Page 1
1
Synthesis of new porphyrin/4-quinolone conjugates and evaluation of their efficiency in the photoinactivation of Staphylococcus aureus
Pedro N. Batalhaa,b, Ana T. P. C. Gomesa, Luana S. M. Forezib, Liliana Costac, Maria
Cecília B. V. de Souzab, Fernanda C. S. Boechatb*, Vitor F. Ferreirab, Adelaide
Almeidac, Maria A. F. Faustinoa, Maria G. P. M. S. Nevesa*, José A. S. Cavaleiroa*
a Department of Chemistry and QOPNA, University of Aveiro, 3810-193 Aveiro,
Portugalb Programa de Pós-Graduação em Química, Instituto de Química, Universidade
Federal Fluminense, 24010-141 Niterói, Rio de Janeiro, Brazil.c Department of Biology and CESAM, University of Aveiro 3810-193 Aveiro, Portugal
Supporting Information
Index
Structural characterization:...........................................................................................................31H NMR spectrum of derivative 4a (CDCl3, 300 MHz). ..................................................................613C NMR spectrum of derivative 4a (CDCl3, 75 MHz). ...................................................................7
Expansion of the 13C NMR spectrum of derivative 4a (CDCl3, 75 MHz).........................................81H NMR spectrum of derivative 4b (CDCl3, 300 MHz). ..................................................................8
Expansion of the 1H NMR spectrum of derivative 4b (CDCl3, 300 MHz). ......................................9
Expansion of the 1H NMR spectrum of derivative 4b (CDCl3, 300 MHz). ......................................913C NMR spectrum of derivative 4b (CDCl3, 125 MHz). ...............................................................10
Expansion of the 13C NMR spectrum of derivative 4b (CDCl3, 125 MHz). ...................................101H NMR spectrum of derivative 4c (CDCl3, 300 MHz)..................................................................11
Expansion of the 1H NMR spectrum of derivative 4c (CDCl3, 300 MHz)......................................11
Expansion of the 1H NMR spectrum of derivative 4c (CDCl3, 300 MHz)......................................1213C NMR spectrum of derivative 4c (CDCl3, 125 MHz).................................................................12
Expansion of the 13C NMR spectrum of derivative 4c (CDCl3, 125 MHz). ....................................13
Expansion of the 13C NMR spectrum of derivative 4c (CDCl3, 125 MHz). ....................................13
Electronic Supplementary Material (ESI) for RSC Advances.This journal is © The Royal Society of Chemistry 2015
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2
1H NMR spectrum of derivative 6a (CDCl3, 300 MHz). ................................................................14
Expansion of the 1H NMR spectrum of derivative 6a (CDCl3, 300 MHz)......................................1413C NMR spectrum of derivative 6a (CDCl3, 125 MHz). ...............................................................15
Expansion of the 13C NMR spectrum of derivative 6a (CDCl3, 125 MHz).....................................151H NMR spectrum of derivative 6b (CDCl3, 300 MHz). ................................................................16
Expansion of the 1H NMR spectrum of derivative 6b (CDCl3, 300 MHz). ....................................1613C NMR spectrum of derivative 6b (CDCl3, 125 MHz). ...............................................................17
Expansion of the 13C NMR spectrum of derivative 6b (CDCl3, 125 MHz). ...................................171H NMR spectrum of derivative 6c (CDCl3/CD3OD, 300 MHz). ....................................................18
Expansion of the 1H NMR spectrum of derivative 6c (CDCl3/CD3OD, 300 MHz). ........................18
Expansion of the 1H NMR spectrum of derivative 6c (CDCl3/CD3OD, 300 MHz). ........................1913C NMR spectrum of derivative 6c (CDCl3/CD3OD, 125 MHz). ...................................................19
Expansion of the 13C NMR spectrum of derivative 6c (CDCl3/CD3OD, 125 MHz). .......................20
Expansion of the 13C NMR spectrum of derivative 6c (CDCl3/CD3OD, 125 MHz). .......................201H NMR spectrum of derivative 8a (CDCl3, 300 MHz). ................................................................21
Expansion of the 1H NMR spectrum of derivative 8a (CDCl3, 300 MHz)......................................2113C NMR spectrum of derivative 8a (CDCl3, 75 MHz). .................................................................22
Expansion of the 13C NMR spectrum of derivative 8a (CDCl3, 75 MHz).......................................221H NMR spectrum of derivative 8b (CDCl3, 500 MHz). ................................................................23
Expansion of the 1H NMR spectrum of derivative 8b (CDCl3, 500 MHz). ....................................2313C NMR spectrum of derivative 8b (CDCl3, 75 MHz). .................................................................24
Expansion of the 13C NMR spectrum of derivative 8b (CDCl3, 75 MHz). .....................................24
UV-Vis spectra of derivatives 4a, 6a and 8a in DMF:H2O (9:1) ………………………………………..25
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3
Structural characterization:
The structures of new derivatives 4a-c, 6a-c and 8a-b were confirmed and
assigned according to their 1H and 13C NMR spectra and their molecular
formulae were confirmed by HRMS. 2D NMR spectra (COSY, HSQC and
HMBC) were also used in order to unequivocally identify the proton and carbon
resonances.
