Synthesis of novel macrocyclic peptido-calix[4]arenes and peptido-pyridines as precursors for potential molecular metallacages, chemosensors and biologically active candidates

Article abstract of DOI:10.1515/znb-2006-1104

Novel macrocyclic dipicolinic acid acylated peptides based on upper rim bridged peptido-calix[4]arenes, peptido-pyridines or hybrid structures of both, were synthesized as potential molecular metallacages and chemosensors. While conventional azide or mixed anhydride (ethyl chloroformate) peptide couplings served well for assembling the L-tyrosine or L-ornithine peptide backbones, the acid chloride of pyridine-2,6-dicarboxylic acid (dipicolinic acid) acid served as the complementary acylating agent. The structure assignment of the new compounds was based on chemical and spectroscopic evidences. Some of these compounds exhibit antimicrobial activities.

Full text of DOI:10.1515/znb-2006-1104

Synthesis of Novel Macrocyclic Peptido-calix[4]arenes and Peptido-
pyridines as Precursors for Potential Molecular Metallacages,
Chemosensors and Biologically Active Candidates
Abd El-Galil El-Sayed Amra, Mohamed Abo-Ghaliab, and Mohamed M. Abdalahc
a Applied Organic Chemistry Department, National Research Centre, Dokki, Cairo, Egypt
b Peptide Chemistry Department, National Research Centre, Dokki, Cairo, Egypt
c Research Units, Hi-Care Pharmaceutical Co., Cairo, Egypt
Reprint requests to Dr. A. E.-G. E. Amr. E-mail: aamr1963@yahoo.com
Z. Naturforsch. 61b, 1335 – 1345 (2006); received March 15, 2006
Novel macrocyclic dipicolinic acid acylated peptides based on upper rim bridged peptido-
calix[4]arenes, peptido-pyridines or hybrid structures of both, were synthesized as potential molecu-
lar metallacages and chemosensors. While conventional azide or mixed anhydride (ethyl chlorofor-
mate) peptide couplings served well for assembling the L-tyrosine or L-ornithine peptide backbones,
the acid chloride of pyridine-2,6-dicarboxylic acid (dipicolinic acid) acid served as the complemen-
tary acylating agent. The structure assignment of the new compounds was based on chemical and
spectroscopic evidences. Some of these compounds exhibit antimicrobial activities.
Key words: Pyridine-2,6-bisamino Acid, Chiral Macrocycles, Peptido-calix[4]arenes,
Antimicrobial Agents
Introduction
The synthesis of chemosensors has become an in-
teresting trend in different fields of analytical chem-
istry [1 – 3]. Calixarenes present a unique class of
molecular structures having a three-dimensional cav-
ity that can host other atoms or molecules. The struc-
tures possess a wide rim with hydrocarbon function-
alities and a narrow rim with phenolic groups. Syn-
thetic modifications at these rims can be carried out
to introduce desired molecular properties, namely, ex-
tra ligands [4, 5]. Accordingly, for versatile applica- Fig. 1. Anticancer cyclic pentapeptide X [19].
tions, narrow rim metal-narrow rim, narrow rim metal-
Recently, we have demonstrated the significance of
2,6-di-substituted pyridine derivatives as biologically
active congeners [14 – 18]. We have, accordingly, re-
ported a potent anti-cancer activity for the cyclic
peptido-pyridines X (Fig. 1) [19].
In view of these findings, we synthesized some
novel macrocyclic 2,6-peptido-pyridines, as well as
the corresponding peptido-calix[4]arenes, as precur-
sors for molecular chemo-sensors and potentially bi-
ologically active compounds.
wide rim or wide rim metal-wide rim cages could
be assembled for hosting organic molecules [6 – 8].
Analogously, for the analytical aspects of hazardous
heavy metal contaminants, chemosensors based on
calixarene metallacage architectures seemed particu-
larly attractive [5]. In this context, we reported the
synthesis of chemosensors as an interesting approach
providing accurate analytical tools in different ana-
lytical fields. In particular, 2,6-peptido-pyridines ex-
hibited a general ionophoric potency [9] and were
used for inventing novel thiocyanate-selective mem-
brane sensors [10]. Furthermore, macrocyclic peptido-
calixarenes have been synthesized and investigated as
Results and Discussion
In the present work we report the synthesis and a
analogues of vancomycin-type antibiotics [11– 13]. preliminary antimicrobial screening of several macro-
c
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A. E.-G. E.-S. Amr et al. · Synthesis of Novel Macrocyclic Peptido-calix[4]arenes
Scheme 1. Synthetic routes
for compounds 3 – 6.
cyclic derivatives based on 2,6-pyridinedicarbonyl ever, when 5,17-diamino-25,26,27,28-tetrakis[(eth-
dichloride (1), 2,6-pyridine-dicarboxylic acid dihydr- oxy-carbonyl)-methoxy]calix[4]arene was allowed to
azide (3), N,N-bis-[1-carboxy-2-(p-hydroxybenzyl)]- react with the same diacid dichloride or diazide, the
2,6-(diaminocarbonyl)-pyridine (5) and the corre- chiral macrocyclic calix[4]arene 7’ was not formed,
sponding hydrazide (6) which was obtained from the presumably due to steric hinderance (Scheme 2).
corresponding ester (4), according to published proce-
dures [20 – 24]. The synthesis of compounds 1 – 6 is
presented in Scheme 1.
