1
Identification of noreremophilane-based inhibitors of
angiogenesis using zebrafish assays†
Kalai Mangai Muthukumarasamy,‡a Kishor L. Handore,‡bd Dipti N. Kakade,b Madhuri V.
Shinde,b Shashi Ranjan,a Naveen Kumar,c Seema Sehrawat,c Chetana Sachidanandanad* and
D. Srinivasa Reddybd*
aCSIR-Institute of Genomics & Integrative Biology, South Campus, New Delhi, 110025, India
bCSIR-National Chemical Laboratory, Division of Organic Chemistry, Dr. Homi Bhabha Road,
Pune, 411008, India
cDepartment of Life Sciences, School of Natural Sciences, Shiv Nadar University, NH 91, UP,
India
dAcademy of Scientific and Innovative Research (AcSIR),110025, New Delhi
‡ These authors contributed equally to the work.
* Corresponding authors
Dr. ChetanaSachidanandan
CSIR-Institute of Genomics & Integrative Biology
South Campus, New Delhi, 110025, India
E-mail: [email protected]
and
Dr. D. Srinivasa Reddy
CSIR-National Chemical Laboratory,
Division of Organic Chemistry,
Dr. HomiBhabha Road,
Pune 411008, Maharashtra, India.
E-mail: [email protected]
Tel. +91 20 25902445. Fax. +91 20 2590262
Electronic Supplementary Material (ESI) for Organic & Biomolecular Chemistry.This journal is © The Royal Society of Chemistry 2015
2
Supporting Information:
Table of contents Page
Synthesis of cis-hydrindanes and 11 3
Synthesis of enantiopurecis-hydrindanes 4
Figures 6
S1 Single X-ray Crystal Structure of 31 6
S2 Developmental effects 7
S3 Effect on early markers 8
Copies of 1H and 13CNMR Spectra 9-63
3
Synthesis of cis-hydrindanes and 11
Scheme S1
4
Synthesis of enantiopure cis-hydrindanes
Scheme S2
5
Single X-ray Crystal Structure of 31
(R)-4,4-dimethyl-2-oxotetrahydrofuran-3-yl(3aS,4R,7aR)-2-((E)-1-(2-(2,4-
dinitrophenyl)hydrazono)ethyl)-3a,4,5,7a-tetrahydro-1H-indene-4-carboxylate (31): To a
solution of 29b(20 mg, 0.062 mmol) in EtOH (3.0 mL) was added 2,4-dinitrophenylhydrazine
(21 mg, 0.106 mmol) followed by 1 drops of conc. HCl at room temperature. The mixture was
allowed to stir at room temperature for overnight. Solvent was removed in vacuo. Ether (5
mL) was added followed by 10% NaHCO3 (2 mL). The aqueous layer was extracted by ether
(2 x 5 mL) and dried over anhydrous Na2SO4. Purification by flash chromatography (2.0:8.0%
EtOAc: Petroleum Ether) to afford 31 (21 mg, 67%) as orange colored solid. 1H NMR (400
MHz, CDCl3) δ 11.2 ( s, 1H), 9.11 (d, J = 2.4 Hz, 1H), 8.30 (dd, J = 9.5, 2.4 Hz, 1H), 7.95 (d, J
= 9.5 Hz, 1H), 6.54 (s, 1H), 5.88–5.78 (m, 2H), 5.45 (s, 1H), 4.07 (s, 2H), 3.16–3.01 (m, 3H),
2.56–2.52 (m, 1H), 2.47–2.41 (m, 1H), 2.34–2.32 (m , 2H), 2.23 (s, 3H), 1.22 (s, 3H), 1.11 (s,
3H); 13C NMR (100 MHz, CDCl3) δ 174.3, 172.5, 151.0, 144.9, 143.6, 138.2, 137.5, 130.1,
130.0, 129.7, 124.3, 123.6, 116.8, 76.4, 75.2, 46.4, 42.5, 40.3, 37.9, 37.5, 27.0, 23.1, 20.1,
12.9.
Single crystals of compounds 31 (KLH-E-23) were obtained from ethyl acetate-hexane
mixture. X-ray intensity data were collected on a Bruker SMART APEX II CCD diffractometer
with graphite-monochromatized (Mo Kα=0.71073 Å) radiation at room temperature 296(2) K.