The HRMS-ESI+ of the porphyrin/quinolone conjugates 4a and 4b show
protonated molecular ions [M+H]+ at m/z 929.27363 and 971.32075 confirming
the success of the Buchwald-Hartwig coupling between porphyrin 1 and the
bromo-quinolones 2a and 2b, respectively. The 1H NMR spectra of these
conjugates show similar patterns considering the resonances of the porphyrin and
quinolone protons.
In the 1H NMR spectrum of conjugate 4a, it was possible to identify three AB
systems at 8.70 and 8.68 ppm, 8.66 and 8.63 ppm and 8.59 and 8.55 ppm with the
same coupling constant (J = 4.7 Hz) that are related with the resonances of six β-
pyrrolic protons. The three singlets at 8.43, 8.35 and 6.48 ppm were assigned to
the resonances of H-2’, H-3 and NH, respectively. The meso-phenyl protons
appeared as three sets of multiplets at 8.02-7.94 ppm, due to the ortho protons of
5,10,15-Ph; at 7.90-7.80 ppm, due to the ortho protons of 20-Ph and at 7.74-7.60
ppm due to the meta and para protons of 5,10,15,20-Ph. The protons of
quinolone core were unequivocally assigned according with the signal
multiplicity and the correlations observed in the COSY spectrum. The resonance
of quinolone H-5’ appeared as a doublet at 7.74 ppm (J = 2.6 Hz) as confirmed
by the correlation with the double doublet at 7.55 ppm (J = 9.0 and 2.6 Hz),
assigned to the resonance of H-7’. The correlation observed between the
resonance of H-7’ and the doublet at 7.32 ppm (J = 9.0 Hz) allowed the
assignment of this signal to H-8’ proton.
A careful analysis of the HMBC spectrum of 4a allowed the unequivocal
assignment of the carbonyl carbon resonances at 166.3 and 173.9 ppm. The
resonance of H-2’ correlates with both carbonyl signals but the resonance of
protons H-5’ only correlates with the signal at 173.9 ppm allowing its assignment
to the C-4’ resonance; the other signal at 166.3 ppm was assigned to the ester
Page 4
4
carbonyl group. The correlation observed between the quartet at 4.43 ppm (J =
7.0 Hz) and the signal at 166.3 ppm confirms the assignment of this quartet to the
methylene protons resonance of the ester group and therefore the triplet at 1.45
ppm (J = 7.1 Hz) is assigned to the CH3 of the same group. The protons
resonance of the ethyl group attached to N-1 were identified as the quartet at 4.21
ppm (J = 7.1 Hz) due to CH2 protons and the triplet at 1.53 ppm (J = 7.1 Hz) due
to the CH3 protons.
Considering the 1H NMR of conjugate 4b, the main difference when compared
with the spectrum of 4a relies on the resonance of the aliphatic pentyl group
attached to N-1. COSY spectrum allowed to identify unequivocally the
correlation between the multiplet at 1.95-1.80 ppm due to the resonance of H-2’’
with the triplet at 4.12 ppm (J = 7.0 Hz) assigned to the two H-1’’ protons.
Additionally, the correlation of the multiplet at 1.95-1.80 ppm with the one at
1.42-1.32 ppm allowed to assign the resonances due to the four remaining
methylene protons (H-3’’ and H-4’’); the correlation of this last multiplet with
the triplet at 0.93 ppm (J = 7.0 Hz) allowed to identify the H-5’’protons.
In the case of the porphyrin/ribonucleoside conjugate 4c the expected m/z
value at 1345.36388 ([M+H]+) observed in the HRMS-ESI+ spectrum confirmed
its molecular formulae. The 1H NMR spectrum of this compound show a more
complex pattern due to the presence of the protected ribose unit, although the
resonances due to the protons of the porphyrinic core and quinolone unit show
similar features to the ones observed for conjugates 4a and 4b. In this case, the
2D NMR spectra were fundamental for the unequivocal assignment of all sugar
protons and of their protecting groups. The multiplets at 8.02-7.95 ppm and at
7.95-7.85 ppm assigned to the resonances of the ortho protons of 5,10,15-Ph and
20-Ph, respectively, also include the resonances of two ortho OBz protons each.
The remaining two ortho OBz protons appear as a multiplet at 8.77-8.05 ppm.
The most prominent difference observed for the quinolone proton resonances was
related to H-2’ which appeared as a singlet at lower field than in conjugates 4a
and 4b (8.92 ppm versus 8.43 ppm); this fact can be justified by the electron-
withdrawing character of the ribofuranosyl group attached to N-1’. The resonance
of quinolone protons H-5’, H-7’ and H-8’ maintained the same chemical shift
pattern observed for derivative 4a. The doublet at 7.77 ppm (J = 2.5 Hz) was
assigned to H-5’ resonance which correlates with the multiplet at 7.60-7.34 ppm
Page 5
5
due to the resonances of H-7’ and H-8’ along with the nine meta and para OBz
protons. The amino bridge proton (NH) was also identified as the singlet at 6.41
ppm. The COSY spectrum analysis was essential for the unequivocal assignments
of the ribose moiety protons (Figure 2). The low field doublet in the aliphatic
region at 6.48 ppm (J = 4.6 Hz) was attributed to the H-1’’. The correlation
between this signal with the triplet at 6.01 ppm (J = 4.6 Hz) allowed its
assignment to H-2’’. The H-3’’ resonance was identified as the triplet at 5.90
ppm (J = 4.6 Hz) due to the correlations observed with H-2’’ and with the
multiplet at 4.96-4.82 ppm, assigned to the resonances of H-4’’and H-5’’.
