Synthesis and antibacterial evaluation of a new series of N-Alkyl-2-alkynyl/(E)-alkenyl-4-(1H)-quinolones

Article abstract of DOI:10.3390/molecules17078217

To gain further insight into the structural requirements of the aliphatic group at position 2 for their antimycobacterial activity, some N-alkyl-4-(1H)-quinolones bearing position 2 alkynyls with various chain length and triple bond positions were prepared and tested for in vitro antibacterial activity against rapidly-growing strains of mycobacteria, the vaccine strain Mycobacterium bovis BCG, and methicillin-resistant Staphylococcus aureus strains, EMRSA-15 and -16. The compounds were also evaluated for inhibition of ATP-dependent MurE ligase of Mycobacterium tuberculosis. The lowest MIC value of 0.5 mg/L (1.2-1.5 μM) was found against M. fortuitum and M. smegmatis. These compounds displayed no or only weak toxicity to the human lung fibroblast cell line MRC-5 at 100 μM concentration. The quinolone derivatives exhibited pronounced activity against the epidemic MRSA strains (EMRSA-15 and -16) with MIC values of 2-128 mg/L 5.3-364.7 μM), and M. bovis BCG with an MIC value of 25 mg/L (66.0-77.4 μM). In addition, the compounds inhibited the MurE ligase of M. tuberculosis with moderate to weak activity showing IC50 values of 200-774 μM. The increased selectivity towards mycobacterial bacilli with reference to MRC-5 cells observed for 2-alkynyl quinolones compared to their corresponding 2-alkenyl analogues serves to highlight the mycobacterial specific effect of the triple bond. Exploration of a terminal bromine atom at the side chain of N-alkyl-2-(E)-alkenyl-4-(1H)-quinolones showed improved antimycobacterial activity whereas a cyclopropyl residue at N-1 was suggested to be detrimental to antibacterial activity.

Full text of DOI:10.3390/molecules17078217

Molecules 2012, 17, 8217-8240; doi:10.3390/molecules17078217
OPEN ACCESS
molecules
ISSN 1420-3049
www.mdpi.com/journal/molecules
Article
Synthesis and Antibacterial Evaluation of a New Series of
N-Alkyl-2-alkynyl/(E)-alkenyl-4-(1H)-quinolones
Abraham Wube 1, Juan-David Guzman 2,3, Antje Hüfner 4, Christina Hochfellner 1,
Martina Blunder 1, Rudolf Bauer 1, Simon Gibbons 3, Sanjib Bhakta 2 and Franz Bucar 1,*
1
Department of Pharmacognosy, Institute of Pharmaceutical Sciences, Karl-Franzens-University Graz,
Universitätsplatz 4, A-8010 Graz, Austria; E-Mails: abraham.wube@uni-graz.at (A.W.);
christina.hochfellner@uni-graz.at (C.H.); martina.blunder@uni-graz.at (M.B.);
rudolf.bauer@uni-graz.at (R.B.)
2
Department of Biological Sciences, Institute of Structural and Molecular Biology, Birkbeck,
University of London, Malet Street, London WC1E 7HX, UK;
E-Mails: talauma@gmail.com (J.-D.G.); s.bhakta@bbk.ac.uk (S.B.)
3
Department of Pharmaceutical and Biological Chemistry, UCL School of Pharmacy,
29-39 Brunswick Square, London WC1N 1AX, UK; E-Mail: simon.gibbons@ucl.ac.uk
4
Department of Pharmaceutical Chemistry, Institute of Pharmaceutical Sciences,
Karl-Franzens-University Graz, Universitätsplatz 1, A-8010 Graz, Austria;
E-Mail: antje.huefner@uni-graz.at
* Author to whom correspondence should be addressed; E-Mail: franz.bucar@uni-graz.at;
Tel.: +43-316-380-5531; Fax: +43-316-380-9860.
Received: 10 April 2012; in revised form: 12 June 2012 / Accepted: 15 June 2012 /
Published: 9 July 2012
Abstract: To gain further insight into the structural requirements of the aliphatic group at
position 2 for their antimycobacterial activity, some N-alkyl-4-(1H)-quinolones bearing
position 2 alkynyls with various chain length and triple bond positions were prepared and
tested for in vitro antibacterial activity against rapidly-growing strains of mycobacteria, the
vaccine strain Mycobacterium bovis BCG, and methicillin-resistant Staphylococcus aureus
strains, EMRSA-15 and -16. The compounds were also evaluated for inhibition of
ATP-dependent MurE ligase of Mycobacterium tuberculosis. The lowest MIC value of
0.5 mg/L (1.2–1.5 µM) was found against M. fortuitum and M. smegmatis. These
compounds displayed no or only weak toxicity to the human lung fibroblast cell line MRC-5
at 100 µM concentration. The quinolone derivatives exhibited pronounced activity against
the epidemic MRSA strains (EMRSA-15 and -16) with MIC values of 2–128 mg/L

Molecules 2012, 17
(5.3–364.7 µM), and M. bovis BCG with an MIC value of 25 mg/L (66.0–77.4 µM). In
8218
addition, the compounds inhibited the MurE ligase of M. tuberculosis with moderate to
weak activity showing IC50 values of 200–774 µM. The increased selectivity towards
mycobacterial bacilli with reference to MRC-5 cells observed for 2-alkynyl quinolones
compared to their corresponding 2-alkenyl analogues serves to highlight the mycobacterial
specific effect of the triple bond. Exploration of a terminal bromine atom at the side chain
of N-alkyl-2-(E)-alkenyl-4-(1H)-quinolones showed improved antimycobacterial activity
whereas a cyclopropyl residue at N-1 was suggested to be detrimental to antibacterial activity.
Keywords: N-alkyl-2-alkynyl/(E)-alkenyl-4(1H)-quinolone; antimycobacterial; MRSA;
cytotoxicity
1. Introduction
Since the discovery of the bacterial ethiology of tuberculosis (TB) by Robert Koch in 1882, various
attempts have been devised to treat infections caused by Mycobacterium tuberculosis. Streptomycin
and p-aminosalicylic acid were the first drugs introduced to treat TB, followed by isoniazid,
pyrazinamide, rifampicin and ethambutol, which were championed as the first-line agents [1,2].
However, the emergence of strains that are resistant to these weapons necessitated further drug options
that led to the second- and third-generation antibiotics. In the early 1990’s fluoroquinolones were
introduced to tackle the ever increasing danger of bacterial resistance and have enjoyed great success
against both Gram-positive and Gram-negative bacteria and certain anaerobes. Unfortunately,
mutations resulting from spontaneous chromosomal alterations, and their widespread overuse and
misuse have contributed significantly to quinolone resistance [3]. Currently there is increasing concern
regarding the paucity of new antimycobacterial therapeutic agents that are coming onto the market in
spite of increased mycobacterial resistance. Therefore, new drugs with new modes of action are
required in view of worldwide-emerging multi drug resistant (MDR-) and extensively drug resistant
(XDR) M. tuberculosis strains [4].
Cell wall biosynthesis is one of the prime targets for drug discovery, because it is an essential
process in the bacterial life cycle. Enzymes involved in the later stage of bacterial cell wall
biosynthesis have been the targets of successfully marketed antibacterial drugs such as β-lactams
including pencillins, carbapenems and cephalosporins, and vancomycin as well as several promising
agents in advanced clinical study such as telavancin and capuramycin [5]. In contrast, except for
fosfomycin no successful antibacterial agents targeting the first cytoplasmic steps of murein
biosynthesis have appeared on the market.
Mur ligases are enzymes that catalyze in a stepwise fashion the early cytoplasmic reactions of
bacterial cell wall biosynthesis, and one of these are the ATP-dependent amide forming ligases of
M. tuberculosis, being MurE the enzyme responsible for catalyzing the addition of meso-diaminopimelic
acid (m-DAP) to the nucleotide precursor UDP-MurNAc-L-Ala-D-Glu to form UDP-MurNAc-L-Ala-D-
Glu-m-DAP [6]. We have recently disclosed N-methyl-4(1H)-quinolones bearing an alkenyl moiety at
position 2 as a novel class of M. tuberculosis MurE inhibitors [7]. The compounds inhibited

Molecules 2012, 17
8219
M. tuberculosis MurE in vitro in the micromolar range, making them successful anti-TB scaffolds for
further rounds of medicinal chemistry improvement.
Our investigations into the design and synthesis of this class of antimycobacterials started with the
initial discovery of the potent antimycobacterial properties of quinolone alkaloids isolated from the
Chinese medicinal plant Euodia rutaecarpa [8,9]. In recent reports, we have described the synthesis,
antimycobacterial and cytotoxicity evaluation of a series of N-methyl-2-alkenyl-4(1H)-quinolones [10,11].
We observed that quinolones having alkenyl moieties at C-2 displayed superior inhibitory effect
compared to their alkyl analogues and subsequent mechanistic investigations indicated that the
antimycobacterial activity of these compounds is related to inhibition of M. tuberculosis MurE ligase [7].
We are particularly interested in the synthesis and biological evaluation of other N-alkyl-4(1H)-
quinolones, since the antimycobacterial mechanism of action is assumed to be different from the
currently used anti-TB agents, and also different from DNA gyrase/topoisomerase inhibition of
clinically-used fluoroquinolones that have an essential free carboxylic group at position C-3 [12],
absent in active N-alkyl-4(1H)-quinolones. Based on these findings we decided to continue our
investigation on modification of the aliphatic side chain at position 2, and the present study was
principally undertaken to explore the impact of the triple bond on the antibacterial and cytotoxic
activities of the quinolones at C-2. Here we report the synthesis and biological evaluation of N-alkyl-
4(1H)-quinolones having alkynyls at C-2 with the intent of elucidating the role that these groups may
play in the in vitro antibacterial profile against fast and slow growing strains of mycobacteria, and
methicillin-resistant S. aureus strains keeping in mind potential correlations with the M. tuberculosis
MurE enzyme inhibitory data. In addition, the influence of a terminal bromine atom at the side chain
on the antimycobacterial activity was evaluated. The cytotoxicity profile of the synthetic derivatives
was assessed against the human lung fibroblast cell line MRC-5.
2. Results and Discussion
2.1. Synthesis
As shown in Scheme 1, our rational design for the synthesis of N-alkyl-2-alkynyl-4(1H)-quinolones
was based on the replacement of the double bond with a triple bond, while maintaining the basic
quinolone skeleton. The synthetic route targeting compounds 8a–t is given in Scheme 1. The key
intermediates, 3a–e and 6a–d, which are essential for the synthesis of our target quinolone derivatives
were obtained in two ways. Pd(OAc)2 and PMe3 mediated direct coupling of methyl vinyl ketone (2)
and 1-alkynes 1 in acetone afforded 5-alkynyl-2-ones 3a–e [13]. The 3-alkynyl-2-ones 6a–d, on the
other hand, were prepared according to the Xing and O’Doherty procedure [14] by treatment of the
terminal alkynes 1 with acetaldehyde in the presence of n-BuLi in THF and subsequent oxidation with
MnO2. The synthesis of this series of 4(1H)-quinolones was accomplished by the well-known
condensation of methyl alkynyl ketones 3a–e and 6a–d with N-alkyl isatoic acid anhydrides 7a–d in
presence of LDA in THF at −78 °C.

Molecules 2012, 17
8220
Scheme 1. Synthesis of N-alkyl-2-alkynyl-4(1H)-quinolones.
O
ii
R'
+
HO
H
O
R'
R'
(1)
(5a). R' = octyl,
(5b). R' = decyl,
(5c). R' = dodecyl,
(5d). R' = tridecyl
(4)
+
(1)
(2)
I
O
iii
(7a). R = methyl,
(7b). R = ethyl,
(7c). R = n-propyl,
(7d). R = n-butyl
O
O
R'
+
(7a). R = methyl,
(7b). R = ethyl,
(7c). R = n-propyl,
(7d). R = n-butyl
O
O
O
N
R
O
(3a). R' = hexyl,
R' +
(3b). R' = heptyl,
(3c). R' = octyl,
(3d). R' = decyl,
(3e). R' = dodecyl
N
R
O
(6a). R' = octyl,
(6b). R' = decyl,
(6c). R' = dodecyl,
(6d). R' = tridecyl
iv
iv
O
R'
N
n
R
(8a). n = 2, R = methyl, R' = hexyl; (8k). n = 2, R = ethyl, R' = octyl;
(8b). n = 2, R = ethyl, R' = hexyl; (8l). n = 2, R = n-propyl, R' = octyl;
(8c). n = 2, R = n-propyl, R' = hexyl; (8m). n = 2, R = n-butyl, R' = octyl;
(8d). n = 2, R = methyl, R' = heptyl; (8n). n = 0, R = methyl, R' = decyl;
(8e). n = 2, R = ethyl, R' = heptyl;
(8f). n = 2, R = n-propyl, R' = heptyl; (8p). n = 2, R = ethyl, R' = decyl;
(8g). n = 0, R = ethyl, R' = octyl; (8q). n = 2, R = n-propyl, R' = decyl;
(8o). n = 2, R = methyl, R' = decyl;
(8h). n = 0, R = n-propyl, R' = octyl; (8r). n = 0, R = methyl, R' = dodecyl;
(8i). n = 0, R = n-butyl, R' = octyl; (8s). n = 2, R = methyl, R' = dodecyl;
(8j). n = 2, R = methyl, R' = octyl; (8t). n = 0, R = methyl, R' = tridecyl;
Reagents and conditions: (i) Pd(OAc)2, PMe3, acetone, 60 °C; (ii) n-BuLi, THF, −78 °C; (iii) MnO2,
THF; (iv) LDA, THF, −78 °C.
In order to study the biological effect of the cyclopropyl moiety which is the constituent of many
therapeutically important fluoroquinolones, we explored the possibility of introducing a cyclopropyl
group at N-1. N-Cyclopropyl isatoic acid anhydride (12) was synthesized from 2-iodobenzoic acid (9)
as shown in Scheme 2. Copper (0) mediated substitution of the iodine of 9 with cyclopropyl amine in
DMF gave 2-(cyclopropylamino)benzoic acid (11). Compound 11 was then treated with NEt3 and
bis-(trichloromethyl) carbonate in the presence of catalytic N,N-dimethylaminopyridine in CH2Cl2 to
afford 12.
The intermediate, ω-bromo-α,β-unsaturated methyl alkenyl ketones 17a–b, required to incorporate
a terminal bromine atom in the aliphatic side chain, were prepared from commercially available
α,ω-diols 13a–b. Bromination of the diols with HBr and subsequent oxidation with pyridinium
chlorochromate (PCC) gave ω-bromoaldehydes 15a–b. Compounds 17a–b were prepared as
previously reported [11] by refluxing 15a–b with the ylide methylcarbonylmethylenephosphorane (16)
obtained from chloroacetone and triphenylphosphine.
To further assess the biological role of the cyclopropyl moiety at N-1 and compare their
antimycobacterial properties with our previously reported N-methyl-2-(E)-alkenyl-4(1H)-quinolones,
compounds 22a–b were prepared from the commercially available aldehydes 20a–b following the

