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S1 Supporting Information Enantioselective bacterial hydrolysis of amido esters and diamides derived from ()-trans-cyclopropane-1,2-dicarboxylic acid Katharina G. Hugentobler and Francisca Rebolledo* Table of Contents: I. Procedures for the synthesis of racemic compounds implied in the determination of the enantiomeric excesses: 1. Mixed benzyl methyl trans-cyclopropane-1,2-dicarboxylate (8) (S1) 2. General procedure for the preparation of racemic amido esters (±)-12a-c (S2) 3. General procedure for the preparation of racemic carbamates (±)-14a-c (S3) 4. General procedure for the derivatization of the carboxylic acids with diazomethane (S3) Scheme I (S3) II. Chiral HPLC conditions and chromatograms: 1. Biotransformation of methyl (±)-trans-2-carbamoylcyclopropanecarboxylate ((±)-4b) (S4) 2. Biotransformation of benzyl (±)-trans-2-carbamoylcyclopropanecarboxylate ((±)-4c) (S5) 3. Biotransformation of (±)-trans-N-benzylcyclopropane-1,2-dicarboxamide ((±)-10a) (S6) 4. Biotransformation of (±)-trans-N-allylcyclopropane-1,2-dicarboxamide ((±)-10b) (S7) 5. Biotransformation of (±)-trans-N-allyl-N-methylcyclopropane-1,2-dicarboxamide(±)-10c (S8) 6. The Curtius rearrangement of 11a-c: carbamates 14a-c (S9) III. 1 H and 13 C NMR spectra for new compounds (S10 to S34) I. Procedures for the synthesis of racemic compounds implied in the determination of the enantiomeric excesses 1. Mixed benzyl methyl trans-cyclopropane-1,2-dicarboxylate (8). The required racemic mixed benzyl methyl ester ()-8 could not be prepared by conventional methods. A nearly racemic sample was obtained as follows: first, ()-4b (43 mg) was incubated with a bacterial suspension (A 650 = 1.5) in fresh 0.10 M phosphate buffer pH 7.0 (43 mL) and ethanol (0.43 mL), at 28 ºC during 25 h. The resulting, nearly racemic, product 6b (ee = 8%, 37 mg, 0.26 mmol) was converted into the acid chloride as described for its analogue (±)-6a in the first step corresponding to the synthesis of (±)-9a-c. Then, the crude acid chloride was dissolved in anhydrous THF (2.0 mL) and treated with benzyl alcohol (53 L, 0.51 mmol) and triethylamine (72 mL, 0.51 mmol) at room temperature. After 12 h, aqueous 1 M HCl was added and the mixture extracted with AcOEt. Evaporation of organic solvents and purification of the residue by flash chromatography (n-hexane/ethyl acetate 15:1 as the eluent) yielded product 8 (ee = 8% chiral- HPLC analysis) as a colourless liquid. H (300.13 MHz, CDCl 3 ) 7.43-7.30 (5 H, m, Ph), AB system Electronic Supplementary Material (ESI) for Organic & Biomolecular Chemistry This journal is © The Royal Society of Chemistry 2013

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Page 1: Supporting Information Enantioselective bacterial ... · Supporting Information Enantioselective bacterial hydrolysis of amido esters and diamides derived from ( )-trans-cyclopropane-1,2-dicarboxylic

S1

Supporting Information

Enantioselective bacterial hydrolysis of amido esters and diamides derived from

()-trans-cyclopropane-1,2-dicarboxylic acid

Katharina G. Hugentobler and Francisca Rebolledo*

Table of Contents:

I. Procedures for the synthesis of racemic compounds implied in the determination of the

enantiomeric excesses:

1. Mixed benzyl methyl trans-cyclopropane-1,2-dicarboxylate (8) (S1)

2. General procedure for the preparation of racemic amido esters (±)-12a-c (S2)

3. General procedure for the preparation of racemic carbamates (±)-14a-c (S3)

4. General procedure for the derivatization of the carboxylic acids with diazomethane (S3)

Scheme I (S3)

