Supplementary informationLipophilic teicoplanin pseudoaglycon derivatives active against vancomycin and

teicoplanin resistant enterococci

Zsolt Szűcs,a Ilona Bereczki,a Magdolna Csávás,a Erzsébet Rőth,a Anikó Borbás,a Gyula

Batta,b Eszter Ostorházi,c Réka Szatmári,c Pál Herczegha

aDepartment of Pharmaceutical Chemistry, University of Debrecen, Egyetem tér 1, H-4032

Debrecen, Hungary,bDepartment of Organic Chemistry, University of Debrecen, Egyetem tér 1, H-4032

Debrecen, Hungary,cDepartment of Medical Microbiology, Semmelweis University, Mária u. 41, H-1085

Budapest, Hungary*Correspondence: Pál Herczegh, Department of Pharmaceutical Chemistry, University of Debrecen, Egyetem tér 1, H-4010, Debrecen, HungaryE-mail: herczeghp@gmail.com;

Table of contents:

Antibacterial study................................................................................................2

MALDI-TOF spectra of teicoplanin derivatives 1, 2, 4, 5 and 6..........................3

Figures and tables for NMR assignment...............................................................8

NMR Spectra.......................................................................................................12

References...........................................................................................................24

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Antibacterial studyThe antibacterial activity of 1-9 was tested against a panel of Gram-positive bacteria using

broth microdilution method as previously described.1

Table S1. Antibacterial activity of compounds 1-9

Teicoplanin

1 2 3 4 5 6 7 8 9

(MIC, µg/ml)

Bacillus subtilis ATCC 6633

0.5 1.0 1.0 4.0 1.0 2.0 16 0.5 1.0 2.0

Staphylococcus aureus MSSA ATCC 29213

0.5 1.0 4.0 4.0 0.5 0.5 1.0 0.5 0.5 0.5

Staphylococcus aureus MRSA ATCC 33591

0.5 0.5 1.0 4.0 0.5 0.5 1.0 0.5 0.5 0.5

Staphylococcus epidermidis biofilm forming ATCC 35984

4.0 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.156

Enterococcus faecalis ATCC 29212

1.0 0.5 0.5 0.5 2.0 4.0 1.0 0.5 1.0 4.0

Staphylococcus epidermidis mecA

16 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.312

Enterococcus faecalis 15376 VanA

256 0.5 4.0 0.5 1.0 4.0 2.0 1.0 2.0 4.0

Enterococcus faecalis ATCC 51299 VanB

0.5 1.0 4.0 0.5 1.0 4.0 2.0 1.0 1.0 4.0

MIC: Minimum Inhibitory Concentration; ATCC: American Type Culture Collection; MSSA: Methicillin Sensitive Staphylococcus; MRSA: Methicillin Resistant Staphylococcus aureus; mecA: mecA gene expression in Staphylococcu;, vanA +: vanA gene positive; vanB +: vanB gene positive.

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MALDI-TOF spectra of teicoplanin derivatives 1, 2, 4, 5 and 6

Compound 1

S3

Compound 2

S4

1726.7800

10001200

14001600

18002000

2200m/z

0

100

200

300

Intens.

Compound 4

S5

Compound 5

S6

Compound 6

S7

1123.95

1165.27

1200.90

1228.28

1258.72

1298.65

1345.331371.42

1408.27

1461.33

1522.08

1581.60

1633.89

1680.41

1780.26

1825.24

1858.45

1893.421914.44

1935.47

1980.39

2075.21

2110.402134.62

2156.53

2202.31

2243.94

2271.47

2424.57

2817.54

12001400

16001800

20002200

24002600

2800m/z

0

500

1000

1500

2000

2500

3000

Intens.

Figures and tables for NMR assignment

Fig. S1. Structure and atom numbering for the NMR assignment of compounds 1, 2, 4, 5, 6.

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Fig. S2. Atom numberings of the lipophilic substituents (R in Fig. S1) for the NMR assignment.

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Table S2. 2D NMR data for compounds 1 and 2.

