Table 3.

Broad-spectrum antiviral activity against EV-D68, EV-A71, CVB3, and mouse microsomal stability.

Anti-EV-D68 in RD cells (μM)	Anti-EV-A71 in RD cells (μM)	Anti-CVB3 in Vero C1008 cells (μM)	Mouse microsomal stability T_1/2 (mins)	CL_int(liver) (ml/min/kg)
US/KY/14–18953 EC₅₀ = 0.4 ± 0.2 CC₅₀ = 73.7 ± 19.1 SI₅₀ = 184 US/MO/14–18947 EC₅₀ = 0.2 ± 0.07 CC₅₀ = 73.7 ± 19.1 SI₅₀ = 369	Tainan EC₅₀ = 4.2 ± 0.5 CC₅₀ = 73.7 ± 19.1 SI₅₀ = 18 US/AK EC₅₀ > 10	EC₅₀ = 0.3 ± 0.2 CC₅₀ = 116.2 ± 56.3 SI₅₀ = 387	22.4	245.5
US/KY/14–18953 EC₅₀ = 0.9 ± 0.3 CC₅₀ = 47.8 ± 3.6 SI₅₀ = 53 US/MO/14–18947 EC₅₀ = 0.6 ± 0.3 CC₅₀ = 47.8 ± 3.6 SI₅₀ = 80	Tainan EC₅₀ = 7.8 CC₅₀ = 47.8 ± 3.6 SI₅₀ = 6 US/AK EC₅₀ > 10	EC₅₀ = 1.1 ± 0.2 CC₅₀ > 30.0 SI₅₀ > 27	60.2	91.2
US/KY/14–18953 EC₅₀ = 0.5 ± 0.3 CC₅₀ = 65.8 ± 16.8 SI₅₀ = 132 US/MO/14–18947 EC₅₀ = 0.3 ± 0.1 CC₅₀ = 65.8 ± 16.8 SI₅₀ = 219	Tainan EC₅₀ = 18.3 ± 4.2 CC₅₀ = 65.8 ± 16.8 SI₅₀ = 4 US/AK EC₅₀ = 11.7 ± 3.0 CC₅₀ = 65.8 ± 16.8 SI₅₀ = 6	EC₅₀ = 2.0 ± 0.2 CC₅₀ = 185.9 ± 13.9 SI₅₀ = 93.0	N.T.	N.T.
US/KY/14–18953 EC₅₀ = 0.3 ± 0.2 CC₅₀ = 80.6 ± 3.3 SI₅₀ = 269 US/MO/14–18947 EC₅₀ = 0.2 ± 0.1 CC₅₀ = 80.6 ± 3.3 SI₅₀ = 403	Tainan EC₅₀ = 5.2 ± 0.2 CC₅₀ = 80.6 ± 3.3 SI₅₀ = 16 US/AK EC₅₀ = 4.9 ± 0.2 CC₅₀ = 80.6 ± 3.3 SI₅₀ = 16	EC₅₀ = 0.4 ± 0.1 CC₅₀ = 128.8 ± 25.8 SI₅₀ = 322.0	23.6	232.2
US/KY/14–18953 EC₅₀ = 0.1 ± 0.1 CC₅₀ = 36.2 ± 3.1 SI₅₀ = 362 US/MO/14–18947 EC₅₀ = 0.05 ± 0.02 CC₅₀ = 36.2 ± 3.1 SI₅₀ = 724	Tainan EC₅₀ = 2.2 ± 0.1 CC₅₀ = 36.2 ± 3.1 SI₅₀ = 17 US/AK EC₅₀ = 2.3 ± 0.1 CC₅₀ = 36.2 ± 3.1 SI₅₀ = 16	EC₅₀ = 0.06 ± 0.04 CC₅₀ = 48.8 ± 17.7 SI₅₀ = 813	34.2	160.5
US/KY/14–18953 EC₅₀ = 0.2 ± 0.1 CC₅₀ = 32.4 ± 4.1 SI₅₀ = 162 US/MO/14–18947 EC₅₀ = 0.1 ± 0.02 CC₅₀ = 32.4 ± 4.1 SI₅₀ = 324	Tainan EC₅₀ = 3.1 ± 1.0 CC₅₀ = 32.4 ± 4.1 SI₅₀ = 11 US/AK EC₅₀ = 3.6 ± 0.2 CC₅₀ = 32.4 ± 4.1 SI₅₀ = 9	EC₅₀ = 0.2 ± 0.2 CC₅₀ = 40.1 ± 13.2 SI₅₀ = 200.5	114.7	47.9
US/KY/14–18953 EC₅₀ = 0.4 ± 0.1 CC₅₀ = 48.2 ± 2.3 SI₅₀ = 121 US/MO/14–18947 EC₅₀ = 0.3 ± 0.2 CC₅₀ = 48.2 ± 2.3 SI₅₀ = 161	Tainan EC₅₀ = 9.0 ± 0.3 CC₅₀ = 48.2 ± 2.3 SI₅₀ = 5 US/AK EC₅₀ = 14.0 ± 2.1 CC₅₀ = 48.2 ± 2.3 SI₅₀ = 3	EC₅₀ = 0.4 ± 0.1 CC₅₀ = 52.3 ± 5.0 SI₅₀ = 131	114.1	48.1
