European Journal of Medicinal Chemistry 156 (2018) 580e586

Contents lists available at ScienceDirect

European Journal of Medicinal Chemistry

journal homepage: [Link]

Research paper

Design, synthesis and antimicrobial evaluation of propylene-tethered

ciprofloxacin-isatin hybrids
Ruo Wang a, *, Xueyang Yin b, Yaohuan Zhang a, Weitao Yan a
a
College of Chemistry, Fuzhou University, Fuzhou, Fujian 350116, China
b
Shanghai Key Laboratory of Green Chemistry and Chemical Processes, Department of Chemistry, School of Chemistry and Molecular Engineering, East
China Normal University, Shanghai 200241, China

a r t i c l e i n f o a b s t r a c t

Article history: Twelve novel propylene-tethered ciprofloxacin-isatin hybrids 3a-f and 4a-f were designed, synthesized
Received 30 January 2018 and characterized by MS, HRMS, 1H NMR and 13C NMR. All hybrids were evaluated for their in vitro
Received in revised form antimicrobial activities against representative Gram-positive, Gram-negative and mycobacterial patho-
7 May 2018
gens, cytotoxicity in VERO cell line as well as metabolic stability and in vivo pharmacokinetic (PK)
Accepted 9 July 2018
Available online 17 July 2018
properties. The preliminary results indicated that all mono-isatin-ciprofloxacin hybrids exhibited
excellent antibacterial activities with MIC ranging from
0.03 to 0.5 mg/mL against most of the tested
strains. In particular, ciprofloxacin-isatin hybrid 3d was highly potent against all tested Gram-positive
Keywords:
Ciprofloxacin
and Gram-negative strains including clinically important drug-resistant pathogens, which was compa-
Isatin rable to or more potent than the parent ciprofloxacin and reference levofloxacin. Whereas, conjugate 3b
Hybrids (MIC: 0.10 and 0.5 mg/mL) was 4- and 8-fold more active than ciprofloxacin (MIC: 0.78 mg/mL) and
Antimicrobial rifampicin (MIC: 0.39 mg/mL) against MTB H37Rv, and 4->256 times more potent than the three refer-
Antibacterial ences ciprofloxacin (MIC: 2.0 mg/mL), rifampicin (MIC: 32 mg/mL) and isoniazid (>128 mg/mL) against
Antimycobacterial MDR-TB. Both hybrid 3b and 3d with low cytotoxicity (CC50: 64 and 256 mg/mL) also showed acceptable
Structure-activity relationship metabolic stability and in vivo PK properties, could act as leads for further optimization.
© 2018 Elsevier Masson SAS. All rights reserved.

1. Introduction develop novel antimicrobial agents active against both drug-

sensitive and drug-resistant bacterial infections.
Bacterial infections, which are caused by Gram-positive, Gram- Fluoroquinolones (FQs), are a family of synthetic broad spec-
negative and mycobacterial pathogens, are responsible for the trum antibiotics, and are the second widest used antibiotics in
majority of hospital-acquired infections, and lead to extensive clinical practice for various bacterial infections including upper and
mortality and burden on global healthcare systems [1,2]. It's esti- lower respiratory infections, and their value and role in the treat-
mated that infectious diseases result in around 10 million deaths ment of bacterial infections continue to expand [1]. FQs predomi-
every year (over 15% of all deaths), and tuberculosis (TB) is the nately act by binding two type II bacterial topoisomerase enzymes,
ninth leading cause of death throughout the world and the leading DNA gyrase (main target for Gram-negative bacteria) and topo-
cause from a single infectious agent according to the latest World isomerase IV (primary target for Gram-negative bacteria) [5,6], and
Health Organization (WHO) report [3,4]. The emergency and a wide DNA gyrase is deemed as the only type II topoisomerase present in
spread of drug-resistant organisms such as methicillin-resistant MTB and is thus the only target for FQs action [7,8]. Besides their
S. aureus (MRSA), methicillin-resistant S. epidermidis (MRSE), typical antibacterial activities, FQs also demonstrated various
vancomycin-resistant S. aureus (VRSA), extended-spectrum b-lac- atypical biological properties such as antimalarial [9,10], antitumor
tamase (ESBL)-producing E. coli and drug-resistant TB (DR-TB) has [11] and anti-TB activities [12e15], play a pivotal role in new drug
already increased up to alarming level in the recent decades and is discovery.
associated with considerable mortality [3,4]. Thus, it's imperative to However, as other antimicrobial agents, the resistance of path-
ogens to fluoroquinolones develops rapidly and spreads widely
mainly attribute to the long-term, broad, inappropriate use and
* Corresponding author. even abuse, makes FQs more and more ineffective. Therefore,
E-mail address: wangruo1201@[Link] (R. Wang). enhancing the potency of fluoroquinolones has become

[Link]
0223-5234/© 2018 Elsevier Masson SAS. All rights reserved.
 R. Wang et al. / European Journal of Medicinal Chemistry 156 (2018) 580e586 581

