S1

Highly efficient asymmetric bioreduction of 1-aryl-2-(azaaryl)ethanones. Chemoenzymatic synthesis of lanicemine

Ramón Liz,a Elisa Liardoa,b and Francisca Rebolledoa

aDepartamento de Química Orgánica e Inorgánica, Universidad de Oviedo, 33006 Oviedo, Spain.bCurrent address: EVOTEC, Center of Drug Discovery and Development, 37135, Verona, Italy.

frv@uniovi.es

SUPPORTING INFORMATION

Table of contents

1. General information (p. S2)

2. Synthesis of ketone substrates 1 (p. S2)

3. Enzymatic screening for the reduction of ketones 1 (p. S3)

4. HPLC and GC analytical data for C and ee determinations (p. S7)

5. Copy of chiral HPLC chromatograms (p. S8)

6. Assignment of the absolute configuration of the optically active alcohols 2 from their MTPA

ester derivatives 3 (p. S14)

7. Copy of NMR spectra of Mosher ester derivatives 3 (p. S15)

8. Copy of 1H- and 13C-NMR spectra of compounds 2a-f, 5, 6, and 7 (p. S24)

Electronic Supplementary Material (ESI) for Organic & Biomolecular Chemistry.This journal is © The Royal Society of Chemistry 2019

S2

1. General information

Enzymes

Codex® KRED Screening Kit was purchased from Codexis.1

General methods1H-NMR and proton-decoupled 13C-NMR spectra (CDCl3) were obtained using a Bruker DPX-300

(1H, 300.13 MHz and 13C, 75.5 MHz) spectrometer using the δ scale (ppm) for chemical shifts. Some 1H-NMR spectra of MTPA ester derivatives were measured in a 600 or 400 MHz spectrometer.

Calibration was made on the signal of the solvent (1H: residual CHCl3, 7.26; residual CHD2OD, 3.31; 13C: CDCl3, 76.95; CD3OD, 49.00). The selective-1D TOCSY experiments were performed using

standard Bruker pulse program (selmlgp). HPLC analyses were performed on a Hewlett Packard 1100

LC liquid chromatograph. GC analyses were performed on a Hewlett Packard 6890 Series II

chromatograph using the column CP-ChiraSil-DEX CB (25 m 0.25 nm 0.25 μm, 12.2 psi N2).

Optical rotations were measured using a Perkin-Elmer 241 polarimeter and are quoted in units of

10-1 · deg · cm2 · g-1.

2. Synthesis of ketone substrates 1Ketone substrates were prepared following the method of Wolfe et al.2 Over a THF solution of the

corresponding (pyridinylmethyl)lithium [or (quinolin-2-ylmethyl)lithium] another 1.2 M THF

solution of the corresponding amide, Ar-CO-NEt2 (2.5 mmol), was dropped (–70 ºC to rt).

Organolithium reagents were previously obtained by dropping (–70 ºC to rt) a 2.5 M hexane solution

of butyllithium (2.5 mmol) over a 1.0 M THF solution of 2-methylpyridine or 2-methylquinoline (2.5

mmol); however, 3- and 4-methylpyridine had to be deprotonated with a THF solution of LDA, to

avoid the nucleophilic attack of BuLi to the pyridine ring (to C-2 of 4-Me-Py or to C-2/C-6 of 3-Me-

Py). When the reaction was terminated (TLC control, hexane:ethyl acetate 1:1), the same protocol to

that described by Wolfe et al.2 was followed, except that the crude was purified by flash column

chromatography. The spectroscopic data of these substrates were in good agreement with those

previously published. The ratio of keto-enol tautomers measured in the 1H-NMR spectra of ketones

1a-e in CDCl3 were 63:37 (for 1a and 1b), 87:13 (for 1c), >99:<1 (for 1d), and 98:2 (for 1e).

Quinoline derivative 1f consisted into a 7:93 mixture of keto-enaminone tautomers (Scheme S1) such

as was established by analysis of the 1H- and 13C-NMR spectra and by comparison with the published

data.3

1 General Information for this set of engineered enzymes is available from the web page of Codexis: https://www.codexis-estore.com/product-page/codex-ketoreductase-kred-screening-kit.2 R. P. Cassity, L. T. Taylor, J. F. Wolfe, J. Org. Chem., 1978, 43, 2286-2288.

