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0013588527
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note
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Enzyme assay; The inhibitory effects of each compound on the enzymatic activities of five serine proteases were evaluated using the purified enzymes and chromogenic substrates. The enzymes and those substrates used here were as follow, N-succinyl-Ala-Ala-Pro-Phe-pNA (1.5 mM) for human recombinant chymase and bovine pancreatic α-chymotrypsin (Sigma Type I-S; 47 ng/mL); N-succinyl-Ala-Ala-Pro-Phe-pNA (3 mM) for human leukocyte cathepsin G (Elastin Products Co., Inc.; 2.5 U/mL); N-succinyl-Ala-Ala-Ala- pNA (1 mM) for porcine pancreatic elastase (Sigma Type III; 0.8 mg/mL); Bz-L-Arg-pNA (1 mM) for porcine pancreatic trypsin (Wako Pure Chemicals; 1 U/mL). All experiments were carried out using 50 mM HEPES buffer (pH 7.4) containing 0.1 mM NaCl, except for the buffer used for chymase, which contained 50 mM Tris-HCl (pH 8.0) and 1 M KCl.
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13
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0013617749
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note
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Each enantiomer of 2,2-dimemyl-3-(N-benzoyl)amino-5-phenylpentanoic acid was isolated by HPLC using chiral column (CHIRALCEL OD, 10 × 250 mm) eluted with a mixture of hexane and 2-propanol (3:1) and was used for the anhydride formation according to the procedure shown in Scheme 1.
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14
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0013590717
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note
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D = +51.0° (c 0.25, MeOH).
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15
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0030053083
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