A General Strategy for the Synthesis of Large Peptides: r1~he Combined Solid-Phase and Solution Approach.

Riniker, B.; Flörsheimer, Andreas; Fretz, Heinz; Sieber, Peter; Kamber, B.

doi:10.1016/0040-4020(93)80017-n

Cited by 82 publications

(43 citation statements)

References 32 publications

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“…There are many reports of the syntheses of peptide libraries (16,17), and many applications for these libraries have been reported, such as determination of substrate specificity (18), protease inhibitors (19,20), ligand-binding activity (21,22), mapping the S' subsites of serine proteases (23), and optimization of enzyme substrates (24). We decided to explore the use of libraries…”

mentioning

confidence: 99%

Inhibitors of human heart chymase based on a peptide library.

Bastos

Maeji

Abeles

1995

Proc. Natl. Acad. Sci. U.S.A.

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We have synthesized two sets of noncleavable peptide-inhibitor libraries to map the S and S' subsites of human heart chymase. Human heart chymase is a chymotrypsin-like enzyme that converts angiotensin I to angiotensin II. The first library consists of peptides with 3-fluorobenzylpyruvamides in the P1 position. (Amino acid residues of substrates numbered P1, P2, etc., are toward the N-terminal direction, and P;, P2, etc., are toward the C-terminal direction from the scissile bond.) The Pi and P' positions were varied to contain each one of the 20 naturally occurring amino acids and P3 was kept constant as an arginine. The second library consists of peptides with phenylalanine keto-amides at P1, glycine in PN, and benzyloxycarbonyl (Z)-isoleucine in P4. The P2 and P3 positions were varied to contain each ofthe naturally occurring amino acids, except for cysteine and methionine. The peptides of both libraries are attached to a solid support (pins). The peptides are evaluated by immersing the pins in a solution of the target enzyme and evaluating the amount of enzyme absorbed. The pins with the best inhibitors will absorb most enzyme. The libraries select the best and worst inhibitors within each group of peptides and provide an approximate ranking of the remaining peptides according to Ki. Through this library, we determined that Z-Ile-Glu-Pro-Phe-CO2Me and (F)-Phe-CO-Glu-Asp-ArgOMe should be the best inhibitors of chymase in this collection of peptide inhibitors. We synthesized the peptides and found K1 values were 1 nM and 1 ,uM, respectively. The corresponding K1 values for chymotrypsin were 10 nM and 100 ,uM. The use of libraries of inhibitors has advantages over the classical method of synthesis of potential inhibitors in solution: the libraries are reusable, the same libraries can be used with a variety of different serine proteases, and the method allows the screening of hundreds of compounds in short periods of time.Inhibitors of proteolytic enzymes generally have the following

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confidence: 99%

Inhibitors of human heart chymase based on a peptide library.

Bastos

Maeji

Abeles

1995

Proc. Natl. Acad. Sci. U.S.A.

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“…To improve the cleavage yield by concurrent lose of cleavage effectiveness, thiol scavengers are added. 18 Resins and linkers of the trityl type have the additional advantage to suppress the piperazine formation at the dipeptide step of the synthesis. 8 This side reaction is especially pronounced in peptides, which contain Pro or Gly at their C-terminus.…”

Section: Solid Phase Synthesis Of Protected Segmentsmentioning

confidence: 99%

“…9 Preparative reverse phase high performance liquid chromatography (RP-HPLC) using acetonitrile/water systems as the eluent is best suited for purifying Fmoc/tBu-protected segments. 18,40 Diphenyl derivatized materials are best suited for the purification of very lypophylic fragments. Segments of medium polarity are effectively purified by C8-HPLC.…”

Section: Purification Of Protected Segmentsmentioning

confidence: 99%

“…If a permanent protecting group is required, the fragment is tert-butylated by tert-butyl trichloroacetimidate 54 (TBTA). This combined solid-phase and solution approach was exemplified during the synthesis 18 of calcitonin and neuropeptide Y by the application of the HMPB-linker 5. The esterification of long segments is easily performed by treatment with 2-chlorotritylchloride and DIPEA in DMF/DCM mixtures.…”

Section: Lypophilic Segment Coupling-phase Change Synthesismentioning

confidence: 99%

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9-Fluorenylmethyloxycarbonyl/tbutyl-based convergent protein synthesis

Barlos

Gatos

1999

Biopolymers

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Besides linear solid phase peptide synthesis, segment condensation in solution and chemical ligation, convergent peptide synthesis (CPS) was developed in order to enable the efficient preparation of complex peptides and small proteins. According to this synthetic strategy, solid phase synthesized and suitably protected peptide fragments corresponding to the entire peptide/protein‐sequence are condensed on a solid support or in solution, to the target protein. This review summarizes CPS performed utilizing the mild 9‐fluorenylmethyloxycarbonyl/tbutyloxycarbonyl‐based protecting scheme for the amino acids. © 1999 John Wiley & Sons, Inc. Biopoly 51: 266–278, 1999

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“…Even though SPPS can be troublesome in practice for peptides of more than 10 residues, in some cases peptides of about 50 residues have been synthesized successfully by this system (Patarroyo et al 1988) and several peptides from 30 to 50 residues synthesized by SPPS are now in the market (Bruckdorfer et al 2004). Commonly, peptides of less than 30 residues are produced entirely by solid phase sequential synthesis (Lloyd-Williams and Giralt, 2000), while larger peptides (up to 60 residues) must be produced by convergent synthesis in which protected peptide fragments are synthesized by SPPS and then combined by liquid phase synthesis (Riniker et al 1993;Barlos and Gatos, 1999). Proteins are preferably produced by chemoselective ligation in which all the unprotected linked fragments have been previously synthesized by SPPS.…”

Section: Large Scale Chemical Synthesis Of Peptidesmentioning

confidence: 99%

Peptide synthesis: chemical or enzymatic

Guzmán

Barberis

Illanes

2007

Electron. J. Biotechnol.

178

139

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Abbreviations:CD 280synthesis is probably the way to go, since the good properties of each technology can be synergistically used in the context of one process objective.Peptides are heteropolymers composed by amino acid residues linked by peptidic bonds between the carboxyl group of one amino acid residue and the α-amino group of the next one. The definition is rather vague in terms of chain length, peptides ranging from two to a few dozens residues. Its lower limit of molecular mass has been set rather arbitrarily in 6000 Da; molecules larger than that are considered proteins. Peptides are molecules of paramount importance in several fields, especially in health care and nutrition. The case of the hormone insulin (51 residues, 5773 Da) and the non-caloric sweetener aspartame (a dipeptide of aspartic acid and esterified phenylalanine) are relevant examples of those fields of application and the range of molecular size. Medium to small size peptides are, however, the most relevant for such applications.

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A General Strategy for the Synthesis of Large Peptides: r1~he Combined Solid-Phase and Solution Approach.

Cited by 82 publications

References 32 publications

Inhibitors of human heart chymase based on a peptide library.

Inhibitors of human heart chymase based on a peptide library.

9-Fluorenylmethyloxycarbonyl/tbutyl-based convergent protein synthesis

Peptide synthesis: chemical or enzymatic

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