Oligopeptide Synthesis Research Articles

Mechanistic Insight into the TBHP-Mediated Decarboxylative Condensation of α-Ketoacids: Reaction Development and Application to Oligopeptide Synthesis.

With the aim of achieving the convergent elongation of peptide chains, an amide bond formation reaction that enables a peptide fragment coupling has long been pursued. The decarboxylative amidation recently reported by our group is a potential solution to this problem. In this article, a mechanistic analysis of the t-butyl hydroperoxide (TBHP) mediated-decarboxylative amidation of α-ketoacids that results in a significant advance in convergent peptide synthesis is described. Despite the observation of epimerization with low bulk substrates in preliminary studies, a systematic examination and understanding of the reaction mechanism enabled the development of a modified epimerization-free reaction whereby peptide fragment couplings using peptide α-ketoacids were successfully achieved.

A practical approach for oligopeptide synthesis via synergistic photoredox, cobaloxime and organophosphorus triple catalysis

A catalytic oxidation–reduction condensation strategy for the synthesis of oligopeptides is reported. The triple catalytic system is compatible with all proteinogenic amino acids and applicable for multigram scale peptide synthesis.

Unnatural Lysines with Reduced Sidechain N‐Basicity: Synthesis of N‐trifluoroethyl Substituted Lysine and Homologs

AbstractThe syntheses are reported of Nϵ‐(2,2,2‐trifluoroethyl)‐D, L‐lysine (tFK) and Nζ‐(2,2,2‐trifluoroethyl)‐D, L‐homolysine (tFK+1) from amino alcohols HO−(CH2)n−NH2. The syntheses involve reductive amination, Appel bromination, and the stereoselective bond formation between Cα of the amino acid and the fluorinated alkyl chain in the Schöllkopf bislactim amino acid synthesis. The methyl esters of the fluorinated amino acids are the relevant substrates for oligopeptide synthesis. With the R‐Schöllkopf reagent, we stereoselectively generated methyl Nϵ‐boc‐Nϵ‐(2,2,2‐trifluoroethyl)‐L‐lysinate and methyl Nζ‐boc‐Nζ‐(2,2,2‐trifluoroethyl)‐L‐homolysinate. Products and intermediates were characterized by 1H NMR, 13C NMR, COSY, HSQC, and LCMS. A variety of N‐functionality may be introduced by reacting hemiacetals with different appendages. This fluorine modification reduces the sidechain N‐basicity by combined ‐I effect of the three fluorines. This effect increases the [amine]/[ammonium ion] ratio of the sidechain amine in lysine to facilitate carbamylation at lower pH conditions.

Protecting group principles suited to late stage functionalization and global deprotection in oligosaccharide synthesis

Chemical synthesis is a powerful tool to access homogeneous complex glycans, which relies on protecting group (PG) chemistry. However, the overall efficiency of chemical glycan assembly is still low when compared to oligonucleotide or oligopeptide synthesis. There have been many contributions giving rise to collective improvement in carbohydrate synthesis that includes PG manipulation and stereoselective glycoside formation and some of this chemistry has been transferred to the solid phase or adapted for programmable one pot synthesis approaches. However, after all glycoside bond formation reactions are completed, the global deprotection (GD) required to give the desired target OS can be challenging. Difficulties observed in the removal of permanent PGs to release the desired glycans can be due to the number and diversity of PGs present in the protected OSs, nature and structural complexity of glycans, etc. Here, we have reviewed the difficulties associated with the removal of PGs from densely protected OSs to obtain their free glycans. In particularly, this review focuses on the challenges associated with hydrogenolysis of benzyl groups, saponification of esters and functional group interconversion such as oxidation/reduction that are commonly performed in GD stage. More generally, problems observed in the removal of permanent PGs is reviewed herein, including benzyl, acyl (levulinoyl, acetyl), N-trichloroacetyl, N-2,2,2-trichloroethoxycarbonyl, N-phthaloyl etc. from a number of fully protected OSs to release the free sugar, that have been previously reported in the literature.

Two-Component Redox Organocatalyst for Peptide Bond Formation.

