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Synthesis and biological evaluation of 2,4,5-trisubstituted thiazoles as antituberculosis agents effective against drug-resistant tuberculosis
Uttam B. Karalea,b, Vagolu Siva Krishnac, E. Vamshi Krishnab,d, Amit S. Choudharie, Manjulika Shuklaf, Vikas R. Gaikwada,g, B. Mahizhavenih, Sidharth Choprab,f, Sunil Misrab,d
Dhiman Sarkare, Dharmarajan Sriramc, V.N. Azger Dusthackeerh and Haridas B. Rode*a,b
a Department of Organic Synthesis and Process Chemistry, CSIR-Indian Institute of Chemical Technology, Tarnaka, Hyderabad-500007, India
[email protected] Academy of Scientific and Innovative Research (AcSIR), Ghaziabad, Uttar Pradesh- 201 002, India
c Department of Pharmacy, Birla Institute of Technology & Science-Pilani, Hyderabad Campus, Jawahar Nagar, Shameerpet mandal, R.R. District, Hyderabad- 500078, India
d Department of Applied Biology, CSIR-Indian Institute of Chemical Technology, Tarnaka, Hyderabad-500007, India
e Department of Biology, CSIR-National Chemical Laboratory, Pashan road, Pune- 411008, Indiaf Department of Microbiology, CSIR-Central Drug Research Institute, Lucknow-226021, Uttar Pradesh, India
g Department of Medicinal Chemistry, National Institute of Pharmaceutical Education and Research, Balanagar, Hyderabad - 500 037, India
h Department of Bacteriology, National Institute for Research in Tuberculosis, Chennai- 600031, India
1. Experimental and Methods
1.1. General
1H and 13C NMR spectra were recorded on a Bruker Avance 300 or 400 or 500 spectrometer and are
referred to the residual solvent signal: CDCl3: (7.26) for 1H and (77.16) for 13C. Chemical shifts (δ) are
given in ppm and coupling constants (J) were measured in Hz. The following abbreviations are used;
s-singlet, d-doublet, dd-doublet of doublet, t-triplet, td-triplet of doublet, tt- triplet of triplet, ddd-
doublet of doublet of doublet, dt-doublet of triplet, q-quartet, dq- doublet of quartet, qd-quartet of
doublet, qt-quartet of triplet, q-quintet, sept- septet, br-broad. HRMS spectra were recorded using
Xevo G2-XS QTof (Waters); values are given in m/z. Column chromatography was carried out using
silica gel (60–120 mesh) packed in glass columns. Analytical TLC was carried out on Macherey-
Nagel 60 F245aluminium-backed silica gel plates. Anhydrous solvents and chemical materials were
purchased from commercial sources.
1.2. In-vitro Mycobacterium tuberculosis H37Rv assay
Briefly, the inoculum was prepared from fresh LJ medium re-suspended in 7H9-S medium (7H9
broth, 0.1% casitone, 0.5% glycerol, supplemented oleic acid, albumin, dextrose, and catalase
[OADC]), adjusted to a McFarland tube No. 1, and diluted 1:20; 100 µl was used as inoculum. Each
drug stock solution was thawed and diluted in 7H9-S at four-fold the final highest concentration
tested. Serial two-fold dilutions of each drug were prepared directly in a sterile 96-well microtiter
plate using 100 µl 7H9-S. A growth control containing no antibiotic and a sterile control were also
prepared on each plate. Sterile water was added to all perimeter wells to avoid evaporation during the
incubation. The plate was covered, sealed in plastic bags and incubated at 37 oC in normal
atmosphere. After 7 days incubation, 30 µl of alamar blue solution was added to each well, and the
plate was re-incubated overnight. A change in colour from blue (oxidized state) to pink (reduced)
indicated the growth of bacteria, and the MIC was defined as the lowest concentration of drug that
prevented this change in colour.
1.3. In-vitro Mycobacterium tuberculosis H37Ra assay
1.3.1.Cultivation of mycobacteria
Microbial strains of Mycobacterium tuberculosis H37Ra (ATCC 25177) was obtained from
AstraZeneca, India. The stock culture was maintained at -80 oC and subcultured once in a liquid
medium before inoculation into an experimental culture. Cultures were grown in Dubos media
(enrichment media) at 37 °C and 150 RPM. For antimycobacterial assay, M. pheli medium (minimal
essential medium) was used. It contains 0.5g KH2PO4, 0.25g trisodium citrate, 60mg MgSO4, 0.5g
aspargine and 2ml glycerol in distilled water (100ml) followed by pH adjustment to 6.6. It takes at
least 8-10 days to reach OD 1 at 620 nm.
1.3.2. Anti-mycobacterial activity
The antitubercular activity for each synthesized compound was determined by measuring inhibition of
growth against avirulent strain of M. tuberculosis H37Ra (ATCC 25177) using an established XTT
Reduction Menadione assay (XRMA) method1. For screening against dormant stage bacilli, Mtb
H37Ra culture was treated with synthesized compounds at different concentration (10-0.312µg/ml)
and incubated for 12 days. Briefly, 2.5 µl of the compound was added to the 247.5 µl of M.
tuberculosis culture in M. Pheli medium. Post incubation, 40µl XTT (1.25mM) was added to each
well and incubated for 20 mins. Next, 2.5µl of Menadione (6mM) was added to each well and read at
470 nm using a micro plate reader (Spectramax plus384 plate reader, Molecular Devices Inc).
Rifampicin served as positive control. Cells treated with the vehicle alone were considered as control.
Cell-free wells were used as a blank.
1.4. Cytotoxicity study
Estimation of cytotoxicity efficacy (IC50 values ) of the compounds were carried out as per our earlier
report2. Normal Cell line CHO-K1-Chinese hamster ovary cells (ATCC® CCL-61™), B16-F10
Mouse skin melanoma (ATCC® CRL 6475™) and MCF7 Human Breast Adenocarcinoma
(ATCC® HTB-22™) were obtained from ATCC (Bethesda, MD, USA). All the cell lines were
maintained in DMEM supplemented with 10% FBS, 2 mM l-glutamine, 100 U/ml penicillin, and 100
μg/ml streptomycin at 37 °C in a 5 % CO2 incubator. Cultured cells were loaded into 96 well culture
plate and allowed to attach properly. Test compounds of different concentrations ranging from 1 to 10
µM were added in triplicates. The cells were incubated for 24 hrs and followed by 0.5 mg/mL of
MTT (3-(4, 5-Dimethyl-2-thiazolyl)-2, 5-diphenyl-2H-tetrazolium bromide) was added to each well
and incubated for 3 hr. To dissolve the insoluble formazan crystals, 100 µl DMSO was added to each
well and finally the absorbance of the plates was measured using a Synergy H1 Multi-mode Plate
Reader, USA. IC50 values were calculated based on concentration versus percentage death of cells.