S3

N NO OH

Scheme S1

3. Enzymatic screening for the reduction of ketones 13.1 General procedure for the bioreduction

Each reaction was carried out in a 1.5 mL Eppendorf tube using 11 mol of ketone 1 and 2 mg of

KRED. The solvent was added in such amount that a final 20 mM substrate concentration was

achieved. This solvent consisted in a mixture of propan-2-ol (17.5% v/v), DMSO (if necessary to

completely dissolve 1, up to a maximum of 3.5% v/v), and 125 mM phosphate buffer at pH 7.0 [also

containing 1.25 mM MgSO4 and the cofactor NADP+ (1.0 mM)]. The order of addition to the

Eppendorf tube was: (1) KRED, (2) the buffer solution, and (3) solution of substrate in the organic

solvent(s). The resulting reaction mixture was shaken at 250 rpm and 30 °C during 24 h, except 48 h

for ketone 1f. After this time, the mixture was extracted with ethyl acetate (2 × 500 μL), the organic

layers were separated by centrifugation (90 s, 13000 rpm), combined, and the solvents evaporated.

The degree of conversion and enantiomeric excess of the corresponding alcohol was determined by

chiral GC and HPLC (see Section 4).

3.2 About the results of the screening

The complete Codexis KREDs kit was tested with all the ketones. However, in some cases, we have

only included in the corresponding Table those reactions in which a degree of conversion higher than

25% was achieved.

3 J. Bai, P. Wang, W. Cao, X. Chen, J. Mol. Struct., 2017, 1128, 645-652.

S4

Table S1. Enzymatic reduction of 1-phenyl-2-(pyridin-2-yl)ethanone (1a) using KREDs following

the general procedure of section 3.1.

Propan-2-olPhosphate buffer, pH 7.0250 rpm, 30°C, 24 h

KRED, NADP+

*

N NO OH

1a 2a

KREDa C (%)b ee (%)c

P1-B02 69 >99 (S)P1-B05 >99 66 (S)P1-B10 >99 >99 (S)P1-B12 >99 >99 (S)P1-C01 >99 14 (S)P2-B02 >99 68 (R)P2-C02 >99 70 (R)P2-D03 >99 42 (R)P2-D12 >99 56 (S)P2-G03 >99 >99 (R)

aTo facilitate the understanding of the information here included, results of the KREDs showing a R preference have been highlighted in grey. bDegree of conversion (C) is determined by GC analysis (see section 4.1). cEnantiomeric excess (ee) of 2a is determined by chiral-HPLC analysis (see section 4.1). The absolute configuration of the major stereoisomer of the alcohol is indicated between brackets.

Table S2. Enzymatic reduction of 1-(4-fluorophenyl)-2-(pyridin-2-yl)ethanone (1b) using KREDs

following the general procedure of section 3.1.

Propan-2-olPhosphate buffer, pH 7.0250 rpm, 30°C, 24 h

KRED, NADP+

*

N NO OH

F F1b 2b

KREDa C (%)b ee (%)c

P1-B02 84 >99 (S)P1-B05 84 98 (R)P1-B10 90 >99 (S)P1-B12 95 >99 (S)P2-B02 85 95 (R)P2-C02 90 72 (R)P2-D03 >99 68 (R)P2-D12 95 28 (S)P2-G03 >99 >99 (R)

aTo facilitate the understanding of the information here included, results of the KREDs showing a R preference have been highlighted in grey. bDegree of conversion (C) is determined by GC analysis (see section 4.2). cee of 2b is determined by chiral-HPLC analysis (see section 4.2). The absolute configuration of the major stereoisomer of the alcohol is indicated between brackets.

S5

Table S3. Enzymatic reduction of 1-(4-methoxyphenyl)-2-(pyridin-2-yl)ethanone (1c) using KREDs

following the general procedure of section 3.1.

Propan-2-olPhosphate buffer, pH 7.0250 rpm, 30°C, 24 h

KRED, NADP+

*

N NO OH

MeO MeO1c 2c

KRED C (%)a ee (%)a

P1-B05 81 97 (R)P2-B02 >99 87 (R)P2-C02 >99 >99 (R)P2-D03 94 43 (R)P2-G03 >99 >99 (R)

aDegree of conversion (C) and ee of 2c are determined by chiral-HPLC analysis (see section 4.3). The absolute configuration of the major stereoisomer of the alcohol is indicated between brackets.

Table S4. Enzymatic reduction of 1-phenyl-2-(pyridin-3-yl)ethanone (1d) using KREDs following the general procedure of section 3.1.