Molecules 2012, 17
8221
same methods described before [10]. Similarly, analogue 23 containing cyclopropyl and undecynyl
groups at positions 1 and 2, respectively (Scheme 2), was prepared from compounds 3b and 12 in
order to examine their impact on biological potency. The identity of the synthetic intermediates and the
4(1H)-quinolone derivatives was confirmed by analysis of 1D and 2D-NMR spectroscopic and
LC-ESI-MS data. The compounds 8a–8t, 18a–b, 19a–b, 22a–b and 23 are reported here for the first time.
Scheme 2. Synthesis of N-cyclopropyl-2-(E)-alkenyl-4(1H)-quinolones and N-cyclo-
propyl-2-alkynyl-4(1H)-quinolones.
O
CO2H
NH2
CO2H
O
i
ii
+
NH
N
O
I
(9)
(10)
(11)
(12)
O
iii
iv
HO
OH
PPh3
n
HO
Br
n
HOC
Br
+
m
(13a). n = 11,
(13b). n = 12
(14a). n = 11,
(14b). n = 12
(15a). m = 10,
(15b). m = 11
(16)
v
O
O
O
O
Br +
12
Br
m
+
m
N
O
(17a). m = 10,
(17b). m = 11
(17a). m = 10,
(17b). m = 11
vi
CH3
vi
(7a)
O
O
Br
N
m
Br
N
m
CH3
(19a). m = 10,
(19b). m =11
(18a). m = 10,
(18b). m = 11
O
N
O
v
vi
+
CHO
16
n
+
12
n
n
(20a). n = 10,
(20b). n = 11
(21a). n = 10,
(21b). n = 11
(22a). n = 10,
(22b). n = 11
O
N
O
vi
+
12
(3b)
(23)
Reagents and conditions: (i) Cu, pyridine, DMF; (ii) NEt3, (CCl3)2CO3, DMAP, CH2Cl2; (iii) HBr;
(iv) PCC, NaOAc, celite, CH2Cl2; (v) THF, reflux; (vi) LDA, THF, −78 °C.
2.2. Biological Evaluation
All of the novel synthetic compounds were screened for their in vitro antimycobacterial activity
against M. smegmatis using a microbroth dilution method [10] and the most active compounds were

Molecules 2012, 17
8222
further tested against M. fortuitum, M. phlei, M. smegmatis mc2155 and M. bovis BCG. Enzymatic
inhibition of the MurE ligase of M. tuberculosis was also determined.
In our microtiter broth dilution assay, the quinolone derivatives displayed significant inhibitory
effect against M. smegmatis, with MIC values ranging from 0.5 to 128 mg/L (1.2–350.7 µM). The
most active compounds were 8n, 8r, 19a and 19b, with an MIC value of 0.5 mg/L (1.2–1.5 µM).
As shown in Table 1, compound 8a with the shortest aliphatic chain (3-decynyl and methyl groups
at position 2 and 1, respectively), displayed the lowest growth inhibition (MIC value of 32 mg/L).
As the aliphatic chain length increased, an increase in potency was observed with maximum activity
achieved for 1-tetradecynyl (compound 8r) and 1-dodecynyl (compound 8n) groups. Further
elongation of the aliphatic chain resulted in a gradual loss of activity. The antimycobacterial data in
Table 1 also indicated that the potency of the quinolone derivatives was related to chain length of the
aliphatic group at position 1 and 2. As shown in Table 2, among the rapidly-growing strains of
mycobacteria, M. smegmatis mc2155 was found to be the most susceptible to the test compounds. In
the spot culture growth inhibition assay (SPOTi), the tested compounds were found to be moderate
growth inhibitors of M. bovis BCG (MIC value of 25 mg/L). Against the epidemic strains of MRSA
(EMRSA-15 and -16) the most active quinolone derivatives, 8l, 8m and 8n displayed pronounced
growth inhibition with MIC values ranging from 2 to 8 mg/L (5.5–22.8 µM).
Table 1. MIC values against M. smegmatis and cytotoxicity against MRC-5 cells of the
N-alkyl-2-alkynyl/(E)-alkenyl-4(1H)-quinolone derivatives.
O
O
R´
N
N
R
R´
n
R
18a- b, 19a- b, 22a- b
8a-t, 23
MIC against
Substituent
M. smegmatis
Metabolic active MRC-5 cells (%)
Comp.
(ATCC 14468)
n
R
R'
mg/L µM
100 µM
60 µM
30 µM
10 µM
8a
8b
8c
8d
8e
8f
2 –CH3
2 –C2H5
2 –C3H7
2 –CH3
2 –C2H5
2 –C3H7
0 –C2H5
0 –C3H7
0 C4H9
2 –CH3
2 –C2H5
2 –C3H7
2 –C4H9
hexyl
hexyl
hexyl
heptyl
heptyl
heptyl
octyl
octyl
octyl
octyl
octyl
octyl
octyl
32
16
8
108.5 89.3 ± 2.94 97.5 ± 0.74 98.4 ± 1.07 100 ± 1.05
51.8 85.7 ± 1.96 98.4 ± 0.77 100 ± .73 102 ± 0.85
24.8 100 ± 1.45 100 ± 0.55 102 ± 0.42 103 ± 1.22
25.9 97.3 ± 0.66 98.7 ± 0.8 99.8 ± 0.52 101 ± 1.08
24.8 100 ± 1.05 100 ± 0.56 101 ± 1.42 102 ± 1.05
11.9 70.9 ± 8.50 91.7 ± 2.77 101 ± 1.28 100 ± 1.05
6.5 12.1 ± 4.79 56.5 ± 2.03 98.7 ± 2.97 101 ± 0.97
12.4 14.1 ± 7.83 39.5 ± 3.59 101 ± 2.47 103 ± 1.38
11.9 28.7 ± 8.95 67.2 ± 9.90 101 ± 0.81 103 ± 3.13
12.4 95.84 ± 0.96 95.1 ± 0.81 96.7 ± 1.49 97.6 ± 1.03
11.9 74.8 ± 1.99 94.7 ± 0.92 95.1 ± 1.04 97.6 ± 1.69
8
8
4
8g
8h
8i
2
4
4
8j
4
8k
8l
4
1
2.8
5.5
9.0 ± 6.85 69.1 ± 8.04 99.1 ± 2.32 100 ± 0.31
0.4 ± 0.02 67.5 ± 3.19 89.7 ± 2.46 94.9 ± 3.18
8m
2

Molecules 2012, 17
8223
Table 1. Cont.
MIC against
Substituent
M. smegmatis
(ATCC 14468)
mg/L µM
Metabolic active MRC-5 cells (%)
Compound
n
R
R'
100 µM
60 µM
30 µM
10 µM
8n
8o
0 –CH3
2 –CH3
2 –C2H5
2 –C3H7
0 –CH3
2 –CH3
0 –CH3
decyl
0.5
2
1.5
100 ± 0.82 100 ± 0.81 100 ± 0.72 101 ± 0.85
decyl
5.7 90.7 ± 3.48 95.8 ± 2.54 97.9 ± 0.88 100 ± 0.73
2.7 8.75 ± 9.35 93.3 ± 1.13 95.4 ± 0.76 97.3 ± 1.37
8p
decyl
1
8q
decyl
1
2.6
0.2 ± 0.09 19.8 ± 9.85 67.9 ± 2.65 92.7 ± 4.62
8r
dodecyl
dodecyl
tridecyl
0.5
1.5 95.7 ± 1.14 97.1 ± 1.69 98.1 ± 1.65 99.7 ± 2.03
8s
128 337.7 41.5 ± 9.49 58.9 ± 3.65 92.2 ± 0.70 93.9 ± 0.61
128 350.7 93.8 ± 0.96 98.8 ± 1.28 100 ± 1.38 100 ± 1.01
8t
18a
18b
19a
19b
22a
22b
23
-
-
Cp bromodecyl
Cp bromoundecyl
8
8
18.6
18.1
1.2
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
- –CH3 bromodecyl
0.5
- –CH3 bromoundecyl 0.5
1.2
-
-
Cp
Cp
Cp
-
undecyl
128 350.7
128 337.8
dodecyl
2
-
hexyl
8
4
2
24.3
29.2
9.8
Isoniazid
-
-
Ethambutol -
-
Vinblastin *
51.5 ± 2.36
ND: not determined; * 0.12 µM.
Table 2. In vitro biological activity of selected N-alkyl-4(1H)-quinolone derivatives and
positive controls.
MIC mg/L(µM)
M. bovis S. aureus
IC50 (µM)
Compound
M. smegmatis
mc2155
4 (12.9)
2 (5.7)
2 (5.5)
1 (3.1)
1 (2.8)
1 (2.7)
1 (2.6)
1 (2.8)
1 (2.5)
1 (2.4)
4 (29.2)
4 (19.6)
ND
S. aureus
M. fortuitum M. phlei
Mtb MurE
BCG EMRSA-15 EMRSA-16
8g
8l
8m
8n
8o
8p
8q
8r
19a
8 (25.9)
2 (5.7)
2 (5.5)
0.5 (1.5)
2 (5.7)
4 (10.9)
2 (5.3)
0.5 (1.5)
1 (2.5)
1 (2.4)
2 (14.6)
8 (39.2)
ND
4 (12.9)
ND
ND
ND
8 (22.8)
4 (10.9)
4 (12.4)
8 (22.8)
ND
ND
263
235
251
774
ND
200
631
ND
ND
ND
ND
ND
ND
4 (11.4) 25 (71.2)
4 (10.9) 25 (68.5)
1 (3.1) 25 (77.4)
2 (5.7)
2 (5.5)
2 (6.2)
2 (5.7) 25 (71.2) 8 (22.8)
2 (5.5)
ND
ND
2 (5.3) 25 (66.0)
2 (5.3)
128 (337.7)
2 (5.7) 25 (71.2) 16 (45.6) 128 (364.7)
1 (2.5)
1 (2.4)
8 (58.3) 0.1 (0.7)
4 (19.6)
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
19b
Isoniazid
Ethambutol
Norfloxacin
Tetracycline
1-Methyl-2-[(5Z)-
tetradecenyl]-
4(1H)-quinolone
ND
ND
ND
0.5 (1.6) 128 (400.9)
0.25 (0.6) 0.25 (0.6)
ND
ND
ND
ND
ND
ND
ND
ND
36 [7]
ND: Not determined.

Molecules 2012, 17
8224
When assayed against M. tuberculosis ATP-dependent MurE using a colorimetric method the tested
compounds displayed moderate to low activity, with IC50 values ranging from 200 µM to 774 µM. Our
biological results further strengthen our previous finding that optimal activity is achieved when the
4(1H)-quinolones possess an aliphatic chain at C-2 which is 12–14 carbons in length. When the length
of the aliphatic side chain at position 2 of the quinolone nucleus was further increased, for instance
compounds 8s and 8t, a marked reduction of the antimycobacterial activity was noted. A significant
decrease in antimycobacterial potency was also encountered upon introduction of a cyclopropyl group
at N-1. However, it should be pointed out that compounds 8c and 23, both having a dec-3-ynyl group
at position 2 of the quinolone nucleus and n-propyl and cyclopropyl groups at N-1, respectively, were
found to be equipotent against M. smegmatis. The most active compounds 8n and 8r are of particular
interest because they are devoid of toxicity to the human diploid embryonic lung cell line (MRC-5) up
to a concentration of 100 µM.
The antimycobacterial activity of compounds having the same substituent at N-1 and an alkynyl
group with the same chain length at position 2, but differing on the position of the triple bond, such as
8b and 8g (N-Et and decynyl), 8c and 8h (N-propyl and decynyl), 8j and 8n (N-Me and dodecynyl)
and 8o and 8r (N-Me and tetradecynyl), was compared in order to assess the effect of the position of
the triple bond on the activity. It was realized that compounds 8g, 8h, 8n and 8r exhibited two to four
fold greater potencies than 8b, 8c, 8j and 8o. In general, a higher antimycobacterial activity was
observed for compounds having the triple bond in position α,β to the quinolone nucleus compared to
compounds having the triple bond in a position distant from the quinolone nucleus. However, this
tendency was inverted for N-propyl or N-cyclopropyl substitution, where compounds having the triple
bond distant from the quinolone 8l and 8m were slightly more active than compounds having the triple
bond in position α,β, such as 8h and 8i. This SAR suggests that antimycobacterial potency depends on
different structural parameters in a non-linear way and a relation valid for a set of compounds might
not be valid for a different set, and thus point out on a complementary approach for obtaining
meaningful SAR.
Compounds 18a–b and 19a–b were prepared in order to explore the effects of a terminal bromine
atom on the antimycobacterial activity. It is worth mentioning that compound 19b showed a two-fold
increase in antimycobacterial potency against M. smegmatis compared to its analogue lacking a
bromine atom reported in our previous work [10,11]. This result is quite interesting and we are aiming
to explore the possibility of the same effect for other active anti-TB scaffolds having long aliphatic
chains. The presence of a bromine at the terminal position may increase the probability of covalent
bonding to a particular protein by nucleophilic attack, an hypothesis which deserves further attention.
On the other hand, compounds 18a–b and 22a–b, bearing a cyclopropyl moiety at N-1 appeared to
be less potent than their analogues 19a–b having a methyl group at N-1, again suggesting that a
cyclopropyl group is detrimental to the activity.
In order to assess the cytotoxicity of these novel quinolone derivatives, the effect on the viability of
a human lung fibroblast cell line MRC-5, was measured using XTT proliferation assay. The
compounds displayed little or no toxicity at 100 µM test concentration. By comparing the cytotoxicity
results of this study with that of our previous reports [10,11], one can notice a remarkable reduction in
cytotoxicity to the MRC-5 cell line, indicating that the triple bond was important in reducing the
toxicity of the quinolones. This finding finds support in other studies which revealed that 5-decynyl,