II. Chiral HPLC conditions and chromatograms:

1. Biotransformation of methyl (±)-trans-2-carbamoylcyclopropanecarboxylate ((±)-4b) (S4)

2. Biotransformation of benzyl (±)-trans-2-carbamoylcyclopropanecarboxylate ((±)-4c) (S5)

3. Biotransformation of (±)-trans-N-benzylcyclopropane-1,2-dicarboxamide ((±)-10a) (S6)

4. Biotransformation of (±)-trans-N-allylcyclopropane-1,2-dicarboxamide ((±)-10b) (S7)

5. Biotransformation of (±)-trans-N-allyl-N-methylcyclopropane-1,2-dicarboxamide(±)-10c (S8)

6. The Curtius rearrangement of 11a-c: carbamates 14a-c (S9)

III. 1H and

13C NMR spectra for new compounds (S10 to S34)

I. Procedures for the synthesis of racemic compounds implied in the determination of the

enantiomeric excesses

1. Mixed benzyl methyl trans-cyclopropane-1,2-dicarboxylate (8).

The required racemic mixed benzyl methyl ester ()-8 could not be prepared by conventional

methods. A nearly racemic sample was obtained as follows: first, ()-4b (43 mg) was incubated

with a bacterial suspension (A650 = 1.5) in fresh 0.10 M phosphate buffer pH 7.0 (43 mL) and

ethanol (0.43 mL), at 28 ºC during 25 h. The resulting, nearly racemic, product 6b (ee = 8%, 37 mg,

0.26 mmol) was converted into the acid chloride as described for its analogue (±)-6a in the first step

corresponding to the synthesis of (±)-9a-c. Then, the crude acid chloride was dissolved in

anhydrous THF (2.0 mL) and treated with benzyl alcohol (53 L, 0.51 mmol) and triethylamine (72

mL, 0.51 mmol) at room temperature. After 12 h, aqueous 1 M HCl was added and the mixture

extracted with AcOEt. Evaporation of organic solvents and purification of the residue by flash

chromatography (n-hexane/ethyl acetate 15:1 as the eluent) yielded product 8 (ee = 8% chiral-

HPLC analysis) as a colourless liquid. H (300.13 MHz, CDCl3)7.43-7.30 (5 H, m, Ph), AB system

Electronic Supplementary Material (ESI) for Organic & Biomolecular ChemistryThis journal is © The Royal Society of Chemistry 2013

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centered to 5.13 ppm (|JA,B| = 12.5 Hz, 2H, CH2-Ph,), 3.70 (3 H, s, OMe), 2.38-2.17 (2 H, m, 2 x

CH), 1.52-1.40 (m, 2H, CH2); C (75.5 MHz, CDC13)172.0 (C=O), 171.5 (C=O), 135.4 (C), 128.5

(CH), 128.3 (CH), 128.2 (CH), 66.8 (CH2), 55.1 (CH3), 22.3 (CH), 22.2 (CH), 15.4 (CH2). MS

(ESI+) m/z 257.0 ((M+Na)+, 100%).

2. General procedure for the preparation of racemic amido esters (±)-12a-c.

A mixture of dimethyl (±)-trans-cyclopropane-1,2-dicarboxylate ((±)-3b) (0.16 g, 1.0 mmol) and

the corresponding amine (1.0 mmol) was heated in a sealed tube nearly to the boiling point of the

amine during 20 h (for 12a) or 60 h (for 12b,c). Then aqueous 1 M HCl (5 mL) and CH2Cl2 (8 mL)

were added. The organic phase was separated and the aqueous layer again extracted with CH2Cl2.

The organic phases were combined and washed with brine. Evaporation of solvents and subsequent

flash column chromatography of the residue (n-hexane/ethyl acetate 2:1 as the eluent) yielded the

corresponding amido esters (±)-12a-c.