Peptides are fundamental therapeutic modalities whose sequence-specific synthesis can be automated. Yet, modern peptide synthesis remains atom uneconomical and requires an excess of coupling agents and protected amino acids for efficient amide bond formation. We recently described the rational design of an organocatalyst that can operate on Fmoc amino acids─the standard monomers in automated peptide synthesis (J. Am. Chem. Soc. 2019, 141, 15977). The catalytic cycle centered on the conversion of the carboxylic acid to selenoester, which was activated by a hydrogen bonding scaffold for amine coupling. The selenoester was generated in situ from a diselenide catalyst and stoichiometric amounts of phosphine. Although the prior system catalyzed oligopeptide synthesis on solid phase, it had two significant requirements that limited its utility as an alternative to coupling agents─it depended on stoichiometric amounts of phosphine and required molecular sieves as dehydrating agent. Here, we address these limitations with an optimized method that requires only catalytic amounts of phosphine and no dehydrating agent. The new method utilizes a two-component organoreductant/organooxidant-recycling strategy to catalyze amide bond formation.

Electrochemical Oligopeptide Synthesis Assisted By a Soluble Tag Method

Recently, peptides have attracted much attention as the medicinal candidates with higher specificity and lower side effects than small molecular medicines. Peptides have been synthesized using stoichiometric amount of coupling reagents. While these reagents enable quantitative reaction, stoichiometric byproducts from coupling reagents cause poor atom economy. In this context, “General methods for catalytic/ sustainable (direct) amide or peptide formation” is selected as one of the top research area in green chemistry. [1] To solve the problem, we are aiming to establish a method of electrochemical oligopeptide synthesis and recycle coupling reagents. Inspired by the report of Uribe’s group, [2] we selected triphenylphosphine (TPP) as the coupling reagent in electrochemical peptide bond formation. Although triphenylphosphine oxide (TPPO) is generated as the stoichiometric byproduct, reduction of TPPO to TPP are achieved in various ways. [3] Therefore, TPP can be a recyclable coupling reagent in peptide synthesis and this lead to reduce the amount of waste. In addition, we employed soluble tags as protecting group of carboxylic acids, which are consisted of benzyl group with long alkyl chain and precipitate in polar solvents, to make purification easier. [4] Until now, we have succeeded quantitative electrochemical peptide bond formation. Notably, we found that soluble tag serves to prevent undesired oxidation of amino acids maybe because long alkyl chain of soluble tag keep amino acids away from electrode. Work to synthesis bioactive oligopeptides is in progress.Reference[1] M. C. Bryan, P. J. Dunn, D. Entwistle, F. Gallou, S. G. Koenig, J. D. Hayler, M. R. Hickey, S. Hughes, M. E. Kopach, G. Moine, P. Richardson, F. Roschangar, A. Steven and F. J. Weiberth, Green Chem., 2018, 20, 5082-5103.[2] A. Palma, J. Cardenas and B. A. Frontana-Uribe, Green Chem., 2009, 11, 283–293.[3] Y. Okada, H. Suzuki, T. Nakae, S. Fujita, H. Abe, K. Nagano, T. Yamada, N. Ebata, S. Kim, and K. Chiba, J. Org. Chem., 2013, 78, 320−327.[4] D. Hérault, D. H. Nguyen, D. Nuel and G. Buono, Chem. Soc. Rev., 2015, 44 , 2508-2528. Figure 1

Toward Automated Enzymatic Synthesis of Oligosaccharides.

Oligosaccharides together with oligonucleotides and oligopeptides comprise the three major classes of natural biopolymers. Automated systems for oligonucleotide and oligopeptide synthesis have significantly advanced developments in biological science by allowing nonspecialists to rapidly and easily access these biopolymers. Researchers have endeavored for decades to develop a comparable general automated system to synthesize oligosaccharides. Such a system would have a revolutionary impact on the understanding of the roles of glycans in biological systems. The main challenge to achieving automated synthesis is the lack of general synthetic methods for routine synthesis of glycans. Currently, the two main methods to access homogeneous glycans and glycoconjugates are chemical synthesis and enzymatic synthesis. Enzymatic glycosylation can proceed stereo- and regiospecifically without protecting group manipulations. Moreover, the reaction conditions of enzyme-catalyzed glycosylations are extremely mild when compared to chemical glycosylations. Over the past few years methodology toward the automated chemical synthesis of oligosaccharides has been developed. Conversely, while automated enzymatic synthesis is conceptually possible, it is not as well developed. The goal of this survey is to provide a foundation on which continued technological advancements can be made to promote the automated enzymatic synthesis of oligosaccharides.

Catalytic Oligopeptide Synthesis.

Waste-free catalytic assembly of α-amino acids is fueled by a multiboron catalyst that features a characteristic B3NO2 heterocycle, providing a versatile catalytic protocol wherein functionalized natural α-amino acid units are accommodated and commonly used protecting groups are tolerated. The facile dehydrative conditions eliminate the use of engineered peptide coupling reagents, exemplifying a greener catalytic alternative for peptide coupling. The catalysis is sufficiently robust to enable pentapeptide synthesis, constructing all four amide bond linkages in a catalytic fashion.