Molecules 2012, 17
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5-dodecynyl and 5-tetradecynyl pyrimidine nucleoside derivatives have enhanced antimycobacterial
activity while being devoid of cytotoxic activity up to a high concentration [15]. In addition, the
position of the triple bond in the side chain seems to have a significant influence on cytotoxcity which
is increased in compounds with a triple bond more distant to the quinolone nucleus. This can be
concluded by comparing results of 8l, 8n, 8p, and 8r having similar antimycobacterial activity on
M. smegmatis but different cytotoxcity depending on triple bond position.
3. Experimental
3.1. General
The strains M. smegmatis (ATCC 14468), M. smegmatis mc2155, M. fortuitum (ATCC 6841),
M. phlei (ATCC 11758) and M. bovis BCG (ATCC 35734) were obtained from American Type
Culture Collection or German Collection of Microorganisms and Cell Culture (DSMZ). EMRSA-15
and 16 were gifts from Dr Paul Stapleton (UCL School of Pharmacy, UK). M. tuberculosis MurE
ligase was over-expressed in E. coli BL21(DE3)pLysS cells (New England Biolabs, Hitchin, UK) and
purified according to our previous report [7].
All chemicals were purchased from Sigma-Aldrich (Munich, Germany). Reactions were carried out
using oven-dried glassware under an atmosphere of argon. THF was distilled from sodium and stored
in molecular sieve (4 Å). DMF was distilled from CaH2. Melting points were recorded using a
KOFLER hot plate microscope and are uncorrected. IR spectra obtained on a Perkin-Elmer 281 B
1
13
spectrometer, were recorded in KBr unless otherwise noted. H and C-NMR spectra were recorded
on a Varian 400 MHz spectrometer (at 400 and 100 MHz, respectively) using deuterated chloroform as
solvent with TMS as internal standard. Mass spectra were obtained by LC-ESI-MS analysis in positive
mode on a Thermo Finnigan LCQ Deca XP Plus mass spectrometer connected to a Surveyor
LC-system (Thermo-Finnigan, West Palm Beach, FL, USA). Precoated Si gel 60 F254 plates (Merck,
Darmstadt, Germany) were used to monitor the progress of the reactions and column fractions. Spots
were detected by UV/254 nm and spraying with molybdato-phosphoric acid and subsequent heating.
Compounds were purified by column chromatography on silica gel 60 (0.063–0.200 mm) using
cyclohexane/ethyl acetate mixtures as eluent.
3.2. Synthesis
3.2.1. General Procedure for Synthesis of Alkynones 3a–e
Compounds 3a–e were obtained according to the procedure described previously [13] with some
modifications. To a stirred solution of Pd(OAc)2 (0.05 equiv.) in P(CH3)3 (1.0 M in toluene, 0.2 equiv.)
at 60 °C, acetone (~2 mL) and a mixture of 1-alkyne 1 (1 equiv.) and methyl vinyl ketone (2,
3.0 equiv.) were added. The reaction mixture was stirred for 24 h at 60 °C. Then the catalyst was
filtered off on celite, extracted with ether, dried over Na2SO4 and concentrated. The crude product was
purified by silica gel CC eluting with cyclohexane/ethyl acetate = 95:5 to afford 3a–e.
5-Dodecyn-2-one (3a) was prepared from 1-octyne (5.0 g, 45.5 mmol), 2 (9.6 g, 136.4 mmol),
Pd(OAc)2 (0.51 g, 2.3 mmol) and P(CH3)3 (9.1 mL, 9.1 mmol) as a colourless oil (75%) [16].

Molecules 2012, 17
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5-Tridecyn-2-one (3b) was prepared from 1-nonyne (5.0 g, 40.3 mmol), 2 (8.5 g, 121 mmol),
1
Pd(OAc)2 (0.45 g, 2.0 mmol) and P(CH3)3 (8.1 mL, 8.1 mmol) as a colourless oil (82%). H-NMR δ:
2.60 (t, J = 7.6 Hz, 2H, H-3), 2.39 (t, J = 7.6 Hz, 2H, H-4), 2.16 (s, 3H, H-1), 2.10 (t, J = 7.6 Hz, 2H,
H-7), 1.43 (quint, J = 6.8 Hz, 2H, H-8), 1.30–1.25 (m, 8H, H-9-12), 0.86 (t, J = 6.8 Hz, 3H, H-13).
13C-NMR δ: 207.2 (C-2), 80.7 (C-6), 78.4 (C-5), 43.0 (C-3), 31.8, 29.4, 29.3, 29.1, 28.9, 22.6, 18.6,
14.0, 13.5.
5-Tetradecyn-2-one (3c) was prepared from 1-decyne (5.0 g, 36.2 mmol), 2 (7.6 g, 108.7 mmol),
Pd(OAc)2 (0.41 g, 1.8 mmol) and P(CH3)3 (7.2 mL, 7.2 mmol) as a colourless oil (79%) [13].
5-Hexadecyn-2-one (3d) was prepared from 1-dodecyne (5.0 g, 30.1 mmol), 2 (6.3 g, 90.4 mmol),
1
Pd(OAc)2 (0.34 g, 1.5 mmol) and P(CH3)3 (6.0 mL, 6.0 mmol) as a colourless oil (73%). H-NMR δ:
2.61 (t, J = 7.2 Hz, 2H, H-3), 2.40 (t, J = 7.2 Hz, 2H, H-4), 2.15 (s, 3H, H-1), 2.09 (t, J = 7.2 Hz, 2H,
H-7), 1.43 (quint, J = 6.8 Hz, 2H, H-8), 1.30–1.23 (m, 14H, H-9-15), 0.87 (t, J = 6.8 Hz, 3H, H-16).
13C-NMR δ: 207.1 (C-2), 80.9 (C-6), 78.3 (C-5), 42.9 (C-3), 31.9, 29.8, 29.5, 29.5, 29.3, 29.1, 29.0,
28.8, 22.6, 18.6, 14.0, 13.5.
5-Octadecyn-2-one (3e) was prepared from 1-tetradecyne (4.0 g, 20.6 mmol), 2 (4.3 g, 61.9 mmol),
1
Pd(OAc)2 (0.23 g, 1.0 mmol) and P(CH3)3 (4.1 mL, 4.1 mmol) as a colourless oil (73%). H-NMR δ:
2.59 (t, J = 7.6 Hz, 2H, H-3), 2.41 (t, J = 7.6 Hz, 2H, H-4), 2.14 (s, 3H, H-1), 2.08 (t, J = 7.6 Hz, 2H,
H-7), 1.44 (quint, J = 6.8 Hz, 2H, H-8), 1.33–1.21 (m, 18H, H-9-17), 0.85 (t, J = 6.8 Hz, 3H, H-18).
13C-NMR δ: 207.3 (C-2), 81.0 (C-6), 78.5 (C-5), 42.8 (C-3), 31.8, 29.8, 29.8, 29.5, 29.5, 29.4, 29.3,
29.1, 29.0, 28.8, 22.6, 18.6, 14.0, 13.5.
3.2.2. General Procedure for Synthesis of Alkynols 5a–d
To a stirred solution of 1-alkyne (1 equiv.) in THF at −78 °C, n-BuLi (1.6 M in hexane) (1.1 equiv.)
was added dropwise and stirring was continued for 2 h. To this solution acetaldehyde (2.0 equiv.) was
added and further stirred for about 1 h until the temperature dropped to room temperature. The reaction
was quenched by the addition of NH4Cl solution and extracted with CH2Cl2 (3×), washed with brine,
dried over Na2SO4 and concentrated. The crude product was purified by silica gel CC eluting with
cyclohexane/ethyl acetate = 9:1.
3-Dodecyn-2-ol (5a) was prepared from 1-decyne (10.0 g, 72.5 mmol) in THF (150 mL), n-BuLi (49.8 mL,
79.7 mmol), acetaldehyde (6.4 g, 145.1 mmol) as a yellow oil (77%). 1H-NMR δ: 4.46 (qd, J = 6.4, 1.6 Hz,
1H, H-2), 2.14 (td, J = 6.8, 1.6 Hz, 2H, H-5), 1.45 (quint, J = 7.6 Hz, 2H, H-6), 1.37 (d, J = 6.4 Hz,
13
3H, H-1), 1.30–1.21 (m, 10H, H-7-11), 0.85 (t, J = 6.8 Hz, 3H, H-12). C-NMR δ: 84.4 (C-4), 82.2
(C-3), 58.4 (C-2), 31.2, 29.1, 29.0, 28.5, 28.4, 24.6, 22.4, 18.5, 13.9.
3-Tetradecyn-2-ol (5b) was prepared from 1-dodecyne (4.0 g, 24.1 mmol) in THF (75 mL), n-BuLi
(16.6 mL, 26.5 mmol), acetaldehyde (2.1 g, 48.2 mmol) as a yellow oil (69%) [17].
3-Hexadecyn-2-ol (5c) was prepared from 1-tetradecyne (5.0 g, 25.8 mmol) in THF (80 mL), n-BuLi
(17.7 mL, 28.4 mmol), acetaldehyde (2.27 g, 51.6 mmol) as a yellow oil (81%) [18].

Molecules 2012, 17
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3-Heptadecyn-2-ol (5d) was prepared from 1-pentadecyne (4.0 g, 19.2 mmol) in THF (75 mL), n-BuLi
(13.2 mL, 21.2 mmol), acetaldehyde (1.69 g, 38.5 mmol) as a yellow oil (65%). 1H-NMR δ: 4.49 (qd,
J = 6.8, 1.6 Hz, 1H, H-2), 2.17 (td, J = 6.8, 1.6 Hz, 2H, H-5), 1.48 (quint, J = 7.2 Hz, 2H, H-6), 1.40
(d, J = 6.4 Hz, 3H, H-1), 1.33–1.21 (m, 20H, H-7-16), 0.87 (t, J = 6.8 Hz, 3H, H-17). 13C-NMR δ: 84.7
(C-4), 82.2 (C-3), 58.5 (C-2), 31.9, 29.7, 29.6, 29.6, 29.6, 29.5, 29.3, 29.1, 28.8, 28.6, 24.7, 22.6,
18.6, 14.1.
3.2.3. General Procedure for the Synthesis of Alkynones 6a–d
A mixture of 5 (1.0 equiv.) in THF and MnO2 (0.67 equiv.) was stirred at room temperature for 24 h.
The reaction mixture was filtered through celite to give 6.
3-Dodecyn-2-one (6a) was prepared from 5a (10.0 g, 54.9 mmol), in THF (50 mL) and MnO2 (3.2 g,
36.8 mmol) as a colorless oil (79%). 1H-NMR δ: 2.32 (t, J = 7.2 Hz, 2H, H-5), 2.29 (s, 3H, H-1), 1.55
(quint, J = 7.6 Hz, 2H, H-6), 1.30–1.23 (m, 10H, H-7-11), 0.87 (t, J = 6.8 Hz, 3H, H-12). 13C-NMR δ:
184.8 (C-2), 94.1 (C-4), 81.3 (C-3), 32.7, 31.1, 29.3, 29.0, 28.4, 27.6, 22.5, 18.9, 14.0.
3-Tetradecyn-2-one (6b) was prepared from 5b (3.0 g, 14.3 mmol), in THF (15 mL) and MnO2 (0.83 g,
9.6 mmol) as a colourless oil (87%). 1H-NMR δ: 2.33 (t, J = 6.8 Hz, 2H, H-5), 2.30 (s, 3H, H-1), 1.56
(quint, J = 7.2 Hz, 2H, H-6), 1.31–1.23 (m, 14H, H-7-13), 0.87 (t, J = 6.8 Hz, 3H, H-14).
13C-NMR δ: 184.8 (C-2), 94.1 (C-4), 81.4 (C-3), 32.7, 31.8, 29.5, 29.4, 29.2, 29.0, 28.8, 27.6, 22.6,
18.9, 14.0.
3-Hexadecyn-2-one (6c) was prepared from 5c (4.5 g, 18.9 mmol), in THF (20 mL) and MnO2 (1.1 g,
12.7 mmol) as a colourless oil (82%). 1H-NMR δ: 2.32 (t, J = 6.8 Hz, 2H, H-5), 2.29 (s, 3H, H-1), 1.55
13
(quint, J = 7.2 Hz, 2H, H-6), 1.33–1.19 (m, 18H, H-7-15), 0.86 (t, J = 6.8 Hz, 3H, H-16). C-NMR
δ: 184.7 (C-2), 94.0 (C-4), 81.3 (C-3), 32.6, 31.8, 29.6, 29.6, 29.5, 29.4, 29.2, 29.0, 28.8, 27.6, 22.6,
18.9, 14.0.
3-Heptadecyn-2-one (6d) was prepared from 5d (3.0 g, 11.9 mmol), in THF (15 mL) and MnO2 (0.69 g,
7.9 mmol) as a colourless oil (83%). 1H-NMR δ: 2.34 (t, J = 6.8 Hz, 2H, H-5), 2.31 (s, 3H, H-1), 1.57
(quint, J = 7.6 Hz, 2H, H-6), 1.33–1.21 (m, 20H, H-7-16), 0.87 (t, J = 6.8 Hz, 3H, H-17).
13C-NMR δ: 184.9 (C-2), 94.1 (C-4), 81.4 (C-3), 32.7, 31.9, 29.6, 29.6, 29.6, 29.6, 29.4, 29.3, 29.0,
28.8, 27.7, 22.7, 18.9, 14.1.
3.2.4. Preparation of 2-(Cyclopropylamino)benzoic acid (11)
2-Iodobenzoic acid (10.0 g, 40.3 mmol, 1.0 equiv.) dissolved in DMF (50 mL) was poured into an
argon purged two mouth flask containing freshly prepared Cu (512 mg, 8.1 mmol, 0.2 equiv.), pyridine
(4.8 g, 60.5 mmol, 1.5 equiv.) and cyclopropylamine (5.1 g, 88.7 mmol, 2.2 equiv.). The mixture was
stirred for 24 h at room temperature and poured into 500 mL of acidified water (pH 4.5). Filtration and
drying of the slurry gave a white precipitate of 11 (83% yield). 1H-NMR δ:7.98 (d, J = 8.0 Hz, 1H, H-6),
7.72 (bs, 1H, –NH–), 7.44 (t, J = 7.6 Hz, 1H, H-4), 7.17 (d, J = 8.0 Hz, 1H, H-3), 6.68 (t; J = 7.6 Hz,
1H, H-5), 2.49 (sept, J = 3.2 Hz, 1H, N–CH–), 0.83 (m, 2H, N–CH–CH2–), 0.60 (m, 2H, N–CH–CH2–).