Reaction of 5,17-diaminomethyl-25,26,27,28-tetra-
n-propoxycalix[4]arene with the diacid derivative 5
in the presence of ethyl chloroformate (Method A) or
with the diazide of 6 afforded the corresponding chiral
macrocyclic calix[4]arene 8. When 5,17-diamino-
25,26,27,28-tetrakis-[(ethoxycarbonyl)methoxy]calix
[4]arene was allowed to react with the same mixed
anhydride or the diazide, chiral macrocyclic calix[4]
arene 9 was isolated (Scheme 3).
In addition, the synthesis of macrocyclic amide
and peptido-calixarene derivatives was realized by
the use of the potent azide and mixed anhydride
coupling methods [21, 25], which lead mostly to
the formation of optically pure products. The di-
ester derivative 4 was hydrolyzed to the correspond-
ing diacid 5 by 1 N sodium hydroxide. Coupling
Additionally, in the present work, it was found
of 5,17-diaminomethyl-25,26,27,28-tetra-n-propoxy- interesting to extend this course of investigation to
calix[4]arene with 2,6-pyridinedicarbonyl dichloride the synthesis of some new macrocyclic compounds
(1) (Method A) in the presence of triethylamine af- with a chiral cavity formed by bridging 2,6-bis-
forded the macrocyclic pyridino-calix[4]arene deriva- (aminocarbonyl)pyridine with 3,3’-binaphthyldialde-
tive 7. This compound was also obtained by reacting hyde. Thus, condensation of acid hydrazide 6 with
2,6-pyridinedicarboxylic acid hydrazide (3) (as azide, 3,3’-binaphthyldialdehydewas undertaken in refluxing
Method B) with the same diaminocalix[4]arene. How- methanol to afford the corresponding macrocyclic hy-
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A. E.-G. E.-S. Amr et al. · Synthesis of Novel Macrocyclic Peptido-calix[4]arenes
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Scheme 2. Synthetic routes for macrocyclic calix[4]arene 7.
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A. E.-G. E.-S. Amr et al. · Synthesis of Novel Macrocyclic Peptido-calix[4]arenes
Scheme 3. Amidolysis of diaminocalix[4]arenes with diacid 5 or dihydrazide 6. Formation of peptido-calix[4]arenes 8
and 9.
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A. E.-G. E.-S. Amr et al. · Synthesis of Novel Macrocyclic Peptido-calix[4]arenes
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Scheme 4. Synthetic routes for macrocyclic compounds 10 – 12.
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A. E.-G. E.-S. Amr et al. · Synthesis of Novel Macrocyclic Peptido-calix[4]arenes
Scheme 5. Synthetic routes for
macrocyclic bicyclodipyridyl
derivatives 14 – 16.
drazone 10 (Scheme 4). Condensation of the same hy- 3,8,16,21,27,28-hexaaza-2,9,15,22-tetraoxotricyclo-
drazide 6 with selected tetraacid dianhydrides, namely, [3,21,1,110,14]octacosa-1(26),10,11,13,23,25-hexene
1,2,4,5-benzenetetracarboxylic dianhydride or 1,4,5,8- (14) was isolated in a pure form. The latter compound
naphthaline-tetracarboxylic dianhydride in refluxing 14 was reacted with excess hydrazine hydrate in
acetic acid afforded the corresponding macrocyclic boiling methanol to give the corresponding hydrazide
tetraamide pyridine derivatives 11 and 12, respectively 15, which was then treated with aromatic aldehydes,
(Scheme 4).
namely, p-fluorobenzaldehyde or p-chlorobenzalde-
Treatment of 2,6-pyridinedicarbonyl dichloride hyde in refluxing methanol to afford the corresponding
(1) with L-ornithine methyl ester dihydrochloride hydrazones 16a, b, respectively (Scheme 5).
in the presence of triethylamine to afford the ex-
Antimicrobial activity
pected
4-carbomethoxy-2,9-dioxo-3,8,14-triazabi-
was,
cyclo[8,3,1]dodeca-1(13),12,10-triene (13)
The newly synthesized compounds were tested for
however, unsuccessful. Instead, 8,17-dicarbomethoxy- their preliminary antimicrobial activity against differ-
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A. E.-G. E.-S. Amr et al. · Synthesis of Novel Macrocyclic Peptido-calix[4]arenes
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Table 1. Antimicrobial activi-
ties of newly synthesized com-
pounds 5, 7 – 16.
Inhibition zones (cm)
Gram + ve
bacteria
B.
aureus
1.32
1.00
1.95
1.66
1.85
1.86
1.55
1.45
2.00
2.05
1.72
2.10
Gram -ve
bacteria
E.
Yeast
Fungi
Compound
No.
B.
Staph.
aureus
1.55
1.65
2.00
1.85
1.66
1.55
1.95
1.56
1.98
2.00
1.56
2.00
C.
albicans
–
A.
subtilis
1.43
1.75
1.88
1.78
1.54
1.48
1.77
1.98
2.05
1.89
1.46
2.00
coli
niger
1.75
1.65
1.50
1.98
1.42
2.00
1.85
2.35
2.05
2.15
1.50
2.10
5
7
8
9
10
11
12
13
0.65
0.60
0.75
0.88
0.68
0.65
0.55
0.88
0.98
0.90
0.64
0.95
–
0.98
1.05
1.20
–
–
1.00
1.25
0.95
–
15
16a
16b
ChloramphinicolꢀR
–
ent microorganisms representing Gram-positive bac- under reduced pressure and the residue cooled and acidified
with 1 N hydrochloric acid to pH ca. 3. The resulting solid
was filtered off, washed with water and crystallized from
EtOH/H2O to give 5.
teria (Bacillus subtilis, Bacillus aureus and Staphylo-
coccus aureus), Gram-negative bacteria (Escherichia
coli), yeast (Candida albican) and fungi (Aspergillus
niger). The most active compounds were: 14, 15, 16
(all organisms), 8 (B. aureus), 8, 12 (Staph. aureus), 9
(E. coli), 8 – 10 (C. albicans) and 9, 11 (A. niger). The
results are summarized in Table 1.
