Phe-CO-Arg-Val-DPhe

As lowering crude protein (CP) in poultry diets continues to minimize amino acid excess, it is important to understand the limiting order of amino acids and the impact of their deficiencies. Therefore, a pair of experiments were conducted to observe the effects of individual amino acid deletions on growth performance, carcass traits, and nutrient utilization. Both experiments involved 3 control diets based on wheat and soybean meal, including a 210.0 g/kg CP industry control (IC), 186.7 g/kg CP positive control (PC) supplemented with feed-grade amino acids to match the IC amino acid profile, 186.7 g/kg CP negative control (NC) with reducing N corrected apparent metabolizable energy (AMEN) by 0.5 MJ/kg and removing feed-grade amino acids beyond L-Lys-HCl, DL-Met, and L-Thr from PC. Ten deletion diets where the following supplemented amino acids were individually removed from the PC: Val, Ile, Leu, Trp, Arg, His, Phe + Tyr, glycine equivalence (Glyequi), Pro, and Energy (0.5 MJ/kg reduction in AMEN of the PC). All diets were formulated to contain similar concentrations of digestible Lys, total sulfur amino acid (TSAA) and Thr. Experimental diets were offered to broiler chickens from 15 to 22 d post-hatch in a cage study (Exp. 1) to gain digestibility and nutrient utilization data; whereas they were offered from 15 to 35 d post-hatch in a floor-pen study (Exp. 2) to gain performance and carcass yield data. The removal of supplemented Val, Arg, and Ile resulted in reduction on broiler performance (P < 0.05), and the removal of Val, Arg, Ile, and Glyequi negatively influenced carcass traits (P < 0.05). Results from both experiments indicate that Val and Arg are co-limiting in wheat-soybean meal diets, but that Ile and Glyequi may potentially limit breast and thigh development.

Additional structure-activity relationship studies on potent cyclic peptide inhibitors of very late antigen-4 (VLA-4) are reported. The new N- to C-terminal cyclic hexa-, hepta- and octapeptide inhibitors like cyclo(MeIle/MePhe-Leu-Asp-Val-X) (X = 2-4 amino acids containing hydrophobic and/or basic side chains) were synthesized using solid phase peptide synthesis methods. The peptides were evaluated in in vitro cell adhesion assays and in in vivo inflammation models. Many of the peptides like cyclo(MePhe-Leu-Asp-Val-D-Arg-D-Arg) (ZD7349) (17), cyclo(MeIle-Leu-Asp-Val-D-Arg-D-Arg-D-Phe) (20), cyclo(MeIle-Leu-Asp-Val-D-Arg-D-Arg-MePhe) (21) and cyclo(MePhe-Leu-Asp-Val-D-Arg-D-Arg-D-Ala-D-Ala) (23) were potent inhibitors of VLA-4-mediated cell adhesion and inhibited ovalbumin-induced delayed type hypersensitivity (DTH) response in mice. The more potent compounds were highly selective and did not affect U937 cell adhesion to fibronectin (VLA-5), phorbolmyristate acetate or PMA-differentiated U937 cell adhesion to intercellular cell adhesion molecule-1 (ICAM-1)-expressing Chinese hamster ovary cells (LFA-1) and adenosine diphosphate (ADP)-induced platelet aggregation (GPIIb/IIIa). In contrast to the inhibitors like Ac-cyclo(D-Lys-D-Ile-Leu-Asp-Val) and cyclo(CH2CO-Ile-Leu-Asp-Val-Pip-CH2CO-Ile-Leu-Asp-Val-Pip) described earlier, the new compounds were much more compatible with the depot formulations based on poly(DL-lactide-co-glycolide) polymers. The hexapeptide cyclo(MePhe-Leu-Asp-Val-D-Arg-D-Arg) (ZD7349) (17) inhibited MOLT-4 cell adhesion to fibronectin and vascular cell adhesion molecule-1 (VCAM-1) with IC50 values of 260 and 330 nM, respectively, and did not show any significant effect against other integrins (IC50 > 300 microM). ZD7349 inhibited ovalbumin-induced DTH response in mice when administered continuously using a mini-pump (ED50 0.01 mg/kg/day) or when given as an s.c. or i.v. bolus injection at a dose of 1-10 mg/kg. ZD7349 was also active in type II collagen-induced arthritis (CIA) and experimental autoimmune encephalomyelitis (EAE) tests at a dose of 3-10 mg/kg. The peptide was released from some formulations over a period of 10-20 days. ZD7349 is currently undergoing pre-clinical investigation.