Assignment 1 13C 1 1H 2 13C 2 1H

x1 55.9 5.68 56.1 5.65x2 55.0 4.95 55.1 4.94x3 58.4 5.35 58.5 5.35x4 n.d. n.d. 55.3 5.65z6 76.7 5.39 76.7 5.382f 130.7 7.68 130.6 7.663b 110.5 6.36 110.6 6.354b 108.0 5.53 108.0 5.544f 104.9 5.11 104.9 5.115b n.d. n.d. n.d. n.d.6b 128.9 7.88 129.0 7.87GlcNAc 1 99.8 4.40 99.8 4.40

C5 56.4 3.60 56.5 3.60C6 34.4 1.67/1.55 34.5 1.66/1.55C7 40.3 2.41/1.85 40.3 2.42/1.87C8 28.6 1.35 28.7 1.33C9 25.3 1.54 25.4 1.51C10 38.5 3.18/3.13 38.5 3.18/3.13C11 35.0 2.17 35.1 2.15C12 172.5 - 172.3 -C13 - - 38.7 3.01C14 - - 29.3 1.38H3 - - - 7.75

Table S2. (continued) 2D NMR data for compounds 4, 5 and 6.

Assignment 4 13C 4 1H 5 13C 5 1H 6 13C 6 1H

x1 64.3 7.09 64,3 7,05 64.4 7.08x2 55.7 4.86 55,7 4,86 55.7 4.86x3 58.5 5.40 58.5 5,40 58.6 5.40x4 54,9 5,66 54,9 5,66 54.9 5.64z6 75.8 5,36 75.6 5,40 76,0 5,422f 131,9 7,67 131,7 7,67 131,8 7,683b 110.5 6.34 110,5 6,34 110,5 6,334b 108.5 5.55 108,4 5,54 108,4 5,544f 105.0 5.08 104,9 5,07 104.9 5,095b 136.4 7.08 136,4 7,09 136,5 7,116b 129.0 7.87 129.0 7.87 128.9 7.86GlcNAc 1 99.1 4.34 98.8 4,38 99,2 4,39

Triazole 4 144,3 - 144,3 - 142.3 -Triazole 5 124,4 7,73 124,4 7,74 123.2 7,66

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α-Glcp 1 97,4 4,74 97,5 4,71 - -α-Glcp 2 80,4 3,15 80,0 3,07 - -α-Glcp 3 81,6 3,39 81,5 3,24 - -α-Glcp 4 77,4 3,29 70,1 3,12 - -α-Glcp 5 70,0 3,54 71,5 3,42 - -MeO-1 54,6 3,26 54,6 3,26 - -

Butyl 2, 2' 19.0 1.32 18.9 1.31 - -Butyl 3, 3' 32.2 1,46 32.1 1.45 - -Butyl 4, 4' 13.9 0,85 14,0 0,86 - -

Phe 2,6 114.0 6,90 - - - -Phe 3,5 129,7 7,22 - - - -Phe 4 131,1 - - - - -Phe 1 159,2 - - - - -Phe 9 (OCH2) 73.9 4.64 / 4.49 - - - -Phe O-Me 55,3 3,72 - - - -

α-Galp 1 - - - - 96.2 5.39

α-Galp 2-5

- - - - 67.7 3.81- - - - 70.0 4.32- - - - 70.6 4.60- - - - 71.4 4.20

α-Galp 6 - - - - 31.7 3.44 / 3.27α-Galp Cq - - - - 108.5/109.0 -

α-Galp CH3- - - - 25.9 1.29- - - - 24.7 1.23

Maleimide 2,5 - - - - 166.0 -Maleimide 3,4 - - - - 136.0 -Maleimide CH2 - - - - 33.6 4.59

TEG a 63,6 4,43 63,6 4,44 - -TEG bulk 69.5-70.4 3.42-3.49 69.5-70.5 3.43-3.51 - -

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NMR SpectraMethyl 2,3-di-O-n-butyl-4,6-O-p-methoxybenzylidene-α-D-glucopyranoside (14)

S12

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Methyl 2,3-di-O-n-butyl-4-O-p-methoxybenzyl-α-D-glucopyranoside (15)

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Methyl 2,3-di-O-n-butyl-4-O-p-methoxybenzyl-6-O-(2,5,8,11,14-pentaoxaheptadec-16-

ynyl)-α-D-glucopyranoside (16)

S16

S17

Methyl 2,3-di-O-n-butyl-6-O-(2,5,8,11,14-pentaoxaheptadec-16-ynyl)-α-D-

glucopyranoside (17)

S18

S19

Compound 1

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Compound 2

S21

Compound 4

S22

Compound 5

S23

Compound 6

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References1. Sztaricskai, F.; Batta, G., Herczegh, P.; Balázs, A.; Jekő, J.; Rőth, E.; Szabó, P. T.;

Kardos, S.; Rozgonyi, F.; Boda, Z. A new series of glycopeptide antibiotics incorporating a squaric acid moiety. J. Antibiot. 2006, 59, 564-582.

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