Molecules 2012, 17
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13C-NMR δ: 174.0 (–COOH), 152.4 (C-2), 135.4 (C-4), 132.3 (C-6), 115.3 (C-5), 112.9 (C-3), 108.8
(C-1), 24.2 (N–CH–), 7.6 (2 × N–CH–CH2–).
3.2.5. Preparation of N-Cyclopropylisatoic Anhydride (12)
2-(Cyclopropylamine)benzoic acid (4.0 g, 22.6 mmol, 1 equiv.) and N(Et)3 (2.2 g, 21.5 mmol,
0.95 equiv.) were dissolved in CH2Cl2 (50 mL) and cooled to 0 °C with ice. Bis(trichloromethyl)
carbonate (2.0 g, 6.8 mmol, 0.3 equiv.) dissolved in CH2Cl2 (25 mL) was added to the mixture using a
syringe followed by N,N-dimethylaminopyridine (0.4 g, 3.3 mmol, 0.15 equiv.) in CH2Cl2 (15 mL).
After 2 h of stirring at the same temperature the reaction was quenched by adding 25 mL of HCl
(1.0 M), extracted with CH2Cl2, dried over Na2SO4 and concentrated to give a light yellow powder of
12 (94%). 1H-NMR δ: 8.11 (d, J = 8.0 Hz, 1H, H-5), 7.79 (t, J = 7.6 Hz, 1H, H-7), 7.65 (d, J = 8.0 Hz, 1H,
H-8), 7.31 (t, J = 7.6 Hz, 1H, H-6), 2.93 (quint, J = 3.2 Hz, 1H, N–CH–), 1.33 (m, 2H, N–CH–CH2–),
13
0.99 (m, 2H, N–CH–CH2–). C-NMR δ: 158.7 (C-4), 147.7 (C-2), 142.6 (C-8a), 136.8 (C-7), 130.3
(C-5), 124.1 (C-6), 115.4 (C-8), 111.8 (4a), 27.1 (N–CH–), 9.9 (2 × N–CH–CH2–).
3.2.6. Synthesis of ω-Bromoalcohols 14a–b
Treatment of the diols 13a–b with 48% of HBr according to a previously described method [10]
provided 11-bromoundecan-1-ol (14a) [19] and 12-bromododecan-1-ol (14b) [20].
3.2.7. Synthesis of ω-Bromoaldehydes 15a–b
Both 11-bromoundecanal (15a) and 12-bromodecanal were obtained from 14a and 14b,
respectively, by the action of PCC in the presence of NaOAc in CH2Cl2 according to Houghton et al. [21],
spectral data are in accordance with [22].
3.2.8. Synthesis of ω-Bromo-(3E)-ketones 17a–b
14-Bromo-(3E)-tetradecen-2-one (17a) was prepared from 15a and ylide 16 according to Wube et al. [10]
1
as a colorless oil (78% yield). H-NMR δ: 6.81 (dt, J = 16.0, 6.8 Hz, 1H, H-4), 6.06 (d, J = 16.0 Hz,
1H, H-3), 3.41 (t, J = 6.8 Hz, 2H, H-14), 2.25 (s, 3H, H-1), 2.21 (q, J = 7.2 Hz, 2H, H-5), 1.84 (quint,
13
J = 6.8 Hz, 2H, H-13), 1.44 (m, 4H, H-12, 6), 1.32–1.23 (m, 10H, H-7-11). C-NMR δ: 198.6 (C-2),
148.5 (C-4), 131.1 (C-3), 33.9 (C-14), 32.6 (C-13), 32.3 (C-5), 29.2 (C-10), 29.2 (C-9), 29.0 (C-8),
28.6 (C-6), 28.0 (C-7), 27.9 (C-11), 26.7 (C-12), 26.7 (C-1).
15-Bromo-(3E)-pentadecen-2-one (17b) was prepared from 15b and ylide 16 as a colourless oil (84%
yield). 1H-NMR δ: 6.81 (dt, J = 16.0, 6.4 Hz, 1H, H-4), 6.06 (d, J = 16.0 Hz, 1H, H-3), 3.41 (t, J = 6.8 Hz,
2H, H-15), 2.25 (s, 3H, H-1), 2.22 (q, J = 6.8 Hz, 2H, H-5), 1.85 (quint, J = 6.8 Hz, 2H, H-14), 1.43
13
(m, 4H, H-13, 6), 1.32–1.23 (m, 12H, H-7-12). C-NMR δ: 198.9 (C-2), 148.7 (C-4), 131.2 (C-3),
33.9 (C-15), 32.4 (C-14), 32.4 (C-5), 29.4 (C-10), 29.4 (C-9), 29.3 (C-11), 29.3 (C-8), 29.1 (C-7), 28.6
(C-6), 28.0 (C-12), 26.8 (C-13), 26.7 (C-1).

Molecules 2012, 17
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3.2.9. General Procedure for the Synthesis of 8a–t, 18a–b, 19a–b, 22a–b and 23
Compounds 8a–t, 18a–b, 19a–b, 22a–b and 23 were prepared according to procedure described
previously [10] from methyl alkynyl ketones (1.0 equiv.) in THF, LDA (1.8 M in THF/heptane/ethylbenzene)
(1.0 equiv.) and N-alkyl isatoic anhydride (0.75 equiv.) in THF at −78 °C. The purity of the quinolones
was determined by LC-MS (88–95%).
1-Methyl-2-(3′-decynyl)-4(1H)-quinolone (8a) was prepared from 3a (1.0 g, 5.6 mmol) in THF (15 mL),
LDA (3.1 mL, 5.6 mmol) and N-methylisatoic anhydride (7a) (0.74 g, 4.2 mmol) in THF (10 mL) as a
yellow oil (63%). IR (KBr, cm−1): 3406, 2929, 2857, 1627, 1600, 1500, 1469, 1177, 759. 1H-NMR δ:
8.44 (d, J = 8.0 Hz, 1H, H-5), 7.68 (t, J = 7.6 Hz, 1H, H-7), 7.52 (d, J = 8.0 Hz, 1H, H-8), 7.39 (t,
J = 7.2 Hz, 1H, H-6), 6.32 (s, 1H, H-3), 3.78 (s, 3H, N–CH3), 2.95 (t, J = 7.2 Hz, 2H, H-1'), 2.57 (t,
J = 6.8 Hz, 2H, H-2'), 2.12 (t, J = 6.8 Hz, 2H, H-5'), 1.44 (quint, J = 7.2 Hz, 2H, H-6'), 1.32–1.20 (m,
6H, H-7'-9'), 0.86 (t, J = 6.8 Hz, 3H, H-10'). 13C-NMR δ: 177.5 (C-4), 152.6 (C-2), 140.6 (C-8a), 132.2
(C-7), 126.8 (C-5), 126.6 (C-4a), 123.5 (C-6), 115.5 (C-8), 110.7 (C-3), 82.8 (C-4'), 77.2 (C-3'), 35.3
(N–CH3), 32.9 (C-1'), 31.6 (C-8'), 28.6 (C-7'), 28.4 (C-6'), 22.6 (C-9'), 18.7 (C-2'), 18.5 (C-5'), 14.0
(C-10'). ESI-MS m/z (rel. int.): [M+H]+ 296 (100).
1-Ethyl-2-(3'-decynyl)-4(1H)-quinolone (8b) was prepared from 3a (1.0 g, 5.6 mmol) in THF (15 mL),
LDA (3.1 mL, 5.6 mmol) and N-ethylisatoic anhydride (7b) (0.8 g, 4.2 mmol) in THF (10 mL) as a
yellow oil (55%). IR (KBr, cm−1): 3428, 2929, 2854, 1624, 1600, 1490, 1468, 1430, 1308, 759.
1H-NMR δ: 8.46 (d, J = 8.0 Hz, 1H, H-5), 7.63 (t, J = 7.6 Hz, 1H, H-7), 7.55 (d, J = 8.0 Hz, 1H, H-8),
7.37 (t, J = 7.6 Hz, 1H, H-6), 6.34 (s, 1H, H-3), 4.33 (q, J = 7.2 Hz, 2H, N–CH2–CH3), 2.96 (t, J = 7.2 Hz,
2H, H-1'), 2.61 (t, J = 6.8 Hz, 2H, H-2'), 2.13 (t, J = 6.8 Hz, 2H, H-5'), 1.45 (t, J = 7.2 Hz, 3H,
N–CH2–CH3), 1.42 (quint, J = 6.8 Hz, 2H, H-6'), 1.30–1.21 (m, 6H, H-7'-9'), 0.86 (t, J = 6.8 Hz, 3H,
13
H-10'). C-NMR δ: 177.4 (C-4), 152.6 (C-2), 140.5 (C-8a), 132.2 (C-7), 126.9 (C-5), 126.7 (C-4a),
123.4 (C-6), 115.4 (C-8), 110.9 (C-3), 82.7 (C-4'), 77.1 (C-3'), 41.2 (N–CH2–CH3), 33.0 (C-1'), 31.5
(C-8'), 28.7 (C-7'), 28.5 (C-6'), 22.5 (C-9'), 18.8 (C-2'), 18.7 (C-5'), 14.2 (N–CH2–CH3), 14.0 (C-10').
ESI-MS m/z (rel. int.): [M+H]+ 310 (100).
1-(n-Propyl)-2-(3′-decynyl)-4(1H)-quinolone (8c) was prepared from 3a (1.0 g, 5.6 mmol) in THF
(15 mL), LDA (3.1 mL, 5.6 mmol) and N-(n-propyl)isatoic anhydride (7c) (0.85 g, 4.2 mmol) in THF
(10 mL) as a yellow oil (56%). IR (KBr, cm−1): 3420, 2929, 2856, 1627, 1600, 1489, 1468, 1428,
1177, 759. 1H-NMR δ: 8.45 (d, J = 8.0 Hz, 1H, H-5), 7.64 (t, J = 8.0 Hz, 1H, H-7), 7.46 (d, J = 8.4 Hz,
1H, H-8), 7.35 (t, J = 7.6 Hz, 1H, H-6), 6.29 (s, 1H, H-3), 4.13 (t, J = 8.0 Hz, 2H, N–CH2–CH2–CH3),
2.91 (t, J = 7.6 Hz, 2H, H-1'), 2.58 (t, J = 7.6 Hz, 2H, H-2'), 2.12 (t, J = 7.6 Hz, 2H, H-5'), 1.84 (m, 2H,
N–CH2–CH2–CH3), 1.44 (quint, J = 7.6 Hz, 2H, H-6'), 1.35–1.23 (m, 6H, H-7'-9'), 1.08 (t, J = 7.6 Hz,
13
3H, N–CH2–CH2–CH3), 0.85 (t, J = 7.2 Hz, 3H, H-10'). C-NMR δ: 175.4 (C-4), 152.7 (C-2), 140.7
(C-8a), 132.1 (C-7), 126.8 (C-5), 126.6 (C-4a), 123.3 (C-6), 115.5 (C-8), 110.9 (C-3), 82.7 (C-4'), 77.1
(C-3'), 47.8 (N–CH2–CH2–CH3), 33.2 (C-1'), 31.4 (C-8'), 28.7 (C-7'), 28.5 (C-6'), 22.5 (C-9'), 22.1
(N–CH2–CH2–CH3), 18.8 (C-2'), 18.6 (C-5'), 14.0 (C-10'), 11.0 (N–CH2–CH2–CH3). ESI-MS m/z
(rel. int.): [M+H]+ 324 (100).