M. p. 142 – 5 ◦C. – [α]D25 = +10 (MeOH). – IR (film): ν =
˜
3600 – 2617 (broad band, OH and NH), 1732 (C=O, acid),
1657, 1533, 1350 (amide I, II and III) cm−1. – 1H NMR
(270 MHz, DMSO-d6): δ = 3.35 (d, 4H, 2×CH2Ph), 4.40 –
4.50 (m, 2H, 2×CH-N), 7.20 – 7.35 (m, 8H, 2Ph-H), 8.15 –
8.30 (m, 3H, pyr-H), 8.58 (br. s, 2H, 2 × NH, exchange-
able with D2O), 9.86 (br. s, 2H, 2 × OH, exchangeable
with D2O) and 10.40 (br. s, 2H, 2 × OH, exchangeable
with D2O). – 13C {H} NMR (270 MHz, DMSO-d6): δ =
45.66 (CH2Ph), 51.90 (CHNH), 115.31, 124.86, 128.01,
130.23, 138.43, 139.62, 148.53, 156.02 (Ar-C), 123.68,
138.22, 148.15 (pyr-C), 163.24, 172.90 (CO). – MS (EI,
70 eV): m/z (%) = 493 [M+, 15], 459 [M+-2OH, 100], 311
[C16H13N3O4, 80]. – C25H23N3O8 (493.47): calcd. C 60.84,
H 4.69, N 8.51; found C 60.78, H 4.61, N 8.48.
Experimental Section
Melting points were determined in open glass capillaries
using in Electrothermal IA 9000 Series digital melting point
apparatus (Electrothermal, Essex, U.K.) and are uncorrected.
Elemental analyses were performed with all final compounds
with an Elementar, Vario EL, Microanalytical Unit, Na-
tional Research Centre, Cairo Egypt and were in good agree-
ment ( 0.2%) with the calculated values. The IR spectra
(KBr) were recorded on an FT IR-8201 PC spectrophotome-
ter (Schimadzu, Japan). The 1H NMR spectra were mea-
sured with a Jeol 270 MHz spectrometer (FTGNM-EX 270,
Japan) in DMSO-d6 or CDCl3. The chemical shifts were
Synthesis of the macrocyclic calix[4]arene derivative 7
recorded relative to TMS. The Mass spectra (EI) were run M e t h o d A : [ A c i d c h l o r i d e m e t h o d ]
at 70 eV with a Finnegan SSQ 7000 spectrometer (Thermo-
To
a solution of 5,17-diaminomethyl-25,26,27,28-
instrument System Incorporated, USA), m/z values are indi-
cated in Dalton. TLC (Silica gel, aluminum sheets 60 F254
Merck, Darmstadt, Germany) was used for tracing the reac-
tions. The starting materials 1, 3 and 6 were prepared accord-
ing to reported procedures [20 – 24].
tetra-n-propoxycalix[4]arene (0.33 g, 0.5 mmol) in dry
dichloromethane (50 mL), triethylamine (0.101 g, 1 mmol)
was added with stirring at −10 ◦C over 1/2 h, then pyridine-
2,6-dicarbonyl dichloride (1) [22, 23] (0.102 g, 0.5 mmol)
in dry dichloromethane (10 mL) was slowly added. The
reaction mixture was stirred for 3 h at −10 ◦C and left
overnight at r. t., washed with water, 1 N hydrochloric acid,
1 N aqueous sodium bicarbonate and water, and dried over
,
Synthesis of N,N-bis[1-carboxy-2-(p-hydroxybenzyl)]-2,6-
(diaminocarbonyl)pyridine (5)
1 N Sodium hydroxide (25 mL) was added dropwise to a anhydrous sodium sulphate. The solvent was evaporated
stirred solution of the dimethyl ester derivative 4 (1 mmol) under reduced pressure, the product was purified by prepar-
◦
in methanol (20 mL) at −5 C. Stirring was continued for ative chromatography (silica gel, CHCl3:MeOH, 9:1, v/v) to
2 h at −5 ◦C, then for 12 h at r. t. Methanol was distilled off give the macrocyclic calixarene derivative 7.
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A. E.-G. E.-S. Amr et al. · Synthesis of Novel Macrocyclic Peptido-calix[4]arenes
M e t h o d B : [ A z i d e m e t h o d ]
over anhydrous sodium sulphate. The solvent was evaporated
under reduced pressure, and the crude product was purified
A cold mixture (−5 ◦C) of 2,6-pyridinedicarboxylic acid
dihydrazide (3) [24] (0.0.1 g, 0.5 mmol) in 5 N hydrochlo-
ric acid (0.6 mL), glacial acetic acid (1.2 mL) and water
(5 mL) was stirred for 10 min, then aqueous sodium nitrite
(0.069 g, 1 mmol in 3 mL of water) was added in one por-
tion and the mixture stirred for 30 min. The obtained azide
was extracted with dichloromethane (60 mL), washed with
cold water, 3% aqueous sodium bicarbonate, followed by
cold water and dried over anhydrous sodium sulphate. The
obtained azide solution was added in one portion to a cold
by preparative chromatography (silica gel, CHCl3:MeOH,
8.5:1.5, v/v) to give the corresponding macrocyclic deriva-
tives 8 and 9, respectively.
