Since the identification of the first antimicrobial peptide (AMP), cecropin, from silk moths (Hyalophora cecropia) in the early 1980s, approximately 2000 AMPs, have been isolated from various biological sources from insects to humans. The unique therapeutic features of AMPs, such as nonspecific mode of action and broad-spectrum activity, have attracted the attention of many research groups around the world. Many reports have proposed that AMPs exert their action by binding to the cell membrane, causing cell death by direct membrane disruption or translocation into the cytosol where potential internal targets are affected. AMPs are known to exert microbial killing via a unique, multifaceted mode of action, which makes it very difficult for pathogens to develop resistance against them. In addition, the different mode of action of AMPs compared to the conventional antibiotics, which specifically target cellular components at various levels, explains their broad spectrum of activity, including against drug-resistant pathogens. Consequently, extensive efforts have been made to develop AMPs as novel antimicrobial agents. In addition to the native lipoand glycopeptide antibiotics such as daptomycin, polymyxin B, and vancomycin, which are already available on the market, many synthetic AMPs are still in the drug development phase, yet to reach clinical application. Despite being recognized as a promising class of molecules for many years, AMPs have not made a substantial impact in the treatment of infections caused by resistant bacteria and fungi. This limitation is mainly due to clinical challenges, such as cytotoxicity and poor pharmacokinetics associated with AMPs, mainly attributed to their comparatively large molecular size and susceptibility to protease degradation. The past two decades have witnessed the emergence of macrocycles, including macrocyclic peptides, as a promising yet underexploited class of de novo drug candidates. Macrocyclic molecules represent a unique chemical class, with features such as structural compactness and high stability against enzymatic degradation, making them highly suitable for drug development. The therapeutic potential of macrocycles is further evident from the discovery of various naturally occurring peptides-based anti-infectives, such as daptomycin, polymyxin B, vancomycin, amphotericin B, and gramicidin S, all of which possess a welldefined cyclic structural framework. Accordingly, over the years, to address the clinical drawbacks associated with AMPs, numerous chemical modification strategies have been implemented to devise macrocyclic molecules with structural rigidity and enzymatic stability. Previous studies have revealed the importance of the orientation of the side chain functionalities in facilitating effective electrostatic and/or hydrophobic interactions with target membranes. Thus, backbone stereochemistry could greatly influence the membrane interaction properties of peptide molecules.