Based on heteronuclear (1H-13C) HSQC spectra it was possible to assign the
signals at 90.3, 74.4 and 70.8 to the sugar C-1’’, C-2’’ and C-3’’ resonances
respectively. The resonances of C-4’’ and C-5’’ were attributed to the signals at
80.8 and 63.5 ppm, although their differentiation was not possible. The HMBC
spectrum allowed the unequivocal identification of carbonyl C-4’ resonance at
173.8 ppm and of CO2Et at 165.0 ppm. The signals at 166.1, 165.1 and 164.7
ppm were assigned to the carbonyl resonances of the three benzoyl protecting
groups.
An important feature of the 1H NMR spectra of conjugates 6a and 6b is the
presence of a signal at around -2.60 ppm due to the resonances of the inner NH
protons, which confirms the success of the demetallation step. In the case of
porphyrin/quinolone 6c it is possible to note also the absence of the signals due to
the resonances of the benzoyl groups confirming the presence of deprotected
hydroxyl groups. The HRMS-ESI+ of the porphyrin/quinolone conjugates 6a-c
show the expected [M+H]+ molecular ions at the m/z values 873.35446,
915.40094 and 977.36557 respectively.
The molecular formulae of the intracyclized N-(6-quinolonil)quinolino[2,3,4-
af]porphyrins 8a and 8b were also unambiguously confirmed by HRMS-ESI+
showing the expected [M+H]+ molecular ions at m/z values 871.33852 and
913.38539 respectively.
The main difference between the 1H NMR spectra of conjugates 8a and 8b is
related with the protons’ signals of the aliphatic groups attached to N-1 being the
pattern similar to the one of the corresponding non-cyclized precursors.
Taking into account the spectrum of derivative 8a, the presence of two
doublets at 9.69 ppm (J = 4.7 Hz) and 9.66 ppm (J = 8.2 Hz), due to H-18 and H-
Page 6
6
5’ respectively, is consistent with an intracyclized porphyrinic core.38
Correlations observed on the COSY spectra allowed the identification of the
resonance of H-17 as the doublet at 8.83 ppm (J = 4.7 Hz) and of H-4’ which is
in the multiplet at 7.85-7.49 ppm (Figure 2). The resonances of H-3, H-2’, H-3’,
H-8’’ and of the meta and para protons of 5,10,15-Ph were also assigned to this
multiplet. The remaining β-pyrrole proton resonances appear as four doublets at
8.75 ppm, 8.69 ppm 8.65 ppm and 8.61 ppm with the coupling constant (J = 4.7
Hz). The quinolone proton H-5’’ was identified as the doublet at 9.12 ppm (J =
2.5 Hz) and its correlation with the double doublet at 8.10 ppm (J = 8.6 and 2.5
Hz) allowed the assignment of the latter to the H-7’’. The singlet at 8.67 ppm was
assigned to the quinolone proton H-2’’ resonance. The resonances of the six
ortho protons on 5,10,15-Ph appear as three broad multiplets at 8.32-8.22, 8.20-
8.14 and 8.14-8.04 ppm and the presence of the carboethoxy group was
confirmed by the presence of the expected quartet and triplet at 4.46 ppm and
1.44 ppm. Finally the two internal NH protons appeared as a singlet at -1,32 ppm.
4a - 1H.esp
18 16 14 12 10 8 6 4 2 0 -2 -4Chemical Shift (ppm)
3.2
3.2
2.0
2.0
0.9
1.0
1.1
11.4
1.1
3.1
6.0
0.9
1.0
1.0
5.0
8.69
8.67
8.42
8.36
8.00
7.85
7.69
7.67
7.66
7.65
7.25
6.48
4.47
4.44
4.42
4.40
4.22
4.20
1.55
1.53
1.51
1.47
1.45
1.43
0.00
1H NMR spectrum of derivative 4a (CDCl3, 300 MHz).
N
N N
NNi
HN
N
O
O O
4a
5
10
15
20
2'5'
7'8'
Page 7
7
4a - 1H.esp
8.5 8.0 7.5 7.0 6.5Chemical Shift (ppm)
0.9
1.0
1.1
11.4
1.1
3.1
6.0
0.9
1.0
1.0
5.0
8.70
8.70
8.69
8.68
8.67
8.64
8.62
8.61
8.59
8.54
8.53
8.42
8.36
8.00
8.00
7.99
7.98
7.97
7.96
7.87
7.85
7.84
7.69
7.68
7.67
7.67
7.66
7.65
7.50
7.49
7.47
7.46
7.32
7.29
7.25
6.48
Expansion of the 1H NMR spectrum of derivative 4a (CDCl3, 300 MHz).
4a - 13C.esp
220 200 180 160 140 120 100 80 60 40 20 0 -20Chemical Shift (ppm)
173.
8516
6.32
147.
1814
2.75
139.
1713
3.63
133.
5313
2.24
132.
0812
8.59
127.
7512
7.68
127.
1112
6.88
118.
5711
6.88
113.
