Table 2. In Vitro Activities and Binding of the Reference Compounds Miconazole (1) and Fluconazole and of the Newly Synthesized Compounds 2a, 3a–5a, 3b–6b, 7, and 8b–10b.

Cpd	R	R1	R2	R3	R4	R5	X	stereo configuration	MW (g/mol)	EC50a (μM)	KDb (μM)	PDB ID
2a								mixed	379.33	∼50% inhibition at 10 μM	0.088 ± 0.004	7RKR
3a	H	OCH3		H		CF3		R	390.36	69% inhibition at 50 μM	N/Dc
3b	H	OCH3	H	H	H	CF3	CH	S	390.36	24.0 ± 0.9	N/D
4a	H	F		Cl		Cl		R	379.21	21.50 ± 0.04	0.173 ± 0.006
4b	H	F	H	Cl	H	Cl	CH	S	379.21	4.80 ± 0.05	0.028 ± 0.002	7RKW
5a	Cl	Cl		H		CF3		R	429.22	10.20 ± 0.03	0.161 ± 0.008
5b	Cl	Cl	H	H	H	CF3	CH	S	429.22	4.30 ± 0.05	0.070 ± 0.002	7RKT
6b	Cl	Cl	Cl	H	Cl	H	CH	S	430.11	2.60 ± 0.03	≤0.005d
7	H	F	H	H	H	CF3	N	S	379.31	20.60 ± 0.04	0.087 ± 0.010
8b	H	F		Cl		Cl		S	365.23	2.40 ± 0.04	0.007 ± 0.002
9b	Cl	Cl		H		CF3		S	415.24	1.50 ± 0.02	≤0.005
10b	Cl	Cl		Cl		Cl		S	416.13	3.50 ± 0.01	≤0.005
MICe (1)	Cl	Cl	Cl	H	Cl	H	CH	mixed	416.10	1.40 ± 0.02	≤0.005
FLUf								N/A	306.30	13.9 ± 0.0115	0.141 ± 0.021	6AY4

^aCompound concentration corresponding to 50% growth inhibition determined from dose–response curves: experiments performed in triplicate against axenically cultured N. fowleri trophozoits. Standard deviation for each compound was calculated from three independent experiments.

^bK_D was determined at 0.5 μM NfCYP51; the inhibitor concentration was 0.025–0.5 μM. The standard deviation for each compound was calculated from three independent titrations.

^cN/D = not determined.

^dEstimated from linear dependence of binding from compound concentration.

^eMIC = miconazole.

^fFLU = fluconazole.