(1) Methyl ()-trans-2-(N-benzylcarbamoyl)cyclopropanecarboxylate (()-12a). White solid

(0.12 g, 50%); mp 104-105 ºC; H (300.13 MHz, CDC13) 7.40-7.25 (5 H, m, Ph), 6.14 (1 H, br s,

NH), 4.44 (2 H, d, J = 5.7 Hz, CH2Ph), 3.67 (3 H, s, OCH3), 2.21 (1 H, ddd, J1 = 9.2 Hz, J2 = 5.5

Hz, J3 = 3.7 Hz, CH), 1.94 (1 H, ddd, , J1 = 8.9 Hz, J2 = 5.4 Hz, J3 = 3.7 Hz, CH), 1.49 (1 H, ddd,

J1 = 9.1 Hz, J2 = 5.5 Hz, J3 = 3.6 Hz, CHH), 1.33 (1 H, ddd, J1 = 9.1 Hz, J2 = 5.5 Hz, J3 = 3.6 Hz,

CHH); C (75.5 MHz, CDC13) 173.1 (C=O), 169.9 (C=O), 137.9 (C), 128.7 (CH), 127.8 (CH),

127.6 (CH), 52.0 (CH3), 44.0 (CH2), 24.2 (CH), 21.3 (CH), 14.8 (CH2); MS (ESI+) m/z 256.0

((M+Na)+, 100%).

(2) Methyl ()-trans-2-(N-allylcarbamoyl)cyclopropanecarboxylate (()-12b). White solid (62

mg, 34%); mp 86-87 ºC; H (300.13 MHz, CDC13) 5.84 [2 H, ddt (Jtrans = 17.2 Hz (d), Jcis = 10.2

Hz (d), J = 5.7 Hz (t), CH=CH2) overlapped with br s (NH)], 5.24-5.12 (2 H, m, CH=CH2), 3.90 (2

H, tt, J1 = 5.8 Hz, J2 = 1.5 Hz, NCH2), 3.69 (3 H, s, OCH3), 2.18 (1 H, ddd, J1 = 8.8 Hz, J2 = 5.7

Hz, J3 = 3.8 Hz, CH), 1.94 (1 H, ddd, J1 = 8.6 Hz, J2 = 5.8 Hz, J3 = 3.8 Hz, CH), 1.47 (1 H, ddd, J1

= 8.8 Hz, J2 = 5.8 Hz, J3 = 3.8 Hz, CHH), 1.33 (1 H, ddd, , J1 = 8.7 Hz, J2 = 5.7 Hz, J3 = 3.8 Hz,

CHH); C (75.5 MHz, CDC13) 173.1 (C=O), 169.8 (C=O), 133.8 (CH), 116.7 (CH2), 52.0 (CH3),

42.3 (CH2), 24.3 (CH), 21.3 (CH), 14.7 (CH2); MS (ESI+) m/z 389.1 ((2M+Na)+, 100%; (M+H)

+,

85%).

(3) Methyl ()-trans-2-(N-allyl-N-methylcarbamoyl)cyclopropanecarboxylate (()-12c).

Colorless oil (20 mg, 10%); H (300.13 MHz, CDC13) corresponding to a 47:53 mixture of

rotamers: 5.90-5.61 (1 H, m, CH=CH2), 5.32-5.05 (2 H, m, CH=CH2), 4.17-3.94 (2 H, m, NCH2),

3.70 and 3.68 (2 H, two s, OCH3), 3.09 (3 H for minor rotamer, s, NCH3), 2.94 (3 H for major

Electronic Supplementary Material (ESI) for Organic & Biomolecular ChemistryThis journal is © The Royal Society of Chemistry 2013

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rotamer, s, NCH3), 2.40-2.10 (2 H, m, 2 x CH), 1.50-1.12 (2 H, m, CH2); C (75.5 MHz, CDC13)

corresponding to a mixture of rotamers: 173.3, 173.2, 170.3, 169.8, 132.7, 132.4, 117.4, 116.7,

52.2, 52.0, 50.4, 34.7, 34.2, 21.7, 21.1, 21.0, 15.4, 15.3; MS (ESI+) m/z 220.0 ((M+Na)+, 100%).