The X-ray generator was operated at 50 kV and 30 mA. Diffraction data were collected with a
scan width of 0.5° and at different settings of and 2. The sample-to-detector distance
was fixed at 5.00 cm. The X-ray data acquisition was monitored by APEX II program suite.1
All the data were corrected for Lorentz-polarization and absorption effects using SAINT and
SADABS programs integrated in APEX II program package.1 The structures were solved by
direct method and refined by full matrix least squares, based on F2, using SHELX-972.
Molecular diagrams were generated using XSHELL program integrated in SHELXTL
package. All the H-atoms (except H-atom bound to N atom) were placed in geometrically
idealized position (C-H = 0.93 Å for the phenyl H-atom, C-H = 0.97 Å for the methylene H-
atom, C-H = 0.98 Å for the methine H-atom and C-H = 0.96 Å for the methyl H-atom) and
constrained to ride on their parent atoms [Uiso(H) = 1.2Ueq(C) for the phenyl, methylene and
methine group and Uiso(H) = 1.5 Ueq(C) for the methyl group]. The H-atom attached to N-
atom in 31 (KLH-E-23) is located in difference Fourier and refined isotropically.
Crystallographic data for 31 (KLH-E-23). (C24H26N4O8): M = 498.49, Crystal dimensions
0.53 x 0.49 x 0.30 mm3, monoclinic, space group P 21,a = 5.9435(4), b = 12.4041(8), c =
16.5108(11) Å, = 92.519(4)°, V = 1216.06(14) Å3, Z = 2, calcd = 1.361 gcm–3, µ (Mo-K) =
0.104 mm–1, F(000) = 524, 2max = 56.64, T = 296(2) K, 23510 reflections collected, 5326
unique, 4403 observed (I > 2 (I)) reflections, 332 refined parameters, R value 0.0545, wR2
= 0.1114, (all data R = 0.0702, wR2 = 0.1181), S = 1.123, minimum and maximum
6
transmission 0.947 and 0.970; maximum and minimum residual electron densities +0.20 and
–0.21 e Å-3.
Figures:
Figure S1. ORTEP of Compound 31 (KLH-E-23)
Crystallographic data for compound31 (KLH-E-23)deposited with the Cambridge
Crystallographic Data Centre as supplementary publication no. CCDC 1009597.
7
Figure S2. Noreremophilane scaffold elicits pleiotropic effects in zebrafish embryos.
Compared to a vehicle treated 2-day old control embryo (a) embryos treated with
25µMNER11 (b) has curved tail, necrosis in the brain (arrowhead) and pericardial edema
(arrow). Two day old control transgenic Tg(myl7:GAL4-VP16) embryos show looped heart
with two chambers, atrium (A) and ventricle (V) in green (c, arrowhead) while compound
treated embryos show a tube shaped heart (d, arrowhead).
V A
8
Figure S3. Early markers of development are unaffected by treatment with NER11. RNA in
situ hybridization was performed on 10 somite stage embryos treated with DMSO (a,c,e,g,i,k)
or 11(25µM) (b, d, f, h, j, l). (a-f) dorsal views with anterior to the left. (g-l) are lateral views
with anterior to the left and dorsal to the top.