Mitomycin C was used as positive control for the comparison.
1.5. Anti-bacterial activity
1.5.1.Bacterial strains
Ivacaftor was screened against a bacterial panel consisting of ESKAPE pathogens namely Escherichia
coli (ATCC 25922), Klebsiella pneumoniae (BAA-1705), Acinetobacter baumannii (BAA-1605),
Pseudomonas aeruginosa (ATCC 27853) and Staphylococcus aureus (ATCC 29213). The bacteria
were routinely cultivated on Mueller-Hinton Agar (MHA). Before starting the experiment, a single
colony was picked from MHA plate, inoculated in Mueller-Hinton cation supplemented broth (MHB)
and incubated overnight at 37 °C with shaking for 18-24 h to get the starter culture.
1.5.2. Antibiotic susceptibility testing
Antibiotic susceptibility testing was carried out according to the CLSI guidelines for broth
microdilution assay3. 10 mg/mL stock solutions of test compounds were prepared in
Dimethylsulfoxide (DMSO). Bacterial cultures were inoculated in MHB and optical density (OD) of
the cultures was measured at the 600 nm wavelength, followed by dilution to achieve ~10 5 CFU/ml.
The compounds were tested ranging from 64-0.5 mg/L in two-fold serial diluted fashion with 2.5 μL
of each concentration added to each well of a 96-well round bottom microtiter plate. Later, 97.5 μL of
bacterial suspension was added to each well containing the test compound along with appropriate
controls. The plates were incubated at 370C for 18-24 h following which the growth was enumerated
and MIC was identified. The MIC is defined as the lowest compound concentration where there is no
visible growth. For each compound, MIC determinations were carried independently 3 times using
duplicate samples.
Table 1: Effect of thiazole analogous against various bacteria
Com. no. E.coli
ATCC
25922
S.aureus
ATCC 29213
K.pneumoniae
BAA 1705
A.baumannii
BAA 1605
P.aeruginosa
ATCC 27853
13d >50 >50 >50 >50 >50
13h >50 >50 >50 >50 >50
13n >50 50 >50 >50 >50
13o >50 50 >50 >50 >50
13p >50 50 >50 >50 >50
Levofloxacin
(µM/mL)
<0.03 0.25 64 8 1
MIC values are in µM unless otherwise stated.
1.6. Molecular modeling study
1.6.1. Molecular docking
In silico analysis were performed in Dell Precision T7610 workstation (8 processors; 8 GB RAM;
ZOTAC 3GB graphics; Maestro 9.8, Schrodinger, New York, U.S.A) workstation running on Redhat
6.1 Linux environment.
Molecular docking calculations were conducted using Glide application implemented in the Maestro
9.8 software package (Schrodinger, LLC, New York, 2015). Target structure was prepared by adding
hydrogens, followed by an energy minimisation to a convergence of RMSD 0.30 Å using OPLS_2003
as force field. Ligands were energetically minimised and ionization considered at pH 7 using ionizer
subprogram of LigPrep. The ‘Generate grid’ sub application of the Glide tool was utilised for the
generation of grid by selecting the ACP binding site residues (CTP synthase PyrG, PDB ID 4ZDK).
The generated grid was then utilised as a receptor for docking of thiazole derivatives using the ‘extra
precision’ (XP) flexible docking method, located in the Glide tool.4,5 Docked protein and ligand
complexes were visualized and molecular surface complex pictures were generated using Maestro.
1.6.2 Docking results
Compounds were docked in the active site of CTP synthase PyrG. The table 2 describe the docking score of
compounds and their MIC against M. tuberculosis H37Rv.
Table 2: Docking score and MIC of compounds.
Compound Name
XP G
Score
(kcal/mol) MIC (µg/mL)
13a -5.376 7.9
13c -5.851 16.3
13d -6.041 4
13e -5.449 8.1
13f -6.988 15
13g -5.74 17
13h -5.651 1.8
13i -5.263 8.9
13j -5.694 19.3
13k -5.808 4.5
13l -6.274 4.5
13m -5.597 4.5
13n -5.636 4.2
13o -5.656 4.2
13P -5.641 2.1
13q -5.732 13.6
13r -5.533 14.5
14a -6.317 18.7
14b -6.78 9.3
14c -6.433 5.4
15a -7.105 4.9
15b -6.506 4.9
15c -7.554 5.6
15e -6.822 10.6
15f -7.519 9.5
16a -6.189 20.7
16b -6.551 10.3
16c -6.456 5.9
Figure 1: Graph of docking score Vs MIC of compounds against M. tuberculosis H37Rv
0 5 10 15 20 25
-8
-7
-6
-5
-4
-3
-2
R² = 2.46995930163196E-05
MIC values (µg/mL)
Doc
king
scor
e (k
cal/m
ol)
No correlation observed between the docking score and MIC of compounds.
1.6 NMR spectra of the compounds
-0.50.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.0
8.35
1.09
2.31
1.00
1.38
1.39
1.41
1.42
3.38
3.40
3.41
3.42
3.44
3.45
3.46
4.39
4.40
4.42
4.43
8.05
1H NMR of 12a
0102030405060708090100110120130140150160170180
14.0
22.9
33.1
60.9
126.
1
146.
2
161.
1
178.
5
13C NMR of 12a
0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.5
1.31
3.95
2.26
1.07
2.03
2.23
0.98
2.00
0.90
1.21
1.22
1.24
1.32
1.34
1.35
1.37
1.40
1.42
1.43
1.45
1.45
1.47
1.48
1.72
1.79
1.79
1.80
1.81
1.82
1.83
2.10
2.11
2.13
2.13
3.03
3.03
3.04
3.05
3.06
3.06
3.07
3.08
3.09
4.35
4.36
4.38
4.39
8.01
1H NMR of 12b
0102030405060708090100110120130140150160170180
14.3
25.5
25.9
33.7
42.6
61.2
126.