Propan-2-olPhosphate buffer, pH 7.0250 rpm, 30°C, 24 h

KRED, NADP+

*

N NO OH

1d 2d

KREDa C (%)b ee (%)b

P1-B02 >99 >99 (S)P1-B05 >99 98 (S)P1-B10 >99 >99 (S)P1-B12 >99 >99 (S)P1-C01 >99 97 (S)P1-H08 >99 98 (S)P2-B02 >99 5 (R)P2-C02 >99 <5 (R)P2-D03 >99 90 (S)P2-D11 >99 >99 (S)P2-D12 >99 94 (S)P2-G03 >99 94 (R)

aTo facilitate the understanding of the information here included, results of the KREDs showing a R preference have been highlighted in grey. bDegree of conversion (C) and ee of 2d are determined by chiral-HPLC analysis (see section 4.4). The absolute configuration of the major stereoisomer of the alcohol is indicated between brackets.

S6

Table S5. Enzymatic reduction of 1-phenyl-2-(pyridin-4-yl)ethanone (1e) using KREDs following

the general procedure of section 3.1.

Propan-2-olPhosphate buffer, pH 7.0250 rpm, 30°C, 24 h

KRED, NADP+

*

N NO OH

1e 2e

KREDa C (%)b ee (%)b

P1-B02 >99 >99 (S)P1-B05 >99 97 (S)P1-B10 >99 >99 (S)P1-B12 >99 >99 (S)P1-C01 >99 96 (S)P1-H08 >99 96 (S)P2-B02 >99 36 (S)P2-C02 >99 45 (S)P2-D03 >99 90 (S)P2-D11 >99 >98 (S)P2-D12 >99 96 (S)P2-G03 >99 91 (R)P2-H07 33 90 (S)P3-G09 36 58 (R)

aTo facilitate the understanding of the information here included, results of the KREDs showing a R preference have been highlighted in grey. bDegree of conversion (C) and ee of 2e are determined by chiral-HPLC analysis (see section 4.5). The absolute configuration of the major stereoisomer of the alcohol is indicated between brackets.

Table S6. Enzymatic reduction of 1-phenyl-2-(quinolin-2-yl)ethanone (1f) using KREDs following

the general procedure of section 3.1.

Propan-2-olPhosphate buffer, pH 7.0250 rpm, 30°C, 48 h

KRED, NADP+

*N N

O OH

1f 2f

KRED C (%)a ee (%)a

P1-B02 >99 >99 (S)P1-B12 27 >99 (S)P2-B02 93 22 (S)P2-C02 25 61 (S)

aDegree of conversion (C) and ee of 2f are determined by chiral-HPLC analysis (see section 4.6). The absolute configuration of the major stereoisomer of the alcohol is indicated between brackets.

S7

4. HPLC and CG analytical data for C and ee determinations

4.1. Bioreductions of 1a: Degree of conversion (C) was analyzed by GC: CP-ChiraSil; program:

160/5/1/180/20/200 [(initial T (ºC) / time (min) / ramp (ºC/min) / T (ºC) / ramp (°C/min) / final T

(°C)]. Retention times (min): 15.80 (ketone 1a); 17.00 and 17.33 (alcohol 2a).

The ee of 2a was determined by HPLC: Chiralcel OJ-H, hexane:propan-2-ol 90:10, 25 ºC, 0.8

mL/min, 214 nm. Retention times (min): 14.27 (R) and 16.60 (S); RS = 4.0.

4.2. Bioreductions of 1b: C was analyzed by GC: CP-ChiraSil; program: 160/5/1/175/20/180 [(initial

T (ºC)/time (min)/ramp (ºC/min) / T (ºC) / ramp (°C/min) / final T (°C)]. Retention times (min): 14.92

(ketone 1b); 17.39 and 17.93 (alcohol 2b).

The ee of 2b was determined by HPLC: Chiralcel OJ-H, hexane:propan-2-ol 95:5, 30 ºC, 0.8 mL/min,

214 nm. Retention times (min): 16.61 (R) and 17.66 (S); RS = 1.8.

4.3. Bioreductions of 1c: Degree of conversion and ee of 2c were determined by HPLC: Chiralpak

AD-H, hexane:propan-2-ol 90:10, 35 ºC, 0.8 mL/min, 214 nm. Retention times (min): 17.75 (ketone

1c); 20.96 [(R)-2c] and 22.54 [(S)-2c], RS = 1.5.

