A PEGA resin for use in the solid-phase chemical–enzymatic synthesis of glycopeptides

Meldal, Morten; Auzanneau, France‐Isabelle; Hindsgaul, Ole; Palcic, Monica M.

doi:10.1039/c39940001849

Cited by 125 publications

(81 citation statements)

References 10 publications

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“…In a previous study it was demonstrated that enzymes up to 50 kDa can enter the PEGA 1900 polymer matrix [31,41]. In order to develop selective enzyme inhibitors for MMP-9 it became necessary to increase the apparent pore size of PEGA supports since MMP-9 which in its proform has a MW of 92 kDa, even after cleavage of its propeptide by activation, exist in forms of approx.…”

Section: Synthesis Of Improved Pega Supportsmentioning

confidence: 99%

PEGA supports for combinatorial peptide synthesis and solid-phase enzymatic library assays

et al. 1998

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Permeable resins cross-linked with long PEG chains were synthesized for use in solid-phase enzyme library assays. High molecular weight bis-amino-polyethylene glycol (PEG) 4000, 6000, 8000 were synthesized by a three-step reaction starting from PEG-bis-OH. Macromonomers were synthesized by partial or di-acryloylation of bis-amino-PEG derivatives. Bis/mono-acrylamido-PEG were copolymerized along with acrylamide by inverse suspension copolymerization to yield a less cross-linked resin (Type I, compounds 6-9). Furthermore, acryloyl-sarcosin ethyl ester was co-polymerized along with bis-acrylamido PEG to obtain more crosslinked capacity resin (Type II, compounds 13-19). N,N-Dimethylacrylamide was used as a co-monomer in some cases. The polymer was usually obtained in a well-defined beaded form and was easy to handle under both wet and dry conditions. The supports showed good mechanical properties and were characterized by studying the swelling properties, size distribution of beads, and by estimating the amino group capacity. Depending on the PEG chain length, the monomer composition and the degree of cross-linking the PEGA supports showed a high degree of swelling in a broad range of solvents, including water, dichloromethane, DMF, acetonitril, THF and toluene: no swelling was observed in diethyl ether. The PEGA resins (Type I) with an amino acid group capacity between 0.07 and 1.0 mmol/g could be obtained by variation of the monomer composition in the polymerization mixture. Fluorescent quenched peptide libraries were synthesized on the new polymer using a multiple column library synthesizer and incubated with the matrix metalloproteinase MMP-9 after it had been activated by 4-aminophenyl mercuric acetate resulting in 67/83 kDa active enzyme. The bright beads were separated manually under a fluorescence microscope and sequenced to obtain peptide substrates for MMP-9. After treatment with ethylene diamine, high-loaded resins (Type II) have been employed in continuous flow peptide synthesis to yield peptides in excellent yield and purity.

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Section: Synthesis Of Improved Pega Supportsmentioning

confidence: 99%

PEGA supports for combinatorial peptide synthesis and solid-phase enzymatic library assays

et al. 1998

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“…While parallel synthesis of arrays of glycopeptides is readily achieved by implementation of the building-block approach (Scheme 3 a), [125] glycopeptide library synthesis in a combinatorial manner by the split and mix method has yet to prove [44,148] Lg leaving group, Pg protecting group.…”

Section: Preparation and Analysis Of Solid-phase Glycopeptide Librariesmentioning

confidence: 99%

“…PEG-based resins swell tremendously in aqueous media, thus allowing biomolecules access to the entire bead. [43,44] These resins are, therefore, suitable for protein ligation, [45] enzymatic reactions, and screening for enzyme activity and inhibition. [42, 46±51] Since PEG-based polymers confer the advantage of a support that is amenable to both synthesis as well as screening of enzyme activity and protein binding, efforts have been made to generate such polymers with superior properties in both areas.…”

Section: Supports For Solid-phase Librariesmentioning

confidence: 99%

Glycopeptide and Oligosaccharide Libraries

Hilaire

Meldal

2000

Angew. Chem. Int. Ed.

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Despite the burgeoning interest in the various biological functions and consequent therapeutic potential of the vast number of oligosaccharides found in nature on glycoproteins and cell surfaces, the development of combinatorial carbohydrate chemistry has not progressed as rapidly as expected. The reason for this imbalance is rooted in the difficulty of oligosaccharide assembly and analysis that renders synthesis a rather cumbersome endeavor. Parallel approaches that generate series of analogous compounds rather than real libraries have therefore typically been used. Since generally low affinity is obtained for interactions between carbohydrate receptors and modified oligosaccharides designed as mimetics of natural carbohydrate ligands, glycopeptides have been explored as alternative mimics. Glycopeptides have been proven in many cases to be superior ligands with higher affinity for a receptor than the natural carbohydrate ligand. High-affinity glycopeptide ligands have been found for several types of receptors including the E-, P-, and L-selectins, toxins, glycohydrolases, bacterial adhesins, and the mannose-6-phosphate receptor. Furthermore, the assembly of glycopeptides is considerably more facile than that of oligosaccharides and the process can be adapted to combinatorial synthesis with either glycosylated amino acid building blocks or by direct glycosylation of peptide templates. The application of the split and combine approach using ladder synthesis has allowed the generation of very large numbers of compounds which could be analyzed and screened for binding of receptors on solid phase. This powerful technique can be used generally for the identification and analysis of the complex interaction between the carbohydrates and their receptors.