M e t h o d B : [ A z i d e m e t h o d ]
A cold mixture (−5 ◦C) of the dihydrazide 6 [21] (0.521 g,
1 mmol) in 5 N hydrochloric acid (1.2 mL) with glacial acetic
acid (2.4 mL) and water (10 mL) was stirred for 10 min, then
aqueous sodium nitrite (0.138 g, 2 mmol in 6 mL of water)
was added in one portion and the mixture stirred for 30 min.
The obtained azide was extracted with dichloromethane
(120 mL), washed with cold water, 3% aqueous sodium bi-
carbonate followed by cold water, and dried over anhydrous
sodium sulphate. The obtained azide solution was added in
one portion to a cold solution (−5 ◦C) of the diaminocalix-
arene derivatives, namely, 5,17-diaminomethyl-25,26,27,28-
tetra-n-propoxycalix[4]arene or 5,17-diamino-25,26,27,28-
tetrakis[(ethoxycarbonyl)methoxy]calix[4]arene (1 mmol) in
dry dichloromethane (25 mL). The reaction mixture was
stirred at the same temperature for 3 h, then overnight at r. t.,
washed with 0.5 N hydrochloric acid, 3% aqueous sodium bi-
carbonate, water and dried over anhydrous sodium sulphate.
The solvent was distilled off under reduced pressure, and
the crude product was purified by preparative chromatog-
raphy (silica gel, CHCl3 : MeOH, 8.5 : 1.5, v/v) to give the
corresponding macrocyclic derivatives 8 and 9, respectively.
The products were identified by their m. p. and Rf-values in
comparison with authentic samples previously obtained by
Method A.
◦
solution (−5 C) of 5,17-diaminomethyl-25,26,27,28-tetra-
n-propoxycalix[4]arene (0.33 g, 0.5 mmol) in dry dichloro-
methane (15 mL). The reaction mixture was stirred at the
same temperature for 3 h, then for 15 h at r. t., washed with
0.5 N hydrochloric acid, water, 3% aqueous sodium bicar-
bonate, and water, and dried over anhydrous sodium sul-
phate. The solvent was distilled off under reduced pressure to
dryness, the residue was purified by preparative chromatog-
raphy using silica gel (CHCl3:MeOH, 9:1, v/v) to give the
macrocyclic calixarene derivative 7 as a white powder.
◦
˜
M. p. 80 – 2 C. – IR (film): ν = 3344 (NH, amide), 1660,
1529, 1260 (amide I, II and III) cm−1. – 1H NMR (270 MHz,
DMSO-d6): δ = 0.90,– 1.00 (t, 12H, 4 × CH3), 1.95 – 2.10
(m, 8H, 4 × CH2), 3.62 (t, 8H, 4 × CH2O), 4.18 (d, 4H,
2 × CH2N), 3.20, 4.42 (2d, 8H, 4 × Ar-CH2-Ar) [26], 6.62
(s, 4H, calix-H), 7.08 – 7.20 (m, 6H, calix-H), 8.10 – 8.22
(m, 3H, pyr-H) and 8.45 (t, 2H, 2NH, exchangeable with
D2O). – 13C {H} NMR (270 MHz, DMSO-d6): δ = 14.12,
14.35 (Me), 31.18, 31.22 (OCH2CH2), 40.54 (CH2NH),
61.00 (OCH2), 71.94 (Ar-CH2-Ar), 123.72, 138.18, 148.10
(pyr-C), 124.82, 128.62, 129.84, 130.50, 133.05, 151.62,
153.22, 158.32, 169.32 (all calix-C), 163.92 (CONH). – MS
(EI, 70 eV): m/z (%) = 782 [M+, 100], 753 [M+-C2H5,
50], 710 (70), 627 (24), 452 (18), 121 (60), 105 (45) and
78 (45). – C49H55N3O6 (781.99): calcd. C 75.25, H 7.08,
N 5.37; found: C 75.18, H 6.98, N 5.32.