2
146.
2
161.
4
178.
0
13C NMR of 12b
0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.0
3.14
3.08
2.14
1.00
1.36
1.37
1.39
2.74
4.37
4.38
4.40
4.41
8.01
1H NMR of 12c
0102030405060708090100110120130140150160
14.4
19.4
61.4
127.
3
146.
9
161.
4
166.
8
13C NMR of 12c
0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.5
3.00
6.06
0.96
1.96
2.08
0.88
1.81
1.24
1.26
1.28
1.40
1.42
3.33
3.34
3.36
3.38
3.40
3.41
3.43
4.28
4.29
4.31
4.33
6.00
6.81
6.83
6.91
6.92
6.93
6.94
6.95
6.95
6.95
6.97
1H NMR of 13a
0102030405060708090100110120130140150160170180
14.2
23.4
33.7
61.3
101.
5
108.
111
0.5
124.
012
4.4
139.
6
145.
114
7.4
148.
4
162.
4
176.
1
13C NMR of 13a
0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.5
3.25
6.12
1.07
1.97
2.07
1.89
0.99
1.00
1.25
1.27
1.29
1.41
1.43
3.34
3.35
3.37
3.39
3.41
3.42
4.28
4.30
4.31
4.33
4.65
6.96
7.02
7.07
7.07
7.09
7.09
8.76
1H NMR of 13b
0102030405060708090100110120130140150160170180f1 (ppm)
14.3
23.4
33.8
61.5
67.4
116.
611
7.7
125.
512
5.9
126.
2
139.
8
144.
3
162.
416
5.7
165.
7
176.
6
13C NMR of 13b
0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.59.09.5
3.06
6.32
0.95
2.05
0.97
1.00
1.00
1.01
1.01
1.00
0.74
0.76
0.78
1.48
1.49
3.44
3.46
3.48
3.49
3.51
3.97
3.99
4.01
4.02
7.42
7.44
7.62
7.64
7.66
7.69
7.69
7.70
7.71
8.03
8.06
8.49
8.50
9.30
1H NMR of 13c
0102030405060708090100110120130140150160170180
13.6
23.4
33.8
61.1
118.
2
126.
412
8.3
128.
412
8.9
132.
313
5.1
140.
714
2.8
143.
9
152.
8
161.
5
178.
1
13C NMR of 13c
0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.59.0
3.07
6.02
1.00
1.96
1.01
0.95
1.02
0.98
0.89
0.82
1.11
1.13
1.15
1.42
1.44
3.36
3.38
3.40
3.41
3.43
3.45
3.47
4.21
4.23
4.25
4.27
7.53
7.53
7.54
7.55
7.55
7.56
7.57
7.70
7.70
7.71
7.72
7.72
7.74
7.74
7.79
7.81
8.09
8.11
8.22
8.22
8.94
8.94
1H NMR of 13d
0102030405060708090100110120130140150160170180f1 (ppm)
14.1
23.3
33.7
61.4
124.
512
7.0
127.
312
8.0
129.
313
0.3
136.
614
0.9
141.
3
147.
615
0.8
161.
9
177.
5
13C NMR of 13d
0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.59.0
3.06
6.22
0.98
2.12
0.99
1.00
0.88
2.03
1.18
1.20
1.22
1.46
1.48
3.42
3.44
3.45
3.47
3.49
4.27
4.29
4.30
4.32
7.88
7.88
7.90
7.90
8.13
8.15
8.21
8.21
8.88
1H NMR of 13e
102030405060708090100110120130140150160170180
14.2
23.4
33.9
61.6
129.
213
0.7
132.
213
3.2
140.
714
2.5
143.
014
3.4
145.
714
5.8
162.
1
177.
6
13C NMR of 13e
0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.5
3.10
6.11
0.96
2.00
1.12
0.78
0.99
0.93
1.15
1.16
1.18
1.44
1.46
3.38
3.39
3.41
3.43
3.45
3.46
3.48
4.22
4.24
4.26
4.28
7.46
7.46
7.47
7.48
7.49
7.49
7.51
7.51
7.53
7.53
7.60
7.61
7.62
7.63
7.64
7.64
7.73
7.73
7.75
7.75
1H NMR of 13f
102030405060708090100110120130140150160170180
14.0
23.3
33.8
61.5
113.
9
117.
4
129.
213
1.2
132.
313
2.9
135.
2
139.
814
2.4
161.
5
178.
2
13C NMR of 13f
0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.0
3.01
6.02
0.92
1.94
1.00
1.83
0.85
1.21
1.23
1.25
1.43
1.45
3.37
3.39
3.40
3.42
3.44
3.46
3.47
4.26
4.28
4.30
4.31
7.50
7.52
7.54
7.68
7.68
7.68
7.69
7.70
7.70
7.70
7.71
7.72
7.72
7.76
7.77
7.77
1H NMR of 13g
0102030405060708090100110120130140150160170180
14.2
23.4
33.8
61.6
112.
6
118.
3
129.
113
2.4
132.
613
3.5
134.
4
140.
814
2.2
161.
9
177.
6
13C NMR of 13g
0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.5f1 (ppm)
3.31
6.16
1.00
2.11
1.99
2.10
1.22
1.23
1.25
1.43
1.45
3.38
3.39
3.41
3.42
3.44
3.45
3.46
4.27
4.28
4.30
4.31
7.57
7.59
7.69
7.70
1H NMR of 13h
0102030405060708090100110120130140150160170180f1 (ppm)
14.1
23.3
33.7
61.6
112.
6
118.
4
130.
813
1.8
135.
9
140.
814
2.6
161.
9
177.
8
13C NMR of 13h
0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.0
3.27
5.52
1.13
2.23
2.14
1.09
2.05
2.05
1.09
2.00
1.24
1.26
1.28
1.29
1.31
1.38
1.38
1.39
1.41
1.41
1.42
1.44
1.47
1.47
1.50
1.50
1.73
1.83
1.84
1.84
1.86
1.87
1.88
2.15
2.19
3.02
3.03
3.04
3.05
3.06
3.07
3.08
3.09
3.09
4.28
4.29
4.31
4.33
6.00
6.81
6.83
6.92
6.93
6.94
6.95
6.95
6.95
1H NMR of 13i
102030405060708090100110120130140150160170
14.2
25.6
26.0
33.9
42.8
61.1
101.