6010
9.47
60.9
2
48.9
7
14.7
114
.48
-0.0
2
13C NMR spectrum of derivative 4a (CDCl3, 75 MHz).
N
N N
NNi
HN
N
O
O O
4a
5
10
15
20
2'5'
7'8'
N
N N
NNi
HN
N
O
O O
4a
5
10
15
20
2'5'
7'8'
Page 8
8
4a - 13C.esp
145 140 135 130 125 120 115 110Chemical Shift (ppm)
147.
1814
5.20
143.
0714
2.89
142.
7514
2.59
142.
0814
1.58
141.
2114
0.93
140.
6314
0.58
140.
1513
9.17
133.
6313
3.60
133.
5313
2.85
132.
5513
2.24
132.
0813
1.93
131.
6113
1.54
128.
5912
7.75
127.
6812
7.65
127.
1112
6.88
120.
3912
0.06
118.
57
116.
8811
6.22
115.
81
113.
60
112.
27
109.
47
Expansion of the 13C NMR spectrum of derivative 4a (CDCl3, 75 MHz).
4b - 1H.esp
18 16 14 12 10 8 6 4 2 0 -2 -4Chemical Shift (ppm)
3.0
4.2
3.3
2.3
1.9
2.0
0.9
1.0
1.1
9.0
3.3
1.2
2.0
6.0
0.9
0.9
1.9
4.0
8.69
8.67
8.40
8.35
7.99
7.98
7.97
7.67
7.67
7.66
7.25
6.49
4.45
4.42
4.12
1.88
1.47
1.45
1.43
1.40
1.39
1.38
1.37
0.96
0.93
0.00
1H NMR spectrum of derivative 4b (CDCl3, 300 MHz).
N
N N
NNi
HN
N
O
O O
4a
5
10
15
20
2'5'
7'8'
N
N N
NNi
HN
N
O
O O
4b
5
10
15
20
2'5'
7'8'
1''
2''
3''
4''
5''
Page 9
9
4b - 1H.esp
9.0 8.5 8.0 7.5 7.0 6.5Chemical Shift (ppm)
0.9
1.0
1.1
9.0
3.3
1.2
2.0
6.0
0.9
0.9
1.9
4.0
8.70
8.70
8.69
8.68
8.67
8.66
8.64
8.60
8.59
8.58
8.40
8.35
8.02
8.00
8.00
7.99
7.98
7.97
7.97
7.94
7.73
7.72
7.67
7.67
7.66
7.65
7.54
7.53
7.33
7.30
7.25
6.49
Expansion of the 1H NMR spectrum of derivative 4b (CDCl3, 300 MHz).
4b - 1H.esp
4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0Chemical Shift (ppm)
3.0
4.2
3.3
2.3
1.9
2.0
4.47
4.45
4.42
4.40
4.15
4.12
4.10
1.92
1.90
1.88
1.86
1.83
1.47
1.45
1.43
1.40
1.39
1.38
1.37
1.36
0.96
0.93
0.91
Expansion of the 1H NMR spectrum of derivative 4b (CDCl3, 300 MHz).
N
N N
NNi
HN
N
O
O O
4b
5
10
15
20
2'5'
7'8'
1''
2''
3''
4''
5''
N
N N
NNi
HN
N
O
O O
4b
5
10
15
20
2'5'
7'8'
1''
2''
3''
4''
5''
Page 10
10
4b - 13C.esp
220 200 180 160 140 120 100 80 60 40 20 0 -20Chemical Shift (ppm)
173.
7216
6.42
147.
8014
5.41
142.
97
133.
6813
3.59
132.
3613
1.65
128.
6512
7.79
127.
7312
7.13
126.
9411
6.99
113.
8511
2.19
60.8
8
54.1
9
28.7
522
.28
14.5
513
.93
13C NMR spectrum of derivative 4b (CDCl3, 125 MHz).
4b - 13C.esp
150 145 140 135 130 125 120 115 110Chemical Shift (ppm)
147.
80
145.
4114
3.16
142.
9714
2.81
142.
1414
1.63
141.
2714
1.01
140.
7014
0.65
140.
1413
9.31
133.
6813
3.59
132.
9113
2.36
132.
1313
1.97
131.
6513
1.57
130.
9212
9.40
128.
6512
7.79
127.
7312
7.69
127.
1312
6.94
120.
3212
0.12
118.
61
116.
9911
6.27
115.
85
113.
85
112.
19
109.
49
Expansion of the 13C NMR spectrum of derivative 4b (CDCl3, 125 MHz).
N
N N
NNi
HN
N
O
O O
4b
5
10
15
20
2'5'
7'8'
1''
2''
3''
4''
5''
N
N N
NNi
HN
N
O
O O
4b
5
10
15
20
2'5'
7'8'
1''
2''
3''
4''
5''
Page 11
11
4c - 1H.esp
18 16 14 12 10 8 6 4 2 0 -2 -4Chemical Shift (ppm)
3.3
2.0
3.0
0.9
0.8
0.8
0.8
11.4
11.4
1.1
1.2
4.0
7.7
1.9
0.8
1.7
3.7
0.8
8.69
8.60
8.00
7.99
7.98
7.68
7.68
7.66
7.42
7.39
7.26
6.41
5.90
4.93
4.89
4.86
4.22
4.16
1.32
1.29
1.27
1.26
1H NMR spectrum of derivative 4c (CDCl3, 300 MHz).