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“…Para que essa lavagem seja realizada adequadamente, costuma-se usar uma série de solventes com diversos graus de polaridade, os quais exercem um efeito de compactação/descompactação do polímero, favorecendo a expulsão das impurezas do interior da estrutura polimérica 24 . O produto pode então ser isolado e separado da resina por meio de uma hidrólise em meio básico 25 , ácido 1 , enzimática [26][27][28][29] , ou em condições menos comuns (por exemplo, ligantes fotolábeis, que sofrem clivagem quando expostos a determinados comprimentos de onda) 30,31 . O acompanhamento da reação também constitui-se em outra grande diferença quando comparada à síntese clássica 12,21,23 , já que os intermediários da SOFS não podem ser purificados, pelo menos rotineiramente.…”

Section: Síntese Orgânica Em Fase Sólida -Sofsunclassified

A síntese orgânica em fase sólida e seus suportes poliméricos mais empregados

Marquardt¹,

Eifler‐Lima²

2001

Quím. Nova

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Recebido em 26/7/00; aceito em 6/4/01 THE SOLID PHASE ORGANIC SYNTHESIS AND ITS MOST USED POLYMERIC SUPORTS: In the last decade we have seen improved a powerfull tool to medicinal chemistry: the Solid Phase Organic Synthesis (SPOS). This metodology can be used to synthesize a large library of compounds in a short time by combinatorial chemistry, where simple chemical substances can be combinated one to each other building a library of complex compounds. In this work we present the solid phase organic synthesis and their advantage upon the tradicional organic synthesis methodology, as well as the main polimers used in the SPOS technique.Keywords: solid phase organic synthesis; polymer; drug design. Quim. Nova, Vol. 24, No. 6, 846-855, 2001. Divulgação INTRODUÇÃOA descoberta de novas substâncias com atividade terapêuti-ca, até alguns anos atrás, era realizada praticamente por meio de abordagens empíricas e básicas, uma vez que os compostos eram sintetizados, ou obtidos muitas vezes a partir de fontes naturais, e eram testados, tanto in vitro como in vivo, de forma randomizada. Esta abordagem necessita de quantidades significativas de produtos para os testes biológicos, levando à necessidade da realização de sínteses repetidas vezes, tornando todo o processo demorado e muito trabalhoso.Há mais de três décadas, uma inovadora metodologia para a síntese de peptídeos foi apresentada à comunidade científica por Bruce Merrifield 1 . Tal metodologia veio revolucionar a síntese orgânica por suas características peculiares, uma vez que abandonou a tradicional rotina, própria da síntese em solução, e introduziu o uso de polímeros insolúveis como suporte, ligados covalentemente aos substratos (geralmente peptídeos ou aminoácidos) 1,2 . A importância de suas descobertas, a partir de uma idéia aparentemente simples e, portanto, brilhante, o conduziu ao Prêmio Nobel de química em 1984, com a síntese da bradicinina 3 . A partir deste primeiro estudo seguiram-se muitos outros, com o objetivo de esclarecer, aprimorar e estender a outros setores da pesquisa os princípios utilizados por Merrifield. Com isso, dezenas de novos polímeros foram sintetizados e modificados, reações foram testadas e otimizadas, técnicas de análi-ses quali e quantitativas foram adaptadas. Concomitante a isso, a técnica em fase sólida era aplicada a novas classes de molé-culas: peptídeos 2 , nucleotídeos 4 , sacarídeos 5,6 e, mais recentemente, sobretudo a partir da segunda metade da década de 80, moléculas de natureza não polimérica 7-9 , de menor peso molecular, incluindo nesta última categoria, moléculas de origem natural 10 . Atualmente, devido à grande diversidade das reações químicas existentes, estas pesquisas tornaram-se mais freqüentes e o volume de trabalhos publicados cresce progressivamente, atestando a alta potencialidade dessa metodologia 11,12 , que está se tornando ferramenta de rotina usada pela comunidade científica internacional 13 . Neste contexto, a indústria farmacêutica não poderia ignorar as inúmeras vantagens desta metodologia para a...

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A PEGA resin for use in the solid-phase chemical–enzymatic synthesis of glycopeptides

Cited by 125 publications

References 10 publications

PEGA supports for combinatorial peptide synthesis and solid-phase enzymatic library assays

PEGA supports for combinatorial peptide synthesis and solid-phase enzymatic library assays

Glycopeptide and Oligosaccharide Libraries

A síntese orgânica em fase sólida e seus suportes poliméricos mais empregados

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