Facile folding of insulin variants bearing a prosthetic C-peptide prepared by α-ketoacid-hydroxylamine (KAHA) ligation

Boross, Gábor N.; Shimura, Satomi; Besenius, Melissa; Tennagels, Norbert; Rossen, Kai; Wagner, Michael R.; Bode, Jeffrey W.

doi:10.1039/c8sc03738h

Cited by 24 publications

(33 citation statements)

References 30 publications

(27 reference statements)

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“…Our group recently reported a convergent, modular synthesis for the homoserine (Hse) variants of insulins (ThrB27Hse), which allows the on-demand incorporation of any additional functionality into the amino acid sequence. [26] Importantly, ThrB27Hse replacement showed no detriment to the biological activity across a number of different insulin variants.…”

Section: Synthesis Of Glargine M2 Insulin With a Photoprotected Ornitmentioning

confidence: 97%

Chemoselective Derivatization of Folded Synthetic Insulin Variants with Potassium Acyltrifluoroborates (KATs)

Boross

Schauenburg

Bode

2019

Helvetica Chimica Acta

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Synthetic folded insulin variants containing an ornithine‐hydroxylamine residue are readily modified in aqueous buffers by amide‐forming ligations with potassium acyltrifluoroborates (KATs). The synthetic insulin analogs were prepared by Fmoc‐SPPS, α‐ketoacid‐hydroxylamine (KAHA) ligation, and a prosthetic C‐peptide that delivers the correct disulfide pattern and allows facile incorporation at the B0 position of Glargine M2 of a new ornithine hydroxylamine protected with a photolabile group. The folded insulin is readily modified by photo‐deprotection followed by amide‐forming KAT ligation to give insulin variants labeled with dyes, lipids, and PEGs, as well as the formation of a covalent dimer.

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Section: Synthesis Of Glargine M2 Insulin With a Photoprotected Ornitmentioning

confidence: 97%

Chemoselective Derivatization of Folded Synthetic Insulin Variants with Potassium Acyltrifluoroborates (KATs)

Boross

Schauenburg

Bode

2019

Helvetica Chimica Acta

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“…Recently, Kent's group further evaluated several methods in constructing the GluA4-ThrB30 ester intermediate to demonstrate that native chemical ligation was the optimal synthetic route [21]. Bode and coworkers have applied their KAHA (α-ketoacidhydroxylamine) ligation chemistry to the syntheses of the M2 metabolite of glargine as well as mouse, guinea pig and human insulin [22]. The site chosen for the keto-acid/5-oxaproline KAHA ligation was TyrB26-ThrB27 resulting in a homoserine (Hse) residue at B27 following the concomitant O->N acyl shift and traceless removal of the prosthetic C-peptide under alkaline treatment.…”

Section: Approachmentioning

confidence: 99%

Recent Advances in the Chemical Synthesis of Insulin and Related Peptides

2020

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“…Two main synthetic strategies are often applied for the formation of disulfide bridges in peptides and proteins . The first one relies on subjecting the reduced peptide to freely oxidative folding conditions using a buffer containing redox reagents (e.g., Cys/Cystine) to form the correct connectivity under thermodynamic control ,,. The main limitations of this approach are the slow rate of the reaction (hours–days) and the requirement for careful optimization of the desired ratio between the redox reagents for each system …”

Section: Figurementioning

confidence: 99%

Palladium‐Mediated Direct Disulfide Bond Formation in Proteins Containing S‐Acetamidomethyl‐cysteine under Aqueous Conditions

et al. 2019

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One of the applied synthetic strategies for correct disulfide bond formation relies on the use of orthogonal Cys protecting groups. This approach requires purification before and after the deprotection steps, which prolongs the entire synthetic process and lowers the yield of the reaction. A major challenge in using this approach is to be able to apply one‐pot synthesis under mild conditions and aqueous media. In this study, we report the development of an approach for rapid disulfide bond formation by employing palladium chemistry and S‐acetamidomethyl‐cysteine [Cys(Acm)]. Oxidation of Cys(Acm) to the corresponding disulfide bond is achieved within minutes in a one‐pot operation by applying palladium and diethyldithiocarbamate. The utility of this reaction was demonstrated by the synthesis of the peptide oxytocin and the first total chemical synthesis of the protein thioredoxin‐1. Our investigation revealed a critical role of the Acm protecting group in the disulfide bond formation, apparently due to the generation of a disulfiram in the reaction pathway, which significantly assists the oxidation step.

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Facile folding of insulin variants bearing a prosthetic C-peptide prepared by α-ketoacid-hydroxylamine (KAHA) ligation

Abstract: Ester forming KAHA-ligation and a traceless prosthetic C-peptide provides a modular platform for the synthesis and folding of insulin variants.

Cited by 24 publications

References 30 publications

Chemoselective Derivatization of Folded Synthetic Insulin Variants with Potassium Acyltrifluoroborates (KATs)

Chemoselective Derivatization of Folded Synthetic Insulin Variants with Potassium Acyltrifluoroborates (KATs)

Recent Advances in the Chemical Synthesis of Insulin and Related Peptides

Palladium‐Mediated Direct Disulfide Bond Formation in Proteins Containing S‐Acetamidomethyl‐cysteine under Aqueous Conditions

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