Template-Assisted Enzymatic Synthesis of Oligopeptides from a Polylactide Chain.

Peptides are often attached to polymer materials, as bioactive components, for the control of interactions between the material and its surrounding proteins and cells. However, synthesizing peptides and attaching them to polymers can be challenging and laborious. Herein, we describe the grafting of oligopeptides to an aliphatic polyester, using a one-step chemo-enzymatic synthesis with papain as the biocatalyst. To enable enzyme-mediated functionalization of the polyester, ethyl hept-6-enoylalaninate (grafter) was synthesized and attached to polylactide chains using thiol-ene click reactions. The oligopeptides were grafted onto the polylactide chains using two different synthetic routes: the grafting from strategy, in which the grafter was attached to the polyester prior to oligopeptide synthesis, or the grafting to strategy, in which oligopeptides were synthesized on the grafter first, then attached to the polymer chain. The final products were analyzed and their structures were confirmed using nuclear magnetic resonance (NMR). The peptide attachment was evaluated using size exclusion chromatography (SEC), contact angle measurement and energy-dispersive X-ray spectroscopy-scanning electron microscopy (EDS-SEM). Furthermore, the mechanistic aspects of the synthesis of the oligopeptides on the grafter were studied using molecular dynamics (MD) simulations. The simulation revealed that hydrogen bonding (between the P1 amide nitrogen of the grafter backbone and the carbonyl oxygen of D158 in the papain) maintain the grafter in a productive conformation to stabilize the transition state of nitrogen inversion, a key step of the biocatalytic mechanism. Apart from being biologically relevant, both experimental and computational results suggest that the designed grafter is a good template for initiating chemo-enzymatic synthesis. The results also showed that the grafting to strategy was more successful compared to the grafting from strategy. Overall, a successful synthesis of predefined peptide functionalized polylactide was prepared, where the oligopeptides were grafted in an easy, time efficient, and environmentally friendly way.

Synthesis of Acylborons by Ozonolysis of Alkenylboronates: Preparation of an Enantioenriched Amino Acid Acylboronate

AbstractA concise synthesis of acylborons was achieved by ozonolysis of alkenyl MIDA (N‐methyliminodiacetic acid) boronates. This reaction exhibits excellent functional‐group tolerance and is applicable to various acyl MIDA boronates and potassium acyltrifluroborates (KATs) which could not be synthesized by previous methods. In addition, α‐amino acylborons, which would be essential for peptide ligations, were prepared for the first time. The acylboron of l‐alanine was obtained in high enantiopurity and found to be configurationally stable. Oligopeptide synthesis between the α‐amino KATs and amino acid in dilute aqueous media was studied.

Synthesis of Acylborons by Ozonolysis of Alkenylboronates: Preparation of an Enantioenriched Amino Acid Acylboronate.

A concise synthesis of acylborons was achieved by ozonolysis of alkenyl MIDA (N-methyliminodiacetic acid) boronates. This reaction exhibits excellent functional-group tolerance and is applicable to various acyl MIDA boronates and potassium acyltrifluroborates (KATs) which could not be synthesized by previous methods. In addition, α-amino acylborons, which would be essential for peptide ligations, were prepared for the first time. The acylboron of l-alanine was obtained in high enantiopurity and found to be configurationally stable. Oligopeptide synthesis between the α-amino KATs and amino acid in dilute aqueous media was studied.

General, Mild, and Metal-Free Synthesis of Phenyl Selenoesters from Anhydrides and Their Use in Peptide Synthesis.

A mild, practical, and simple procedure for phenyl selenoesters synthesis from several anhydrides and diphenyl diselenide was developed. This transition-metal-free method provides a straightforward entry to storable Fmoc-amino acid selenoesters which are effective chemoselective acylating reagents. An application to oligopeptide synthesis was illustrated.

Synthesis of peptides with narrow molecular weight distributions via exopeptidase-catalyzed aminolysis of hydrophobic amino-acid alkyl esters

This is the first report on exopeptidase-catalyzed oligopeptide synthesis. Oligo(l-leucine) was synthesized using the exopeptidase carboxypeptidase Y while preventing enzymatic hydrolysis. A high yield of the polypeptide with a narrow polydispersity was successfully obtained by limiting polypeptide cleavage during aminolysis.

