Table 1 Initial SAR to differentiate TLR inhibition from ER binding affinity.

IC₅₀ and EC₅₀values for TLR9 were obtained using RAW264.7 cells. Values are presented as means ± SD of three independent experiments.

Entry	Compound	R₁	R₃	R₄	R₅	TLR9	TLR9	RBA*
Entry	Compound	R₁	R₃	R₄	R₅	IC₅₀ (μM)	Toxicity EC₅₀ (μM)	ERα	ERβ
1	MPP	OH		CH₃	BSC	2.11 ± 0.64	13.2 ± 2.4	12 ± 2	0.05 ± 0.01
2	TSI-13-03	OH		Et		32.1 ± 1.43	43 ± 6.4	8.92 ± 0.53	0.324 ± 0.01
3	TSI-13-30	OCH₃		CH₃	BSC	1.97 ± 1.72	33.2 ± 2.7	0.013 ± 0.001	<0.001
4	TSI-13-27	OH		H	BSC	1.03 ± 0.8	12.3 ± 3.7	5.92 ± 0.13	0.021 ± 0.007
5	TSI-13-04	OH		Et	BSC	2.84 ± 0.12	11.6 ± 0.9	11.5 ± 1.5	0.49 ± 0.27
6	TSI-13-19	OH			BSC	28.2 ± 0.5	35.7 ± 4.7	17.8 ± 5.1	0.044 ± 0.02
7	TSI-13-06	OH		BSC	Et	3.12 ± 0.103	11.2 ± 1.1	7.34 ± 0.036	0.467 ± 0.13
8	TSI-13-07	OH		BSC	n-Pr	2.96 ± 0.2	12.9 ± 0.3	8.54 ± 2.1	1.45 ± 0.05
9	TSI-13-05	OH	BSC	Et		3.33 ± 0.2	7.4 ± 0.4	0.132^†	0.010^†

*RBA indicates relative binding affinity values, where estradiol = 100.

†Single determinations.