4
107.
911
0.4
123.
912
4.3
139.
3
145.
014
7.3
148.
2
162.
3
175.
0
13C NMR of 13i
0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.59.0
3.75
5.77
1.39
2.41
2.26
1.16
2.31
1.98
2.06
1.04
1.00
1.25
1.27
1.28
1.36
1.38
1.39
1.39
1.40
1.41
1.42
1.44
1.47
1.47
1.49
1.50
1.74
1.76
1.84
1.87
2.16
2.18
3.04
3.04
3.05
3.06
3.07
3.07
3.08
3.09
3.10
4.28
4.30
4.31
4.33
4.65
6.97
6.98
6.99
7.06
7.07
7.08
7.08
8.97
1H NMR of 13j
0102030405060708090100110120130140150160170180
14.3
25.8
26.1
34.0
43.0
61.4
67.3
116.
611
7.7
125.
512
5.9
126.
2
139.
7
144.
214
4.3
162.
4
165.
8
175.
6
13C NMR of 13j
0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.59.09.5
3.10
1.50
2.41
2.41
1.19
2.34
2.27
1.12
2.22
1.06
1.15
1.08
1.10
1.09
1.00
0.76
0.78
0.79
1.24
1.24
1.27
1.28
1.29
1.30
1.31
1.32
1.34
1.39
1.41
1.42
1.43
1.45
1.45
1.46
1.48
1.48
1.49
1.51
1.52
1.54
1.55
1.57
1.58
1.60
1.61
1.74
1.75
1.75
1.76
1.76
1.78
1.78
1.78
1.79
1.79
1.86
1.87
1.88
1.89
1.90
1.91
2.23
2.24
2.26
2.27
3.12
3.12
3.13
3.15
3.15
3.16
3.17
3.18
3.98
4.00
4.01
4.03
7.42
7.43
7.62
7.64
7.66
7.68
7.69
7.70
7.71
8.03
8.03
8.03
8.05
8.05
8.05
8.49
8.50
9.30
1H NMR of 13k
0102030405060708090100110120130140150160170180
13.6
25.7
26.1
33.9
43.0
61.0
118.
2
126.
412
8.3
128.
913
2.2
135.
0
140.
514
2.7
143.
8
152.
8
161.
5
177.
0
13C NMR of 13k
0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.59.0
3.25
1.14
2.42
2.30
1.29
2.11
2.17
1.09
2.20
1.10
1.08
1.09
1.09
1.08
1.00
1.16
1.17
1.19
1.28
1.28
1.29
1.30
1.31
1.32
1.33
1.34
1.39
1.39
1.40
1.41
1.42
1.42
1.44
1.44
1.45
1.46
1.47
1.47
1.50
1.51
1.53
1.53
1.55
1.56
1.57
1.58
1.75
1.76
1.76
1.77
1.78
1.79
1.86
1.87
1.87
1.89
1.89
1.90
2.21
2.21
2.23
2.24
3.10
3.11
3.12
3.13
3.13
3.14
3.15
3.16
3.16
4.26
4.27
4.29
4.30
7.58
7.58
7.59
7.61
7.61
7.75
7.75
7.76
7.76
7.76
7.78
7.78
7.84
7.85
8.13
8.14
8.26
8.26
8.97
8.97
1H NMR of 13l
0102030405060708090100110120130140150160170180
14.2
25.8
26.1
34.0
43.1
61.6
124.
712
7.1
127.
412
8.2
129.
513
0.5
136.
714
1.0
141.
3
147.
7
151.
0
162.
1
176.
7
13C NMR of 13l
0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.59.0
3.01
1.27
2.34
2.36
1.05
2.07
2.00
0.97
2.00
0.96
0.93
0.84
1.82
1.19
1.20
1.22
1.29
1.32
1.41
1.42
1.43
1.44
1.45
1.45
1.53
1.54
1.55
1.56
1.87
1.88
1.89
1.90
2.22
2.24
3.10
3.11
3.11
3.12
3.13
3.14
3.15
3.15
3.16
4.28
4.29
4.30
4.32
7.88
7.88
7.90
7.90
8.12
8.14
8.20
8.21
8.88
1H NMR of 13m
0102030405060708090100110120130140150160170180
14.2
25.8
26.1
34.0
43.1
61.6
129.
113
0.6
132.
213
3.2
140.
614
2.6
143.
014
3.3
145.
714
5.8
162.
2
176.
7
13C NMR of 13m
0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.5
2.92
1.16
2.16
1.99
1.00
1.93
1.89
1.00
1.91
0.88
0.94
0.92
0.85
1.17
1.18
1.19
1.27
1.27
1.28
1.29
1.30
1.30
1.32
1.38
1.39
1.40
1.41
1.41
1.41
1.42
1.43
1.44
1.45
1.46
1.46
1.48
1.49
1.51
1.51
1.53
1.54
1.56
1.56
1.74
1.74
1.75
1.75
1.75
1.76
1.77
1.77
1.77
1.78
1.85
1.86
1.87
1.88
1.88
1.89
2.21
2.21
2.23
2.24
3.09
3.10
3.10
3.11
3.12
3.13
3.14
3.14
4.24
4.26
4.27
4.29
7.46
7.48
7.50
7.50
7.51
7.51
7.53
7.53
7.61
7.61
7.63
7.63
7.64
7.64
7.74
7.74
7.76
7.76
1H NMR of 13n
0102030405060708090100110120130140150160170180
14.1
25.7
26.1
33.9
43.0
61.5
114.
0
117.
4
129.
213
1.2
132.
313
3.0
135.
313
9.7
142.
3
161.
6
177.
3
13C NMR of 13n
0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.0
3.00
1.01
2.07
1.99
1.06
2.03
2.04
0.97
2.00
1.00
1.95
0.87
1.21
1.23
1.25
1.27
1.30
1.39
1.40
1.40
1.42
1.43
1.44
1.46
1.48
1.49
1.51
1.52
1.74
1.77
1.84
1.85
1.86
1.88
1.88
1.89
2.17
2.18
2.21
3.06
3.07
3.08
3.09
3.10
3.10
3.12
3.12
3.13
4.26
4.28
4.29
4.31
7.50
7.52
7.54
7.67
7.67
7.68
7.69
7.69
7.69
7.71
7.71
7.71
7.76
7.76
7.76
1H NMR of 13o
0102030405060708090100110120130140150160170180
14.2
25.7
26.1
34.0
43.0
61.6
112.