4.4. Bioreductions of 1d: Degree of conversion was analyzed by HPLC: Chiralcel OJ-H,

hexane:propan-2-ol 80:20, 30 ºC, 0.8 mL/min, 214 nm. Retention times (min): 10.92 and 11.24

(alcohol 2d); 15.74 (ketone 1d).

The ee of 2d was determined by HPLC: Chiralpak AD-H, hexane:propan-2-ol 90:10, 30 ºC, 0.8

mL/min, 214 nm. Retention times (min): 18.05 (S) and 20.51 (R); RS = 2.4.

4.5. Bioreductions of 1e: Degree of conversion and ee of 2e were determined by HPLC: Chiralpak

AD-H, hexane:propan-2-ol 90:10, 30 ºC, 0.8 mL/min, 214 nm. Retention times (min): 16.99 [(R)-2e]

and 18.48 [(S)-2e], RS = 1.7; 22.00 (ketone 1e).

4.6. Bioreductions of 1f: Degree of conversion and ee of 2f were determined by HPLC: Chiralpak

AD-H, hexane:propan-2-ol 70:30, 30 ºC, 0.8 mL/min, 214 nm. Retention times (min): 8.71 (ketone

1f); 12.40 [(S)-2f] and 14.08 [(R)-2f], RS = 2.5.

4.7. 2-(2-Azido-2-phenylethyl)pyridine (5) and 1-phenyl-2-(pyridin-2-yl)ethanamine (6)

The ee of 5 was determined after hydrogenation and transformation of the resulting amine 6 into its

N-Boc derivative 7 (see the following section 4.8).

4.8. tert-Butyl [1-phenyl-2-(pyridin-2-yl)ethyl]carbamate 7

The ee of 7 was determined by HPLC: Chiralcel OD, hexane:propan-2-ol 98:2, 25 ºC, 0.8 mL/min,

214 nm. Retention times (min): 19.83 [(R)-7] and 22.94 [(S)-7], RS = 1.8.

S8

5. Copy of chiral HPLC chromatogramsRacemic 1-phenyl-2-(pyridin-2-yl)ethanol 2a

(R)-2a (ee > 99%) isolated from the bioreduction of 1a with KRED-P2-G03

(S)-2a (ee > 99%) isolated from the bioreduction of 1a with KRED-P1-B12.

OH N

S9

Racemic 1-(4-fluorophenyl)-2-(pyridin-2-yl)ethanol 2b

(R)-2b (ee > 99%) isolated from the bioreduction of 1b with KRED-P2-G03

(S)-2b (ee > 99%) isolated from the bioreduction of 1b with KRED-P1-B12

OH N

F

S10

Racemic 1-(4-methoxyphenyl)-2-(pyridin-2-yl)ethanol 2c

(R)-2c (ee > 99%) isolated from the bioreduction of 1c with KRED-P2-G03

OH N

MeO

S11

Racemic 1-phenyl-2-(pyridin-3-yl)ethanol 2d

(S)-2d (ee > 99%) isolated from the bioreduction of 1d with KRED-P1-B02

(R)-2d (ee = 94%) isolated from the bioreduction of 1d with KRED-P2-G03

OHN

S12

Racemic 1-phenyl-2-(pyridin-4-yl)ethanol 2e

(R)-2e (ee = 91%) isolated from the bioreduction of 1e with KRED-P2-G03

(S)-2e (ee > 99%) isolated from the bioreduction of 1d with KRED-P1-B02

OH N

S13

Racemic 1-(phenyl)-2-(quinolin-2-yl)ethanol 2f

(S)-2f (ee >99%) isolated from the bioreduction of 1f with KRED-P1-B02

OH N

S14

Enantioenriched (ee = 38%) sample of tert-butyl (R)-[1-phenyl-2-(pyridine-2-yl)ethyl]carbamate 7

(S)-7 (ee = 97%) derived from (S)-6 and (S)-5, in the strategy starting from (R)-2a (ee > 99%)

(R)-7 (ee = 97%) derived from (R)-6 and (R)-5, in the strategy starting from (S)-2a (ee > 99%)

NH NBoc

S15

6. Assignment of the absolute configuration of the optically active alcohols 2 from their MTPA ester derivatives 3

General procedure for the preparation of esters 3.

A solution of the corresponding racemic or enantioenriched alcohol 2 (75 mol) in anhydrous THF

(1.5 mL) was treated with 4-(dimethylamino)pyridine (150 mol) and (S)-MTPA-Cl (100 mol).