9
1H and 13C NMR Spectra
9.0 8.5 8.0 7.5 7.0 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0.0 -0.5
3.35 3.342.222.112.00 1.061.00
1.2
4
1.2
6
1.2
8
2.1
8
2.2
1
2.2
3
2.2
5
2.2
7
2.2
9
2.3
2
2.3
5
2.3
6
2.7
7
2.7
9
2.8
3
2.8
7
3.0
9
3.1
1
3.1
1
4.1
4
4.1
5
4.1
7
4.1
9
5.7
4
6.7
0
7.2
6
10
1 H NMR of Compound 11 (CDCl3; 400MHz)
210 200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10 0
14.3
6
26.6
1
27.1
8
36.4
7
37.6
5
41.9
9
46.0
8
60.7
1
76.8
5
77.1
6
77.4
8
124.6
8
129.4
1
145.5
1
145.6
5
174.7
8
197.0
4
11
13 C NMR of Compound 11 (CDCl3; 100MHz)
9.0 8.5 8.0 7.5 7.0 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0.0
4.10 3.302.322.12 1.051.00
1.1
1
1.2
6
1.2
8
1.2
9
2.1
6
2.2
0
2.2
2
2.2
9
2.3
1
2.3
5
2.4
6
2.4
7
2.4
8
2.4
9
2.8
3
2.8
4
2.8
6
2.8
7
4.1
1
4.1
3
4.1
5
4.1
7
4.1
9
4.2
1
5.6
9
5.7
1
5.7
3
5.7
6
6.7
6
7.2
6
12
1 H NMR of Compound 12 (CDCl3; 400MHz)
210 200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10 0
14
.46
19
.78
24
.54
26
.49
36
.56
44
.85
47
.45
48
.26
60
.49
76
.90
77
.16
77
.41
12
4.2
8
12
7.9
9
14
2.8
1
15
2.4
1
17
3.8
0
19
7.4
2
13
13 C NMR of Compound 12 (CDCl3; 100MHz)
9.0 8.5 8.0 7.5 7.0 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0.0
2.962.06 1.071.021.00
2.2
5
2.2
6
2.2
7
2.2
8
2.2
9
2.3
1
2.3
3
2.4
6
2.4
7
2.8
3
2.8
7
2.9
3
2.9
5
3.1
6
3.1
6
3.1
8
5.7
7
6.7
7
7.2
6
14
1 H NMR of Compound 13 (CDCl3; 400MHz)
210 200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10 0
26
.73
26
.88
36
.56
37
.52
41
.75
45
.84
76
.85
77
.16
77
.47
12
4.4
4
12
9.6
1
14
5.3
7
14
5.7
6
18
0.7
4
19
7.2
7
15
13 C NMR of Compound 13 (CDCl3; 100MHz)
9.0 8.5 8.0 7.5 7.0 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0.0 -0.5
3.43 3.032.15 1.06 1.051.00
1.1
8
2.1
9
2.2
6
2.2
7
2.3
0
2.3
2
2.3
4
2.5
4
2.5
5
2.5
6
2.5
8
2.8
5
2.8
7
2.8
9
2.9
1
5.7
1
5.7
4
5.7
7
5.8
0
6.8
4
7.2
6
16
1 H NMR of Compound 14 (CDCl3; 400MHz)
200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10 0
19
.92
24
.46
26
.64
36
.68
44
.71
47
.52
48
.19
76
.85
77
.16
77
.48
12
4.0
0
12
8.1
7
14
3.1
5
15
1.9
4
17
9.5
9
19
7.6
1
17
13 C NMR of Compound 14 (CDCl3; 100MHz)
9.0 8.5 8.0 7.5 7.0 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0.0 -0.5
3.353.163.042.13 1.111.00
1.1
2
2.1
7
2.2
1
2.2
3
2.2
6
2.3
0
2.3
2
2.4
9
2.5
0
2.5
1
2.5
2
2.8
3
2.8
5
2.8
7
2.8
8
3.7
0
5.6
9
5.7
1
5.7
2
5.7
4
5.7
6
6.7
5
7.2
6
18
1 H NMR of Compound 15 (CDCl3; 400MHz)
210 200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10 0
19
.88
24
.66
26
.58
36
.63
44
.83
47
.44
48
.28
51
.71
76
.91
77
.16
77
.42
12
4.2
3
12
8.1
1
14
2.9