6
118.
3
129.
013
2.3
132.
613
3.5
134.
4
140.
714
2.1
161.
9
176.
6
13C NMR of 13o
0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.03.
001.
082.
032.
07
1.07
2.01
2.05
0.95
1.93
1.84
2.04
1.22
1.24
1.25
1.27
1.30
1.40
1.40
1.41
1.42
1.43
1.44
1.49
1.49
1.51
1.52
1.85
1.86
1.86
1.88
1.88
1.89
2.17
2.18
2.18
2.21
3.07
3.08
3.08
3.09
3.10
3.11
3.12
3.12
3.13
4.27
4.29
4.30
4.32
7.57
7.59
7.68
7.70
1H NMR of 13p
0102030405060708090100110120130140150160170180
14.2
25.7
26.0
33.9
43.0
61.5
112.
6
118.
4
130.
813
1.8
135.
914
0.6
142.
5
161.
9
176.
7
13C NMR of 13p
0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.0
3.21
3.17
2.17
2.17
1.00
1.96
1.26
1.28
1.29
2.71
4.29
4.30
4.31
4.33
6.00
6.81
6.82
6.92
6.92
6.93
6.94
6.95
6.95
1H NMR of 13q
0102030405060708090100110120130140150160170
14.2
19.3
61.3
101.
5
108.
111
0.5
123.
912
4.0
139.
7
146.
314
7.4
148.
4
162.
016
4.0
13C NMR of 13q
0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.5
3.84
3.00
2.08
2.04
1.96
1.25
1.26
1.27
2.78
4.29
4.30
4.32
4.33
7.57
7.59
7.69
7.71
1H NMR of 13r
0102030405060708090100110120130140150160170
14.1
19.4
61.6
112.
7
118.
4
130.
813
1.8
135.
5
140.
914
3.7
161.
6
165.
7
13C NMR of 13r
0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.0
3.09
3.02
3.14
1.02
1.09
1.01
2.07
2.00
2.03
2.10
1.00
710.
7471
2.36
717.
6471
9.27
786.
5779
3.50
1631
.74
1638
.65
1645
.55
1652
.46
1659
.38
2594
.43
2601
.39
2608
.55
2609
.53
2616
.65
2623
.62
3633
.80
3639
.11
3640
.82
3642
.43
3666
.03
3667
.77
3673
.96
3681
.25
3684
.43
3685
.64
3692
.40
3821
.57
3830
.12
3845
.18
3853
.76
1H NMR of 14a
0102030405060708090100110120130140150160170180
22.2
23.1
33.4
48.6
112.
3
118.
7
126.
312
7.4
128.
813
1.1
131.
713
5.8
139.
814
2.0
143.
3
160.
5
176.
4
13C NMR of 14a
0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.5
5.99
3.08
0.96
1.02
1.20
1.94
1.92
1.97
2.00
1.00
1.42
1.43
1.44
1.44
1.57
1.59
3.26
3.28
3.29
3.31
3.32
5.19
5.20
5.22
5.22
5.23
5.25
7.25
7.26
7.26
7.27
7.27
7.28
7.28
7.28
7.33
7.33
7.34
7.35
7.36
7.36
7.37
7.37
7.38
7.39
7.66
7.69
1H NMR of 14b
0102030405060708090100110120130140150160170180
22.2
23.1
33.4
48.6
112.
3
118.
7
126.
312
7.4
128.
813
1.1
131.
613
5.8
139.
814
2.0
143.
3
160.
5
176.
4
13C NMR of 14b
0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.0
6.00
2.07
1.03
2.11
3.01
3.02
1.04
1.02
1.03
1.02
2.04
2.02
1.40
1.41
2.82
2.83
2.85
3.22
3.23
3.24
3.26
3.57
3.58
3.60
3.84
3.86
6.75
6.75
6.76
6.76
6.78
6.78
6.80
6.82
7.62
7.62
7.64
7.65
7.67
7.68
7.70
1H NMR of 14c
0102030405060708090100110120130140150160170180
23.0
33.4
35.6
40.8
55.9
56.0
111.
511
2.1
112.
3
118.
712
0.8
131.
113
1.6
131.
713
5.8
139.
614
2.1
147.
814
9.1
161.
4
176.
2
13C NMR of 14c
0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.5
6.05
2.00
1.14
2.02
2.00
2.00
2.10
2.00
1.42
1.43
3.23
3.24
3.25
3.28
3.29
3.30
3.32
3.33
3.34
3.36
3.43
3.44
3.44
3.68
3.69
3.70
3.75
3.76
3.77
7.61
7.63
7.67
7.69
1H NMR of 14d
-100102030405060708090100110120130140150160170180
23.1
33.7
42.6
47.4
66.7
66.8
112.
3
118.
4
129.
113
2.8
133.
513
5.2
144.
8
164.
2
178.
6
13C NMR of 14d
0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.0
5.95
1.00
2.24
2.09
0.98
1.02
0.99
2.02
2.08
1.13
1.40
1.42
3.23
3.24
3.26
3.28
3.30
4.46
4.47
5.94
6.75
6.77
6.79
6.79
6.81
6.81
6.83
6.83
7.66
7.68
7.71
7.73
7.78
7.80
7.81
1H NMR of 14e
0102030405060708090100110120130140150160170
23.0
33.4
43.0
101.
1
108.
310
8.5
112.
3
118.
712
1.1
131.
113
1.6
132.
213
5.7
139.
914
1.9
147.
014
7.9
161.
2
176.
4
13C NMR of 14e
0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.5
6.03
1.02
2.00
1.24
3.04
2.06
1.01
1.00
1.06
1.41
1.42
3.24
3.25
3.27
3.28
3.29
4.58
4.59
7.25
7.25
7.27
7.66
7.68
7.70
7.72
7.92
7.93
7.94
8.52
8.53
8.60
1H NMR of 14f
0102030405060708090100110120130140150160170180
22.9
33.3
40.6
112.
3
118.
5
123.
5
131.
013
1.6
134.
013
5.5
140.
114
1.5
148.
814
9.1
161.
4
176.