Molecules 2012, 17
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1-Methyl-2-(3'-undecynyl)-4(1H)-quinolone (8d) was prepared from 3b (1.0 g, 5.2 mmol) in THF (15 mL),
LDA (2.9 mL, 5.2 mmol) and N-methylisatoic anhydride (7a) (0.69 g, 3.9 mmol) in THF (10 mL) as a
yellow semi solid (58%). IR (KBr, cm−1): 3420, 2928, 2855, 1628, 1600, 1500, 1469, 1177, 759.
1H-NMR δ: 8.43 (d, J = 8.0 Hz, 1H, H-5), 7.65 (t, J = 8.0 Hz, 1H, H-7), 7.49 (d, J = 8.4 Hz, 1H, H-8),
7.36 (t, J = 7.6 Hz, 1H, H-6), 6.27 (s, 1H, H-3), 3.76 (s, 3H, N–CH3), 2.92 (t, J = 7.2 Hz, 2H, H-1'),
2.55 (t, J = 6.8 Hz, 2H, H-2'), 2.11 (t, J = 6.8 Hz, 2H, H-5'), 1.44 (quint, J = 7.2 Hz, 2H, H-6'),
13
1.33–1.21 (m, 8H, H-7'-10'), 0.86 (t, J = 6.8 Hz, 3H, H-11'). C-NMR δ: 177.4 (C-4), 153.1 (C-2),
141.8 (C-8a), 132.2 (C-7), 126.6 (C-5), 126.4 (C-4a), 123.5 (C-6), 115.4 (C-8), 111.1 (C-3), 82.8
(C-4'), 76.9 (C-3'), 34.4 (N–CH3), 34.0 (C-1'), 31.7 (C-9'), 29.0 (C-8'), 28.8 (C-7'), 28.7 (C-6'), 22.6
(C-10'), 18.6 (C-2'), 18.5 (C-5'), 14.1 (C-11'). ESI-MS m/z (rel. int.): [M+H]+ 310 (100).
1-Ethyl-2-(3'-undecynyl)-4(1H)-quinolone (8e) was prepared from 3b (1.0 g, 5.2 mmol) in THF (15 mL),
LDA (2.9 mL, 5.2 mmol) and N-ethylisatoic anhydride (7b) (0.74 g, 3.9 mmol) in THF (10 mL) as a
yellow semi solid (62%). IR (KBr, cm−1): 3434, 2930, 2851, 1621, 1600, 1490, 1468, 1431, 1309, 759.
1H-NMR δ: 8.45 (d, J = 8.0 Hz, 1H, H-5), 7.65 (t, J = 8.0 Hz, 1H, H-7), 7.51 (d, J = 8.4 Hz, 1H, H-8),
7.35 (t, J = 7.2 Hz, 1H, H-6), 6.28 (s, 1H, H-3), 4.27 (q, J = 7.6 Hz, 2H, N–CH2–CH3), 2.91 (t, J = 7.2 Hz,
2H, H-1'), 2.59 (t, J = 6.8 Hz, 2H, H-2'), 2.12 (t, J = 6.8 Hz, 2H, H-5'), 1.44 (t, J = 7.2 Hz, 2H,
N–CH2–CH3), 1.42 (quint, J = 6.8 Hz, 2H, H-6'), 1.31–1.21 (m, 8H, H-7'-10'), 0.85 (t, J = 6.8 Hz, 3H,
13
H-11'). C-NMR δ: 177.1 (C-4), 151.9 (C-2), 141.1 (C-8a), 132.1 (C-7), 126.9 (C-5), 126.7 (C-4a),
123.2 (C-6), 115.4 (C-8), 110.8 (C-3), 82.4 (C-4'), 77.4 (C-3'), 41.3 (N–CH2–CH3), 33.0 (C-1'), 31.6
(C-9'), 29.0 (C-8'), 28.8 (C-7'), 28.6 (C-6'), 22.6 (C-10'), 18.8 (C-2'), 18.7 (C-5'), 14.2 (N–CH2–CH3),
14.0 (C-14'). ESI-MS m/z (rel. int.): [M+H]+ 324 (100).
1-(n-Propyl)-2-(3'-undecynyl)-4(1H)-quinolone (8f) was prepared from 3b (1.0 g, 5.2 mmol) in THF
(15 mL), LDA (2.9 mL, 5.2 mmol) and N-(n-propyl)isatoic anhydride (7c) (0.79 g, 3.9 mmol) in THF
(10 mL) as a yellow oil (59%). IR (KBr, cm−1): 3426, 2929, 2856, 1627, 1600, 1488, 1468, 1427,
1177, 759. 1H-NMR δ: 8.46 (d, J = 8.0 Hz, 1H, H-5), 7.66 (t, J = 7.6 Hz, 1H, H-7), 7.48 (d, J = 8.0 Hz,
1H, H-8), 7.37 (t, J = 7.6 Hz, 1H, H-6), 6.36 (s, 1H, H-3), 4.13 (t, J = 8.0 Hz, 2H, N–CH2–CH2–CH3),
2.93 (t, J = 7.6 Hz, 2H, H-1'), 2.61 (t, J = 7.2 Hz, 2H, H-2'), 2.13 (t, J = 7.2 Hz, 2H, H-5'), 1.85 (sext,
J = 8.0 Hz, 2H, N–CH2–CH2–CH3), 1.45 (quint, J = 7.2 Hz, 2H, H-6'), 1.33–1.21 (m, 8H, H-7'-10'),
1.08 (t, J = 7.2 Hz, 2H, N–CH2–CH2–CH3), 0.86 (t, J = 6.8 Hz, 3H, H-11'). 13C-NMR δ: 177.3 (C-4),
152.7 (C-2), 140.5 (C-8a), 132.3 (C-7), 126.7 (C-5), 126.5 (C-4a), 123.2 (C-6), 115.6 (C-8), 111.0 (C-3),
82.5 (C-4'), 77.0 (C-3'), 47.7 (N–CH2–CH2–CH3), 32.9 (C-1'), 31.5 (C-9'), 29.0 (C-8'), 28.7 (C-7'),
28.4 (C-6'), 22.6 (C-10'), 22.3 (N–CH2–CH2–CH3), 18.7 (C-2'), 18.6 (C-5'), 14.0 (C-11'), 11.1
(N–CH2–CH2–CH3). ESI-MS m/z (rel. int.): [M+H]+ 338 (100).
1-Ethyl-2-(1'-decynyl)-4(1H)-quinolone (8g) was prepared from 6a (1.0 g, 5.6 mmol) in THF (15 mL),
LDA (3.1 mL, 5.6 mmol) and N-ethylisatoic anhydride (7b) (0.80 g, 4.2 mmol) in THF (10 mL) as a
light yellow semi solid (54%). IR (KBr, cm−1): 3372, 2928, 2855, 2234, 1625, 1598, 1488, 1421, 758.
1H-NMR δ: 8.45 (d, J = 8.0 Hz, , 1H, H-5), 7.69 (t, J = 7.6 Hz, 1H, H-7), 7.50 (d, J = 8.0, Hz, 1H,
H-8), 7.38 (t, J = 7.6 Hz, 1H, H-6), 6.58 (s, 1H, H-3), 4.53 (q, J = 7.2 Hz, 2H, N–CH2–CH3), 2.53 (t,
J = 7.2 Hz, 2H, H-3'), 1.67 (quint, J = 7.2 Hz, 2H, H-4'), 1.49 (t, J = 7.2 Hz, 2H, N–CH2–CH3), 1.33–1.22

Molecules 2012, 17
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(m, 10H, H-5'-9'), 0.89 (t, J = 7.2 Hz, 3H, H-10'). 13C-NMR δ: 177.1 (C-4), 141.3 (C-8a), 137.5 (C-2),
132.5 (C-7), 126.8 (C-4a), 126.6 (C-5), 123.5 (C-6), 115.5 (C-8), 115.1 (C-3), 111.3 (C-2'), 74.5 (C-1'),
42.4 (N–CH2–CH3), 31.8 (C-8'), 29.2 (C-7'), 29.1 (C-6'), 29.0 (C-5'), 28.1 (C-4'), 22.6 (C-9'), 19.3 (C-3'),
14.2 (N–CH2–CH3), 14.0 (C-10'). ESI-MS m/z (rel. int.): [M+H]+ 310 (100).
1-(n-Propyl)-2-(1'-decynyl)-4(1H)-quinolone (8h) was prepared from 6a (1.0 g, 5.6 mmol) in THF
(15 mL), LDA (3.1 mL, 5.6 mmol) and N-(n-propyl)isatoic anhydride (7c) (0.85 g, 4.2 mmol) in THF
(10 mL) as a light yellow semi solid (51%). IR (KBr, cm−1): 3430, 2927, 2855, 2235, 1624, 1598,
1
1488, 1420, 1177, 758. H-NMR δ: 8.45 (d, J = 8.0 Hz, 1H, H-5), 7.66 (t, J = 7.2 Hz, 1H, H-7), 7.44
(d, J = 8.0 Hz, 1H, H-8), 7.36 (t, J = 7.2 Hz, 1H, H-6), 6.56 (s, 1H, H-3), 4.40 (t, J = 7.6 Hz, 2H,
N–CH2–CH2–CH3), 2.51 (t, J = 6.8 Hz, 2H, H-3'), 1.86 (m, 2H, N–CH2–CH2–CH3), 1.65 (quint,
J = 6.8 Hz, 2H, H-4'), 1.33–1.21 (m, 10H, H-5'-9'), 1.03 (t, J = 7.2 Hz, 3H, N–CH2–CH2–CH3), 0.89 (t,
J = 6.8 Hz, 3H, H-10'). 13C-NMR δ: 177.1 (C-4), 141.0 (C-8a), 138.2 (C-2),132.5 (C-7), 126.7 (C-4a),
126.5 (C-5), 123.7 (C-6), 115.4 (C-8), 115.2 (C-3), 111.0 (C-2'), 75.1 (C-1'), 47.9 (N–CH2–CH2–CH3),
31.7 (C-8'), 29.1 (C-7'), 29.0 (C-6'), 29.0 (C-5'), 28.1 (C-4'), 22.6 (C-9'), 22.3 (N–CH2–CH2–CH3),
19.8 (C-3'), 14.0 (C-10'), 11.0 (N–CH2–CH2–CH3). ESI-MS m/z (rel. int.): [M+H]+ 324 (100).
1-(n-Butyl)-2-(1'-decynyl)-4(1H)-quinolone (8i) was prepared from 6a (1.0 g, 5.6 mmol) in THF (15 mL),
LDA (3.1 mL, 5.6 mmol) and N-(n-butyl)isatoic anhydride (7d) (0.91 g, 4.2 mmol) in THF (10 mL) as
a light yellow oil (54%). IR (KBr, cm−1): 3428, 2928, 2856, 2233, 1622, 1597, 1490, 1466, 1422, 759.
1H-NMR δ: 8.43 (d, J = 8.0 Hz, 1H, H-5), 7.66 (t, J = 7.6 Hz, 1H, H-7), 7.45 (d, J = 8.0, Hz, 1H, H-8),
7.33 (t, J = 7.6 Hz, 1H, H-6), 6.58 (s, 1H, H-3), 4.38 (t, J = 7.6 Hz, 2H, N–CH2–CH2–CH2–CH3), 2.53
(t, J = 6.8 Hz, 2H, H-3'), 1.78 (quint, J = 7.2 Hz, 2H, N–CH2–CH2–CH2–CH3), 1.65 (quint, J = 6.8 Hz,
2H, H-4'), 1.48 (m, 2H, N–CH2–CH2–CH2–CH3), 1.31–1.24 (m, 10H, H-5'-9'), 1.04 (t, J = 7.6 Hz, 3H,
N–CH2–CH2–CH2–CH3), 0.87 (t, J = 6.8 Hz, 3H, H-10’). 13C-NMR δ: 177.1 (C-4), 141.3 (C-8a),
137.3 (C-2),132.5 (C-7), 126.7 (C-4a), 126.5 (C-5), 123.7 (C-6), 115.7 (C-8), 115.0 (C-3), 111.0
(C-2'), 74.4 (C-1'), 48.4 (N–CH2–CH2–CH2–CH3), 31.8 (C-8'), 31.4 (N–CH2–CH2–CH2–CH3), 29.1
(C-7'), 29.0 (C-6'), 29.0 (C-5'), 28.1 (C-4'), 22.6 (C-9'), 20.9 (N–CH2–CH2–CH2–CH3), 19.3 (C-3'),
14.3 (N–CH2–CH2–CH2–CH3), 14.0 (C-10'). ESI-MS m/z (rel. int.): [M+H]+ 338 (100).
1-Methyl-2-(3'-dodecynyl)-4(1H)-quinolone (8j) was prepared from 3c (1.0 g, 4.8 mmol) in THF (15 mL),
LDA (2.7 mL, 4.8 mmol) and N-methylisatoic anhydride (7a) (0.64 g, 3.6 mmol) in THF (10 mL) as a
light yellow semi solid (60%). IR (KBr, cm−1): 3420, 2927, 2855, 1628, 1600, 1499, 1469, 759.
1H-NMR δ: 8.38 (dd, J = 8.0, 1.6 Hz, 1H, H-5), 7.61 (t, J = 8.4 Hz, 1H, H-7), 7.45 (d, J = 8.4 Hz, 1H,
H-8), 7.32 (t, J = 7.6 Hz, 1H, H-6), 6.19 (s, 1H, H-3), 3.71 (s, 3H, N–CH3), 2.87 (t, J = 7.6 Hz, 2H,
H-1'), 2.52 (t, J = 7.6 Hz, 2H, H-2'), 2.07 (t, J = 7.6 Hz, 2H, H-5'), 1.43 (quint, J = 7.2 Hz, 2H, H-6'),
13
1.31–1.20 (m, 10H, H-7'-11'), 0.84 (t, J = 6.8 Hz, 3H, H-12'). C-NMR δ: 177.6 (C-4), 152.9 (C-2),
141.8 (C-8a), 132.1 (C-7), 126.5 (C-5), 126.3 (C-4a), 123.3 (C-6), 115.4 (C-8), 111.1 (C-3), 82.6
(C-4'), 77.0 (C-3'), 34.3 (N–CH3), 33.9 (C-1'), 31.8 (C-10'), 29.1 (C-9'), 29.1 (C-8'), 29.0 (C-7'), 28.8
(C-6'), 22.6 (C-11'), 18.6 (C-2'), 18.4 (C-5'), 14.0 (C-12'). ESI-MS m/z (rel. int.): [M+H]+ 324 (100).
1-Ethyl-2-(3'-dodecynyl)-4(1H)-quinolone (8k) was prepared from 3c (1.0 g, 4.8 mmol) in THF (15 mL),
LDA (2.7 mL, 4.8 mmol) and N-ethylisatoic anhydride (7b) (0.69 g, 3.6 mmol) in THF (10 mL) as a