9
15
2.1
5
17
4.2
9
19
7.4
8
19
13 C NMR of Compound 15 (CDCl3; 100MHz)
9.0 8.5 8.0 7.5 7.0 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0.0 -0.5
3.343.23 2.451.14 1.121.00
1.1
2
1.4
1
1.4
5
1.4
9
1.5
2
1.5
4
1.5
7
1.6
4
1.6
6
1.7
2
1.7
6
2.0
3
2.3
2
2.4
1
2.4
4
2.4
8
2.5
0
2.5
2
3.7
0
6.8
3
7.2
6
20
1 H NMR of Compound 5 (CDCl3; 400MHz)
210 200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10
19
.46
20
.90
23
.21
24
.02
26
.52
33
.37
46
.58
48
.67
48
.90
51
.59
76
.85
77
.16
77
.48
14
2.9
2
15
4.2
8
17
4.9
9
19
7.9
5
21
13 C NMR of Compound 5 (CDCl3; 100MHz)
9.0 8.5 8.0 7.5 7.0 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0.0 -0.5
3.28 3.232.362.201.101.041.00
1.2
0
1.4
2
1.4
5
1.4
7
1.5
0
1.5
9
1.6
6
1.6
9
1.7
6
1.7
8
2.0
7
2.3
4
2.3
7
2.4
3
2.4
5
2.5
0
2.5
1
2.5
3
6.9
3
7.2
7
22
1 H NMR of Compound 6 (CDCl3; 400MHz)
210 200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10 0
19
.28
20
.75
22
.88
23
.87
26
.50
33
.21
46
.60
48
.52
48
.69
76
.84
77
.16
77
.47
14
3.0
0
15
4.2
7
18
0.7
0
19
8.1
7
23
13 C NMR of Compound 6 (CDCl3; 100MHz)
9.0 8.5 8.0 7.5 7.0 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0.0 -0.5
6.583.692.412.122.11 1.091.031.00
0.9
4
0.9
6
1.9
2
1.9
5
1.9
8
2.2
2
2.2
4
2.2
5
2.3
1
2.3
7
2.4
0
2.4
3
2.7
9
2.8
5
2.9
2
3.1
2
3.1
4
3.1
6
3.8
9
3.9
0
3.9
1
3.9
2
3.9
3
5.7
6
6.7
2
7.2
6
24
1 H NMR of Compound 16 (CDCl3; 400MHz)
200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10 0
19
.23
26
.64
27
.26
27
.92
36
.52
37
.67
42
.12
46
.15
70
.91
76
.84
77
.16
77
.47
12
4.7
2
12
9.4
9
14
5.7
6
17
4.9
0
19
7.1
3
25
13 C NMR of Compound 16 (CDCl3; 100MHz)
9.0 8.5 8.0 7.5 7.0 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0.0 -0.5
3.683.383.072.62 2.552.372.161.00 1.00
0.9
2
0.9
4
0.9
6
1.3
6
1.3
8
1.4
0
1.4
2
1.6
1
1.6
3
1.6
4
2.1
9
2.2
0
2.2
2
2.2
3
2.3
0
2.3
7
2.3
8
2.4
0
2.4
1
2.8
5
2.9
3
3.1
0
3.1
2
3.1
4
4.1
1
4.1
2
4.1
4
4.1
5
5.7
5
5.7
6
6.7
1
7.2
6
26
1 H NMR of Compound 17 (CDCl3; 400MHz)
200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10 0
13
.83
19
.31
26
.63
27
.24
30
.80
36
.49
37
.69
42
.07
46
.12
64
.66
76
.85
77
.16
77
.48
12
4.7
2
12
9.4
5
14
5.5
6
14
5.7
6
17
4.9
4
19
7.1
3
27
13 C NMR of Compound 17 (CDCl3; 100MHz)
9.0 8.5 8.0 7.5 7.0 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0.0
5.54 3.852.46 2.382.02 1.211.001.00
2.1
9
2.2
1
2.2
4
2.2
5
2.2
6
2.3
0
2.3
1
2.4
0
2.4
4
2.7
7
2.8
3
2.8
7
2.9
3
3.1
2
3.1
5
5.1
2
5.1
5
5.1
9
5.2
3
5.7
6
5.7
8
6.6
1
7.2
6
7.3
3
7.3
5
7.3
6
7.3
7
28
1 H NMR of Compound 18 (CDCl3; 400MHz)
210 200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10 0