3. General procedure for the preparation of racemic carbamates (±)-14a-c

Racemic carbamates (±)-14a-c were obtained from the corresponding racemic carboxylic acid (±)-

11a-c as described for the optically active samples. In turn, racemic carboxylic acids (±)-11a-c were

prepared as follows: the racemic ethyl amido ester (±)-9a-c (0.50 mmol) was dissolved in ethanol

(0.50 mL) and then aqueous 1 M NaOH (0.50 mL) was added. The mixture was heated at 80 ºC

during 2 h. After this time, ethanol was removed, water (5 mL) and aqueous 1 M NaOH (0.2 mL)

were added, and the mixture extracted with CH2Cl2. The organic phase was discarded. The aqueous

phase was acidified with conc. HCl and extracted with ethyl acetate (2 × 10 mL). The combined

organic phases were washed with brine and dried with Na2SO4. Evaporation of solvents yielded the

corresponding racemic carboxylic acid. Spectroscopic data for (±)-11a-c matched those of the

optically active samples.

(±)-11a: Yield, 95%; mp 172-173 ºC.

(±)-11b: Yield, 90%; mp 152-153 ºC.

(±)-11c: Yield, 91%; mp 73-75 ºC.

4. General procedure for the derivatization of the carboxylic acids with diazomethane.

An excess of a solution of diazomethane in a mixture of diethyl ether/methanol (40 mM) was added

to a sample of 1.0 mg of the corresponding carboxylic acid. After 2-3 min. solvents were

evaporated and the resulting methyl ester was analysed by chiral GC or HPLC. The optically active

carboxylic acids (isolated from the enzymatic reactions) and the methyl esters derivatives are

collected in Scheme I.

Scheme I. Methyl esters derivatives

Electronic Supplementary Material (ESI) for Organic & Biomolecular ChemistryThis journal is © The Royal Society of Chemistry 2013

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II. Chiral HPLC conditions and chromatograms:

1. Biotransformation of methyl (±)-trans-2-carbamoylcyclopropanecarboxylate ((±)-4b) (Table

1 of the manuscript, entry 4)

Remaining substrate (1R,2R)-4b was isolated with ee > 99%.

HPLC conditions: Chiralcel OD; hexane/propan-2-ol 90:10, 0.8 mL/min, 210 nm, 20 C; tR = 13.8

(1R,2R) and 15.9 (1S,2S) min; RS = 2.1.

(±)-4b (1R,2R)-4b with ee > 99%

Product (1S,2S)-6b was isolated with ee = 91%. It was transformed into (1S,2S)-3b (Scheme I)

GC conditions for (±)-3b: RTBetaDEXse chiral column, heating at 50 ºC during 5 min, and then, a

ramp of 1 ºC/min; tR = 56.8 (1R,2R) and 58.2 (1S,2S) min; RS = 2.9.

(±)-3b (1S,2S)-3b with ee = 91%

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2. Biotransformation of benzyl (±)-trans-2-carbamoylcyclopropanecarboxylate ((±)-4c):

Remaining substrate (1R,2R)-4c was isolated with ee = 65%.

HPLC conditions: Chiralpak IA; n-hexane/propan-2-ol 93:7, 0.8 mL/min, 20 C, 215 nm; tR = 20.4

(1S,2S) and 23.1 (1R,2R) min; RS = 2.2.

(±)-4c (1R,2R)-4b with ee > 99%

Product (1S,2S)-6c was isolated with ee = 94%. For the chiral-HPLC analysis, 6c was transformed

into 8 (Scheme I).

HPLC conditions: Chiralpak IA; n-hexane/propan-2-ol 98:2, 0.8 mL/min, 20 C, 215 nm; tR = 9.2

(1R,2R) and 9.8 (1S,2S) min; RS = 1.3.