Molecules 2012, 17
8232
white solid (55%). m.p. 66–68 °C. IR (KBr, cm−1): 3440, 2919, 2850, 1621, 1600, 1490, 1468, 1308,
769. 1H-NMR δ: 8.43 (d, J = 8.0 Hz, 1H, H-5), 7.64 (t, J = 8.0 Hz, 1H, H-7), 7.50 (d, J = 8.4 Hz, 1H,
H-8), 7.34 (t, J = 7.2 Hz, 1H, H-6), 6.28 (s, 1H, H-3), 4.26 (q, J = 7.2 Hz, 2H, N–CH2–CH3), 2.90 (t,
J = 7.6 Hz, 2H, H-1'), 2.57 (t, J = 7.2 Hz, 2H, H-2'), 2.10 (t, J = 6.8 Hz, 2H, H-5'), 1.43 (t, J = 7.6 Hz,
N–CH2–CH3), 1.41 (quint, J = 7.2 Hz, 2H, H-6'), 1.31–1.20 (m, 10H, H-7'-11'), 0.85 (t, J = 6.8 Hz, 3H,
13
H-12'). C-NMR δ: 177.5 (C-4), 152.8 (C-2), 140.5 (C-8a), 132.1 (C-7), 126.9 (C-5), 126.7 (C-4a),
123.2 (C-6), 115.4 (C-8), 110.9 (C-3), 82.7 (C-4'), 77.0 (C-3'), 41.2 (N–CH2–CH3), 33.0 (C-1'), 31.7
(C-10'), 29.1 (C-9'), 29.0 (C-8'), 28.8 (C-7'), 28.8 (C-6'), 22.5 (C-11'), 18.8 (C-2'), 18.6 (C-5'), 14.1
(N–CH2–CH3), 14.0 (C-12'). ESI-MS m/z (rel. int.): [M+H]+ 338 (100).
1-(n-Propyl)-2-(3'-dodecynyl)-4(1H)-quinolone (8l) was prepared from 3c (1.0 g, 4.8 mmol) in THF
(15 mL), LDA (2.7 mL, 4.8 mmol) and N-(n-propyl)isatoic anhydride (7c) (0.74 g, 3.6 mmol) in THF
(10 mL) as a light yellow semi solid (51%). IR (KBr, cm−1): 3426, 2928, 2855, 1628, 1600, 1488,
1
1467, 1427, 759. H-NMR δ: 8.44 (d, J = 8.0 Hz, 1H, H-5), 7.66 (t, J = 8.0 Hz, 1H, H-7), 7.47 (d,
J = 8.0 Hz, 1H, H-8), 7.36 (t, J = 7.6 Hz, 1H, H-6), 6.28 (s, 1H, H-3), 4.16 (t, J = 8.0 Hz, 2H,
N–CH2–CH2–CH3), 2.90 (t, J = 7.6 Hz, 2H, H-1'), 2.59 (t, J = 7.2 Hz, 2H, H-2'), 2.13 (t, J = 7.6 Hz,
2H, H-5'), 1.85 (sext, J = 7.2 Hz, 2H, N–CH2–CH2–CH3), 1.43 (quint, J = 7.6 Hz, 2H, H-6'), 1.33–1.21
(m, 10H, H-7'-11'), 1.07 (t, J = 7.2 Hz, 3H, N–CH2–CH2–CH3), 0.86 (t, J = 6.8 Hz, 3H, H-12').
13C-NMR δ: 175.8 (C-4), 152.5 (C-2), 140.9 (C-8a), 132.2 (C-7), 126.7 (C-5), 126.5 (C-4a), 123.1
(C-6), 115.4 (C-8), 110.8 (C-3), 82.6 (C-4'), 77.1 (C-3'), 47.8 (N–CH2–CH2–CH3), 33.1 (C-1'), 31.6
(C-10'), 29.1 (C-9'), 29.0 (C-8'), 29.0 (C-7'), 28.6 (C-6'), 22.6 (C-11'), 22.0 (N–CH2–CH2–CH3), 18.8
(C-2'), 18.6 (C-5'), 14.0 (C-14’), 10.9 (N–CH2–CH2–CH3). ESI-MS m/z (rel. int.): [M+H]+ 352 (100).
1-(n-Butyl)-2-(3'-dodecynyl)-4(1H)-quinolone (8m) was prepared from 3c (1.0 g, 4.8 mmol) in THF
(15 mL), LDA (2.7 mL, 4.8 mmol) and N-(n-butyl)isatoic anhydride (7d) (0.79 g, 3.6 mmol) in THF
(10 mL) as a yellow oil (58%). IR (KBr, cm−1): 3425, 2927, 2855, 1628, 1600, 1488, 1467, 1427, 759.
1H-NMR δ: 8.44 (dd, J = 8.0, 1.6 Hz, 1H, H-5), 7.65 (td, J = 8.0, 1.6 Hz, 1H, H-7), 7.48 (d, J = 8.4 Hz,
1H, H-8), 7.35 (t, J = 7.6 Hz, 1H, H-6), 6.27 (s, 1H, H-3), 4.13 (t, J =8.0 Hz, 2H, N–CH2–(CH2)2–CH3),
2.91 (t, J = 7.2 Hz, 2H, H-1'), 2.58 (t, J = 7.2 Hz, 2H, H-2'), 2.11 (m, 2H, H-5'), 1.80 (quint,
J = 7.2 Hz, 2H, N–CH2–CH2–CH2–CH3), 1.46 (m, 2H, N-(CH2)2–CH2–CH3), 1.41 (quint, J = 7.2 Hz,
2H, H-6'), 1.32–1.23 (m, 10H, H-7'-11'), 1.03 (t, J = 7.2 Hz, 3H, N–(CH2)3–CH3), 0.85 (t, J = 6.8 Hz,
3H, H-12'). 13C-NMR δ: 177.4 (C-4), 152.8 (C-2), 140.7 (C-8a), 132.1 (C-7), 126.8 (C-5), 126.6
(C-4a), 123.3 (C-6), 115.6 (C-8), 110.8 (C-3), 82.7 (C-4'), 77.1 (C-3'), 47.6 (N–CH2–(CH2)2–CH3),
33.2 (C-1'), 31.8 (C-10'), 30.8 (N–CH2–CH2–CH2–CH3), 29.1 (C-9'), 29.0 (C-8'), 28.8 (C-7'), 28.8
(C-6'), 22.6 (C-11'), 20.1 (N–(CH2)2–CH2–CH3), 18.8 (C-2'), 18.6 (C-5'), 14.1 (C-14’), 13.8
(N–(CH2)3–CH3). ESI-MS m/z (rel. int.): [M+H]+ 366 (100).
1-Methyl-2-(1'-dodecynyl)-4(1H)-quinolone (8n) was prepared from 6b (1.5 g, 7.2 mmol) in THF (25 mL),
LDA (4.0 mL, 7.2 mmol) and N-methylisatoic anhydride (7a) (0.96 g, 5.4 mmol) in THF (15 mL) as a
light yellow semi-solid (51%). IR (KBr, cm−1): 3421, 2925, 2854, 2234, 1625, 1599, 1496, 1469, 757.
1H-NMR δ: 8.41 (d, J = 8.0 Hz, 1H, H-5), 7.66 (t, J = 7.6 Hz, 1H, H-7), 7.44 (d, J = 8.0, Hz, 1H, H-8),
7.36 (t, J = 7.6 Hz, 1H, H-6), 6.55 (s, 1H, H-3), 3.94 (s, 3H, N–CH3), 3.17 (t, J = 7.2 Hz, 2H, H-1'),

Molecules 2012, 17
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2.51 (t, J = 6.8 Hz, 2H, H-3'), 1.64 (quint, J = 6.8 Hz, 2H, H-4'), 1.46 (m, 2H, H-5'), 1.34–1.23
(m, 12H, H-6'-11'), 0.87 (t, J = 6.8 Hz, 3H, H-12'). 13C-NMR δ: 177.0 (C-4), 141.1 (C-8a), 137.4
(C-2),132.4 (C-7), 126.8 (C-4a), 126.7 (C-5), 123.6 (C-6), 115.6 (C-8), 115.0 (C-3), 102.2 (C-2'), 74.6
(C-1'), 36.7 (N–CH3), 31.8 (C-10'), 29.5 (C-9'), 29.4 (C-8'), 29.2 (C-7'), 29.0 (C6'), 29.0 (C5'), 28.0
(C-4'), 22.6 (C-11'), 19.6 (C-3'), 14.0 (C-12'). ESI-MS m/z (rel. int.): [M+H]+ 324 (100).
1-Methyl-2-(3'-tetradecynyl)-4(1H)-quinolone (8o) was prepared from 3d (1.0 g, 4.2 mmol) in THF
(15 mL), LDA (2.4 mL, 4.2 mmol) and N-methylisatoic anhydride (7a) (0.57 g, 3.2 mmol) in THF (10 mL)
as a light yellow solid (60%). m.p. 43–45 °C. IR (KBr, cm−1): 3422, 2922, 2854, 1632, 1598, 1469,
1444, 760. 1H-NMR δ: 8.39 (d, J = 8.0 Hz, 1H, H-5), 7.62 (t, J = 8.0 Hz, 1H, H-7), 7.46 (d, J = 8.4 Hz,
1H, H-8), 7.33 (t, J = 7.2 Hz, 1H, H-6), 6.21 (s, 1H, H-3), 3.72 (s, 3H, N–CH3), 2.88 (t, J = 7.2 Hz, 2H,
H-1'), 2.50 (t, J = 7.6 Hz, 2H, H-2’), 2.10 (t, J = 7.2 Hz, 2H, H-5'), 1.41 (quint, J = 6.8 Hz, 2H, H-6'),
13
1.34–1.21 (m, 14H, H-7'-13'), 0.85 (t, J = 6.8 Hz, 3H, H-14'). C-NMR δ: 177.6 (C-4), 152.9 (C-2),
141.8 (C-8a), 132.1 (C-7), 126.5 (C-5), 126.3 (C-4a), 123.3 (C-6), 115.4 (C-8), 111.1 (C-3), 82.7
(C-4'), 77.1 (C-3'), 34.3 (N–CH3), 33.9 (C-1'), 31.8 (C-12'), 29.2 (C-11'), 29.1 (C-10'), 29.1 (C-9'), 29.0
(C-8'), 29.0 (C-7'), 28.8 (C-6'), 22.6 (C-13'), 18.6 (C-2'), 18.4 (C-5'), 14.0 (C-14'). ESI-MS m/z
(rel. int.): [M+H]+ 352 (100).
1-Ethyl-2-(3'-tetradecynyl)-4(1H)-quinolone (8p) was prepared from 3d (1.0 g, 4.2 mmol) in THF (15 mL),
LDA (2.4 mL, 4.2 mmol) and N-ethylisatoic anhydride (7b) (0.61 g, 3.2 mmol) in THF (10 mL) as
white needles (56%). m.p. 74–76 °C. IR (KBr, cm−1): 3424, 2917, 2850, 1621, 1600, 1468, 1430,
1308, 760. 1H-NMR δ: 8.44 (d, J = 8.0 Hz, 1H, H-5), 7.63 (t, J = 8.0 Hz, 1H, H-7), 7.50 (d, J = 8.4 Hz,
1H, H-8), 7.35 (t, J = 7.2 Hz, 1H, H-6), 6.29 (s, 1H, H-3), 4.24 (q, J = 7.2 Hz, 2H, N–CH2–CH3), 2.91
(t, J = 7.2 Hz, 2H, H-1'), 2.58 (t, J = 7.2 Hz, 2H, H-2'), 2.09 (t, J = 6.4 Hz, 2H, H-5'), 1.42 (t, J = 7.2 Hz,
N–CH2–CH3), 1.40 (quint, J = 7.2 Hz, 2H, H-6'), 1.31–1.21 (m, 14H, H-7'-13'), 0.85 (t, J = 6.8 Hz, 3H,
13
H-14'). C-NMR δ: 177.5 (C-4), 152.7 (C-2), 140.8 (C-8a), 132.1 (C-7), 126.6 (C-5), 126.4 (C-4a),
123.3 (C-6), 115.4 (C-8), 111.1 (C-3), 82.7 (C-4'), 77.0 (C-3'), 41.3 (N–CH2–CH3), 33.1 (C-1'), 31.8
(C-12'), 29.2 (C-11'), 29.2 (C-10'), 29.1 (C-9'), 29.0 (C-8'), 29.0 (C-7'), 28.8 (C-6'), 22.6 (C-13'), 18.8
(C-2'), 18.7 (C-5'), 14.2 (N–CH2–CH3), 14.0 (C-14'). ESI-MS m/z (rel. int.): [M+H]+ 366 (100).
1-(n-Propyl)-2-(3'-tetradecynyl)-4(1H)-quinolone (8q) was prepared from 3d (1.0 g, 4.2 mmol) in
THF (15 mL), LDA (2.4 mL, 4.2 mmol) and N-(n-propyl)isatoic anhydride (7c) (0.66 g, 3.2 mmol) in
THF (10 mL) as a light yellow semi-solid (55%). IR (KBr, cm−1): 3430, 2926, 2854, 1629, 1600, 1488,
1
1467, 1227, 759. H-NMR δ: 8.48 (d, J = 8.0 Hz, 1H, H-5), 7.67 (t, J = 7.6 Hz, 1H, H-7), 7.49 (d,
J = 8.4 Hz, 1H, H-8), 7.38 (t, J = 7.6 Hz, 1H, H-6), 6.43 (s, 1H, H-3), 4.17 (t, J = 8.0 Hz, 2H,
N–CH2–CH2–CH3), 2.94 (t, J = 7.6 Hz, 2H, H-1'), 2.59 (t, J = 7.2 Hz, 2H, H-2'), 2.11 (t, J = 7.2 Hz,
2H, H-5'), 1.86 (sext, J = 7.6 Hz, 2H, N–CH2–CH2–CH3), 1.43 (quint, J = 7.2 Hz, 2H, H-6'), 1.33–1.22
(m, 14H, H-7'-13'), 1.09 (t, J = 7.2 Hz, 2H, N–CH2–CH2–CH3), 0.88 (t, J = 6.8 Hz, 3H, H-14').
13C-NMR δ: 175.4 (C-4), 152.5 (C-2), 141.0 (C-8a), 132.1 (C-7), 126.7 (C-5), 126.5 (C-4a), 123.3
(C-6), 115.3 (C-8), 111.0 (C-3), 82.6 (C-4'), 77.0 (C-3’), 47.6 (N–CH2–CH2–CH3), 33.1 (C-1'), 31.5
(C-12'), 29.3 (C-11'), 29.2 (C-10'), 29.2 (C-9'), 29.1 (C-8'), 29.0 (C-7'), 28.4 (C-6'), 22.6 (C-13'), 22.1

