Rapid High-Fidelity Assembly of Challenging Peptide Sequences

PROJECT SUMMARY Peptide therapeutics combine high potency and selectivity with fewer side effects than traditional small moleculesThis has fueled interest in peptides filling the gap betweenbiologicsand small molecules as compounds that have the key desired properties from both the other classesWorldwide peptide drug sales surpassed $billion inof which nearly $billion was in the USAwith projected growth ofper yearSynthesis and purification of long peptides continue to be major challenges for manufacturersSolid phase peptide synthesisSPPSis the most versatile approach to the synthesis of peptides containing fewer thanamino acid residuesSPPS enables the incorporation of andquot unnaturalandquotelements with desired functionalitye gincreases in conformational and proteolytic stability and potency efficacy at target receptorsWhile current SPPS methods routinely yield high efficiencies per coupling stepfor amer peptide aindividual step coupling efficiency would yield onlyof full length peptide andof peptides lacking at least one amino acid residueandquot deletionsandquotPurification generally involves chromatographic separationwhich is generally viewed as thebottleneckfor peptide synthesisMoreoverdeletions that differ from the full sequence by only a single residue are frequently very challenging to separate using chromatographyThe Kumar group has developed an original andquot F cappingandquotSPPS methodology to simplify and accelerate the process of purificationBrieflyat difficult steps of peptide assemblytruncation sequences are terminated by inert fluorocarbon segmentsandquot F capsandquotand thus are prevented from growing into intractable contaminantsWe will synthesize long and challenging peptide sequences rapidly and in high purityWe aim to reduce the time to synthesize a typicalamino acid long sequence in the laboratory from a week to four hours or lessat the same time eliminating deletionsTo this end we will combine the high temperature Fast Flow technology with our F Capping technologyFast Flow is known for shortened cycle times of minutes vshoursbut it does not address deletionsF Capping suppresses deletions via minute long treatments with afluorousreagentThe tandem use of these technologies is expected to afford candidate peptide drugs at a fraction of the time and cost PUBLIC HEALTH RELEVANCE STATEMENT This proposed work develops a platform technology that allows the synthesis of long and otherwise challenging peptideswith minimal if any need for chromatographyThe peptides we target have relevance to virtually every noncommunicable disease area including metabolic disfunctionsinflammationtraumatic brain injuryAlzheimer sand others