Table 2.

The inhibition effect of prepared bisamides on bacterial adenylyl cyclase (ACT) and cell viability in J744A.1 murine macrophages and their effects on mammalian ACs isoforms in HEK293 cells stably transfected with AC1, AC2, and AC5.

Compound	ACT inhibition	Viability	Response of Control [%]c
	IC50 [μM]a	[%]b	AC1	AC2	AC5
4	2.12 ± 0.58	88	ND	ND	ND
5	14.2 ± 4.3	86	ND	ND	ND
10	>10	83	74 ± 3	143 ± 11	113 ± 1
13 *	4.68 ± 1.56	54	61 ± 9	101 ± 10	147 ± 9
18	>10	72	73 ± 18	185 ± 32	108 ± 13
21a *	1.82 ± 0.58	112	48 ± 3	82 ± 21	177 ± 18
21b	2.62 ± 0.62	99	37 ± 8	136 ± 25	99 ± 13
21c *	0.45 ± 0.17	99	36 ± 10	62 ± 13	129 ± 4
21d	0.68 ± 0.08	81	54 ± 7	156 ± 2	197 ± 14
21e	1.72 ± 0.38	44	38 ± 9	150 ± 26	169 ± 26
21f *	1.16 ± 0.01	57	29 ± 4	160 ± 31	159 ± 7
21g	1.43 ± 0.04	86	32 ± 6	119 ± 13	172 ± 16
21h	1.62 ±0.36	35	25 ±7	110 ± 9	140 ± 29
21i	>10	115	39 ± 3	134 ± 26	117 ± 4
21j	0.26 ± 0.05	72	38 ± 9	104 ± 7	106 ± 8
21k	8.45 ± 0.81	106	57 ± 3	109 ± 5	78 ± 4
21l	0.66 ± 0.18	99	77 ± 6	12 ± 27	73 ± 5
21m	4.30 ± 0.29	99	75 ± 12	122 ± 41	101 ± 13
21n	>10	94	64 ± 16	97 ± 28	77 ± 3
23	0.93 ± 0.19	36	23 ± 4	165 ± 41	131 ± 4
2	0.15 ± 0.03	93	64 ± 9	200 ± 45	99 ± 8
ST034307d	NDg	ND	14 ± 13	398 ± 96	196 ± 31
SKF83566e	ND	ND	78 ± 8	(−)7 ± 11	94 ± 5
SQ22536f	ND	ND	46 ± 2	27 ± 12	45 ± 15

^aCompound concentration that causes a 50% decrease in ACT-induced cAMP accumulation in J774A.1 macrophages; data are the mean ± SD of at least three independent experiments.

^bPercent cell viability in J774A.1 cells (n = 3) at a fixed prodrug concentration (10 μM) versus untreated control.

^cData are the mean ± SEM relative to the control response (100%) of at least two independent experiments at 30 μM concetration.

^dST034307 is a selective inhibitor of AC1.

^eSKF83566 is a selective inhibitor of AC2.

^fSQ22536 is a nonselective P-site inhibitor.

^gND: not determined.

^*<60% cell viability in HEK-AC1 cells (n = 3).