4c - 1H.esp
9.0 8.9 8.8 8.7 8.6 8.5 8.4 8.3 8.2 8.1 8.0 7.9 7.8 7.7 7.6 7.5 7.4 7.3 7.2Chemical Shift (ppm)
11.4
11.4
1.1
1.2
4.0
7.7
1.9
0.8
1.7
3.7
0.8
8.93
8.71
8.70
8.69
8.68
8.67
8.66
8.66
8.65
8.60
8.59
8.36
8.09
8.07
8.06
8.00
7.99
7.98
7.97
7.93
7.90
7.77
7.76
7.72
7.70
7.69
7.68
7.68
7.67
7.66
7.63
7.63
7.60
7.57
7.51
7.49
7.44
7.42
7.39
7.37
7.35
7.26
Expansion of the 1H NMR spectrum of derivative 4c (CDCl3, 300 MHz).
N
N N
NNi
HN
N
O
O O
4c
5
10
15
20
2'5'
7'8' O
O
O
O
O
O
O1'' 2''
3''4''
5''
N
N N
NNi
HN
N
O
O O
4c
5
10
15
20
2'5'
7'8' O
O
O
O
O
O
O1'' 2''
3''4''
5''
Page 12
12
4c - 1H.esp
6.5 6.0 5.5 5.0 4.5 4.0Chemical Shift (ppm)
2.0
3.0
0.9
0.8
0.8
0.8
6.49
6.47
6.41
6.03
6.01
6.00
5.92
5.90
5.88
4.94
4.93
4.92
4.91
4.89
4.88
4.86
4.85
4.28
4.25
4.24
4.22
4.19
4.18
4.16
4.13
4.12
4.11
4.10
Expansion of the 1H NMR spectrum of derivative 4c (CDCl3, 300 MHz).
4c -13C.esp
220 200 180 160 140 120 100 80 60 40 20 0 -20Chemical Shift (ppm)
173.
8216
6.10
165.
1116
4.96
142.
8114
0.64
133.
7313
3.68
133.
6012
9.90
129.
8312
8.67
128.
5912
6.94
126.
9212
0.07
118.
6411
3.32
90.3
2
80.8
374
.43
70.7
8
63.4
660
.72
14.3
4
0.02
13C NMR spectrum of derivative 4c (CDCl3, 125 MHz).
N
N N
NNi
HN
N
O
O O
4c
5
10
15
20
2'5'
7'8' O
O
O
O
O
O
O1'' 2''
3''4''
5''
N
N N
NNi
HN
N
O
O O
4c
5
10
15
20
2'5'
7'8' O
O
O
O
O
O
O1'' 2''
3''4''
5''
Page 13
13
4c -13C.esp
150 145 140 135 130 125 120 115 110Chemical Shift (ppm)
145.
0114
3.14
142.
9314
2.81
142.
4714
2.17
141.
6914
1.34
140.
9014
0.69
140.
6413
9.33
134.
1013
3.90
133.
7313
3.68
133.
6013
2.31
129.
9012
9.83
128.
7212
8.67
128.
6312
8.59
128.
5512
7.73
127.
1512
7.06
126.
94
120.
3112
0.07
118.
64
116.
4911
6.11
115.
89
113.
5611
3.32
110.
79
Expansion of the 13C NMR spectrum of derivative 4c (CDCl3, 125 MHz).
4c -13C.esp
90 85 80 75 70 65 60 55Chemical Shift (ppm)
90.3
2
80.8
3
74.4
3
70.7
8
63.4
6
60.7
2
Expansion of the 13C NMR spectrum of derivative 4c (CDCl3, 125 MHz).
N
N N
NNi
HN
N
O
O O
4c
5
10
15
20
2'5'
7'8' O
O
O
O
O
O
O1'' 2''
3''4''
5''
N
N N
NNi
HN
N
O
O O
4c
5
10
15
20
2'5'
7'8' O
O
O
O
O
O
O1'' 2''
3''4''
5''
Page 14
14
6a - 1H.esp
18 16 14 12 10 8 6 4 2 0 -2 -4Chemical Shift (ppm)
1.9
3.2
3.8
2.0
2.0
0.8
0.0
1.0
1.3
9.0
1.1
2.2
1.2
8.0
0.9
0.9
0.9
2.9
2.1
8.75
8.45
8.22
8.21
8.19
8.18
8.17
7.77
7.75
7.74
7.26
6.73
4.46
4.44
4.42
4.25
4.23
4.21
1.58
1.56
1.53
1.49
1.47
1.44
0.00
-2.6
0
1H NMR spectrum of derivative 6a (CDCl3, 300 MHz).
6a - 1H.esp
8.5 8.0 7.5 7.0Chemical Shift (ppm)
0.8
0.0
1.0
1.3
9.0
1.1
2.2
1.2
8.0
0.9
0.9
0.9
2.9
2.1
8.84
8.83
8.80
8.79
8.77
8.75
8.62
8.60
8.45
8.36
8.22
8.22
8.21
8.20
8.19
8.18
8.17
8.16
7.99
7.88
7.85
7.83
7.81
7.80
7.77
7.75
7.74
7.74
7.73
7.67
7.41
7.38
7.26
6.73
Expansion of the 1H NMR spectrum of derivative 6a (CDCl3, 300 MHz).