Peptidocalixarene 8
M. p. 123 – 5 C. – [α]D25 = +25 (MeOH). – IR (film):
◦
˜
ν = 3595 – 3340 (broad band, OH and NH), 1667, 1525,
1304 (amide I, II and III) cm−1. – 1H NMR (270 MHz,
DMSO-d6): δ = 0.90 – 1.00 (t, 12H, 4 × CH3), 1.90 – 2.08
(m, 8H, 4 × CH2), 3.45 (d, 4H, 2 × CH2Ph), 3.55 (t, 8H,
4 × CH2O), 4.16 (d, 4H, 2 × CH2N), 3.25, 4.35 (2d, 8H,
4×Ar-CH2-Ar), 4.52 – 4.56 (m, 2H, 2×CH-N), 6.58 (s, 4H,
calix-H), 7.05 – 7.15 (m, 6H, calix-H), 7.20 – 7.42 (m, 8H,
Synthesis of the peptidocalixarines 8 and 9
M e t h o d A : [ M i x e d a n h y d r i d e m e t h o d ]
A cold mixture (−30 ◦C) of the diacid derivative 5 2×Ph-H), 8.05 – 8.15 (m, 3H, pyr-H), 8.55 (t, 2H, 2×NH,
(0.493 g, 1 mmol) in dry dichloromethane (50 mL) with ethyl exchangeable with D2O), 9.24 (d, 2H, 2 × NH, exchange-
chloroformate (0.216 g, 2 mmol) and triethylamine (0.202 g, able with D2O) and 10.50 (br. s, 2H, 2 × OH, exchange-
2 mmol) was stirred for 10 min, then, the diaminocalixarene able with D2O). – 13C {H} NMR (270 MHz, DMSO-d6):
derivatives, namely, 5,17-diamino-methyl-25,26,27,28-tetra- δ = 14.15, 14.42 (Me), 31.10, 31.25 (OCH2CH2), 40.64
n-propoxycalix[4]arene or 5,17-di-amino-25,26,27,28-tetra- (CH2NH), 42.44 (CH2Ph), 52.15 (CHNH), 59.98 (OCH2),
kis[(ethoxycarbonyl)methoxy]calix[4]-arene (1 mmol) in 72.00 (Ar-CH2-Ar), 123.75, 137.97, 148.15 (pyr-C), 124.68,
dry dichloromethane (25 mL) were added dropwise. The re- 127.18, 128.00, 128.38, 128.64, 129.82, 130.54, 133.10,
action mixture was stirred at the same temperature for 3 h 134.65, 151.58, 153.26, 158.38, 169.36 (all Ar-C), 163.92,
and for 12 h at r. t., washed with water, 1 N hydrochloric 172.10 (CONH). – MS (EI, 70 eV): m/z (%) = 1108 [M+,
acid, 1 N aqueous sodium bicarbonate, and water, and dried 35], 936 [M+-4C3H9, 65], 459 [C25H21N3O6, 100], 297
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A. E.-G. E.-S. Amr et al. · Synthesis of Novel Macrocyclic Peptido-calix[4]arenes
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[C16H13N2O4, 85], 163 [C9H9NO2, 78]. – C67H73N5O10 (EI, 70 eV): m/z (%) = 855 [M+, 12], 793 [M+ − 2OCH3,
(1108.36): calcd. C 72.60, H 6.64, N 6.32; found C 72.57, 84], 459 [C25H21N3O6, 100], 297 [C16H13N2O4, 18], 163
H 6.61, N 6.30.
[C9H9NO2, 14]. – C49H41N7O8 (855.90): calcd. C 68.75,
H 4.83, N 11.45; found C 68.69, H 4.78, N 11.40.
Peptidocalixarene 9
Synthesis of macrocyclic octaamide tetraimide pyridine
derivatives 11 and 12
◦
M. p. 123 – 5 C. – [α]D25 = +32 (MeOH). – IR (film):
˜
ν = 3598 – 3350 (broad band, OH and NH), 1758 (C=O,
ester), 1667, 1525, 1304 (amide I, II and III) cm−1. –
1H NMR (270 MHz, DMSO-d6): δ = 1.25 (t, 12H, 4×CH3),
3.56 (d, 4H, 2 × CH2Ph), 4.25 – 4.30 (m, 8H, 4 × CH2O),
3.32, 4.60 (2d, 8H, 4 × Ar-CH2-Ar), 4.56 – 4.60 (m, 2H,
2×CH-N), 4.75 (s, 8H, 4×OCH2CO), 6.68 (s, 4H, calix-H),
7.10 – 7.18 (m, 6H, calix-H), 7.24 – 7.35 (m, 8H, 2×Ph-H),
8.12 – 8.24 (m, 3H, pyr-H), 8.75 (d, 2H, 2×NH, exchange-
able with D2O), 9.86 (s, 2H, 2 × NH, exchangeable with
D2O) and 10.48 (br. s, 2H, 2 × OH, exchangeable with
D2O). – 13C {H} NMR (270 MHz, DMSO-d6): δ = 17.15,
18.42 (Me), 31.65, 31.68 (COOCH2), 42.44 (CH2Ph), 52.28
(CHNH), 61.65 (OCH2CO), 72.16 (Ar-CH2-Ar), 123.86,
138.16, 148.12 (pyr-C), 124.70, 127.21, 127.98, 128.42,
128.66, 129.85, 130.56, 133.05, 134.75, 151.60, 153.26,
158.25, 168.96 (all Ar-C), 164.10, 172.25 (CONH), 172.94
(CO-ester). – MS (EI, 70 eV): m/z (%) = 1256 [M+,
24], 907 [M+-4CH2COOC2H5, 54], 459 [C25H21N3O6,
100], 297 [C16H13N2O4, 82], 163 [C9H9NO2, 66]. –
C69H69N5O18 (1256.34): calcd. C 65.96, H 5.54, N 5.57;
found C 65.90, H 5.51, N 5.53.
A suspension of the hydrazide derivative 6 (0.521 g,
1 mmol) and 1,2,4,5-benzenetetracarboxylic dianhydride or
1,4,5,8-naphthalinetetracarboxylic dianhydride (1 mmol) in
acetic acid (50 mL) was refluxed for 7 h. The solid was col-
lected by filtration, washed with acetic acid and crystallized
from dimethylformamide to give the corresponding macro-
cyclic octaamide dipyridyl derivatives 11 and 12, respec-
tively.
