TABLE 1

Summary of hit activities from the primary high-throughput screen, orthogonal, and counter screen assays

Chemical Groups	Number	Compound Identification	DNPEP				ENPEP	Δ Activity Score ^{^a}
			Asp-AMC Hydrolysis		Ang II Hydrolysis		Glu-AMC Hydrolysis
			Activity Score (12.3 µM)	IC₅₀ (µM)	Activity Score (10 µM)	IC₅₀ (µM)	Activity Score (10 µM)
A	1	93293^{^b}	−95.0 ± 1.4	1.4	−93.9 ± 0.6	0.1	−77.8 ± 9.7	−16.1
	2	392425^{^b}	−87 ± 1.4	5.0	−89.6 ± 0.3	0.81	−87.4 ± 2.5	−2.2
B	3	897833^{^c}	−83 ± 0.2	1.7	−55.6 ± 11.3	12	6.5 ± 11.1	−62.1
C	4	98897^{^b}	−95.7 ± 0.4	0.54	−94.3 ± 0.9	1.3	−71.9 ± 2.2	−22.4
	5	769078^{^b}	−63.8 ± 2.4	5.8	−92.7 ± 2.4	0.40	−84.4 ± 1.1	−8.3
	6	702610	−84.4 ± 1.0	2.4	−93.3 ± 0.6	2.0	−45 ± 7.2	−48.3
D	7	241320^{^c}	−74.5 ± 5.2	4.4	−91.8 ± 5.2	0.26	−28.8 ± 8.7	−63.0
	8	705521^{^c}	−82.5 ± 0.9	0.16	−78.9 ± 4.5	5.0	4.5 ± 3.9	−83.4
	9	936427^{^b}	−83 ± 0.7	0.83	−93.0 ± 1.3	1.7	−89 ± 0.8	−4.0
	10	374081^{^c}	−87.4 ± 1.2	2.7	−92.2 ± 2.3	4.7	5.5 ± 3.4	−97.7
	11	405279	−73.6 ± 1.4	2.3	−91.0 ± 2.4	3.6	−45.8 ± 4.9	−45.2
	12	914971	−81.8 ± 0.2	3.1	−50.1 ± 6.5	8.3	−22.4 ± 3.1	−27.7
	13	920521^{^b}	−68.1 ± 2.5	10.2	−59.3 ± 8.9	18	−43.2 ± 2.9	−16.1
E	14	897927^{^c}	−74.4 ± 3.1	5.7	−91.7 ± 3.2	5.7	−17.3 ± 2.5	−74.4
	15	251431^{^c}	−74.7 ± 0.7	1.6	−80.5 ± 4.6	3.6	55.5 ± 9.6	−136.0
F	16	347749^{^b}	−72.3 ± 0.8	2.0	−90.7 ± 3.6	1.4	−72.8 ± 8.9	−17.9
	17	252185^{^c}	−81.7 ± 0.7	3.2	−56.0 ± 9.7	17	−0.2 ± 7.5	−55.8
G	18	505901^{^b}	−93.4 ± 0.2	0.84	−94.0 ± 1.1	2.1	−83.9 ± 2.0	−10.1
	19	923254	−74.6 ± 1.2	7.7	−93.0 ± 2.4	4.2	−44.8 ± 5.3	−48.2
	20	773587	−62.8 ± 2.3	8.0	−88.9 ± 2.5	6.2	−61 ± 4.2	−27.9
H	21	406365^{^c}	−78.8 ± 0.8	5.7	−75.3 ± 4.0	3.0	−0.8 ± 6.6	−74.5
I	22	899189	−77.4 ± 1.5	3.2	−90.7 ± 4.3	6.0	−64.9 ± 3.8	−25.8
	23	312792^{^b}	−77.8 ± 1.0	3.2	−87.2 ± 8.7	2.8	−92.7 ± 1.0	5.5

Ang II, angiotensin II; Asp-AMC, L-aspartic acid 7-amido-4-methylcoumarin; DNPEP, aspartyl aminopeptidase; ENPEP, glutamyl aminopeptidase; Glu-AMC, L-glutamic acid 7-amido-4-methylcoumarin.
↵a Δ Activity score = activity score_DNPEP − activity score_ENPEP, where activity score_DNPEP is for Ang II hydrolysis.
↵b Dual inhibitors of DNPEP and ENPEP.
↵c DNPEP-selective inhibitors.