N
NH N
HN
HN
N
O
O O
6a
5
10
15
20
2'5'
7'8'
N
NH N
HN
HN
N
O
O O
6a
5
10
15
20
2'5'
7'8'
Page 15
15
6a - 13C.esp
220 200 180 160 140 120 100 80 60 40 20 0 -20Chemical Shift (ppm)
173.
7616
6.38
147.
3214
2.19
141.
9713
4.56
134.
4113
2.98
129.
6112
8.74
127.
6912
6.89
126.
7612
6.66
120.
6811
6.87
114.
3210
9.82
60.9
1
48.9
3
14.7
414
.56
0.02
13C NMR spectrum of derivative 6a (CDCl3, 125 MHz).
6a - 13C.esp
145 140 135 130 125 120 115 110Chemical Shift (ppm)
147.
32
142.
7014
2.19
141.
9714
0.57
140.
3014
0.25
134.
5613
4.41
134.
2513
4.17
132.
9813
2.74
132.
6013
0.74
129.
6112
8.74
127.
7512
7.69
127.
4312
7.15
126.
8912
6.76
126.
66
121.
4812
0.68
119.
97
117.
9211
6.92
116.
8711
6.60
114.
32
110.
8610
9.82
Expansion of the 13C NMR spectrum of derivative 6a (CDCl3, 125 MHz).
N
NH N
HN
HN
N
O
O O
6a
5
10
15
20
2'5'
7'8'
N
NH N
HN
HN
N
O
O O
6a
5
10
15
20
2'5'
7'8'
Page 16
16
6b - 1H.esp
18 16 14 12 10 8 6 4 2 0 -2 -4Chemical Shift (ppm)
1.6
3.2
4.3
3.4
2.2
2.0
2.1
0.8
1.0
1.3
9.2
0.9
2.2
1.3
7.8
0.9
0.9
0.9
2.7
1.9
8.75
8.40
8.21
8.21
8.20
8.18
8.18
7.77
7.75
7.74
7.26
6.73
5.30
4.46
4.43
1.91
1.49
1.46
1.44
1.41
1.40
1.39
0.97
0.94
0.92
0.00
-2.6
1
1H NMR spectrum of derivative 6b (CDCl3, 300 MHz).
6b - 1H.esp
9.0 8.5 8.0 7.5 7.0 6.5Chemical Shift (ppm)
0.8
1.0
1.3
9.2
0.9
2.2
1.3
7.8
0.9
0.9
0.9
2.7
1.9
8.84
8.83
8.80
8.79
8.77
8.75
8.62
8.61
8.40
8.36
8.24
8.23
8.21
8.21
8.20
8.19
8.18
7.99
7.97
7.89
7.87
7.82
7.81
7.77
7.75
7.74
7.73
7.64
7.63
7.37
7.33
7.26
6.73
Expansion of the 1H NMR spectrum of derivative 6b (CDCl3, 300 MHz).
N
NH N
HN
HN
N
O
O O
6b
5
10
15
20
2'5'
7'8'
1''
2''
3''
4''
5''
N
NH N
HN
HN
N
O
O O
6b
5
10
15
20
2'5'
7'8'
1''
2''
3''
4''
5''
Page 17
17
6b - 13C.esp
220 200 180 160 140 120 100 80 60 40 20 0 -20Chemical Shift (ppm)
173.
82
166.
4114
7.85
142.
1914
1.97
134.
5613
4.42
133.
0012
9.63
128.
7512
7.69
126.
8912
6.76
126.
6611
9.97
116.
9911
4.29
109.
51
60.9
1
54.1
8
28.7
422
.28
14.5
513
.93
0.01
13C NMR spectrum of derivative 6b (CDCl3, 125 MHz).
6b - 13C.esp
150 145 140 135 130 125 120 115 110Chemical Shift (ppm)
147.
85
142.
6914
2.19
141.
9714
0.59
140.
3014
0.25
138.
0213
7.33
134.
5613
4.42
134.
2513
4.17
133.
7813
3.00
132.
60
129.
6312
8.75
127.
7512
7.69
127.
6612
7.15
126.
8912
6.76
126.
66
121.
4812
0.68
119.
97
117.
9211
6.99
116.
60
114.
29
110.
76
109.
51
Expansion of the 13C NMR spectrum of derivative 6b (CDCl3, 125 MHz).
N
NH N
HN
HN
N
O
O O
6b
5
10
15
20
2'5'
7'8'
1''
2''
3''
4''
5''
N
NH N
HN
HN
N
O
O O
6b
5
10
15
20
2'5'
7'8'
1''
2''
3''
4''
5''
Page 18
18
6c - 1H.esp
18 16 14 12 10 8 6 4 2 0 -2 -4Chemical Shift (ppm)
3.0
1.3
1.0
3.2
1.8
0.8
0.3
1.0
15.3
1.1
5.9
0.8
0.9
1.0
1.0
3.0
0.8
9.13
8.69
8.67
8.62
8.60
8.36
7.99
7.97
7.71
7.68
7.66
7.34
6.06
6.06
4.41
4.38
4.27
4.27
4.24
3.47
3.38
1.45
1.43
1.41
0.00
1H NMR spectrum of derivative 6c (CDCl3/CD3OD, 300 MHz).
