans) template structure possesses two β-turns of type II along Ala6-d-Ala1 and Ala3-Ala4 and is only found for one peptide with two N-methyl groups at d-Ala1 and Ala6 [(NMe(1,6)]. The second (single-cis) template possesses a characteristic cis peptide bond preceding Ala5, which results in type VI β-turn geometry along Ala4-Ala5. Although the second template structure is found in seven peptides carrying N-methyl groups on Ala5, high Caco-2 permeability is only found for a subgroup of two of them [NMe(1,5) and NMe(1,2,4,5)], suggesting that N-methylation of d-Ala1 is a prerequisite for high permeability of the second template structure. The structural similarity of the second template structure with the orally bioavailable somatostatin analog cyclo(-Pro-Phe-NMe-d-Trp-NMe-Lys-Thr-NMe-Phe-), and the striking resemblance with both β-turns of the orally bioavailable peptide cyclosporine A, suggests that the introduction of bioactive sequences on the highly Caco-2 permeable templates may result in potent orally bioavailable drug candidates.