26
.57
27
.11
36
.39
37
.72
42
.09
46
.23
66
.54
76
.85
77
.16
77
.48
12
4.6
3
12
8.3
1
12
8.4
9
12
8.7
6
12
9.4
1
13
6.0
1
14
5.5
7
17
4.6
5
19
7.1
0
29
13 C NMR of Compound 18 (CDCl3; 100MHz)
9.0 8.5 8.0 7.5 7.0 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0.0
3.062.122.122.01 1.061.011.00
2.2
2
2.2
4
2.2
6
2.2
8
2.3
0
2.3
2
2.4
9
2.5
0
2.5
1
2.5
3
2.8
3
2.8
7
2.9
2
2.9
4
3.1
4
3.1
6
4.4
3
4.4
5
4.4
7
4.4
8
4.5
0
4.5
6
4.5
8
4.5
9
4.6
1
5.7
3
5.7
7
5.8
0
6.6
8
7.2
6
30
1 H NMR of Compound 19 (CDCl3; 400MHz)
200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10 0
26
.57
26
.84
36
.43
37
.69
41
.75
46
.06
59
.88
60
.24
60
.61
60
.97
76
.85
77
.16
77
.48
12
1.6
9
12
4.2
3
12
9.5
3
14
4.5
7
14
6.0
4
17
3.3
0
19
6.9
9
31
13 C NMR of Compound 19 (CDCl3; 100MHz)
9.0 8.5 8.0 7.5 7.0 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0.0 -0.5
4.032.17 2.15 2.092.082.08 1.091.011.00
0.2
6
0.2
7
0.2
9
0.3
0
0.5
4
0.5
6
0.5
8
0.5
9
1.1
1
1.1
2
1.1
3
1.1
5
1.1
6
2.1
9
2.2
1
2.2
1
2.2
3
2.2
5
2.3
0
2.3
3
2.3
8
2.3
9
2.8
0
2.8
4
2.8
8
2.9
0
3.0
9
3.1
1
3.1
3
3.9
2
3.9
3
3.9
5
3.9
7
3.9
8
4.0
0
5.7
2
5.7
5
5.7
8
6.7
3
7.2
6
32
1 H NMR of Compound 20 (CDCl3; 400MHz)
210 200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10 0
3.3
3
9.9
7
26
.60
27
.20
36
.45
37
.74
42
.08
46
.21
69
.44
76
.84
77
.16
77
.47
12
4.7
3
12
9.3
5
14
5.4
9
14
5.7
4
17
4.9
5
19
7.0
6
33
13 C NMR of Compound 20 (CDCl3; 100MHz)
9.0 8.5 8.0 7.5 7.0 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0.0 -0.5
4.974.963.97 2.852.16 2.122.04 1.061.00 1.00
-0.0
1
0.9
3
0.9
6
0.9
9
1.0
2
1.2
0
1.2
3
1.2
4
1.4
5
1.4
7
1.7
1
1.7
4
2.2
0
2.2
2
2.2
3
2.2
6
2.2
8
2.2
9
2.3
0
2.3
7
2.3
8
2.8
0
2.8
2
2.8
4
2.9
0
3.0
9
3.1
1
3.4
2
3.4
4
3.9
2
3.9
4
5.7
5
5.7
5
6.7
1
7.2
6
34
1 H NMR of Compound 21 (CDCl3; 400MHz)
200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10 0
25
.78
25
.96
26
.44
27
.26
29
.80
36
.49
37
.28
37
.70
40
.62
42
.12
46
.17
68
.90
69
.95
76
.84
77
.16
77
.36
77
.47
12
4.7
2
12
9.4
3
14
5.5
5
14
5.8
1
17
4.9
4
19
7.1
2
35
13 C NMR of Compound 21 (CDCl3; 100MHz)
9.0 8.5 8.0 7.5 7.0 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0.0
4.022.162.112.00 1.02 1.011.00
2.2
1
2.2
3
2.2
5
2.2
5
2.2
9
2.3
0
2.4
2
2.4
3
2.4
4
2.4
5
2.8
1
2.8
5
2.8
9
2.9
2
3.1
4
3.1
4
5.2
4
5.2
7
5.3
0
5.3
5
5.7
6
5.8
9
5.9
0
5.9
2
5.9
4
5.9
5
5.9
6
6.7
1
7.2
6
36
1 H NMR of Compound 22 (CDCl3; 400MHz)
210 200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10 0
26
.65
27
.18
36
.49
37
.67
42
.02
46
.11
65
.38
76
.84
77
.16
77
.47
11