After completion (TLC control, hexane:ethyl acetate 1:1), aqueous saturated NaHCO3 (8 mL) was

added to the mixture and then it was extracted with ethyl acetate (3 × 10 mL). The joined organic

phases were washed with brine (10 mL) and dried with anh. Na2SO4. Evaporation of the solvent and

subsequent flash column chromatography purification (hexane-ethyl acetate mixtures as the eluent)

yielded the MTPA ester derivatives 3 (Scheme S2).

DMAPAzaAr

OH

RF3C

O

ClPh OMe

THF

*+ F3C

O

OPh OMe

HAzaAr

R

2 3

R AzaArEster3a H 2-Py3b F 2-Py3c OMe 2-Py3d H 3-Py3e H 4-Py3f H 2-Q

Scheme S2

1H-NMR spectra (CDCl3) of the diastereomeric mixture of MTPA esters 3 and those obtained from

the enantioenriched samples are collected bellow. The residual signal at 4.12 ppm of some spectra

corresponds to CH2 of residual ethyl acetate.

S16

7. Copy of NMR spectra of Mosher ester derivatives 37.1.1. 1H-NMR spectra (300 MHz) of the diastereomeric mixture of MTPA esters 3a prepared from racemic alcohol 2a.

3.03.54.04.55.05.56.06.57.07.58.08.5f1 (ppm)

7.26

00

8.458.508.558.608.658.70f1 (ppm)

8.51

768.

5197

8.53

348.

5355

8.60

188.

6042

8.61

798.

6202

3.163.183.203.223.243.263.283.303.323.34f1 (ppm)

3.22

653.

2304

3.29

923.

3029

7.1.2. 1H-NMR spectrum (300 MHz) of (R,R)-3a obtained from (R)-2a.

2.83.03.23.43.63.84.04.24.44.64.85.05.25.45.65.86.06.26.46.66.87.07.27.47.67.88.08.28.48.68.8f1 (ppm)

3.29

883.

3031

7.26

00

8.51

628.

5193

8.53

288.

5356

(R,R)-3a

F3CO

O H

PhPh

OMeN F3C

O

O H

PhPh

OMe

(R,S)-3a

8.53

8.61

3.30 3.23 N

S17

7.2.1. 1H-NMR spectra (300 MHz) of the diastereomeric mixture of MTPA esters 3b prepared from racemic alcohol 2b.

2.83.03.23.43.63.84.04.24.44.64.85.05.25.45.65.86.06.26.46.66.87.07.27.47.67.88.08.28.48.68.8f1 (ppm)

7.26

00

8.58.6f1 (ppm)

8.51

638.

5304

8.59

328.

5957

8.60

938.

6117

3.203.253.30f1 (ppm)

3.22

823.

2321

3.29

233.

2961

7.2.2. 1H-NMR spectrum (300 MHz) of (R,R)-3b obtained from (R)-2b.

2.83.03.23.43.63.84.04.24.44.64.85.05.25.45.65.86.06.26.46.66.87.07.27.47.67.88.08.28.48.68.8f1 (ppm)

3.28

963.

2937

7.26

00

8.51

028.

5133

8.52

608.

5294

(R,R)-3b

F3CO

O H

Ph OMeN F3C

O

O H

Ph OMe

(R,S)-3b

8.52

3.29 3.23

FN

8.61

F

S18

7.3.1. 1H-NMR spectra (300 MHz) of the diastereomeric mixture of MTPA esters 3c prepared from racemic alcohol 2c.

3.03.54.04.55.05.56.06.57.07.58.08.5f1 (ppm)

7.26

00

8.508.558.608.65f1 (ppm)

8.50

348.

5063

8.51

968.

5225

8.58

608.

5889

8.60

228.

6050

3.183.223.263.30f1 (ppm)

3.21

723.

2211

3.28

803.

2919

7.3.2. 1H-NMR spectrum (300 MHz) of (R,R)-3c obtained from (R)-2c.

2.83.03.23.43.63.84.04.24.44.64.85.05.25.45.65.86.06.26.46.66.87.07.27.47.67.88.08.28.48.68.8f1 (ppm)

3.29

013.

2945

7.26

00

8.50

788.

5104

8.52

398.

5265

(R,R)-3c

F3CO

O H

Ph OMeN F3C

O

O H

Ph OMe

(R,S)-3c

8.51

3.29 3.22

OMeN

8.60

OMe

S19

7.4.1. 1H-NMR spectra (600 MHz) of the diastereomeric mixture of MTPA esters 3d prepared from racemic alcohol 2d.