Nearly racemic 8 (ee = 8%) (1S,2S)-8 with ee = 94%

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3. Biotransformation of (±)-trans-N-benzylcyclopropane-1,2-dicarboxamide ((±)-10a) (Table 2

of the manuscript, entry 1):

Remaining substrate (1R,2R)-10a was isolated with ee = 75%.

HPLC conditions: Chiralpak IA; n-hexane/propan-2-ol 95:5, 0.8 mL/min, 20 C, 215 nm; tR = 29.7

(1S,2S) and 34.5 (1R,2R) min; RS = 1.4.

(±)-10a (1R,2R)-10a with ee = 75%

Product (1S,2S)-11a was isolated with ee = 95%. For the chiral-HPLC analysis, 11a was

transformed into 12a (Scheme I).

HPLC conditions: Chiralpak IA; n-hexane/propan-2-ol 93:7, 0.8 mL/min, 20 C, 215 nm; tR = 17.0

(1S,2S) and 18.7 (1R,2R) min; RS = 2.1.

(±)-12a (1S,2S)-12a with ee = 95%

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4. Biotransformation of (±)-trans-N-allylcyclopropane-1,2-dicarboxamide ((±)-10b) (Table 2

of the manuscript, entry 2):

Remaining substrate (1R,2R)-10b was isolated with ee = 93%.

HPLC conditions: Chiralcel OD; n-hexane/propan-2-ol 94:6, 0.8 mL/min, 30 C, 215 nm; tR = 13.1

(1R,2R) and 14.8 (1S,2S) min; RS = 1.2.

(±)-10b (1R,2R)-10b with ee = 93%

Product (1S,2S)-11b was isolated with ee = 92%. For the chiral-HPLC analysis, 11b was

transformed into 12b (Scheme I).

HPLC conditions: Chiralcel OD; n-hexane/propan-2-ol 97:3, 0.8 mL/min, 20 C, 210 nm; tR = 25.9

(1R,2R) and 29.2 (1S,2S) min; RS = 1.7.

(±)-12b (1S,2S)-12b with ee = 92%

Electronic Supplementary Material (ESI) for Organic & Biomolecular ChemistryThis journal is © The Royal Society of Chemistry 2013

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5. Biotransformation of (±)-trans-N-allyl-N-methylcyclopropane-1,2-dicarboxamide ((±)-10c)

(Table 2 of the manuscript, entry 3):

Remaining substrate (1R,2R)-10c was isolated with ee = 97%.

HPLC conditions: Chiralpak IA; n-hexane/propan-2-ol 95:5, 0.8 mL/min, 20 C, 215 nm; tR = 32.6

(1S,2S) and 36.7 (1R,2R) min; RS = 1.3.

(±)-10c (1R,2R)-10c with ee = 97%

Product (1S,2S)-11c was isolated with ee = 99%. For the chiral-HPLC analysis, 11c was

transformed into 12c (Scheme I).

HPLC conditions: Chiralcel OD; n-hexane/propan-2-ol 95:5, 0.8 mL/min, 20 C, 215 nm; tR = 11.5

(1R,2R) and 14.1 (1S,2S) min; RS = 3.5.

(±)-12c (1S,2S)-12c with ee = 99%

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6. The Curtius rearrangement of 11a-c: carbamates 14a-c

14a. HPLC conditions: Chiralcel OD; n-hexane/propan-2-ol 80:20, 0.8 mL/min, 20 C, 215 nm; tR

= 15.3 (1S,2S) and 17.8 (1R,2R) min; RS = 1.7.

(±)-14a (1S,2S)-14a with ee = 95%

14b. HPLC conditions: Chiralcel OD; n-hexane/propan-2-ol 80:20, 0.8 mL/min, 20 C, 210 nm; tR

= 10.1 (1S,2S) and 12.1 (1R,2R) min; RS = 2.3.