8.4
9
12
4.6
4
12
9.4
8
13
2.1
6
14
5.5
7
14
5.6
2
17
4.4
6
19
7.0
9
37
13 C NMR of Compound 22 (CDCl3; 100MHz)
9.0 8.5 8.0 7.5 7.0 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0.0 -0.5
4.292.08 2.031.00 1.00
2.0
7
2.0
7
2.1
5
2.1
6
2.2
1
2.2
6
2.3
1
2.7
9
2.8
6
2.9
2
3.1
1
3.1
3
3.1
5
5.6
1
5.6
7
5.7
7
5.8
0
5.8
3
6.7
6
7.2
6
38
1 H NMR of Compound 23 (CDCl3; 400MHz)
210 200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10 0
26
.69
28
.23
36
.51
37
.78
43
.46
46
.36
76
.84
77
.16
77
.47
12
4.8
0
12
9.5
1
14
5.6
5
14
5.7
2
17
6.8
9
19
7.2
6
39
13 C NMR of Compound 23 (CDCl3; 100MHz)
9.0 8.5 8.0 7.5 7.0 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0.0 -0.5
5.54 3.892.162.15 1.151.04 1.011.00
2.0
6
2.0
9
2.1
2
2.1
4
2.1
6
2.1
7
2.2
0
2.2
5
2.2
8
2.7
5
2.8
1
2.8
5
2.8
9
3.1
2
3.1
4
3.1
6
4.3
5
4.3
6
4.3
8
4.4
0
4.5
2
4.5
3
4.5
6
4.5
7
5.7
8
6.0
4
6.6
5
7.2
6
7.2
8
7.3
1
7.3
3
7.3
5
40
1 H NMR of Compound 24 (CDCl3; 400MHz)
200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10 0
26
.51
28
.24
36
.36
37
.94
43
.62
44
.35
46
.63
76
.85
77
.16
77
.48
12
5.0
8
12
7.8
5
12
8.9
0
12
9.2
4
13
8.4
7
14
5.5
3
14
6.0
3
17
4.2
7
19
7.2
1
41
13 C NMR of Compound 24 (CDCl3; 100MHz)
9.0 8.5 8.0 7.5 7.0 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0.0 -0.5
6.504.39 3.422.082.04 1.18 1.171.00
1.0
6
1.0
8
1.1
2
1.1
4
1.8
4
2.0
6
2.0
9
2.1
1
2.1
7
2.2
0
2.2
2
2.2
4
2.2
6
2.3
5
2.3
9
2.8
0
2.8
3
2.8
6
3.2
1
3.2
8
3.3
3
3.4
5
3.4
7
3.4
8
3.5
0
5.7
9
5.8
0
5.8
3
6.6
6
7.2
6
42
1 H NMR of Compound 25 (CDCl3; 400MHz)
210 200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10 0
13
.31
15
.21
26
.57
28
.71
36
.43
38
.03
39
.20
40
.59
42
.02
46
.99
76
.85
77
.16
77
.48
12
5.3
9
12
9.1
4
14
5.3
5
14
6.5
9
17
3.7
3
19
7.1
7
43
13 C NMR of Compound 25 (CDCl3; 100MHz)
9.0 8.5 8.0 7.5 7.0 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0.0 -0.5
3.613.183.122.14 1.121.00
1.1
2
1.2
6
1.2
8
1.2
9
1.4
3
1.4
7
1.5
0
1.6
3
1.6
6
1.6
9
1.7
3
2.2
7
2.2
8
2.3
0
2.3
1
2.4
0
2.4
3
2.4
6
2.4
8
4.0
9
4.1
2
4.1
4
4.1
6
4.1
7
4.2
0
6.8
4
7.2
6
44
1 H NMR of Compound 26 (CDCl3; 400MHz)
210 200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10 0
14
.46
19
.30
20
.87
23
.07
23
.96
26
.41
33
.22
46
.57
48
.62
48
.90
60
.35
76
.85
77
.16
77
.48
14
2.7
2
15
4.6
1
17
4.4
7
19
7.9
0
45
13 C NMR of Compound 26 (CDCl3; 100MHz)
9.0 8.5 8.0 7.5 7.0 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0.0
3.70 3.512.49 2.382.001.101.00
0.8
7
1.2
5
1.2
7
1.2
9
1.3
8
1.4
0
1.4
3
1.5
5
1.5
6
1.6
1
1.8
4
2.1
4
2.1
6
2.1
8
2.2
0
2.3
0
2.4
2
2.4
7
2.5
1
2.9