3.03.23.43.63.84.04.24.44.64.85.05.25.45.65.86.06.26.46.66.87.07.27.47.67.88.08.28.48.6f1 (ppm)

7.26

00

8.28.38.48.5f1 (ppm)

8.26

06

8.41

938.

4370

8.44

178.

5002

8.50

55

3.283.323.36f1 (ppm)

3.30

31

3.34

94

Selective 1D TOCSY experiments by excitation of the resonances at 8.26 (left) and 8.50 (right) ppm.

6.16.26.36.46.56.66.76.86.97.07.17.27.37.47.57.67.77.87.98.08.18.28.38.48.58.68.78.88.99.09.19.29.3f1 (ppm)

7.08

457.

0945

7.10

62

7.30

117.

3197

8.26

00

8.44

28

6.16.26.36.46.56.66.76.86.97.07.17.27.37.47.57.67.77.87.98.08.18.28.38.48.58.68.78.88.99.09.19.2f1 (ppm)

7.19

557.

2049

7.21

50

7.47

687.

4920

8.41

94

8.50

30

(R,R)-3d

F3CO

O H

PhPh

OMe

N F3CO

O H

PhPh

OMe

(R,S)-3d

3.35 3.30

8.26

8.447.09

7.31

8.50

8.42 7.48

7.20N

S20

7.4.2. 1H-NMR spectrum (600 MHz) of (R,S)-3d obtained from (S)-2d.

2.83.03.23.43.63.84.04.24.44.64.85.05.25.45.65.86.06.26.46.66.87.07.27.47.67.88.08.28.48.68.8f1 (ppm)

3.29

563.

3000

7.26

00

8.358.408.458.50f1 (ppm)

8.41

258.

4197

8.48

278.

4882

8.49

868.

5041

S21

7.5.1. 1H-NMR spectra (300 MHz) of the diastereomeric mixture of MTPA esters 3e prepared from racemic alcohol 2e.

2.83.03.23.43.63.84.04.24.44.64.85.05.25.45.65.86.06.26.46.66.87.07.27.47.67.88.08.28.48.68.8f1 (ppm)

7.26

00

8.38.48.58.6f1 (ppm)

8.38

46

8.50

75

3.253.303.35f1 (ppm)

3.27

993.

2841

3.33

743.

3418

7.5.2. 1H-NMR spectrum (300 MHz) of (R,R)-3e obtained from (S)-2e.

2.83.03.23.43.63.84.04.24.44.64.85.05.25.45.65.86.06.26.46.66.87.07.27.47.67.88.08.28.48.68.89.0f1 (ppm)

3.28

08

7.26

00

8.49

448.

5111

(R,R)-3e

F3CO

O H

PhPh

OMe

N

F3CO

O H

PhPh

OMe

(R,S)-3e

8.38

3.34 3.28

N 8.51

S22

7.6.1. 1H-NMR spectra (400 MHz) of the diastereomeric mixture of MTPA esters 3f prepared from racemic alcohol 2f.

2.83.03.23.43.63.84.04.24.44.64.85.05.25.45.65.86.06.26.46.66.87.07.27.47.67.88.08.28.4f1 (ppm)

3.20

263.

2060

3.29

963.

3031

7.26

00

7.98.08.1f1 (ppm)

7.89

067.

9116

8.02

658.

0475

8.06

108.

0818

8.10

688.

1280

Selective 1D TOCSY experiments by excitation of the resonances at 7.90 (left) and 8.04 (right) ppm.

7.17.27.37.47.57.67.77.87.9f1 (ppm)

7.08

857.

1059

7.89

107.

9124

7.27.37.47.57.67.77.87.98.08.1f1 (ppm)

7.23

547.

2534

8.02

708.

0486

F3CO

O H

PhPh

OMe3.20 N

(R,R)-3f

F3CO

O H

PhPh

OMeN

(R,S)-3f

8.12

3.30 7.24

8.07

8.04

7.107.90

S23

7.6.2. 1H-NMR spectrum (300 MHz) of (R,S)-3f obtained from (S)-2f.

2.83.03.23.43.63.84.04.24.44.64.85.05.25.45.65.86.06.26.46.66.87.07.27.47.67.88.08.28.4f1 (ppm)

3.19

693.

2011

7.26

00

7.907.958.008.058.108.15f1 (ppm)

8.02

798.

0305

8.05

67

8.10

748.

1113

8.13

558.