(±)-14b (1S,2S)-14b with ee = 92%

14c. HPLC conditions: Chiralcel OD; n-hexane/propan-2-ol 85:15, 0.8 mL/min, 20 C, 215 nm; tR =

17.3 (1S,2S) and 21.1 (1R,2R) min; RS = 2.5.

(±)-14c (1S,2S)-14c with ee = 99%

Electronic Supplementary Material (ESI) for Organic & Biomolecular ChemistryThis journal is © The Royal Society of Chemistry 2013

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1H and

13C NMR spectra of 4b in CDCl3:

Electronic Supplementary Material (ESI) for Organic & Biomolecular ChemistryThis journal is © The Royal Society of Chemistry 2013

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1H and

13C NMR spectra of 4c in CDCl3:

Electronic Supplementary Material (ESI) for Organic & Biomolecular ChemistryThis journal is © The Royal Society of Chemistry 2013

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1H and

13C NMR spectra of 6c in CD3OD:

4.6

89

1

2.0

00

0

1.8

93

0

2.0

22

7

Inte

gra

l

7.3

54

6

5.1

79

45.1

37

85.1

35

25.0

93

6

3.3

10

0

(ppm)

1.01.52.02.53.03.54.04.55.05.56.06.57.07.58.0

R-234-crA h1_wsopt MeOD {C:\Bruker\bacs} Bruker 2

17

4.9

256

17

3.1

148

13

7.3

138

12

9.5

960

12

9.3

328

12

9.2

889

67

.89

36

49

.00

00

23

.04

27

22

.99

49

15

.71

18

(ppm)

2030405060708090100110120130140150160170

R-234-crA c13_swopt MeOD 18-07-11

Electronic Supplementary Material (ESI) for Organic & Biomolecular ChemistryThis journal is © The Royal Society of Chemistry 2013

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1H and

13C NMR spectra of 8 in CDCl3:

Electronic Supplementary Material (ESI) for Organic & Biomolecular ChemistryThis journal is © The Royal Society of Chemistry 2013

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1H and

13C NMR spectra of 9a in CDCl3:

5.0

33

2

0.9

70

3

2.0

00

0

2.0

33

7

0.9

97

2

1.1

32

6

1.0

21

7

4.0

63

7

Inte

gra

l

6.6

07

7

4.4

01

04.3

82

7

4.0

37

74.0

13

9

1.2

39

01.2

15

21.1

91

4

(ppm)

1.01.52.02.53.03.54.04.55.05.56.06.57.07.5

R-255-A h1_wsopt CDCl3 1-03-2013

17

2.8

073

17

0.0

132

13

7.9

156

12

8.5

433

12

7.7

500

12

7.4

060

76

.95

00

60

.85

91

43

.78

54

23

.98

77

21

.30

59

14

.70

66

14

.00

46

(ppm)

0102030405060708090100110120130140150160170180

R-255-A c13.220 CDCl3 1-03-2013

Electronic Supplementary Material (ESI) for Organic & Biomolecular ChemistryThis journal is © The Royal Society of Chemistry 2013

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1H and

13C NMR spectra of 9b in CDCl3:

1.8

59

7

1.9

77

1

2.0

87

1

2.0

00

0

0.9

99

6

0.9

73

8

0.9

67

9

4.1

37

6

Inte

gra

l

5.9

01

25.8

82

35.8

67

75.8

63

45.8

48

25.8

44

55.8

29

35.8

25

05.8

10

45.8

06

15.7

90

95.7

72

0

4.1

71

84.1

48

04.1

24

24.1

00

43.9

20

03.9

15

13.9

09

63.9

00

53.8

95

63.8

90

73.8

81

63.8

76

13.8

71

2

2.1

96

02.1

83

22.1

77

12.1

66

82.1

64

32.1

54

02.1

47

92.1

35

1

1.2

88

41.2

64

61.2

40

8

(ppm)