1
2.9
3
2.9
5
4.1
3
4.1
5
4.1
6
4.1
8
6.7
6
7.2
6
46
1 H NMR of Compound 27 (CDCl3; 400MHz)
200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10 0
14.3
8
20.8
7
26.3
2
26.5
3
27.0
9
33.7
7
37.3
3
44.9
6
46.4
9
60.6
3
76.8
5
77.1
6
77.4
7
145.8
4
147.7
0
175.3
1
197.5
3
47
13 C NMR of Compound 27 (CDCl3; 100MHz)
9.0 8.5 8.0 7.5 7.0 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0.0 -0.5
3.913.35 3.212.882.00 0.95 0.00
-0.0
2
1.2
2
1.2
3
1.2
4
1.2
4
1.2
5
1.2
6
1.5
3
1.5
4
1.7
6
1.7
7
1.7
8
1.8
0
2.1
3
2.1
5
2.1
7
2.1
9
2.9
2
2.9
3
2.9
5
2.9
7
2.9
8
3.0
0
3.0
8
3.1
1
4.0
8
4.1
0
4.1
2
4.1
3
4.1
5
7.2
6
48
1 H NMR of Compound 28 (CDCl3; 400MHz)
220 210 200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10 0
14
.40
20
.48
26
.41
27
.95
29
.18
31
.37
32
.40
38
.62
40
.54
43
.84
50
.40
60
.27
76
.84
77
.16
77
.47
17
6.2
1
21
0.7
8
49
13 C NMR of Compound 28 (CDCl3; 100MHz)
9.0 8.5 8.0 7.5 7.0 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0.0
3.68 3.222.26 2.092.00 1.101.05 1.001.00
0.8
7
1.1
3
1.1
4
1.1
5
1.2
1
1.2
3
1.2
4
1.2
5
1.2
8
1.2
8
1.4
1
2.2
5
2.2
7
2.3
0
2.3
1
2.3
1
2.3
9
2.4
3
2.5
4
2.5
5
2.5
6
2.8
3
2.8
7
2.9
2
2.9
4
3.2
0
3.2
2
4.0
3
4.0
6
4.0
7
4.0
8
4.0
9
5.4
0
5.7
8
5.7
8
6.7
8
7.2
6
50
1 H NMR of Compound 29a (CDCl3; 400MHz)
210 200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10 0
20.1
5
23.2
0
26.6
7
27.2
6
36.5
4
37.6
6
40.2
2
42.2
4
46.0
0
75.2
8
76.3
1
76.8
5
77.1
6
77.4
7
124.4
9
129.5
0
145.0
4
145.9
5
172.2
5
173.5
0
197.0
3
51
13 C NMR of Compound 29a (CDCl3; 100MHz)
9.0 8.5 8.0 7.5 7.0 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0.0 -0.5
3.683.452.452.23 1.131.131.11 1.031.00
0.0
0
1.1
2
1.2
1
2.2
5
2.2
6
2.2
9
2.3
1
2.3
4
2.5
2
2.5
4
2.5
7
2.8
2
2.8
4
2.8
8
2.9
4
2.9
6
3.1
5
3.1
6
3.1
8
4.0
4
4.0
6
4.0
7
4.0
9
5.4
4
5.7
8
5.7
9
5.8
2
6.9
3
7.2
6
52
1 H NMR of Compound 29b (CDCl3; 400MHz)
210 200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10 0
20
.14
23
.14
26
.68
26
.88
36
.40
37
.82
40
.35
41
.90
46
.76
75
.29
76
.39
76
.85
77
.16
77
.37
77
.48
12
4.2
1
12
9.7
4
14
5.4
9
14
5.7
0
17
2.4
5
17
4.0
9
19
7.4
3
53
13 C NMR of Compound 29b (CDCl3; 100MHz)
9.0 8.5 8.0 7.5 7.0 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0.0
3.75 3.262.55 2.262.10 1.031.00
1.2
7
1.2
8
1.3
0
1.5
7
1.5
7
2.1
9
2.2
1
2.2
3
2.2
5
2.3
1
2.3
5
2.3
7
2.4
0
2.7
9
2.8
3
2.8
5
2.8
9
2.9
4
3.1
1
3.1
3
3.1
5
4.1
6
4.1
8
4.2
0
4.2
1
5.7
6
6.7
2
7.2
6
54
1 H NMR of Compound 11a (CDCl3; 400MHz)
200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10 0
14
.41
26
.66
27
.25
36
.55
37
.72
42
.08