1398

S24

8. Copy of 1H- and 13C-NMR spectra of compounds 2a-f, 5, 6, and 7

1-Phenyl-2-(pyridin-2-yl)ethanol 2a

2.83.03.23.43.63.84.04.24.44.64.85.05.25.45.65.86.06.26.46.66.87.07.27.47.67.88.08.28.48.68.8f1 (ppm)

2.17

0.75

1.13

1.00

6.27

1.08

1.00

5.15

315.

1683

5.17

815.

1934

7.10

017.

1268

7.26

00

7.60

307.

6091

7.62

867.

6347

7.65

427.

6604

8.52

738.

5303

8.53

258.

5367

8.54

328.

5473

8.54

948.

5526

OH N

S25

1-(4-Fluorophenyl-2-(pyridin-2-yl)ethanol 2b

2.53.03.54.04.55.05.56.06.57.07.58.08.5f1 (ppm)

2.01

2.00

3.89

1.96

0.99

1.00

5.12

475.

1396

5.14

915.

1647

7.26

00

7.59

957.

6057

7.62

507.

6313

7.65

027.

6562

8.51

258.

5287

35404550556065707580859095100105110115120125130135140145150155160165170f1 (ppm)

45.4

7

72.6

0

76.9

5

114.

8411

5.12

121.

7612

3.80

127.

2912

7.40

136.

9913

9.69

139.

73

148.

29

159.

3316

0.31

163.

55

OH N

F

S26

1-(4-Methoxyphenyl-2-(pyridin-2-yl)ethanol 2c

2.83.03.23.43.63.84.04.24.44.64.85.05.25.45.65.86.06.26.46.66.87.07.27.47.67.88.08.28.48.68.8f1 (ppm)

2.07

3.06

2.13

1.98

1.97

2.00

0.97

1.00

3.03

723.

0492

3.08

703.

0989

3.10

713.

1359

3.15

683.

1857

3.79

07

5.08

615.

0980

5.11

525.

1269

6.85

716.

8862

7.08

487.

1106

7.31

907.

3476

7.58

107.

5872

7.60

677.

6128

7.63

247.

6385

8.50

068.

5054

8.50

888.

5164

8.52

028.

5236

35404550556065707580859095100105110115120125130135140145150155160165f1 (ppm)

45.6

6

55.1

5

72.8

2

76.9

5

113.

57

121.

5912

3.79

126.

91

136.

1713

6.82

148.

31

158.

7015

9.59

OH N

MeO

S27

1-Phenyl-2-(pyridin-3-yl)ethanol 2d

2.42.62.83.03.23.43.63.84.04.24.44.64.85.05.25.45.65.86.06.26.46.66.87.07.27.47.67.88.08.28.4f1 (ppm)

2.20

1.03

0.99

0.96

4.84

1.01

2.00

2.91

532.

9339

2.96

132.

9742

2.98

002.

9990

3.02

023.

0451

4.82

414.

8430

4.84

944.

8678

7.10

987.

1262

7.13

607.

1517

7.26

007.

4103

7.41

677.

4231

7.43

637.

4429

7.44

93

8.25

218.

2600

8.28

478.

2905

8.30

128.

3067

35404550556065707580859095100105110115120125130135140145150155f1 (ppm)

42.8

0

74.6

5

76.9

5

123.

1012

5.82

127.

6812

8.39

133.

85

137.

35

143.

58

147.

27

150.

31

OHN

S28

1-Phenyl-2-(pyridin-4-yl)ethanol 2e

2.62.83.03.23.43.63.84.04.24.44.64.85.05.25.45.65.86.06.26.46.66.87.07.27.47.67.88.08.28.48.6f1 (ppm)

2.07

1.17

1.00

1.95

5.00

2.02

2.92

132.

9390

2.96

662.

9856

3.01

293.

0322

3.05

80

3.60

92

4.87

454.

8921

4.90

034.

9179

7.04

687.

0652

7.26

00

8.29

24

35404550556065707580859095100105110115120125130135140145150155160f1 (ppm)

45.0

6

74.1

4

76.9

5

124.

9512

5.76

127.

6612

8.37

143.

62

147.

8214

8.93

OH N

S29

1-Phenyl-2-(quinolin-2-yl)ethanol 2f

3.03.23.43.63.84.04.24.44.64.85.05.25.45.65.86.06.26.46.66.87.07.27.47.67.88.08.2f1 (ppm)

2.07

1.00

0.86

4.07

2.94

2.01

1.99

5.31

845.

3334

5.34

415.

3591

7.26

007.

7088

7.73

227.

