Synthesis and biological evaluation of aryl phosphoramidate prodrugs of

fosfoxacin and its derivatives

Mathilde Munier, Denis Tritsch, Didier Lièvremont, Michel Rohmer, Catherine Grosdemange-Billiard*

Laboratoire Chimie et Biochimie de Molécules Bioactives – Université de Strasbourg/CNRS, UMR 7177,

Institut Le Bel, 4 rue Blaise Pascal, 67081 Strasbourg, France

* Corresponding author: Tel.: +33-368-851-349; e-mail: grosdemange@unistra.fr

Table of contents

Fig. S1 Stability of the prodrugs 8-16 in buffer (TEOA) 3

Fig. S2 Carboxypeptidase Y mediated cleavage of prodrug 8a, monitored by 31P NMR 4

Fig. S3 Carboxypeptidase Y mediated cleavage of prodrug 16a, monitored by 31P NMR. 5

Fig. S4 Carboxypeptidase Y mediated cleavage of prodrug 16b, monitored by 31P NMR. 6

Fig. S5 Carboxypeptidase Y mediated cleavage of prodrug 8b, monitored by 31P NMR. 7

Fig. S6 Stability of 8a in M. smegmatis and E. coli supernatants. 8

Fig. S7 Stability of 8b in M. smegmatis and E. coli supernatants. 9

Fig. S8 Esterase and phosphatase activities in M. smegmatis crude cell free system tested on 8a 10

Fig. S9 Esterase and phosphatase activities in E. coli crude cell free system tested on 8a 11Fig. S10 Esterase and phosphatase activities in M. smegmatis crude cell free system tested on 8a 12

Fig. S11 Esterase and phosphatase activities in E. coli crude cell free system tested on 8a 13

1H -NMR (400 MHz, CDCl3) : 8a 14

13C -NMR (125.8 MHz, CDCl3) : 8a 15

31P-NMR (162.05MHz, CDCl3) : 8a 16

Mass spectroscopy : 8a 17

1H -NMR (400 MHz, CDCl3) : 8b 18

13C -NMR (125.8 MHz, CDCl3) : 8b 19

1

31P-NMR (162.0 MHz, CDCl3) : 8b 20

Mass spectroscopy : 8b 21

1H -NMR (400 MHz, CDCl3) : 16a 22

13C -NMR (125.8 MHz, CDCl3) : 16a 23

31P-NMR (162.0 MHz, CDCl33) : 16a 24

Mass spectroscopy : 16a 25

1H -NMR (300 MHz, CDCl3) : 16b 26

13C -NMR (125.8 MHz, CDCl3) : 16b 27

31P -NMR (162.0 MHz, CDCl3) : 16b 28

Mass spectroscopy : 16b 29

2

Fig. S1

3

Fig. S2

4

Fig. S3

5

Fig. S4

6

Fig. S5

7

Fig. S6

8

Fig. S7

9

Fig. S8

10

Fig. S9

11

Fig. S10

12

Fig. S11

13

1H-NMR (400 MHz, CDCl3) : 8a

14

13C-NMR (125.8 MHz, CDCl3) : 8a

15

31P-NMR (162.0 MHz, CDCl3) : 8a

16

Masse speectroscopy : 8a

17

1H-NMR (400 MHz, CDCl3) : 8b

18

13C-NMR (125.8 MHz, CDCl3) : 8b

19

31P-NMR (162.0 MHz, CDCl3) : 8b

20

Masse speectroscopy : 8b

21

1H-NMR (400 MHz, CDCl3) : 16a

22

13C-NMR (125.8 MHz, CDCl3) : 16a

23

31P-NMR (162.0 MHz, CDCl3) : 16a

24

Masse speectroscopy : 16a

25

1H-NMR (400 MHz, CDCl3) : 16b

26

13C-NMR (125.8 MHz, CDCl3) : 16b

27

31P-NMR (162.0 MHz, CDCl3) : 16b

28

Masse speectroscopy : 16b

29