46
.15
60
.75
76
.84
77
.16
77
.36
77
.47
12
4.7
4
12
9.4
8
14
5.6
6
17
4.8
4
19
7.0
8
55
13 C NMR of Compound 11a (CDCl3; 100MHz)
9.0 8.5 8.0 7.5 7.0 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0.0
3.192.39 2.202.021.00 1.00
0.0
0
1.2
7
1.2
8
1.3
0
2.1
7
2.1
9
2.2
1
2.2
3
2.2
7
2.3
1
2.3
5
2.3
7
2.7
9
2.8
5
2.8
9
2.9
4
3.1
1
3.1
3
3.1
5
4.1
6
4.1
8
4.2
0
4.2
1
5.7
6
6.7
2
7.2
6
56
1 H NMR of Compound 11b (CDCl3; 400MHz)
200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10 0
14
.41
26
.66
27
.25
36
.55
37
.73
42
.08
46
.15
60
.76
76
.85
77
.16
77
.48
12
4.7
4
12
9.4
8
14
5.6
7
17
4.8
4
19
7.0
9
57
13 C NMR of Compound 11b (CDCl3; 100MHz)
9.0 8.5 8.0 7.5 7.0 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0.0 -0.5
3.853.27 2.472.442.252.21 1.081.00
0.0
0
0.8
8
0.9
4
0.9
6
0.9
7
0.9
8
1.2
5
1.2
8
1.2
9
1.6
5
1.6
7
1.6
8
1.7
0
2.2
1
2.2
4
2.2
5
2.2
7
2.2
9
2.3
1
2.3
6
2.3
9
2.4
0
2.8
2
2.8
5
2.8
7
3.1
1
3.1
4
3.1
6
4.0
7
4.0
9
4.1
1
5.7
7
6.7
2
7.2
6
58
1 H NMR of Compound 30a (CDCl3; 400MHz)
210 200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10 0
10
.58
22
.16
26
.66
27
.27
36
.51
37
.70
42
.08
46
.13
66
.40
76
.84
77
.16
77
.47
12
4.7
3
12
9.4
7
14
5.5
7
14
5.7
6
17
4.9
4
19
7.1
4
59
13 C NMR of Compound 30a (CDCl3; 100MHz)
9.0 8.5 8.0 7.5 7.0 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0.0 -0.5
3.853.27 2.472.442.252.21 1.081.00
0.0
0
0.8
8
0.9
4
0.9
6
0.9
7
0.9
8
1.2
5
1.2
8
1.2
9
1.6
5
1.6
7
1.6
8
1.7
0
2.2
1
2.2
4
2.2
5
2.2
7
2.2
9
2.3
1
2.3
6
2.3
9
2.4
0
2.8
2
2.8
5
2.8
7
3.1
1
3.1
4
3.1
6
4.0
7
4.0
9
4.1
1
5.7
7
6.7
2
7.2
6
60
1 H NMR of Compound 30b (CDCl3; 400MHz)
200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10 0
10
.58
22
.16
26
.66
27
.27
36
.51
37
.70
42
.08
46
.13
66
.40
77
.16
77
.36
77
.47
12
4.7
3
12
9.4
7
14
5.5
7
14
5.7
6
17
4.9
4
19
7.1
4
61
13 C NMR of Compound 30b (CDCl3; 100MHz)
12.0 11.5 11.0 10.5 10.0 9.5 9.0 8.5 8.0 7.5 7.0 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0.0 -0.5
3.303.253.042.272.071.121.111.07 1.051.001.00 1.00
1.1
1
1.2
2
2.2
3
2.3
2
2.3
3
2.3
4
2.4
3
2.4
5
2.4
7
2.5
4
2.5
5
2.5
6
3.0
1
3.0
4
3.0
8
3.1
4
4.0
7
5.4
5
5.7
7
5.8
1
5.8
6
5.8
8
6.5
4
7.2
6
7.9
4
7.9
6
8.2
9
8.3
1
9.1
1
9.1
2
11
.19
62
1 H NMR of Compound 31 (CDCl3; 400MHz)
190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10 0
12
.93
20
.11
23
.14
27
.00
37
.54
37
.94
40
.35
42
.50
46
.44
75
.18
76
.38
76
.85
77
.16
77
.48
11
6.7
9
12
3.5
9
12
4.3
3
12
9.7
4
13
0.0
9
13
7.4
7
13
8.1
9
14
3.6
1
14
4.9
4
15
1.0
2
17
2.5
2
17
4.2
9
63
13 C NMR of Compound 31 (CDCl3; 100MHz)