4
13C NMR of 14f
0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.0
6.01
0.96
2.07
5.03
2.04
2.00
1.00
1.34
1.37
3.14
3.16
3.18
3.20
3.23
3.25
3.27
4.51
4.53
7.20
7.22
7.24
7.25
7.28
7.29
7.60
7.63
7.66
7.69
7.80
1H NMR of 14g
0102030405060708090100110120130140150160170180
23.0
33.4
43.3
112.
3
118.
7
127.
612
7.9
128.
813
1.2
131.
713
5.8
138.
313
9.9
141.
9
161.
3
176.
4
13C NMR of 14g
0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.5
6.01
0.92
2.05
1.00
1.01
0.94
1.97
1.99
0.92
1.42
1.44
3.23
3.25
3.27
3.28
3.30
3.32
3.33
4.60
4.62
7.19
7.20
7.21
7.22
7.34
7.36
7.39
7.39
7.67
7.69
7.71
7.99
8.01
8.02
1H NMR of 14h
0102030405060708090100110120130140150160170180
23.0
33.4
40.7
112.
5
118.
712
7.4
129.
513
0.8
131.
113
1.7
134.
013
4.3
134.
4
140.
214
1.7
161.
4
176.
5
13C NMR of 14h
0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.5
6.02
1.00
2.09
1.98
2.00
2.01
2.05
1.10
1.40
1.43
3.20
3.22
3.24
3.27
3.29
3.31
3.33
4.52
4.54
6.99
7.02
7.05
7.28
7.30
7.31
7.66
7.69
7.71
7.73
7.84
7.86
7.88
1H NMR of 14i
0102030405060708090100110120130140150160170180
23.0
33.4
42.5
112.
411
5.5
115.
711
8.7
129.
512
9.6
131.
113
1.7
134.
113
4.2
135.
714
0.0
141.
8
161.
316
1.3
163.
2
176.
5
13C NMR of 14i
0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.0
5.86
0.94
2.01
4.06
1.87
2.02
1.00
1.40
1.43
3.20
3.22
3.24
3.27
3.29
3.31
3.34
4.52
4.54
7.26
7.28
7.29
7.66
7.69
7.70
7.73
7.87
7.89
1H NMR of 14j
0102030405060708090100110120130140150160170180
23.0
33.4
42.6
112.
4
118.
7
128.
912
9.2
131.
213
1.7
133.
413
5.7
136.
914
0.1
141.
7
161.
4
176.
5
13C NMR of 14j
0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.59.09.5
5.98
0.99
0.96
0.98
2.00
1.94
0.91
1.00
1.47
1.48
3.30
3.31
3.33
3.34
3.36
3.37
3.38
7.08
7.09
7.11
7.43
7.44
7.44
7.45
7.45
7.46
7.46
7.46
7.69
7.71
7.71
7.73
7.84
7.84
7.85
7.86
9.44
1H NMR of 14k
0102030405060708090100110120130140150160170180
23.0
33.5
112.
811
6.5
116.
811
8.6
119.
511
9.6
121.
212
1.4
122.
013
1.1
131.
813
4.4
134.
413
5.4
141.
214
1.4
153.
215
6.5
159.
2
176.
7
13C NMR of 14k
0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.0
2.94
3.01
3.02
0.96
1.03
3.00
2.00
2.04
4.04
1.01
1.42
1.43
1.44
1.44
1.67
1.69
3.24
3.26
3.28
3.30
3.31
3.33
3.35
5.35
5.36
5.38
5.40
5.42
7.44
7.45
7.46
7.46
7.47
7.48
7.48
7.48
7.49
7.49
7.50
7.51
7.51
7.64
7.66
7.69
7.71
7.81
7.82
7.82
7.83
7.83
7.84
7.87
7.89
1H NMR of 14l
0102030405060708090100110120130140150160170180
22.1
23.1
33.5
48.8
112.
3
118.
812
4.7
124.
912
6.0
126.
312
7.7
128.
012
8.6
131.
213
1.7
132.
813
3.5
135.
813
9.9
140.
714
2.0
160.
6
176.
4
13C NMR of 14l
0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.0f1 (ppm)
5.91
3.02
0.95
1.00
1.91
1.90
1.97
2.01
1.02
1.42
1.43
1.44
1.44
1.54
1.56
3.24
3.26
3.28
3.29
3.31
3.33
3.35
5.11
5.13
5.15
5.17
5.18
7.23
7.25
7.44
7.46
7.63
7.66
7.67
7.69
7.76
7.78
1H NMR of 14m
0102030405060708090100110120130140150160170180
22.1
23.1
23.1
33.4
48.2
112.
3
118.
712
1.2
128.
013
1.1
131.
713
1.8
135.
714
0.0
141.
814
2.5
160.
5
176.
5
13C NMR of 14m
0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.0
3.02
3.02
3.04
3.01
0.98
1.00
2.00
2.18
1.99
2.07
0.92
1.41
1.42
1.43
1.43
1.55
1.57
2.33
3.23
3.25
3.26
3.28
3.30
3.32
3.33
5.14
5.16
5.18
5.20
5.22
7.15
7.17
7.26
7.28
7.64
7.66
7.69
7.71
7.75
7.78
1H NMR of 14n
010203040506070809010011012013014015016017018021
.222
.223
.1
33.4
48.4
112.
2
118.
8
126.
212
9.4
131.
213
1.7
135.
813
7.1
139.
714
0.3
142.
1
160.
5
176.
3
13C NMR of 14n
0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.5
3.07
3.04
3.00
1.12
1.15
1.99
1.07
1.06
1.02
1.07
2.05
2.06
1.00
1.39
1.39
1.41
1.41
1.56
1.58
3.20
3.21
3.23
3.24
3.25
3.27
3.28
5.22
5.23
5.25
5.26
5.28
5.97
6.79
6.81
7.10
7.11
7.26
7.33
7.35
7.37
7.39
7.73
7.75
1H NMR of 15a
0102030405060708090100110120130140150160170
22.2
23.1
33.3
48.4
101.
4
107.
9
111.
0
124.
312
4.3
126.
312
7.2
128.
7
140.
714
2.2
143.
614
7.3
148.
2
160.
9
174.