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(N–CH2–CH2–CH3), 18.8 (C-2'), 18.6 (C-5'), 14.0 (C-14'), 11.0 (N–CH2–CH2–CH3). ESI-MS m/z
(rel. int.): [M+H]+ 380 (100).
1-Methyl-2-(1′-tetradecynyl)-4(1H)-quinolone (8r) was prepared from 6c (1.5 g, 6.4 mmol) in THF
(25 mL), LDA (3.5 mL, 6.4 mmol) and N-methylisatoic anhydride (7a) (0.85 g, 4.8 mmol) in THF
(15 mL) as a white solid (57%). M.p. 41–43 °C. IR (KBr, cm−1): 3405, 2923, 2849, 2233, 1622, 1595,
1
1496, 1468, 777. H-NMR δ: 8.38 (dd, J = 8.0, 1.6 Hz, 1H, H-5), 7.63 (td, J = 8.0, 1.6 Hz, 1H, H-7),
7.42 (d, J = 8.0 Hz, 1H, H-8), 7.33 (t, J = 7.6 Hz, 1H, H-6), 6.51 (s, 1H, H-3), 3.92 (s, 3H, N–CH3),
2.49 (t, J = 7.2 Hz, 2H, H-3'), 1.63 (quint, J = 7.2 Hz, 2H, H-4'), 1.44 (m, 2H, H-5'), 1.34–1.21 (m,
13
16H, H-6'-13'), 0.85 (t, J = 6.8 Hz, 3H, H-14'). C-NMR δ: 177.1 (C-4), 141.1 (C-8a), 137.3 (C-2),
132.3 (C-7), 126.8 (C-4a), 126.6 (C-5), 123.6 (C-6), 115.6 (C-8), 115.0 (C-3), 102.0 (C-2'), 74.8
(C-1'), 36.7 (N–CH3), 31.8 (C-12'), 29.6 (C-11'), 29.6 (C-10'), 29.5 (C-9'), 29.4 (C-8'), 29.2 (C-7'), 29.0
(C6'), 28.9 (C5'), 27.9 (C-4'), 22.6 (C-13'), 19.6 (C-3'), 14.0 (C-14'). ESI-MS m/z (rel. int.): [M+H]+
352 (100).
1-Methyl-2-(3′-hexadecynyl)-4(1H)-quinolone (8s) was prepared from 3e (1.5 g, 5.7 mmol) in THF
(25 mL), LDA (3.2 mL, 5.7 mmol) and N-methylisatoic anhydride (7a) (0.76 g, 4.3 mmol) in THF
(15 mL) as a light yellow solid (59%). M.p. 58–61 °C. IR (KBr, cm−1): 3432, 2922, 2853, 1631, 1597,
1
1469, 1444, 761. H-NMR δ: 8.46 (d, J = 8.0 Hz, 1H, H-5), 7.73 (t, J = 7.6 Hz, 1H, H-7), 7.58 (d,
J = 8.0 Hz, 1H, H-8), 7.44 (t, J = 7.6 Hz, 1H, H-6), 6.56 (s, 1H, H-3), 3.85 (s, 3H, N–CH3), 3.00 (t,
J = 7.6 Hz, 2H, H-1'), 2.60 (t, J = 7.6 Hz, 2H, H-2’), 2.12 (t, J = 7.2 Hz, 2H, H-5'), 1.41 (m, 2H, H-6'),
13
1.32–1.19 (m, 18H, H-7'-15'), 0.88 (t, J = 6.4 Hz, 3H, H-16'). C-NMR δ: 175.6 (C-4), 155.3 (C-2),
141.7 (C-8a), 133.3 (C-7), 126.4 (C-5), 126.2 (C-4a), 124.4 (C-6), 115.9 (C-8), 110.5 (C-3), 83.4
(C-4'), 76.5 (C-3'), 35.6 (N–CH3), 34.1 (C-1'), 31.9 (C-14'), 29.7 (C-13'), 29.6 (C-12'), 29.6 (C-11'),
29.5 (C-10'), 29.3 (C-9'), 29.1 (C-8'), 28.9 (C-7'), 28.9 (C-6'), 22.7 (C-15'), 18.8 (C-2'), 18.6 (C-5'),
14.1 (C-16'). ESI-MS m/z (rel. int.): [M+H]+ 380 (100).
1-Methyl-2-(1'-pentadecynyl)-4(1H)-quinolone (8t) was prepared from 6d (1.5 g, 6.0 mmol) in THF
(25 mL), LDA (3.3 mL, 6.0 mmol) and N-methylisatoic anhydride (7a) (0.80 g, 4.5 mmol) in THF (15 mL)
as white needles (48%). M.p. 67–69 °C. IR (KBr, cm−1): 3401, 2915, 2851, 2239, 1618, 1596, 1472,
748. 1H-NMR δ: 8.42 (d, J = 8.4, Hz, 1H, H-5), 7.69 (td, J = 8.0, 1.6 Hz, 1H, H-7), 7.48 (d, J = 8.0 Hz,
1H, H-8), 7.35 (t, J = 7.6 Hz, 1H, H-6), 6.55 (s, 1H, H-3), 3.96 (s, 3H, N–CH3), 2.52 (t, J = 7.2 Hz, 2H,
H-3'), 1.66 (quint, J = 7.6 Hz, 2H, H-4'), 1.46 (quint, J = 7.2 Hz, 2H, H-5'), 1.35–1.21 (m, 18H,
13
H-6'-14'), 0.87 (t, J = 6.8 Hz, 3H, H-15'). C-NMR δ: 177.2 (C-4), 141.0 (C-8a), 136.6 (C-2), 132.2
(C-7), 126.7 (C-4a), 126.6 (C-5), 123.7 (C-6), 115.7 (C-8), 115.1 (C-3), 102.1 (C-2'), 74.9 (C-1'), 36.5
(N–CH3), 31.8 (C-13'), 29.6 (C-12'), 29.5 (C-11'), 29.5 (C-10'), 29.3 (C-9'), 29.3 (C-8'), 29.1 (C-7'),
29.1 (C6'), 28.9 (C5'), 28.0 (C-4'), 22.6 (C-14'), 19.5 (C-3'), 14.1 (C-14'). ESI-MS m/z (rel. int.):
[M+H]+ 366 (100).
1-Cyclopropyl-2-[(E)-12-bromodec-1'-enyl]-4(1H)-quinolone (18a) was prepared from 17a (1.0 g,
3.5 mmol) in THF (15 mL), LDA (1.9 mL, 3.5 mmol) and N-cyclopropylisatoic anhydride (12) (0.52 g,
2.6 mmol) in THF (10 mL) as a light yellow solid (49%). M.p. 85–87 °C. IR (KBr, cm−1): 3425, 2926,
1
2850, 1648, 1616, 1594, 1475, 1416, 1310, 1137, 1034, 966, 888, 760. H-NMR δ: 8.39 (dd, J = 8.0,

Molecules 2012, 17
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1.6 Hz, 1H, H-5), 7.90 (d, J = 8.4 Hz, 1H, H-8), 7.64 (td, J = 8.0, 1.6 Hz, 1H, H-7), 7.35 (t, J = 7.6 Hz,
1H, H-6), 6.67 (d, J = 16.0 Hz, 1H, H-1'), 6.55 (s, 1H, H-3), 6.45 (dt, J = 16.0, 6.8 Hz, 1H, H-2'), 3.41
(t, J = 6.8 Hz, 2H, H-12'), 3.28 (sept, J = 4.0 Hz, 1H, N–CH–(CH2)2), 2.30 (q, J = 6.8 Hz, 2H, H-3'),
1.83 (quint, J = 6.8 Hz, 2H, H-11'), 1.51 (quint, J = 6.8 Hz, 2H, H-4'), 1.41–1.23 (m, 12H, H-5'-10'),
0.90 (m, 4H, N–CH–(CH2)2). 13C-NMR δ: 178.2 (C-4), 153.0 (C-2), 142.2 (C-2'), 139.6 (C-8a), 131.5
(C-7), 126.4 (C-4a), 126.2 (C-5), 124.9 (C-1'), 123.4 (C-6), 117.4 (C-8), 108.2 (C-3), 34.1 (C-12'),
33.1 (C-3'), 32.7 (C-11'), 29.9 (C-8'), 29.4 (C-7'), 29.4 (C-9'), 29.3 (C-6'), 29.1 (C-5'), 28.6 (C-4'), 28.1
(C-10'), 24.7 (N–CH–(CH2)2), 2×12.4 (N–CH–(CH2)2). ESI-MS m/z (rel. int.): [M+H+2]+ 432 (100),
[M+H]+ 430 (94).
1-Cyclopropyl-2-[(E)-13-bromotridec-1'-enyl)-4(1H)-quinolone (18b) was prepared from 17b (1.0 g,
3.3 mmol) in THF (15 mL), LDA (1.8 mL, 3.3 mmol) and N-cyclopropylisatoic anhydride (12) (0.5 g,
2.5 mmol) in THF (10 mL) as a light yellow solid (55%). M.p. 61–63 °C. IR (KBr, cm−1): 3422, 3020,
1
2918, 2851, 1653, 1630, 1596, 1571, 1479, 1463, 1413, 1135, 1034, 973, 760. H-NMR δ: 8.38 (dd,
J = 8.0, 1.6 Hz, 1H, H-5), 7.89 (d, J = 8.4 Hz, 1H, H-8), 7.64 (td, J = 8.0, 1.6 Hz, 1H, H-7), 7.35 (t,
J = 7.6 Hz, 1H, H-6), 6.66 (d, J = 16.0 Hz, 1H, H-1'), 6.50 (s, 1H, H-3), 6.44 (dt, J = 16.0, 6.8 Hz, 1H,
H-2'), 3.41 (t, J = 6.8 Hz, 2H, H-13'), 3.27 (sept, J = 4.0 Hz, 1H, N–CH–(CH2)2), 2.30 (q, J = 6.8 Hz,
2H, H-3'), 1.85 (quint, J = 6.8 Hz, 2H, H-12'), 1.51 (quint, J = 6.8 Hz, 2H, H-4'), 1.43–1.23 (m, 14H,
H-5'-11'), 0.90 (m, 4H, N–CH–(CH2)2). 13C-NMR δ: 178.2 (C-4), 152.9 (C-2), 142.1 (C-2'), 139.5
(C-8a), 131.4 (C-7), 126.4 (C-4a), 126.2 (C-5), 124.9 (C-1'), 123.3 (C-6), 117.4 (C-8), 108.2 (C-3),
34.1 (C-13'), 33.1 (C-3'), 32.7 (C-12'), 29.5 (C-9'), 29.5 (C-8'), 29.3 (C-10'), 29.3 (C-7'), 29.2 (C-6'),
28.7 (C-5'), 28.6 (C-4'), 28.1 (C-11'), 24.9 (N–CH–(CH2)2), 2 × 12.4 (N–CH-(CH2)2). ESI-MS m/z
(rel. int.): [M+H+2]+ 446 (100), [M+H]+ 444 (94).
1-Methyl-2-[(E)-12-bromodec-1'-enyl]-4(1H)-quinolone (19a) was prepared from 17a (1.5 g, 5.2 mmol)
in THF (25 mL), LDA (2.9 mL, 5.2 mmol) and N-methylisatoic anhydride (7a) (0.69 g, 3.9 mmol) in
THF (10 mL) as a yellow semi solid (63%). IR (KBr, cm−1): 3422, 2925, 2852, 1620, 1597, 1467,
1438, 761. 1H-NMR δ: 8.44 (d, J = 8.0 Hz, 1H, H-5), 7.67 (t, J = 7.2 Hz, 1H, H-7), 7.49 (t, J = 8.4 Hz,
1H, H-8), 7.38 (t, J = 7.2 Hz, 1H, H-6), 6.46 (s, 1H, H-3), 6.43 (d, J = 16.0 Hz, 1H, H-1'), 6.37 (dt,
J = 16.0, 6.4 Hz, 1H, H-2'), 3.76 (s, 3H, N–CH3), 3.41 (t, J = 6.4 Hz, 2H, H-12'), 2.28 (q, J = 6.8 Hz,
2H, H-3'), 1.85 (quint, J = 6.8 Hz, 2H, H-11'), 1.50 (quint, J = 6.8 Hz, 2H, H-4'), 1.41 (quint,
J = 6.8 Hz, 2H, H-10'), 1.37–1.23 (m, 10H, H-5'-9'). 13C-NMR δ: 177.9 (C-4), 152.5 (C-2), 142.0
(C-2'), 141.4 (C-8a), 132.2 (C-7), 126.6 (C-4a), 126.5 (C-5), 123.8 (C-1'), 123.5 (C-6), 115.5 (C-8),
109.4 (C-3), 35.5 (N–CH3), 34.1 (C-12'), 33.1 (C-3'), 32.7 (C-11'), 29.4 (C-8'), 29.3 (C-9'), 29.3 (C-7'),
29.1 (C-6'), 28.7 (C-5'), 28.5 (C-4'), 28.1 (C-10'). ESI-MS m/z (rel. int.): [M+H]+ 404 (100), [M+H+2]+
406 (92).
1-Methyl-2-[(E)-13-bromotridec-1'-enyl)-4(1H)-quinolone (19b) was prepared from 17b (1.0 g, 3.3 mmol)
in THF (15 mL), LDA (1.8 mL, 3.3 mmol) and N-methylisatoic anhydride (7a) (0.44 g, 2.5 mmol) in
THF (10 mL) as a yellow semi solid (47%). IR (KBr, cm−1): 3420, 2925, 2852, 1622, 1597, 1496,
1468, 760. 1H-NMR δ: 8.45 (d, J = 8.0 Hz, 1H, H-5), 7.70 (t, J = 7.6 Hz, 1H, H-7), 7.52 (t, J = 8.0 Hz,
1H, H-8), 7.40 (t, J = 7.6 Hz, 1H, H-6), 6.48 (s, 1H, H-3), 6.44 (d, J = 16.0 Hz, 1H, H-1'), 6.37 (dt,