US/KY/14–18953 EC₅₀ = 0.3 ± 0.2 CC₅₀ = 36.7 ± 5.6 SI₅₀ = 122 US/MO/14–18947 EC₅₀ = 0.2 ± 0.1 CC₅₀ = 36.7 ± 5.6 SI₅₀ = 184	Tainan EC₅₀ = 7.9 ± 0.5 CC₅₀ = 36.7 ± 5.6 SI₅₀ = 5 US/AK EC₅₀ > 10	EC₅₀ = 0.4 ± 0.2 CC₅₀ = 92.3 ± 3.2 SI₅₀ = 231	N.T.	N.T.
US/KY/14–18953 EC₅₀ = 0.04 ± 0.02 CC₅₀ = 42.3 ± 8.3 SI₅₀ = 1058 US/MO/14–18947 EC₅₀ = 0.05 ± 0.03 CC₅₀ = 42.3 ± 8.3 SI₅₀ = 846	Tainan EC₅₀ = 2.9 ± 0.2 CC₅₀ = 42.3 ± 8.3 SI₅₀ = 15 US/AK EC₅₀ > 10	EC₅₀ = 0.07 ± 0.0 μM CC₅₀ = 94.5 ± 4.1 SI₅₀ = 1350	26.6	206.6
US/KY/14–18953 EC₅₀ = 0.1 ± 0.1 CC₅₀ = 20.6 ± 0.9 SI₅₀ = 206 US/MO/14–18947 EC₅₀ = 0.1 ± 0.03 CC₅₀ = 20.6 ± 0.9 SI₅₀ = 206	Tainan EC₅₀ = 7.9 ± 0.4 CC₅₀ = 20.6 ± 0.9 SI₅₀ = 3 US/AK EC₅₀ = 8.8 ± 0.8 CC₅₀ = 20.6 ± 0.9 SI50 = 2	EC₅₀ = 0.4 ± 0.2 CC₅₀ = 111.8 ± 7.6 SI₅₀ = 280	N.T.	N.T.
US/KY/14–18953 EC₅₀ = 0.1 ± 0.2 CC₅₀ = 39.0 ± 3.4 SI₅₀ = 390 US/MO/14–18947 EC₅₀ = 0.06 ± 0.03 CC₅₀ = 39.0 ± 3.4 SI₅₀ = 650	Tainan EC₅₀ = 6.3 ± 0.6 CC₅₀ = 39.0 ± 3.4 SI₅₀ = 6 US/AK EC₅₀ = 8.0 ± 0.5 CC₅₀ = 39.0 ± 3.4 SI₅₀ = 5	EC₅₀ = 0.2 ± 0.1 CC₅₀ = 90.3 ± 5.9 SI₅₀ = 452	N.T.	N.T.
US/KY/14–18953 EC₅₀ = 0.3 ± 0.1 CC₅₀ = 103.1 ± 49.5 SI₅₀ = 344 US/MO/14–18947 EC₅₀ = 0.2 ± 0.1 CC₅₀ = 103.1 ± 49.5 SI₅₀ = 516	Tainan EC₅₀ = 8.4 ± 2.6 CC₅₀ = 103.1 ± 49.5 SI₅₀ = 12 US/AK EC₅₀ = 11.5 ± 0.8 CC₅₀ = 103.1 ± 49.5 SI₅₀ = 9	EC₅₀ = 0.3 ± 0.05 CC₅₀ = 70.7 ± 4.8 SI₅₀ = 236	75.4	72.8
US/KY/14–18953 EC₅₀ = 0.4 ± 0.1 CC₅₀ = 42.1 ± 13.4 SI₅₀ = 105 US/MO/14–18947 EC₅₀ = 0.2 ± 0.1 CC₅₀ = 42.1 ± 13.4 SI₅₀ = 211	Tainan EC₅₀ = 38.3 ± 6.5 CC₅₀ = 42.1 ± 13.4 SI₅₀ = 1 US/AK EC₅₀ > 30	EC₅₀ = 0.6 ± 0.2 CC₅₀ = 63.7 ± 3.2 SI₅₀ = 106	100.8	54.5
US/KY/14–18953 EC₅₀ = 1.1 ± 0.4 CC₅₀ = 198.6 ± 9.4 SI₅₀ = 181 US/MO/14–18947 EC₅₀ = 1.1 ± 0.1 CC₅₀ = 198.6 ± 9.4 SI₅₀ = 181	Tainan EC₅₀ = 30.9 ± 1.3 CC₅₀ = 198.6 ± 9.4 SI₅₀ = 6 US/AK EC₅₀ = 20.9 ± 8.2 CC₅₀ = 198.6 ± 9.4 SI₅₀ = 10	EC₅₀ = 1.2 ± 0.1 CC₅₀ > 300.0 SI₅₀ > 250	N.T.	N.T.

Antiviral efficacy was determined in the CPE assay. Cytotoxicity was determined using the neutral red method. The results are the mean ± standard deviation of three repeats. SI = selectivity index (CC₅₀/EC₅₀). T_1/2 was obtained using mouse liver microsomes. N.T. = not tested.