5
13C NMR of 15a
0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.5
3.02
3.01
3.07
1.03
1.11
2.03
1.03
1.01
0.95
1.33
2.00
2.02
1.00
1.39
1.40
1.41
1.41
1.56
1.58
3.19
3.21
3.23
3.24
3.26
3.28
3.29
5.21
5.23
5.25
5.25
5.27
5.29
5.97
6.79
6.81
7.04
7.10
7.11
7.26
7.34
7.36
7.37
7.73
7.75
1H NMR of 15b
010203040506070809010011012013014015016017022
.223
.1
33.3
48.4
101.
4
107.
911
1.0
124.
312
4.3
126.
312
7.2
128.
7
140.
714
2.2
143.
614
7.3
148.
2
160.
9
174.
5
13C NMR of 15b
0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.0
6.02
2.21
1.07
2.22
3.02
3.06
2.03
1.17
0.94
2.03
1.02
0.92
1.00
0.45
1.38
1.39
2.82
2.84
2.85
3.18
3.19
3.20
3.22
3.58
3.59
3.60
3.85
3.86
5.99
6.76
6.78
6.78
6.81
6.82
7.03
7.04
7.10
7.11
7.57
7.58
7.59
1H NMR of 15c
0102030405060708090100110120130140150160170180
22.9
33.2
35.6
40.8
55.9
56.0
101.
4
107.
911
1.0
111.
411
2.1
120.
812
4.2
124.
3
131.
8
140.
814
2.0
147.
314
7.7
148.
214
9.0
161.
8
174.
3
13C NMR of 15c
0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.0
6.14
2.03
0.95
2.09
2.04
2.03
2.09
1.00
1.84
1.40
1.41
3.18
3.19
3.20
3.24
3.26
3.27
3.28
3.30
3.31
3.32
3.33
3.64
3.65
3.66
3.72
3.73
3.74
6.00
6.81
6.81
6.82
6.83
6.95
6.97
6.97
1H NMR of 15d
010203040506070809010011012013014015016017018023
.2
33.5
42.4
47.2
66.6
66.6
101.
6
108.
910
8.9
122.
712
4.2
135.
8
142.
6
148.
314
8.3
164.
9
176.
8
13C NMR of 15d
0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.0
6.06
1.02
1.99
2.00
2.00
1.01
2.08
1.00
1.01
0.87
1.00
1.38
1.39
3.17
3.18
3.20
3.21
3.22
3.24
3.25
4.47
4.48
5.92
5.97
6.74
6.75
6.79
6.80
6.82
6.84
7.05
7.05
7.06
7.06
7.12
7.12
7.78
1H NMR of 15e
0102030405060708090100110120130140150160170180
22.9
33.1
42.9
101.
010
1.3
107.
810
8.2
108.
411
0.9
121.
012
4.1
124.
2
132.
4
140.
514
2.3
146.
814
7.2
147.
814
8.2
161.
5
174.
4
13C NMR of 15e
0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.5
6.10
1.04
2.03
2.04
1.04
1.04
0.93
1.00
1.06
0.96
1.03
1.00
1.38
1.39
2.16
3.18
3.19
3.20
3.22
3.23
3.24
3.26
4.58
4.60
5.98
6.81
6.82
7.05
7.05
7.11
7.12
7.26
7.68
7.68
7.68
7.69
7.70
7.70
8.51
8.52
8.52
8.59
8.60
1H NMR of 15f
0102030405060708090100110120130140150160170
22.9
33.2
40.5
101.
3
107.
911
0.9
123.
612
4.0
124.
2
134.
413
5.7
140.
214
2.6
147.
214
8.2
148.
614
9.1
161.
8
174.
6
13C NMR of 15f
0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.0
1.23
2.39
2.08
3.03
1.17
2.16
2.14
1.01
1.01
1.11
1.94
2.12
1.95
2.07
1.00
1.28
1.28
1.29
1.30
1.31
1.32
1.33
1.33
1.34
1.39
1.39
1.40
1.41
1.42
1.43
1.44
1.47
1.49
1.51
1.51
1.51
1.53
1.54
1.57
1.58
1.75
1.75
1.76
1.76
1.76
1.78
1.78
1.78
1.79
1.79
1.86
1.87
1.88
1.89
1.89
1.90
2.15
2.17
2.93
2.94
2.95
2.96
2.97
2.97
2.98
2.99
2.99
5.20
5.21
5.21
5.23
5.24
7.25
7.25
7.25
7.26
7.27
7.27
7.28
7.28
7.33
7.34
7.34
7.35
7.36
7.36
7.37
7.38
7.38
7.63
7.65
7.68
7.70
7.79
7.81
1H NMR of 16a
0102030405060708090100110120130140150160170180
22.2
25.7
26.0
29.8
33.6
42.6
48.6
112.
2
118.
7
126.
312
7.4
128.
713
1.1
131.
613
5.8
139.
514
1.9
143.
3
160.
5
175.
4
13C NMR of 16a
0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.0
3.04
1.88
2.56
2.45
0.86
1.04
1.08
1.47
2.03
2.04
2.01
2.06
1.00
1.57
1.58
1.75
1.75
1.76
1.77
1.78
1.78
1.79
1.79
1.79
1.87
1.87
1.89
1.90
2.15
2.17
2.17
2.18
2.93
2.94
2.94
2.95
2.96
2.97
2.98
2.99
3.00
5.18
5.19
5.21
5.21
5.23
5.25
7.24
7.24
7.25
7.26
7.27
7.27
7.28
7.28
7.34
7.36
7.37
7.38
7.38
7.39
7.63
7.65
7.68
7.70
7.78
1H NMR of 16b
0102030405060708090100110120130140150160170180
22.3
25.8
26.1
29.8
33.7
42.7
48.7
112.
3
118.
8
126.
312
7.4
128.
813
1.2
131.
713
5.9
139.
614
2.0
143.
4
160.
6
175.
4
13C NMR of 16b
-0.50.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.0
1.21
4.68
1.05
2.11
2.04
1.82
1.00
2.00
6.00
3.00
4.93
1.21
1.36
1.64
1.72
1.82
1.92
2.12
2.15
2.83
2.89
2.98
3.56
3.58
3.60
3.62
3.85
3.86
6.70
6.86
7.56
7.78
1H NMR of 16c
0102030405060708090100110120130140150160170
25.5
25.7
33.2
35.4
40.6
42.2
55.7
55.8
111.