Macrocyclic 11
M. p. 240 – 2 ◦C. – [α]2D5 = +42 (DMF). – IR (film):
˜
ν = 3550 – 3290 (broad band, OH and NH), 1734 (C=O,
anhydride), 1654, 1532, 1290 (amide I, II and III) cm−1. –
1H NMR (270 MHz, DMSO-d6): δ = 3.55 (d, 8H, 4 ×
CH2Ph), 4.35 – 4.38 (m, 4H, 4 × CH-N), 7.25 – 7.45 (m,
16H, 4×Ph-H), 8.10 – 8.22 (m, 6H, 2×pyr-H), 8.64 (t, 4H,
4 × NH, exchangeable with D2O), 9.10 (s, 4H, Ar-H), 9.70
(s, 4H, 4 × NH, exchangeable with D2O) and 10.26 (br.s,
4H, 4 × OH, exchangeable with D2O). – 13C {H} NMR
(270 MHz, DMSO-d6): δ = 42.12 (CH2Ph), 51.96 (CHNH),
124.32, 138.36, 148.45 (pyr-C), 126.62, 128.48, 129.38,
134.58, 138.18, 152.45 (all Ar-C), 164.12, 171.98 (CONH),
174.20 (CON). – MS (EI, 70 eV): m/z (%) = 1407 [M+, 12],
978 [M+ − 4 CH2C6H4OH, 62], 459 [C25H21N3O6, 100],
297 [C16H13N2O4, 8], 163 [C9H9NO2, 4]. – C70H50N14O20
(1407.24): calcd. C 59.74, H 3.58, N 13.93; found C 59.68,
H 3.53, N 13.88.
Synthesis of the macrocyclic binaphthyl hydrazone 10
A mixture of the hydrazide derivative 6 (0.521 g, 1 mmol)
and binaphthyl-3,3’-dialdehyde derivative (0.370 g, 1 mmol)
in absolute methanol (100 mL) was heated under reflux for
6 h. The solvent was evaporated under reduced pressure,
the residue was triturated with ether and the solid was col-
lected by filtration, washed with ether and crystallized from
ethanol to give the corresponding bicyclo-macrocyclic hy-
drazone derivative 10 as orange crystals.
Macrocyclic 12
M. p. > 280 ◦C. – [α]D25 = +15 (DMF). – IR (film):
M. p. 265 – 7 ◦C. – [α]2D5 = +34 (DMF). – IR (film):
˜
˜
ν = 3650 – 3450 (broad band, OH and NH), 1668 (C=N, ν = 3565 – 3300 (broad band, OH and NH), 1732 (C=O,
hydrazone), 1645, 1528, 1265 (amide I, II and III) cm−1. – anhydride), 1656, 1530, 1288 (amide I, II and III) cm−1. –
1H NMR (270 MHz, DMSO-d6): δ = 3.20 (s, 6H, 2 × 1H NMR (270 MHz, DMSO-d6): δ = 3.50 (d, 8H, 4 ×
OCH3), 3.46 (d, 4H, 2 × CH2Ph), 4.50 – 4.55 (m, 2H, 2 × CH2Ph), 4.36 – 4.42 (m, 4H, 4 × CH-N), 7.10 – 7.55 (m,
CH-N), 5.25 (s, 2H, 2 × CH=N), 7.10 – 7.40 (m, 16H, 16H, 4 × Ph-H), 8.10 – 8.20 (m, 6H, 2 × pyr-H), 8.58
Ph-H+naphthyl-H), 8.00 – 8.18 (m, 3H, pyr-H), 8.24 (s, 2H, (t, 4H, 4 × NH, exchangeable with D2O), 8.85 (d, 8H,
naphthyl-H), 8.60 (t, 2H, 2×NH, exchangeable with D2O), naphthyl-H), 9.36 (s, 4H, 4 × NH, exchangeable with D2O)
9.18 (s, 2H, 2 × NH, exchangeable with D2O) and 10.35 and 10.12 (br. s, 4H, 4 × OH, exchangeable with D2O). –
(br. s, 2H, 2 × OH, exchangeable with D2O). – 13C {H} 13C {H} NMR (270 MHz, DMSO-d6): δ = 41.12 (CH2Ph),
NMR (270 MHz, DMSO-d6): δ = 34.75 (OMe), 41.95 51.96 (CHNH), 124.32, 138.36, 148.45 (pyr-C), 125.78,
(CH2Ph), 52.45 (CHNH), 123.76, 138.05, 148.10 (pyr-C), 128.88, 132.49, 135.40, 138.00, 149.45, 153.00 (all Ar-C),
147.84 (CH=N), 124.66, 127.18, 127.52, 128.05, 128.44, 163.98, 169.75 (CONH), 173.88 (CON). – MS (EI, 70 eV):
128.62, 130.05, 130.54, 131.75, 134.26, 134.58, 151.35, m/z (%) = 1507 [M+, 32], 1078 [M+ − 4 CH2C6H4OH,
152.45, 153.10 (all Ar-C), 163.96, 172.60 (CONH). – MS 46], 459 [C25H21N3O6, 100], 297 [C16H13N2O4, 15], 163
Unauthenticated
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1344
A. E.-G. E.-S. Amr et al. · Synthesis of Novel Macrocyclic Peptido-calix[4]arenes
[C9H9NO2, 44]. – C78H54N14O20 (1507.36): calcd. C 62.15, 2 × NH2, exchangeable with D2O), 4.50 – 4.55 (m, 2H, 2 ×
H 3.61, N 13.01; found C 61.99, H 3.55, N 12.99.
