. 2019 Jan 4;10(2):168–174. doi: 10.1021/acsmedchemlett.8b00535

Table 1. Affinity and Inhibitory Activity of Macrocyclic Peptides against Flaviviral Proteases.

		ZIKV (gZiPro)^b		IC₅₀/μM
Cpd.	sequence^a	K_i/μM^c (IC₅₀/μM)	K_D/μM^d	ZIKV (bZiPro)^e	DENVpro^f	WNVpro^g
1	-S-Ac-YWKI^MeY_MeNTLVNIC-NH₂	0.80 ± 0.08	0.02	1.5 ± 0.1	>10	>10
1	-S-Ac-YWKI^MeY_MeNTLVNIC-NH₂	(1.32 ± 0.03)	0.02	1.5 ± 0.1	>10	>10
2	-S–Ac-Y^MeY_MeK^MeFK^MeS^MeY_MeK^MeY_Me^MeY_MeKC-NH₂	0.44 ± 0.03	0.009	0.25 ± 0.01	1.7 ± 0.1	3.9 ± 0.4
2	-S–Ac-Y^MeY_MeK^MeFK^MeS^MeY_MeK^MeY_Me^MeY_MeKC-NH₂	(0.62 ± 0.04)	0.009	0.25 ± 0.01	1.7 ± 0.1	3.9 ± 0.4
3	-S-Ac-YTNFYLYPY^MeY_MeFC-NH₂	2.0 ± 0.2	0.008	2.2 ± 0.1	>20	>20
3	-S-Ac-YTNFYLYPY^MeY_MeFC-NH₂	(3.3 ± 0.2)	0.008	2.2 ± 0.1	>20	>20
4	-S–Ac-Y^MeGIAKYN^MeY_Me^MeY_MeIPC-NH₂	20.4 ± 2.3	0.009	4.6 ± 0.3	≥20	≥20
4	-S–Ac-Y^MeGIAKYN^MeY_Me^MeY_MeIPC-NH₂	(16.5 ± 0.9)	0.009	4.6 ± 0.3	≥20	≥20
5	-S-Ac-YTLPFHN^MeGTFFC-NH₂	>100	0.005	≥50	>20	>20
6	-S-Ac-d-YAII^MeY_MeYNKY^MeLNC-NH₂	3.5 ± 0.4	0.168	3.2 ± 0.4	>20	>20
6	-S-Ac-d-YAII^MeY_MeYNKY^MeLNC-NH₂	(7.1 ± 0.2)	0.168	3.2 ± 0.4	>20	>20

The N-terminal thioether-acyl moiety (−S-Ac−) forms a macrocycle through the side chain of the underlined cysteine residue. ^MeY_Me, N-methyl-4-O-methyl-tyrosine; ^MeF, N-methyl-phenylalanine; ^MeS, N-methyl-serine; ^MeG, N-methyl-glycine; ^MeL, N-methyl-leucine; d-Y, d-tyrosine.

Zika virus NS2B-NS3 protease (linked construct) C80S/C143S (1 nM). Substrate: Bz-Nle-Lys-Lys-Arg-AMC.

Inhibition constants (K_i) were calculated from measurements at four different substrate concentrations using a noncompetitive inhibition model (nonlinear least-squares fits). Half maximal inhibitory concentrations (IC₅₀) for a substrate concentration of 15 μM are reported in parentheses.

Dissociation constants (K_D) were determined by surface plasmon resonance using immobilized Zika virus NS2B-NS3 protease (gZiPro).

Zika virus NS2B-NS3 protease (unlinked construct; 0.5 nM). Substrate: Bz-Nle-Lys-Lys-Arg-AMC. Half maximal inhibitory concentrations (IC₅₀) were calculated for a substrate concentration of 15 μM.

Dengue virus serotype 2 NS2B-NS3 protease (100 nM). Substrate: Abz-Nle-Lys-Arg-Arg-Ser-3-(NO₂)Tyr. Half maximal inhibitory concentrations (IC₅₀) were calculated for a substrate concentration of 50 μM.

West Nile virus NS2B-NS3 protease (150 nM). Substrate: Abz-Gly-Leu-Lys-Arg-Gly-Gly-3-(NO₂)Tyr. Half maximal inhibitory concentrations (IC₅₀) were calculated for a substrate concentration of 50 μM.