6c - 1H.esp
9.0 8.5 8.0 7.5 7.0 6.5 6.0Chemical Shift (ppm)
0.8
0.3
1.0
15.3
1.1
5.9
0.8
0.9
1.0
1.0
3.0
0.8
9.13
8.71
8.69
8.67
8.66
8.63
8.62
8.60
8.58
8.51
8.49
8.36
7.99
7.97
7.90
7.88
7.81
7.78
7.76
7.71
7.68
7.66
7.64
7.62
7.59
7.38
7.36
7.34
6.49
6.06
6.06
Expansion of the 1H NMR spectrum of derivative 6c (CDCl3/CD3OD, 300 MHz).
N
NH N
HN
HN
N
O
O O
6c
5
10
15
20
2'5'
7'8' O
OH
OH
HO
1'' 2''
3''4''
5''
N
NH N
HN
HN
N
O
O O
6c
5
10
15
20
2'5'
7'8' O
OH
OH
HO
1'' 2''
3''4''
5''
Page 19
19
6c - 1H.esp
4.7 4.6 4.5 4.4 4.3 4.2 4.1 4.0 3.9 3.8 3.7Chemical Shift (ppm)
1.3
1.0
3.2
1.8
4.43
4.41
4.38
4.36
4.31
4.29
4.27
4.27
4.26
4.24
4.24
4.23
4.21
4.21
4.12
4.11
4.10
4.08
4.07
3.93
3.91
3.90
3.86
3.86
Expansion of the 1H NMR spectrum of derivative 6c (CDCl3/CD3OD, 300 MHz).
6c - 13C.esp
220 200 180 160 140 120 100 80 60 40 20 0 -20Chemical Shift (ppm)
174.
81
167.
44
143.
0014
0.70
133.
6113
2.95
132.
2313
2.05
131.
7412
8.53
127.
8812
7.26
126.
9912
0.18
115.
8111
1.91
108.
7692
.55
84.6
8
75.7
6
69.0
061
.44
60.3
5
14.2
2
0.00
13C NMR spectrum of derivative 6c (CDCl3/CD3OD, 125 MHz).
N
NH N
HN
HN
N
O
O O
6c
5
10
15
20
2'5'
7'8' O
OH
OH
HO
1'' 2''
3''4''
5''
N
NH N
HN
HN
N
O
O O
6c
5
10
15
20
2'5'
7'8' O
OH
OH
HO
1'' 2''
3''4''
5''
Page 20
20
6c - 13C.esp
145 140 135 130 125 120 115 110Chemical Shift (ppm)
144.
7114
3.20
143.
0014
2.89
142.
7114
2.60
142.
2614
1.78
141.
3914
0.91
140.
7014
0.67
139.
37
133.
7113
3.63
133.
6113
2.95
132.
3013
2.23
132.
0513
1.74
131.
7013
0.95
129.
1912
8.53
127.
8812
7.81
127.
2612
6.99
121.
31
120.
18
118.
76
117.
1511
6.49
115.
81
112.
9011
1.91
108.
76
Expansion of the 13C NMR spectrum of derivative 6c (CDCl3/CD3OD, 125 MHz).
6c - 13C.esp
95 90 85 80 75 70 65 60Chemical Shift (ppm)
92.5
5
84.6
8
75.7
6
69.0
0
61.4
4
60.3
5
Expansion of the 13C NMR spectrum of derivative 6c (CDCl3/CD3OD, 125 MHz).
N
NH N
HN
HN
N
O
O O
6c
5
10
15
20
2'5'
7'8' O
OH
OH
HO
1'' 2''
3''4''
5''
N
NH N
HN
HN
N
O
O O
6c
5
10
15
20
2'5'
7'8' O
OH
OH
HO
1'' 2''
3''4''
5''
Page 21
21
8a - 1H.esp
16 14 12 10 8 6 4 2 0 -2 -4Chemical Shift (ppm)
1.8
2.6
2.8
1.8
1.7
1.1
4.3
4.3
5.4
3.1
2.1
2.0
1.0
1.0
0.8
1.2
1.0
0.9
0.9
1.0
1.0
9.69
9.12
8.69
8.69
8.67
8.66
8.61
8.60
7.79
7.78
7.74
7.73
4.48
4.47
4.45
4.44
4.41
4.39
4.38
1.71
1.69
1.68
1.46
1.44
1.43
0.00
-1.3
3
1H NMR spectrum of derivative 8a (CDCl3, 300 MHz).
8a - 1H.esp
9.5 9.0 8.5 8.0 7.5Chemical Shift (ppm)
1.1
4.3
4.3
5.4
3.1
2.1
2.0
1.0
1.0
0.8
1.2
1.0
0.9
0.9
1.0
1.0
9.70
9.69
9.67
9.65
9.12
9.11
8.84
8.83
8.76
8.75
8.69
8.69
8.67
8.66
8.65
8.61
8.60
8.27
8.17
8.16
8.13
8.11
8.11
8.10
8.09
7.84
7.82
7.79
7.78
7.78
7.74
7.73
7.72
7.72
7.71
7.66
7.62
7.53
7.52
Expansion of the 1H NMR spectrum of derivative 8a (CDCl3, 300 MHz).
