title: A convergent solution-phase synthesis of the macrocycle Ac-Phe-[Orn-Pro-D-Cha-Trp-Arg], a potent new antiinflammatory drug.
abstract: Relatively few cyclic peptides have reached the pharmaceutical marketplace during the past decade, most produced through fermentation rather than made synthetically. Generally, this class of compounds is synthesized for research purposes on milligram scales by solid-phase methods, but if the potential of macrocyclic peptidomimetics is to be realized, low-cost larger scale solution-phase syntheses need to be devised and optimized to provide sufficient quantities for preclinical, clinical, and commercial uses. Here, we describe a cheap, medium-scale, solution-phase synthesis of the first reported highly potent, selective, and orally active antagonist of the human C5a receptor. This compound, Ac-Phe[Orn-Pro-d-Cha-Trp-Arg], known as 3D53, is a macrocyclic peptidomimetic of the human plasma protein C5a and displays excellent antiinflammatory activity in numerous animal models of human disease. In a convergent approach, two tripeptide fragments Ac-Phe-Orn(Boc)-Pro-OH and H-d-Cha-Trp(For)-Arg-OEt were first prepared by high-yielding solution-phase couplings using a mixed anhydride method before coupling them to give a linear hexapeptide which, after deprotection, was obtained in 38% overall yield from the commercially available amino acids. Cyclization in solution using BOP reagent gave the antagonist in 33% yield (13% overall) after HPLC purification. Significant features of the synthesis were that the Arg side chain was left unprotected throughout, the component Boc-d-Cha-OH was obtained very efficiently via hydrogenation of d-Phe with PtO(2) in TFA/water, the tripeptides were coupled at the Pro-Cha junction to minimize racemization via the oxazolone pathway, and the entire synthesis was carried out without purification of any intermediates. The target cyclic product was purified (>97%) by reversed-phase HPLC. This convergent synthesis with minimal use of protecting groups allowed batches of 50-100 g to be prepared efficiently in high yield using standard laboratory equipment. This type of procedure should be useful for making even larger quantities of this and other macrocyclic peptidomimetic drugs.
authors.affiliation: Institute for Molecular Bioscience, The University of Queensland, Brisbane 4072, Australia.
authors.forename: Robert C
authors.initials: RC
authors.lastname: Reid
authors.forename: Giovanni
authors.initials: G
authors.lastname: Abbenante
authors.forename: Stephen M
authors.initials: SM
authors.lastname: Taylor
authors.forename: David P
authors.initials: DP
authors.lastname: Fairlie
mesh_terms: D000893:Anti-Inflammatory Agents / Q000032:analysis / Q000138:chemical synthesis* / Q000494:pharmacology; D020650:Combinatorial Chemistry Techniques; D015936:Complement C5a / Q000378:metabolism; D003500:Cyclization; D006801:Humans; D007202:Indicators and Reagents; D018716:Molecular Mimicry; D010456:Peptides, Cyclic / Q000032:analysis / Q000138:chemical synthesis* / Q000494:pharmacology; D044087:Receptor, Anaphylatoxin C5a / Q000037:antagonists & inhibitors; D012697:Serine Endopeptidases / Q000138:chemical synthesis* / Q000494:pharmacology
chemical_list: C480102:Ac-Phe-(Orn-Pro-D-Cha-Trp-Arg); D000893:Anti-Inflammatory Agents; D007202:Indicators and Reagents; D010456:Peptides, Cyclic; D044087:Receptor, Anaphylatoxin C5a; C041833:complement C5a-inhibitors; D015936:Complement C5a; D012697:Serine Endopeptidases