CH-N), 8.10 – 8.25 (m, 6H, 2×pyr-H), 8.58 (m, 2H, 2×NH,
exchangeable with D2O), 8.72 (m, 2H, 2 × NH, exchange-
able with D2O) and 9.30 (m, 2H, 2×NH, exchangeable with
D2O). – 13C {H} NMR (270 MHz, DMSO-d6): δ = 28.22,
30.48, 38.32 (CH2), 51.96 (CHNH), 125.10, 125.34, 137.45,
137.70, 148.66, 148.86 (pyr-C), 163.65, 169.65 (CONH),
170.05 (CO-hydrazide). – MS (EI, 70 eV): m/z (%) = 554
[M+, 15], 492 [M+ −2 NHNH2, 16], 247 [C13H17N3O, 62],
245 [C11H7N3O4, 100]. – C24H30N10O6 (554.56): calcd.
C 51.97, H 5.45, N 25.26; found C 51.94, H 5.39, N 25.22.
Synthesis of 7,17-dicarbomethoxy-3,8,16,21,27,28-hexa-
aza-2,9,15,22-tetraoxotricyclo-[3,21,1,110,14]octacosa-
1(26),10,12,14,23,24-hexene (14)
To a solution of L-ornithine methyl ester (0.146 g,
1 mmol), [obtained from L-ornithine methyl ester dihydro-
chloride (0.219 g, 1 mmol) and triethylamine (0.202 g,
2 mmol) in dry tetrahydrofuran], 2,6-pyridinedicarbonyl
dichloride (1) (0.204 g, 1 mmol) in dry tetrahydrofuran
(10 mL) was added slowly with stirring. Triethylamine was
added to the reaction mixture to adjust pH ≈ 8. The reaction
mixture was stirred for 3 h at −15 ◦C and for 12 h at r. t. The
triethylamine hydrochloride was filtered off and excess sol-
vent was evaporated under reduced pressure. The residue was
dissolved in dichloromethane (50 mL), washed with water,
Synthesis of 7,17-di[oxo(p-substituted phenyl)carbohydr-
azonylmethyl)-3,8,16,21,27,28-hexaaza-2,9,15,22-tetraoxo-
tricyclo[3,21,1,110,14]octacosa-1(26),10,12,14,23,25-hex-
ene (16a, b)
A mixture of the hydrazide derivative 6 (0.554 g, 1 mmol)
1 N hydrochloric acid, 1 N aqueous sodium bicarbonate, and and the appropriate aldehyde (1 mmol), namely, p-fluoro-
water and dried over anhydrous sodium sulphate. The sol- benzaldehyde or p-chlorobenzaldehyde in absolute ethanol
vent was evaporated under reduced pressure to dryness and (50 mL) was heated under reflux for 6 h. The solvent was
evaporated under reduced pressure and the residue was solid-
phy on (silica gel, benzene: MeOH, 7.5 : 2.5, v/v) to give the ified with ether. The solid was collected by filtration, washed
the product was purified by preparative TLC chromatogra-
tricyclodimethyl ester derivative 14 as a white powder.
M. p. 124 – 6 ◦C. – [α]D25 = +40 (EtOH). – IR (film):
with ether and crystallized from a proper solvent to afford the
corresponding hydrazone derivatives 16a, b, respectively.
˜
ν = 3355 (NH, amide), 1745 (C=O, ester), 1669, 1530,
1310 (amide I, II and III) cm−1. – 1H NMR (270 MHz,
CDCl3): δ = 1.26 – 1.30 (m, 4H, 2 × CH2), 1.55 – 1.62 (m,
4H, 2×CH2), 3.15 – 3.30 (m, 4H, 2×CH2), 3.65 (s, 6H, 2×
OCH3), 4.35 – 4.52 (m, 2H, 2 × CH-N), 8.05 – 8.15 (m, 6H,
2×pyr-H), 8.65 (t, 2H, 2×NH, exchangeable with D2O) and
9.50 (d, 2H, 2 × NH, exchangeable with D2O). – 13C {H}
NMR (270 MHz, CDCl3): δ = 28.24, 30.34, 38.10 (CH2),
52.11 (CHNH), 55.18 (OCH3), 125.15, 125.45, 138.68,
138.76, 148.95, 149.05 (pyr-C), 164.15, 169.85 (CONH),
172.75 (CO-ester). – MS (EI, 70 eV): m/z (%) = 554 [M+,
45], 492 [M+ − 2 OCH3, 76], 247 [C13H17N3O, 35], 245
[C11H7N3O4, 100]. – C26H30N6O8 (554.55): calcd. C 56.31,
H 5.45, N 15.16; found: C 56.25, H 5.42, N 15.12.