N
NH N
HN
8a
5
10
15
20N
N
O O
1'2'
3'4'
5'
2''
5''
7''8''
N
NH N
HN
8a
5
10
15
20N
N
O O
1'2'
3'4'
5'
2''
5''
7''8''
Page 22
22
8a - 13C.esp
220 200 180 160 140 120 100 80 60 40 20 0 -20Chemical Shift (ppm)
173.
64
165.
78
142.
3713
8.99
136.
2413
4.70
134.
4712
7.76
127.
4012
7.21
126.
8512
6.76
123.
3212
2.25
118.
7211
1.94
110.
2810
1.34
61.2
0
49.2
5
14.6
614
.45
13C NMR spectrum of derivative 8a (CDCl3, 75 MHz).
8a - 13C.esp
150 145 140 135 130 125 120 115 110 105 100Chemical Shift (ppm)
149.
07
145.
8114
2.43
142.
3714
2.19
138.
99
136.
2413
5.26
134.
7013
4.63
134.
4713
4.05
132.
5112
9.35
128.
6012
8.54
127.
7612
7.40
127.
2112
6.85
126.
7612
6.71
126.
5512
4.18
123.
3212
2.25
122.
1412
2.12
118.
7211
7.28
116.
7711
6.75
115.
17
111.
94
110.
28
101.
34
Expansion of the 13C NMR spectrum of derivative 8a (CDCl3, 75 MHz).
N
NH N
HN
8a
5
10
15
20N
N
O O
1'2'
3'4'
5'
2''
5''
7''8''
N
NH N
HN
8a
5
10
15
20N
N
O O
1'2'
3'4'
5'
2''
5''
7''8''
Page 23
23
8b - 1H.esp
16 14 12 10 8 6 4 2 0 -2 -4Chemical Shift (ppm)
1.7
2.7
6.8
1.9
1.8
1.6
14.4
2.7
1.9
1.9
1.7
1.0
0.9
0.8
0.8
0.8
0.8
0.9
9.70
9.12
8.76
8.70
8.70
8.67
8.62
8.61
7.79
7.79
7.73
7.72
7.26
4.49
4.48
4.46
4.45
2.04
1.59
1.49
1.48
1.47
1.46
1.44
1.02
1.01
1.00
-1.3
1
1H NMR spectrum of derivative 8b (CDCl3, 500 MHz).
8b - 1H.esp
9.5 9.0 8.5 8.0 7.5Chemical Shift (ppm)
14.4
2.7
1.9
1.9
1.7
1.0
0.9
0.8
0.8
0.8
0.8
0.9
9.71
9.70
9.68
9.66
9.12
9.11
8.85
8.84
8.77
8.76
8.70
8.70
8.67
8.66
8.64
8.62
8.61
8.28
8.18
8.17
8.14
8.13
8.12
8.12
8.10
8.10
7.82
7.80
7.79
7.79
7.78
7.74
7.73
7.73
7.73
7.72
7.67
7.63
7.55
Expansion of the 1H NMR spectrum of derivative 8b (CDCl3, 500 MHz).
N
NH N
HN
8b
5
10
15
20N
N
O O
1'2'
3'4'
5'
2''
5''
7''8''
1'''2'''
3'''
4'''5'''
N
NH N
HN
8b
5
10
15
20N
N
O O
1'2'
3'4'
5'
2''
5''
7''8''
1'''2'''
3'''
4'''5'''
Page 24
24
8b - 13C.esp
220 200 180 160 140 120 100 80 60 40 20 0 -20Chemical Shift (ppm)
173.
6216
5.82
149.
56
142.
3713
9.12
136.
2113
4.71
134.
4812
9.29
128.
5412
7.40
127.
2212
6.85
126.
7112
3.32
118.
8311
1.64
101.
36
61.1
9
54.4
8
28.7
722
.31
14.4
613
.95
13C NMR spectrum of derivative 8b (CDCl3, 75 MHz).
8b - 13C.esp
150 145 140 135 130 125 120 115 110 105 100Chemical Shift (ppm)
149.
56
145.
8114
2.42
142.
3714
2.19
139.
1213
8.02
136.
2113
5.24
134.
7113
4.63
134.
4813
4.06
133.
5513
1.42
129.
2912
8.54
127.
7612
7.40
127.
2212
6.85
126.
7612
6.71
126.
5612
4.17
123.
3212
2.24
122.
1211
8.83
117.
2711
6.75
115.
17
111.
6411
0.29
101.
36
Expansion of the 13C NMR spectrum of derivative 8b (CDCl3, 75 MHz).
N
NH N
HN
8b
5
10
15
20N
N
O O
1'2'
3'4'
5'
2''
5''
7''8''
1'''2'''
3'''
4'''5'''
N
NH N
HN
8b
5
10
15
20N
N
O O
1'2'
3'4'
5'
2''
5''
7''8''
1'''2'''
3'''
4'''5'''
Page 25
25
350 400 450 500 550 600 650 7000.0
0.2
0.4
0.6
0.8
1.0
8a6a4a
wavelenght /nm
Abs
orba
nce
Normalized UV-Vis spectra of derivatives 4a, 6a and 8a in DMF:H2O (9:1)