3-Nitro-l,2,4-triazol-l-yl-tris(pyrrolidin-1-yl)phosphonium hexafluorophosphate (PyNTP) as a condensing reagent for solid-phase peptide synthesis

Solid-phase oligopeptide synthesis has been well developed and most short oligopeptides can now be easily synthesized. However, when a desired oligopeptide forms a secondary structure or includes less reactive amino acids such as aminoisobutyric acid, its terminal amino groups become less reactive and synthesis of the desired oligopeptides becomes difficult. To expand the number of synthetic peptide sequences, we have developed efficient coupling conditions using 3-nitro-l,2,4-triazol-l-yl-tris(pyrrolidin-1-yl)phosphonium hexafluorophosphate (PyNTP) as a highly reactive condensing reagent on an unswellable solid support. PyNTP demonstrated higher reactivity than conventional condensing reagents and the optical purity of the synthesized oligopeptides was sufficiently high for application to general oligopeptide synthesis.

Phenolic Ester Mediated Oligopeptide Synthesis Promoted by HOBt

Although substituted phenolic ester mediated peptide synthesis is an efficient and well established method, the same via totally unsubstituted phenyl ester is not preferred due to the extremely slow rate of aminolysis. We have investigated the scope of the unsubstituted phenyl ester as an intermediate in peptide bond formation and found that it may be useful for the design of chemoselective peptide ligation when HOBt is used as an acyl transfer catalyst. The scope of HOBt catalyzed, oxo ester mediated ligation is explored for the synthesis of oligopeptides containing a cysteine, serine and threonine at the N-terminus of the ligating peptide.

Compatibility Study of Merrifield Linker in Fmoc Strategy Peptide Synthesis

The stability of Merrifield linker in Fmoc deprotection process was quantitatively investigated by establishing working curve of two major decomposition components from two resin bound dipeptide models. By sampling reaction solution and analyzing with RP-HPLC, decomposition rate was determined. The results indicated that either α-amino acid or β-amino acid anchored Merrifield linker was endurable for Fmoc strategy peptide synthesis in common de-Fmoc conditions such as 20% piperidine/DMF and 2% DBU/2% piperidine/DMF under room temperature treatments. However, Fmoc-deprotection with microwave assistance of α-amino acid anchored peptide resin with 20% piperidine/DMF more than 20 times or β-amino acid anchored peptide resin with 2% DBU/2% piperidine/DMF more than 30 times is not recommended. Feasibility of the proposed compatibility was verified by design and synthesis of a thymic humoral factor derived peptide via Fmoc strategy on Merrifield resin. Thus by choosing moderate de-Fmoc protocol, Merrifield resin is feasible for Fmoc strategy oligopeptide synthesis.

Compatibility Study of Merrifield Linker in Fmoc Strategy Peptide Synthesis

The stability of Merrifield linker in Fmoc deprotection process was quantitatively investigated by establishing working curve of two major decomposition components from two resin bound dipeptide models. By sampling reaction solution and analyzing with RP-HPLC, decomposition rate was determined. The results indicated that either α-amino acid or β-amino acid anchored Merrifield linker was endurable for Fmoc strategy peptide synthesis in common de-Fmoc conditions such as 20% piperidine/DMF and 2% DBU/2% piperidine/DMF under room temperature treatments. However, Fmoc-deprotection with microwave assistance of α-amino acid anchored peptide resin with 20% piperidine/DMF more than 20 times or β-amino acid anchored peptide resin with 2% DBU/2% piperidine/DMF more than 30 times is not recommended. Feasibility of the proposed compatibility was verified by design and synthesis of a thymic humoral factor derived peptide via Fmoc strategy on Merrifield resin. Thus by choosing moderate de-Fmoc protocol, Merrifield resin is feasible for Fmoc strategy oligopeptide synthesis.

ChemInform Abstract: Organic Synthesis Without Stoichiometric Reagents: A Guiding Principle for Reaction Development

AbstractReview: 66 refs.

Organic Synthesis without Stoichiometric Reagents: A Guiding Principle for Reaction Development

A common theme of our research program is the development of new organic transformations that operate under catalytic conditions or as ligation reactions that do not require the addition of any reagents or other additives. Our catalysis program features the transient generation of reactive species from alpha-functionalized aldehydes via intramolecular redox reactions using N-heterocyclic carbenes as multifunctional catalysts. This approach makes possible the catalytic generation of enolates, homoenolates, and activated carboxylates and their application to diastereo- and enantioselective transformation. Intermolecular redox couplings are key to a general, highly chemoselective amide-forming ligation reaction and its use for oligopeptide synthesis. The concepts behind these transformation and examples of their use as well as current and future directions of our research program are presented.

ChemInform Abstract: Novel L‐Amino Acid Ligases Catalyzing Oligopeptide Synthesis

AbstractReview: 10 refs.