7373

7.76

027.

7914

7.79

617.

8185

7.82

32

8.07

028.

0852

8.09

848.

1129

35404550556065707580859095100105110115120125130135140145150155160165f1 (ppm)

45.9

9

72.8

8

76.9

5

122.

0212

5.80

126.

1812

7.22

127.

5112

8.27

128.

5212

9.79

136.

83

143.

84

146.

82

160.

40

OH N

S30

2-(2-Azido-2-phenylethyl)pyridine 5

2.83.03.23.43.63.84.04.24.44.64.85.05.25.45.65.86.06.26.46.66.87.07.27.47.67.88.08.28.48.6f1 (ppm)

2.17

1.00

2.06

5.16

1.08

0.98

3.13

583.

1555

3.18

183.

1966

3.20

153.

2260

3.24

283.

2720

5.06

335.

0831

5.09

265.

1123

7.07

597.

0796

7.08

317.

1017

7.10

557.

1092

7.12

597.

1298

7.14

227.

1459

7.15

137.

1551

7.16

757.

1713

7.26

007.

5439

7.55

017.

5695

7.57

567.

5951

7.60

12

8.57

238.

5754

8.57

848.

5816

8.58

868.

5917

8.59

488.

5980

404550556065707580859095100105110115120125130135140145150155160165f1 (ppm)

45.0

7

65.7

5

76.9

5

121.

7412

4.07

126.

7112

8.15

128.

65

136.

31

139.

27

149.

34

157.

35

N3 N

S31

1-Phenyl-2-(pyridin-2-yl)ethanamine 6

1.52.02.53.03.54.04.55.05.56.06.57.07.58.08.59.0f1 (ppm)

2.23

2.29

1.00

2.01

5.01

1.04

0.93

2.99

442.

9975

3.02

423.

0396

3.06

953.

1129

3.12

923.

1583

3.17

45

4.46

484.

4812

4.49

484.

5111

7.26

00

7.17.27.37.47.5f1 (ppm)

2.01

5.01

1.04

7.04

117.

0669

7.10

437.

1079

7.12

077.

1244

7.12

947.

1330

7.14

587.

1494

7.26

00

7.52

687.

5330

7.55

237.

5585

7.57

787.

5840

8.568.588.60f1 (ppm)

0.93

8.56

278.

5656

8.56

858.

5714

8.57

908.

5818

8.58

478.

5876

404550556065707580859095100105110115120125130135140145150155160f1 (ppm)

48.2

3

55.9

6

76.9

5

121.

3312

3.97

126.

3112

6.98

128.

36

136.

20

145.

58

149.

31

159.

27

NH2 N

S32

1-Phenyl-2-(pyridin-2-yl)ethanamine dihydrochloride 6 ·2 HCl

2.83.03.23.43.63.84.04.24.44.64.85.05.25.45.65.86.06.26.46.66.87.07.27.47.67.88.08.28.48.68.89.09.29.4f1 (ppm)

1.27

1.07

6.43

5.21

2.04

1.03

1.00

3.31

00

3.73

193.

7641

3.77

943.

8117

3.95

843.

9795

4.00

624.

0271

4.94

854.

9704

4.98

175.

0027

8.46

878.

4741

8.49

548.

5007

8.52

168.

5271

8.73

558.

7379

8.74

068.

7433

8.75

478.

7573

8.75

988.

7624

3035404550556065707580859095100105110115120125130135140145150155160f1 (ppm)

38.9

7

49.0

0

55.6

0

127.

2812

8.56

129.

7813

0.71

131.

12

135.

85

143.

27

148.

07

152.

28

NH2 N

2 HCl

S33

tert-Butyl [1-phenyl-2-(pyridin-2-yl)ethyl]carbamate 7

0.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.5f1 (ppm)

9.45

2.18

1.07

1.06

1.02

6.51

1.04

1.00

1.35

16

5.07

50

5.97

656.

9611

6.98

727.

1212

7.13

417.

1370

7.14

247.

1464

7.15

957.

1629

7.22

047.

2600

7.50

867.

5147

7.53

437.

5403

7.55

997.

5659

8.52

968.

5339

8.53

738.

5455

8.54

878.

5530

101520253035404550556065707580859095100105110115120125130135140145150155160f1 (ppm)

28.2

2

44.5

8

54.7

6

76.9

579

.15

121.

5212

3.92

126.

0412

6.87

128.

22

136.

33

142.

22

148.

89

155.

13

158.

00

NH NBoc