311
1.9
111.
9
118.
512
0.6
130.
913
1.4
135.
613
9.1
141.
9
147.
514
8.8
161.
2
175.
0
13C NMR of 16c
0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.5
1.03
2.26
2.19
1.09
2.08
2.09
1.00
2.05
2.09
2.05
2.03
1.97
2.00
1.27
1.30
1.40
1.40
1.44
1.44
1.47
1.49
1.50
1.52
1.53
1.55
1.56
1.74
1.77
1.77
1.84
1.85
1.86
1.87
1.88
1.89
2.14
2.15
2.17
2.96
2.97
2.98
2.99
3.00
3.01
3.02
3.03
3.25
3.26
3.43
3.45
3.69
3.70
3.71
3.75
3.76
3.77
7.61
7.63
7.67
7.69
1H NMR of 16d
0102030405060708090100110120130140150160170180
25.7
26.0
33.7
42.5
42.8
47.3
66.6
66.8
112.
2
118.
4
129.
013
2.7
134.
113
5.2
144.
7
164.
2
177.
6
13C NMR of 16d
-0.50.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.0
1.11
2.37
2.38
1.11
2.25
2.27
1.12
2.00
2.00
1.06
1.04
1.06
2.01
2.07
1.06
1.26
1.27
1.30
1.33
1.37
1.40
1.43
1.46
1.47
1.51
1.54
1.74
1.77
1.85
1.88
2.13
2.16
2.90
2.91
2.92
2.93
2.94
2.94
2.96
2.96
2.97
4.45
4.47
5.94
6.75
6.77
6.79
6.81
6.83
7.66
7.68
7.71
7.73
7.77
7.79
7.80
1H NMR of 16e
0102030405060708090100110120130140150160170180
25.8
26.0
33.6
42.6
43.1
101.
2
108.
410
8.6
112.
4
118.
812
1.2
131.
213
1.7
132.
213
5.8
139.
814
1.9
147.
114
8.0
161.
3
175.
5
13C NMR of 16e
0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.59.0
1.13
2.13
2.09
1.06
2.06
2.09
1.00
2.09
1.07
2.89
1.99
1.03
0.94
0.93
1.26
1.28
1.39
1.39
1.40
1.41
1.42
1.42
1.44
1.50
1.51
1.53
1.53
1.73
1.84
1.84
1.85
1.86
1.87
1.88
2.12
2.12
2.15
2.15
2.90
2.91
2.92
2.92
2.93
2.94
2.95
2.95
2.96
4.57
4.58
7.24
7.25
7.25
7.26
7.65
7.67
7.70
7.71
7.92
7.93
8.51
8.51
8.52
8.52
8.58
8.59
1H NMR of 16f
0102030405060708090100110120130140150160170180
25.6
25.8
33.4
40.6
42.4
112.
2
118.
6
123.
5
131.
013
1.6
133.
913
5.5
139.
914
1.4
148.
814
9.2
161.
4
175.
5
13C NMR of 16f
0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.0
3.51
3.25
1.12
1.33
4.11
2.18
2.12
1.00
1.57
1.58
2.70
5.17
5.19
5.20
5.22
5.23
7.24
7.30
7.31
7.40
7.64
7.65
7.68
7.68
7.69
7.69
1H NMR of 17a
0102030405060708090100110120130140150160170
19.2
22.2
48.7
112.
4
118.
7
126.
312
7.5
128.
813
1.2
131.
713
5.5
140.
614
2.4
143.
3
160.
3
164.
5
13C NMR of 17a
0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.0
3.18
3.00
1.00
1.07
1.92
1.89
1.86
1.96
1.01
1.57
1.58
2.70
5.19
5.21
5.22
7.25
7.26
7.28
7.33
7.34
7.36
7.37
7.38
7.64
7.65
7.68
7.69
7.78
7.80
1H NMR of 17b
0102030405060708090100110120130140150160
19.1
22.1
48.6
112.
2
118.
6
126.
212
7.3
128.
713
1.1
131.
613
5.4
140.
514
2.3
143.
2
160.
2
164.
5
13C NMR of 17b
0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.0
2.92
1.98
2.00
5.96
1.92
0.99
0.90
3.72
2.68
2.81
2.83
2.84
3.56
3.58
3.59
3.61
3.85
3.86
6.75
6.77
6.80
6.82
7.58
7.59
7.60
7.65
7.65
7.67
7.68
7.68
7.69
1H NMR of 17c
0102030405060708090100110120130140150160
19.1
35.5
40.8
55.9
56.0
111.
411
2.1
112.
3
118.
612
0.7
131.
113
1.5
131.
613
5.5
140.
414
2.4
147.
714
9.0
161.
2
164.
5
13C NMR of 17c
0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.0
3.00
2.03
2.06
2.06
2.06
1.99
2.06
2.74
3.22
3.23
3.24
3.41
3.42
3.43
3.68
3.68
3.69
3.74
3.75
3.76
7.60
7.61
7.67
7.69
1H NMR of 17d
0102030405060708090100110120130140150160170
19.4
42.5
47.3
66.6
66.8
112.
4
118.
3
129.
013
2.8
134.
913
5.0
145.
1
164.
116
6.9
13C NMR of 17d
0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.59.0
3.00
2.02
2.00
2.99
2.01
2.01
1.09
2.68
4.44
4.46
5.94
6.75
6.77
6.78
6.80
6.82
7.66
7.68
7.70
7.72
7.76
1H NMR of 17e
0102030405060708090100110120130140150160170
19.2
43.2
101.
2
108.
410
8.6
112.
5
118.
712
1.3
131.
213
1.8
132.
113
5.5
140.
814
2.2
147.
114
8.0
161.
0
164.
6
13C NMR of 17e
0.00.51.01.52.02.53.03.54.04.55.05.56.06.57.07.58.08.5
3.00
2.08
1.09
1.04
2.02
2.08
0.98
1.01
0.95
2.69
4.57
4.58
7.24
7.26
7.28
7.66
7.68
7.69
7.71
7.90
8.52
8.53
8.59
1H NMR of 17f
0102030405060708090100110120130140150160170
19.0
40.7
112.
3
118.
5
123.
6
131.
013
1.6
133.
913
5.2
135.
614
1.0
141.
8
148.
814
9.1
161.
2
164.
7
13C NMR of 17f
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