Molecules 2012, 17
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J = 16.0, 6.4 Hz, 1H, H-2'), 3.79 (s, 3H, N–CH3), 3.41 (t, J = 6.8 Hz, 2H, H-13'), 2.29 (q, J = 7.2 Hz,
2H, H-3'), 1.86 (quint, J = 6.8 Hz, 2H, H-12'), 1.52 (quint, J = 6.8 Hz, 2H, H-4'), 1.43 (quint, J = 6.8 Hz,
13
2H, H-11'), 1.37–1.22 (m, 12H, H-5'-10'). C-NMR δ: 177.4 (C-4), 152.3 (C-2), 142.4 (C-2'), 141.4
(C-8a), 132.4 (C-7), 126.7 (C-4a), 126.6 (C-5), 123.9 (C-1'), 123.6 (C-6), 115.5 (C-8), 109.4 (C-3),
35.6 (N–CH3), 34.1 (C-13'), 33.2 (C-3'), 32.8 (C-12'), 29.5 (C-9'), 29.5 (C-8'), 29.3 (C-10'), 29.3 (C-7'),
29.1 (C-6'), 28.7 (C-5'), 28.6 (C-4'), 28.1 (C-11'). ESI-MS m/z (rel. int.): [M+H]+ 418 (100), [M+H+2]+
420 (94).
1-Cyclopropyl-2-[(E)-1′-tridecenyl]-4(1H)-quinolone (22a) was prepared from 21a (1.5 g, 6.7 mmol)
in THF (25 mL), LDA (3.7 mL, 6.7 mmol) and N-cyclopropylisatoic anhydride (12) (1.0 g, 5.0 mmol)
in THF (20 mL) as white needles (56%). M.p. 86–88 °C. IR (KBr, cm−1): 3425, 2922, 2851, 1617,
1595, 1571, 1479, 1465, 1420, 1310, 1136, 1031, 966, 759. 1H-NMR δ: 8.37 (dd, J = 8.0, 1.6 Hz, 1H,
H-5), 7.88 (d, J = 8.4 Hz, 1H, H-8), 7.62 (td, J = 8.0, 1.6 Hz, 1H, H-7), 7.33 (t, J = 8.0 Hz, 1H, H-6),
6.65 (d, J = 16.0 Hz, 1H, H-1'), 6.48 (s, 1H, H-3), 6.42 (dt, J = 16.0, 6.8 Hz, 1H, H-2′), 3.26 (sept,
J = 4.0 Hz, 1H, N–CH–(CH2)2), 2.29 (q, J = 6.8 Hz, 2H, H-3′), 1.52 (quint, J = 6.8 Hz, 2H, H-4'),
1.36–1.24 (m, 16H, H-5'-12'), 0.86–0.90 (m, 7H, H-13', N–CH–(CH2)2). 13C-NMR δ: 178.2 (C-4),
152.8 (C-2), 142.1 (C-8a), 139.3 (C-2'), 131.4 (C-7), 126.5 (C-4a), 126.1 (C-5), 124.9 (C-1'), 123.3
(C-6), 117.4 (C-8), 108.2 (C-3), 33.1 (C-3'), 31.8 (C-11'), 29.8 (N–CH–(CH2)2), 29.6 (C-10'), 29.5
(C-9), 29.5 (C-8'), 29.4 (C-7'), 29.3 (C6'), 29.2 (C5'), 28.6 (C-4'), 22.6 (C-12'), 14.0 (C-13'), 12.3
(N–CH–(CH2)2). ESI-MS m/z (rel. int.): [M+H]+ 366 (100).
1-Cyclopropyl-2-[(E)-1′-tetradecenyl]-4(1H)-quinolone (22b) was prepared from 21b (1.5 g, 6.3 mmol)
in THF (25 mL), LDA (3.5 mL, 6.3 mmol) and N-cyclopropylisatoic anhydride (12) (0.96 g, 4.7 mmol)
in THF (20 mL) as white crystals (50%). M.p. 96–98 °C. IR (KBr, cm−1): 3422, 2919, 2848, 1620,
1598, 1572, 1482, 1465, 1419, 1306, 1132, 1037, 966, 750. 1H-NMR δ: 8.36 (d, J = 8.0 Hz, 1H, H-5),
7.87 (d, J = 8.4 Hz, 1H, H-8), 7.62 (td, J = 8.0, 1.6 Hz, 1H, H-7), 7.32 (t, J = 8.0 Hz, 1H, H-6), 6.65 (d,
J = 16.0 Hz, 1H, H-1'), 6.46 (s, 1H, H-3), 6.42 (dt, J = 16.0, 6.8 Hz, 1H, H-2'), 3.25 (sept, J = 4.0 Hz,
1H, N–CH–(CH2)2), 2.28 (q, J = 6.8 Hz, 2H, H-3'), 1.51 (quint, J = 6.8 Hz, 2H, H-4'), 1.37–1.21 (m,
18H, H-5'-13'), 0.86–0.91 (m, 7H, H-14', N–CH–(CH2)2). 13C-NMR δ: 178.2 (C-4), 152.8 (C-2), 142.1
(C-8a), 139.3 (C-2'), 131.3 (C-7), 126.5 (C-4a), 126.1 (C-5), 124.9 (C-1'), 123.2 (C-6), 117.4 (C-8),
108.3 (C-3), 33.1 (C-3'), 31.8 (C-12'), 29.8 (N–CH–(CH2)2), 29.6 (C-11'), 29.6 (C-10'), 29.6 (C-9),
29.5 (C-8'), 29.4 (C-7'), 29.3 (C6'), 29.2 (C5'), 28.7 (C-4'), 22.6 (C-13'), 14.1 (C-14'), 12.3
(N–CH–(CH2)2). ESI-MS m/z (rel. int.): [M+H]+ 380 (100).
1-Cyclopropyl-2-(3′-undecynyl)-4(1H)-quinolone (23) was prepared from 3b (1.5 g, 7.7 mmol) in THF
(25 mL), LDA (4.3 mL, 7.7 mmol) and N-cyclopropylisatoic anhydride (12) (1.17 g, 5.8 mmol) in
THF (25 mL) as a yellow oil (48%). IR (KBr, cm−1): 3424, 2928, 2855, 1628, 1601, 1553, 1481, 1466,
1420, 1311, 1132, 1044, 759. 1H-NMR δ: 8.34 (dd, J = 8.0, 1.6 Hz, 1H, H-5), 7.87 (d, J = 8.4 Hz, 1H,
H-8), 7.60 (td, J = 8.0, 1.6 Hz, 1H, H-7), 7.31 (t, J = 6.8 Hz, 1H, H-6), 6.25 (s, 1H, H-3), 3.27 (sept,
J = 4.0 Hz, 1H, N–CH–(CH2)2), 3.17 (t, J = 7.2 Hz, 2H, H-1'), 2.54 (m, 2H, H-2'), 2.07 (m, 2H, H-5'),
1.41 (quint, J = 6.8 Hz, 2H, H-6'), 1.26–1.15 (m, 6H, H-7'-9'), 0.93 (m, 4H, N–CH–(CH2)2), 0.85 (t,
J = 6.8 Hz, 3H, H-10'). 13C-NMR δ: 178.1 (C-4), 155.6 (C-2), 142.8 (C-8a), 131.1 (C-7), 126.3 (C-4a),

Molecules 2012, 17
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126.1 (C-5), 123.1 (C-6), 117.6 (C-8), 111.0 (C-3), 82.9 (C-4'), 77.5 (C-3'), 33.0 (C-1'), 31.6 (C-8'),
29.1 (C-7'), 28.7 (C-6'), 28.7 (C-5'), 26.8 (N–CH–(CH2)2), 22.5 (C-9'), 18.6 (C-2'), 14.0 (C-10'), 12.3
(N–CH–(CH2)2). ESI-MS m/z (rel. int.): [M+H]+ 336 (100).
3.3. Biological Evaluation
3.3.1. In Vitro Antibacterial Activities against Fast Growing Strains of Mycobacteria and EMRSA-15
and EMRSA-16
In vitro M. smegmatis, M. fortuitum, M. phlei, EMRSA-15 and -16 inhibitory effect of the
4(1H)-quinolone derivatives was assessed in 96-well plates using the broth dilution assay according to
the previously reported protocols [7,23]. The plates containing test compounds, the antibiotic drugs
and growth media were incubated at 37 °C for 72 h for the rapidly-growing mycobacterial strains and
18 h for EMRSA-15 and -16. A methanolic solution of MTT (0.05%) was used to determine the MIC
by a colour change from yellow to blue. Tests were carried out in triplicate and all MIC values were
determined in separate duplicate experiments. Tetracycline and norfloxacin were used as a positive
control for the EMRSA strains and isoniazid and ethambutol for the mycobacterial strains.
3.3.2. Spot-Culture Growth Inhibition Assay (SPOTi) against M. bovis BCG
A diluted culture (~500 viable cells) of M. bovis BCG was spotted into 10% OADC supplemented
Middlebrook 7H10 agar medium in a 24-well plate having various concentrations of the 4(1H)-quinolone
derivatives and isoniazid. The plates were incubated at 37 °C for two weeks and the MIC values were
determined visually as the minimum concentrations where no growth was observed [24].
3.3.3. M. tuberculosis MurE Inhibition Assay
The M. tuberculosis MurE ligase activity was determined by the phosphate colorimetric detection
method as previously reported [7]. The compounds were tested at concentrations of 1000, 300, 100 and
30 µM and the amount of phosphate released was determined by means of a phosphate calibration
curve using the Pi ColorLock kit reagents (Innova Biosciences, Cambridge, UK). Isoniazid was used
as a negative control and IC50 values were determined by extrapolation from the plot of percent
inhibition vs. concentration.
3.3.4. Cytotoxicity Assay
The cytotoxic activity of the synthetic compounds was determined by means of an XTT Cell
Proliferation Kit II (Roche Diagnostics, Mannheim, Germany) using the human diploid embryonic
lung cell line MRC-5 [11]. The quinolone derivatives were tested at concentrations of 100, 60, 30 and
10 µM in triplicate and the percentage of viable cells at each concentration was determined by
comparing with the control.

Molecules 2012, 17
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4. Conclusions
Using the SAR information gained from our previous studies, a further group of quinolone
derivatives possessing a diverse set of alkynyl/(E)-alkenyl substituents at C-2 of the quinolone nucleus
were synthesized. Although the antimycobacterial potencies are comparable to those of the previously
reported alkenyl analoges, the new series of alkynyl bearing compounds displayed higher selectivity
towards mycobacteria and MRSA strains compared to MRC-5 cells. It is reasonable to conclude that
the improved cytotoxicity of this group of analoges is due to the introduction of a triple bond in the
aliphatic side chain of the quinolone nucleus. The selectivity shown by this class of new quinolones
make them promising leads for synthesizing further compounds with improved antibacterial activity.
Compounds with a terminal bromine atom at the side chain of N-alkyl-2-alkynyl/(E)-alkenyl-4-(1H)-
quinolones with improved antimycobacterial activity will be explored due their increased probability
of covalent bonding to particular target proteins.
Conflict of Interest
The authors declare no conflict of interest.
Acknowledgments
The Austrian Science Fund (FWF) project no. P21152-B18 and P24324-B21 is gratefully
acknowledged for financial support. We thank Andreas Leitner, Department of Pharmaceutical
Chemistry, for measurement of IR spectra.
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Sample Availability: Samples of the compounds 8a–b, 8e–f, 8h, 8j–k, 18a–b, 23 are available from the
authors.
© 2012 by the authors; licensee MDPI, Basel, Switzerland. This article is an open access article
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