Peptide
stapling is traditionally used to lock peptide conformations
into α-helical structures using a variety of macrocyclization
chemistries. In an endeavor to add a diversity-generating tool to
this repertoire, we introduce a multicomponent stapling approach enabling
the simultaneous stabilization of helical secondary structures and
the exocyclic N-functionalization of the side chain-tethering
lactam bridge. This is accomplished by means of a novel solid-phase
methodology comprising, for the first time, the on-resin Ugi reaction-based
macrocyclization of peptide side chains bearing amino and carboxylic
acid groups. The exocyclic diversity elements arise from the isocyanide
component used in the Ugi multicomponent stapling protocol, which
allows for the incorporation of relevant fragments such as lipids,
sugars, polyethylene glycol, fluorescent labels, and reactive handles.
We prove the utility of such exocyclic reactive groups in the bioconjugation
of a maleimide-armed lactam-bridged peptide to a carrier protein.
The on-resin multicomponent stapling proved efficient for the installation
of not only one, but also two consecutive lactam bridges having either
identical or dissimilar N-functionalities. The easy access to helical
peptides with a diverse set of exocyclic functionalities shows prospect
for applications in peptide drug discovery and chemical biology.
The cyclization of peptide side chains has been traditionally used to either induce or stabilize secondary structures (β-strands, helices, reverse turns) in short peptide sequences. So far, classic peptide coupling, nucleophilic substitution, olefin metathesis, and click reactions have been the methods of choice to fold synthetic peptides by means of macrocyclization. This article describes the utilization of the Ugi reaction for the side chain-to-side chain and side chain-to-termini macrocyclization of peptides, thus enabling not only access to stable folded structures but also the incorporation of exocyclic functionalities as N-substituents. Analysis of the NMR-derived structures revealed the formation of helical turns, β-bulges, and α-turns in cyclic peptides cross-linked at i, i + 3 and i, i + 4 positions, proving the folding effect of the multicomponent Ugi macrocyclization. Molecular dynamics simulation provided further insights on the stability and molecular motion of the side chain cross-linked peptides.
MCRs are versatile convergent procedures that incorporate 50 three or more reactants into a final product, with the consequent 51 generation of higher levels of molecular complexity and diversity 52 than other traditional organic reactions using a similar synthetic 53 effort [10][11][12][13] . They have been employed in almost all areas 54 of chemical synthesis [10][11][12][14][15][16] , but their applications in drug 55 discovery and development have unquestionable dominated the 56 interest of the chemical community 10,17,18 . MCRs are especially 57 well suited for the construction of heterocyclic-often medic-58 inally privileged-scaffolds, a task where they are comparable to 59 cycloaddition reactions in terms of scope and chemical effi-60 ciency. Indeed, solution-phase protocols are predominant in 61 both the development of new multicomponent processes and 62 the implementation of known MCRs to synthesize bioactive 63 molecules 10-18 . However, solid-phase synthesis also gained Q5 Q6
Constraining small peptides into specific secondary structures has been a major challenge in peptide ligand design. So far, the major solution for decreasing the conformational flexibility in small peptides has been cyclization. An alternative is the use of topological templates, which are able to induce and/or stabilize peptide secondary structures by means of covalent attachment to the peptide. Herein a multicomponent strategy and structural analysis of a new type of peptidosteroid architecture having the steroid as N-substituent of an internal amide bond is reported. The approach comprises the one-pot conjugation of two peptide chains (or amino acid derivatives) to aminosteroids by means of the Ugi reaction to give a unique family of N-steroidal peptides. The conjugation efficiency of a variety of peptide sequences and steroidal amines, as well as their consecutive head-to-tail cyclization to produce chimeric cyclopeptide-steroid conjugates, that is, macrocyclic lipopeptides, was assessed. Determination of the three-dimensional structure of an acyclic N-steroidal peptide in solution proved that the bulky, rigid steroidal template is capable of both increasing significantly the conformational rigidity, even in a peptide sequence as short as five amino acid residues, and inducing a β-turn secondary structure even in the all-s-trans isomer. This report provides the first evidence of the steroid skeleton as β-turn inducer in linear peptide sequences.
Conjugate vaccines against encapsulated pathogens likeStreptococcus pneumoniae face many challenges, including the existence of multiple serotypes with a diverse global distribution that constantly requires new formulations and higher coverage. Multivalency is usually achieved by combining capsular polysaccharide−protein conjugates from invasive serotypes, and for S. pneumoniae, this has evolved from 7up to 20-valent vaccines. These glycoconjugate formulations often contain high concentrations of carrier proteins, which may negatively affect glycoconjugate immune response. This work broadens the scope of an efficient multicomponent strategy, leading to multivalent pneumococcal glycoconjugates assembled in a single synthetic operation. The bioconjugation method, based on the Ugi fourcomponent reaction, enables the one-pot incorporation of two different polysaccharide antigens to a tetanus toxoid carrier, thus representing the fastest approach to achieve multivalency. The reported glycoconjugates incorporate three combinations of capsular polysaccharides 1, 6B, 14, and 18C from S. pneumoniae. The glycoconjugates were able to elicit functional specific antibodies against pneumococcal strains comparable to those shown by mixtures of the two monovalent glycoconjugates.
NMR and CD studies together with molecular dynamics simulation reveal new insights into the s-cis/s-trans isomerism and the effect of the lactam bridge N-substituent on the secondary structure of stapled peptides.
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