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In this strategy, base-labile main chain protection is combined with side-chain protecting groups and resin linkages sensitive to TFA treatment (Scheme 2), thereby bypassing the hazardous HF cleavage step commonly employed in Boc-SPPS.
All Boc-Nα-protected amino acids with side chain protection: Arg(Tos), Asn Xan), Cys Acm), Tyr 2-Br-Z) were commercially available (Bachem Inc., Tyr 2-Br-ZCA, USA).
By investing in quality control and supply chain protection now, pharmaceutical companies, for example, can protect their patents and their market share by ensuring that their supply chain is pure, that their quality is guaranteed and that their customers are getting a safe medication, even when that reassurance costs more.
For the amino acids Asp and Glu, an OtBu side chain protection was applied, whereas for Trp BOC and for Tyr, tBu protection groups were used.
Final cleavage of the peptide from the resin as well as the Boc side chain protection group removal was accomplished by treatment with TFA/CH2Cl2/Et3SiH/H2O 60 35:2.5/2.5 for 2 h.
SPPS using the orthogonal 9-Fluorenylmethyloxycarbonyl (Fmoc)/tertiary-Butyl (tBu) chemistry (Chang and Meienhofer 1978) allows the use of piperidine and trifluoroacetic acid to remove the N-terminal Fmoc and the side chain protection groups, respectively.
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Side chain protections were as follows: Asp, Glu, Ser, Thr and Tyr: t-Bu; Asn, Gln and His: Trt; Arg: Pbf; Lys and Trp: Boc.
The following permanent, orthogonal, side chain protections were used: 2,2,4,6,7-pentamethyldihydrobenzofuran-5-sulfonyl (Pbf) for Arg, tert-butyl for Asp, benzyl (Bz) for Glu, and benzyloxycarbonyl (Cbz) for Lys.
Briefly, Fmoc-Rink-MBHA resin (0.64 mmol/g) was used as solid support, and side-chain protection was performed with tert-butyloxyrocarbonyl for Lys and Trp, and tert-butyl for Tyr.
The peptides were synthesized by a standard solution peptide synthesis using Boc/ tert-butyl strategy for the Tyr and minimal side-chain protection (hydrochloride salt) for the Arg-NH2 moieties.
We have thus recently proposed a novel strategy for obtaining both Asn and Gln aldehyde synthons based on a controlled side-chain protection approach, as well as a suitable solvent partition procedure affording yields higher than 80%% (Carreño et al. 2011).
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