7,17-Di-[oxo(p-fluorophenyl)carbohydrazonylmethyl)-
3,8,16,21,27,28-hexaaza-2,9,15,22-tetra-oxotricyclo-
[3,21,1,110,14]octacosa-1(26),10,12,14,23,25-hexene (16a)
◦
M. p. 254 – 5 C (EtOH). – [α]2D5 = +44 (DMF). – IR
˜
(film): ν = 3340 (NH, amide), 1645 (C=N), 1668, 1535,
1312 (amide I, II and III) cm−1. – 1H NMR (270 MHz,
DMSO-d6): δ = 1.15 – 1.28 (m, 4H, 2 × CH2), 1.54 – 1.60
(m, 4H, 2 × CH2), 3.24 – 3.30 (m, 4H, 2 × CH2), 4.46 –
4.58 (m, 2H, CH-N), 5.45 (s, 2H, 2 × CH=N), 7.26 – 7.42
(m, 8H, 2 × Ph-H), 8.15 – 8.34 (m, 6H, 2 × pyr-H), 8.55
(m, 2H, 2 × NH, exchangeable with D2O), 8.68 (m, 2H,
2×NH, exchangeable with D2O) and 9.42 (m, 2H, 2 ×NH,
exchangeable with D2O). – 13C {H} NMR (270 MHz,
DMSO-d6): δ = 28.42, 30.56, 38.46 (CH2), 52.00 (CHNH),
147.56 (CH=N), 125.05, 125.30, 137.14, 137.28, 148.56,
148.64 (pyr-C), 124.30, 128.37, 129.24, 139.52 (Ar-C),
163.96, 169.85 (CONH), 172.14 (CO-hydrazone). – MS (EI,
70 eV): m/z (%) = 766 [M+, 5], 524 [M+ − 2 F-C6H4CN,
100], 436 [524 − 2 H2N-CO, 16], 245 [C11H7N3O4, 10]. –
C38H36F2N10O6 (766.76): calcd. C 59.52, H 4.73, N 18.27;
found C 59.48, H 4.68, N 18.22.
Synthesis of 7,17-di(oxohydrazinomethyl)-3,8,16,21,27,28-
hexaaza-2,9,15,22-tetra-oxo-tricyclo[3,21,1,110,14]octa-
cosa-1(26),10,12,14,22,25-hexene (15)
A mixture of the dimethyl ester 14 (0.554 g, 1 mmol) and
hydrazine hydrate (0.4 mL, ∼ 8 mmol) in absolute methanol
(50 mL) was refluxed for 6 h. Excess solvent was evaporated
under reduced pressure to dryness and the obtained solid was
crystallized from ethanol to give the corresponding hydrazide
derivative 15 as a white powder.
7,17-Di[oxo(p-chlorophenyl)carbohydrazonylmethyl)-
3,8,16,21,27,28-hexaaza-2,9,15,22-tetraoxo-tricyclo-
[3,21,1,110,14]octacosa-1(26),10,12,14,23,25-hexene (16b)
M. p. 185 – 6 ◦C. – [α]2D5 = +38 (DMF). – IR (film):
˜
ν = 3650 – 3100 (broad band, NH, NH2), 1665, 1532, 1315
1
◦
(amide I, II and III) cm−1. – H NMR (270 MHz, DMSO-
M. p. 262 – 4 C (AcOH). – [α]2D5 = +46 (DMF). – IR
˜
d6): δ = 1.25 – 1.32 (m, 4H, 2 × CH2), 1.52 – 1.58 (m, 4H, (film): ν = 3335 (NH, amide), 1642 (C=N), 1665, 1540,
2×CH2), 3.18 – 3.26 (m, 4H, 2×CH2), 4.30 – 4.40 (br. s, 4H, 1315 (amide I, II and III) cm−1. – 1H NMR (270 MHz,
Unauthenticated
Download Date | 10/8/15 1:12 PM

A. E.-G. E.-S. Amr et al. · Synthesis of Novel Macrocyclic Peptido-calix[4]arenes
1345
DMSO-d6): δ = 1.18 – 1.30 (m, 4H, 2 × CH2), 1.50 – 1.60 Antimicrobial assay
(m, 4H, 2 × CH2), 3.26 – 3.33 (m, 4H, 2 × CH2), 4.42 –
The antimicrobial activity was measured at a concentra-
tion 50 γ/mL of the tested compounds using the bioassay sen-
sitivity technique of antibiotics specified in the US Pharma-
copoeia [27]. The degree of inhibition is measured in com-
4.55 (m, 2H, 2 × CH-N), 5.48 (s, 2H, 2 × CH=N), 7.25 –
7.38 (m, 8H, 2Ph-H), 8.10 – 8.28 (m, 6H, 2 × pyr-H), 8.58
(m, 2H, 2 × NH, exchangeable with D2O), 8.74 (m, 2H,
2×NH, exchangeable with D2O) and 9.38 (m, 2H, 2×NH,
exchangeable with D2O). – 13C {H} NMR (270 MHz,
DMSO-d6): δ = 27.86, 30.45, 38.44 (CH2), 51.86 (CHNH),
146.96 (CH=N), 125.15, 125.31, 137.22, 137.24, 148.25,
148.44 (pyr-C), 123.89, 127.98, 129.34, 139.45 (Ar-C),
163.64, 169.58 (CONH), 171.95 (CO-hydrazone). – MS (EI,
70 eV): m/z (%) = 799 [M+, 10], 524 [M+ −2 Cl-C6H4CN,
100], 436 [524 − 2 H2N-CO, 6], 245 [C11H7N3O4, 18]. –
C38H36Cl2N10O6 (799.67): calcd. C 57.07, H 4.54, N 17.52;
found C 56.98, H 4.48, N 17.46.
R
parison with that of Chloramphinicolꢀ taken as standard.
Acknowledgement
The kind help of Dr. Ahmed Attif, Department of Natu-
ral and Microbial Products, National Research Center, Cairo,
Egypt, for carrying out the antimicrobial screening is highly
appreciated.
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