Supporting Information

Supplementary data

Tables S1-S3 and Fig. S1-S2 are provided as supplementary data.

Table S1:Nucleoside bisubstrate analogues used inthis study and their actual and predicted activities.

S.N. / Structure / Actual activity (pKIapp) / PHASE
predicted activity / MIC99 (µM) / Actual class / Predicted class
1 / / 8.18 / 8.11 / 0.29 / Active / Active
2 / / 8.43 / 8.3 / 0.19 / Active / Active
3 / / 8.64 / 8.59 / 1.56 / Active / Active
4 / / 8.46 / 8.44 / 25 / Mid-active / Active
5 / / 8.49 / 8.69 / 1.56 / Active / Active
6 / / 6.08 / 6.98 / >200 / Low-active / Low-active
7 / / 7.21 / 7.12 / >200 / Low-active / Low-active
8 / / 4.78 / 4.52 / >200 / Low-active / Low-active

Table S2:Nucleoside bisubstrate analogues used inthis study and their actual and predicted activities.

S.N. / Compound,
B = / R / Actual activity (pKIapp) / PHASE
predicted activity / MIC99 (µM) / Actual class / Predicted class
9 / / X=Y= N / 8.18 / 8.36 / 0.39 / Active / Active
10 / X = N; Y = CH / 7.61 / 7.58 / 6.25 / Mid-active / Active
11 / X = Y = CH / 7.70 / 7.62 / 6.25 / Mid-active / Low-active
12 / / = Ob / 6.10 / 6.27 / >100 / Low-active / Low-active
13 / -NMe2 / 6.42 / 6.34 / 50 / Low-active / Low-active
14 / -NHMe / 8.80 / 8.49 / 0.39 / Active / Active
15 / -NHEt / 8.36 / 8.5 / 0.39 / Active / Active
16 / -NH-n-Pr / 8.46 / 7.21 / 0.39 / Active / Active
17 / -NH-i-Pr / 8.15 / 7.98 / 25 / Mid-active / Mid-active
18 / -NH-i-Bu / 5.94 / 6.03 / >100 / Low-active / Low-active
19 / -NHcyclopropyl / 8.73 / 8.66 / 0.098 / Active / Active
20 / -NHcyclobutyl / 6.91 / 6.81 / >100 / Low-active / Low-active
21 / -NHcyclopentyl / 5.03 / 5.2 / >100 / Low-active / Low-active
22 / -NHBn / 5.08 / 6.07 / >100 / Low-active / Low-active
23 / / -Br / 11.69 / 11.7 / >50 / Low-active / Low-active
24 / -N3 / 10.37 / 10.49 / >50 / Low-active / Low-active
25 / -NH2 / 9.74 / 9.95 / >50 / Low-active / Low-active
26 / / -I / 8.52 / 8.5 / 0.19 / Active / Active
27 / -NHPh / 9.03 / 8.88 / 0.049 / Active / Active
28 / -CCPh / 9.40 / 9.45 / 0.049 / Active / Active
29 / -Ph / 9.57 / 9.68 / 0.049 / Active / Active
30 / -biphen-2-yl / 7.33 / 8.87 / >50 / Low-active / Low-active
31 / -biphen-3-yl / 8.85 / 8.96 / 25-50 / Low-active / Low-active
32 / -biphen-4-yl / 9.00 / 9.04 / 25-50 / Low-active / Low-active

aZ = O for compounds 6, 7, 9-29; Z = CH2 for compound 8.bN-1 of the purine is protonated.

Table S3:Nucleoside bisubstrate analogues used inthis study and their actual and predicted activities.

S.N. / R / Actual activity (pKIapp) / PHASE
predicted activity / MIC99 (µM) / Actual class / Predicted class
33 / hydroxymethyl / 9.05 / 9.04 / 6.25 / Mid-active / Mid-active
34 / methoxycarbonyl / 8.50 / 8.62 / 3.13 / Mid-active / Mid-active
35 / ethoxycarbonyl / 8.50 / 8.67 / 3.13 / Mid-active / Mid-active
36 / n-propyl / 8.80 / 9.02 / 1.56 / Active / Mid-active
37 / n-butyl / 8.72 / 9.01 / 6.25 / Mid-active / Mid-active
38 / n-pentyl / 9.21 / 9.33 / 12.5 / Mid-active / Mid-active
39 / n-hexyl / 9.54 / 9.19 / 25 / Mid-active / Mid-active
40 / n-heptyl / 9.39 / 9.25 / >25 / Low-active / Low-active
41 / n-octyl / 8.89 / 8.62 / >25 / Low-active / Low-active
42 / n-decyl / 8.29 / 8.32 / 50 / Low-active / Low-active
43 / n-dodecyl / 8.58 / 8.63 / >25 / Low-active / Low-active
44 / isobutyl / 8.85 / 8.61 / 6.25 / Mid-active / Mid-active
45 / tert-butyl / 9.02 / 9.55 / 6.25 / Mid-active / Mid-active
46 / cyclopropyl / 9.34 / 9.53 / 3.13 / Mid-active / Mid-active
47 / cyclopentyl / 9.27 / 8.75 / 1.56 / Active / Mid-active
48 / cyclohexyl / 9.49 / 9.71 / 12.5 / Mid-active / Mid-active
49 / cyclohex-1-enyl / 8.82 / 8.69 / 6.25 / Mid-active / Mid-active
50 / phenyl / 8.49 / 8.27 / 3.13 / Mid-active / Active
51 / 2-methylphenyl / 9.23 / 9.24 / 3.13 / Mid-active / Mid-active
52 / 3-methylphenyl / 9.02 / 8.81 / 3.13 / Mid-active / Mid-active
53 / 4-methylphenyl / 9.32 / 9.18 / Ndb / - / -
54 / 2-aminophenyl / 9.11 / 9.17 / 0.78 / Active / Active
55 / 3-aminophenyl / 9.20 / 8.87 / Ndb / - / -
56 / 4-aminophenyl / 9.06 / 9.12 / 0.78 / Active / Active
57 / pyrid-2-yl / 9.02 / 9.21 / 0.78 / Active / Active
58 / pyrid-3-yl / 9.07 / 8.91 / 0.78 / Active / Active
59 / pyrid-4-yl / 8.83 / 8.95 / 0.78 / Active / Active
60 / 2-hydroxyphenyl / 8.95 / 9.03 / 1.56 / Active / Active
61 / 3-hydroxyphenyl / 9.24 / 9.12 / 1.56 / Active / Active
62 / 4-hydroxyphenyl / 9.21 / 9.2 / 1.56 / Active / Active
63 / -N3a / 8.29 / 8.4 / 0.39 / Active / Active

a azide instead of triazole substitution; bNot determined.

Fig. S1 CPH1 based alignment nucleoside bisubstrate analogues reported as MbtA inhibitors used inthis study

Fig. S2 Geometry of the pharmacophore red sphere with vector shows acceptor feature, blue sphere with vector shows donor feature and orange torus shows the aromatic ring feature

a) CPH1 features and distances

b) CPH1 features and angles