1.01.52.02.53.03.54.04.55.05.56.06.57.0

R-Id-B AmidoÚster Et/NAlil h1_wsopt CDCl3 15-02-13

1.8

59

7

1.9

77

1

5.9

01

25.8

82

35.8

67

75.8

63

45.8

48

25.8

44

55.8

29

35.8

25

05.8

10

45.8

06

15.7

90

9

5.7

72

0

5.2

38

65.2

33

15.2

28

25.2

23

3

5.1

81

35.1

75

85.1

71

55.1

67

25.1

63

05.1

58

15.1

38

05.1

33

15.1

28

85.1

24

0

(ppm)

5.005.105.205.305.405.505.605.705.805.906.00

R-Id-B AmidoÚster Et/NAlil Ampliaci¾n CDCl3 15-02-13

1.8

59

7

1.9

77

1

5.9

01

25.8

82

35.8

67

75.8

63

45.8

48

25.8

44

55.8

29

35.8

25

05.8

10

45.8

06

15.7

90

9

5.7

72

0

5.2

38

65.2

33

15.2

28

25.2

23

3

5.1

81

35.1

75

85.1

71

55.1

67

25.1

63

05.1

58

15.1

38

05.1

33

15.1

28

85.1

24

0

(ppm)

5.005.105.205.305.405.505.605.705.805.906.00

R-Id-B AmidoÚster Et/NAlil Ampliaci¾n CDCl3 15-02-13

0.9

99

6

0.9

73

8

0.9

67

9

4.1

37

6

Inte

gra

l

2.1

96

02.1

83

22.1

77

12.1

66

82.1

64

32.1

54

02.1

47

92.1

35

1

1.9

64

41.9

51

61.9

44

91.9

35

11.9

32

11.9

22

31.9

16

21.9

03

4

1.4

88

91.4

76

11.4

69

41.4

59

61.4

57

21.4

46

81.4

40

11.4

27

9

1.3

51

71.3

38

91.3

32

91.3

22

51.3

20

01.3

10

31.3

03

6

(ppm)

1.301.401.501.601.701.801.902.002.102.202.30

R-Id-B AmidoÚster Et/NAlil Ampliaci¾n CDCl3 15-02-13

Electronic Supplementary Material (ESI) for Organic & Biomolecular ChemistryThis journal is © The Royal Society of Chemistry 2013

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S16

1H and

13C NMR spectra of 9c in CDCl3:

Electronic Supplementary Material (ESI) for Organic & Biomolecular ChemistryThis journal is © The Royal Society of Chemistry 2013

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S17

1H and

13C NMR spectra of 10a (in CD3OD and DMSO-d6, respectively):

Electronic Supplementary Material (ESI) for Organic & Biomolecular ChemistryThis journal is © The Royal Society of Chemistry 2013

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S18

1H and

13C NMR spectra of 10b in CD3OD:

0.8

95

9

2.0

00

0

2.3

01

7

2.0

94

5

2.5

19

2

Inte

gra

l5.8

43

65.8

29

95.8

25

25.8

07

85.7

91

05.7

73

1

5.2

15

75.2

10

55.1

58

45.1

53

15.1

23

65.1

18

45.0

89

45.0

84

2

3.8

20

03.8

14

73.8

09

53.8

01

63.7

96

33.7

91

0

3.3

10

0

(ppm)

1.21.62.02.42.83.23.64.04.44.85.25.66.0

R-225-crB

0.8

95

9

5.9

01

05.8

82

65.8

64

75.8

48

35.8

43

65.8

29

95.8

25

25.8

07

85.7

91

05.7

73

1

(ppm)

5.805.90

2.0

00

0

5.2

15

75.2

10

5

5.1

58

45.1

53

15.1

23

65.1

18

45.0

89

45.0

84

2

(ppm)

5.10

2.3

01

7

3.8

20

03.8

14

73.8

09

53.8

01

63.7

96

33.7

91

0

(ppm)

3.803.85

Electronic Supplementary Material (ESI) for Organic & Biomolecular ChemistryThis journal is © The Royal Society of Chemistry 2013

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S19

1H and

13C NMR spectra of 10c in CD3OD:

Electronic Supplementary Material (ESI) for Organic & Biomolecular ChemistryThis journal is © The Royal Society of Chemistry 2013

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S20

1H and

13C NMR spectra of 11a in CD3OD:

Electronic Supplementary Material (ESI) for Organic & Biomolecular ChemistryThis journal is © The Royal Society of Chemistry 2013

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S21

1H and

13C NMR spectra of 11b in CD3OD:

0.3

67

2

0.9

02

5

1.7

04

2

2.0

00

0

1.9

83

8

2.3

85

1

8.4

66

2

5.8

52

05.8

29

45.7

95

25.2

19

45.2

14

25.1

62

05.1

56

85.1

29

95.1

25

25.0

95

75.0

91

0

3.8

32

63.8

20

53.8

15

23.8

08

43.8

02

13.7

96

83.3

10

0

(ppm)

1.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.5

R-225-crA2 h1_wsopt MeOD 13-07-11

17

5.8

173

17

2.7

585

13

5.3

394

11

6.3

208

49

.00

00

43

.00

61

24

.49

40

21

.94

17

14

.65

96

(ppm)

2030405060708090100110120130140150160170

R-225-crA2 c13_swopt MeOD 14-07-11

Electronic Supplementary Material (ESI) for Organic & Biomolecular ChemistryThis journal is © The Royal Society of Chemistry 2013

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S22

1H and

13C NMR spectra of 11c in CD3OD:

Electronic Supplementary Material (ESI) for Organic & Biomolecular ChemistryThis journal is © The Royal Society of Chemistry 2013

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S23

1H and

13C NMR spectra of (±)-12a in CDCl3.

Electronic Supplementary Material (ESI) for Organic & Biomolecular ChemistryThis journal is © The Royal Society of Chemistry 2013

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S24

1H and

13C NMR spectra of (±)-12b in CDCl3.

Electronic Supplementary Material (ESI) for Organic & Biomolecular ChemistryThis journal is © The Royal Society of Chemistry 2013

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S25

1H and

13C NMR spectra of (±)-12c in CDCl3.

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S26

1H NMR spectrum of the crude acyl azide (1S,2S)-13a in CDCl3.

The molar ratio 13a:12a is 96:4.

Electronic Supplementary Material (ESI) for Organic & Biomolecular ChemistryThis journal is © The Royal Society of Chemistry 2013

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S27

1H NMR spectrum of the crude acyl azide (1S,2S)-13b in CDCl3.

The molar ratio 13b:12b is 96:4.

Electronic Supplementary Material (ESI) for Organic & Biomolecular ChemistryThis journal is © The Royal Society of Chemistry 2013

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S28

1H NMR spectrum of the crude acyl azide (1S,2S)-13c in CDCl3.

The molar ratio 13c:12c is 93:7.

Electronic Supplementary Material (ESI) for Organic & Biomolecular ChemistryThis journal is © The Royal Society of Chemistry 2013

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S29

1H-NMR,

13C-NMR, and HSQC spectra of 14a in DMSO-d6:

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S30

Electronic Supplementary Material (ESI) for Organic & Biomolecular ChemistryThis journal is © The Royal Society of Chemistry 2013

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S31

1H-NMR,

13C-NMR, and HSQC spectra of 14b in CDCl3:

Electronic Supplementary Material (ESI) for Organic & Biomolecular ChemistryThis journal is © The Royal Society of Chemistry 2013

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S32

Electronic Supplementary Material (ESI) for Organic & Biomolecular ChemistryThis journal is © The Royal Society of Chemistry 2013

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S33

1H-NMR,

13C-NMR, and HSQC spectra of 14c in CDCl3:

Electronic Supplementary Material (ESI) for Organic & Biomolecular ChemistryThis journal is © The Royal Society of Chemistry 2013

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S34

Electronic Supplementary Material (ESI) for Organic & Biomolecular ChemistryThis journal is © The Royal Society of Chemistry 2013