increasingly urgent. mycobacterial activities against MTB H37Rv and MDR-TB strains
Isatin is an endogenous compound identified in many organ- [34]. The minimum inhibitory concentration (MIC) is defined as the
isms, endows with various biological properties such as antibac- minimum concentration of compound required to give 90% inhi-
terial [1], anticancer [16], anti-HIV [17], antimalarial [10], and anti- bition of bacterial growth and MICs of the compounds are reported
TB activities [12] which may ascribed to its ability to exert non- in Table 1, 2 and 3, respectively.
covalent interactions such as electrostatic interactions and As it can be seen from Table 1, all mono-isatin-CPFX 3a-f dis-
hydrogen bonds etc. Furthermore, many isatin related compounds played promising activities against the tested Gram-positive strains
such as semaxanib and nintedanib have been approved for clinical with MIC ranging from 0.06 to 64 mg/mL. Among them, the most
use. Thus, exploitation of isatin moiety in antimicrobial area will be active hybrid 3d was highly potent against all tested Gram-positive
especially fruitful. strains, especially against clinically important pathogens MSSE,
Various FQs-isatin hybrids tethered with different linkers MRSA, MSSA, MRSA and Enterococcus faecalis with MIC of 0.06,
exhibited excellent in vitro and in vivo potency against diverse 0.06, 0.06 and 0.5 mg/mL, respectively, which was 2e64 folds more
microorganisms [18e35]. The previous studies demonstrated that potent than the references CPFX (MIC: 0.125e64 mg/mL) and lev-
the linkers between FQs and isatin play a crucial role for the anti-TB ofloxacin (MIC: 0.125e4 mg/mL).
activity [28e35], so the linkers are worth to be further optimized. From Table 2, it can be concluded that all mono-isatin-CPFX 3a-f
Ciprofloxacin (CPFX), as the second generation fluoroquinolone, displayed excellent potency against the tested Gram-negative
endows with excellent antimicrobial activity, notable pharmaco- strains with MIC in a range of
0.03e2 mg/mL. Interestingly,
kinetic properties and few side effects, is used widely in clinical hybrid 3d also demonstrated highest potent against all tested
practice for the treatment of various bacterial infections [8]. Gram-negative strains with MIC of
0.03e0.5 mg/mL, respectively,
Moreover, CPFX has been recommended as the second-line agents which was comparable to or slightly more active than the refer-
by the WHO for the treatment of TB mainly in cases involving ences CPFX (MIC:
0.03e2 mg/mL) and levofloxacin (MIC:
resistance or intolerance to first-line anti-TB therapy [8]. Thus,
0.03e2 mg/mL).
ciprofloxacin derivatives have caused continuous interests. The SAR revealed that the antibacterial activity of mono-isatin-
Based on the above considerations, a novel set of propylene- CPFX 3a-f were far more potent than the corresponding bis-isatin-
tethered CPFX-isatin hybrids 3a-f and 4a-f with more flexible CPFX 4a-f (MIC: 4->128 mg/mL) against all tested Gram-positive
propylene as linker was designed, synthesized and examined for and Gram-negative strains, suggesting carboxylic acid at C-3 posi-
their in vitro antimicrobial activities against representative Gram- tion is essential for gyrase binding and bacterial membrane trans-
positive, Gram-negative and mycobacterial pathogens as well as port; introduction of imine at C-3 position of isatin motif could
cytotoxicity in VERO cell line. Our primary objective was to opti- boost up the activity, and the relative contribution order was
mize the potency of these hybrids against clinically important -NOMe > -NOEt > -O; Substituents at C-5 position of isatin moiety
pathogens. A preliminary structure-activity relationship (SAR) have great influence on the activity, and hybrids with electron-
study is also explored to facilitate the further design. The design donating -Me were more potent than the corresponding unsub-
strategy is illustrated in Fig. 1. stituted analogs.
The antimycobacterial results showed that all hybrids exhibited
2. Results and discussion considerable activity with MIC in a range of 0.1e64 mg/mL against
MTB H37Rv and MDR-TB, and all mono-isatin-CPFX hybrids except
The synthetic pathway for propylene-tethered CPFX-isatin hy- 3f were no inferior to the parent CPFX. The SAR indicated that
brids 3a-f and 4a-f is outlined in Scheme 1. Alkylation of C-5 mono-isatin-CPFX hybrids were more potent than the corre-
substituted isatins 1a,b was performed with 1,3-dibromopropane sponding bis-isatin-CPFX hybrids which was in accordance with
in presence of K2CO3 to provide N-(3-bromopropyl)isatins 2a,b, the previous antibacterial study; for mono-isatin-CPFX hybrids,
which were then incorporated into CPFX core to afford the desired introduction of imines at C-3 position and electron-donating -Me at
targets 3a,b and bis-isatin-CPFX hybrids 4a,b. Subsequently, C-5 position of isatin moiety reduced the activity against both of the
condensation of hybrids 3a,b or 4a,b with methoxylamine or tested strains generally. In particular, the most potent hybrid 3b
ethoxylamine hydrochloride in the presence of sodium bicarbonate with MIC of 0.1 and 0.5 mg/mL was 4- and 8-fold more active than
provided other conjugates 3c-f and 4c-f. the parent CPFX (MIC: 0.78 mg/mL) and rifampicin (RIF, MIC:
All propylene-tethered isatin-CPFX hybrids 3a-f and 4a-f were 0.39 mg/mL) against MTB H37Rv, and 4->256 times more potent
assessed for their in vitro antibacterial activities against clinically than the three references CPFX (MIC: 2.0 mg/mL), RIF (MIC: 32 mg/
important Gram-positive, Gram-negative pathogens as well as anti- mL) and isoniazid (INH, >128 mg/mL) against MDR-TB.
The propylene-tethered isatin-CPFX hybrids 3a-f and 4a-f were
subsequently examined for toxicity (CC50) in a mammalian VERO
cell line [36]. After 72 h of exposure, viability was assessed on the
basis of cellular conversion of MTT (3-(4,5-dimethylthiazol-2-yl)-
2,5-diphenyltetrazolium bromide) into a formazan product and the
results are reported in Table 3. All hybrids (CC50: 16e256 mg/mL)
showed acceptable cytotoxicity, and fortunately, the cytotoxicity of
the most potency 3b (CC50: 64 mg/mL) and 3d (CC50: 256 mg/mL)
were comparable to that of CPFX (CC50: 128 mg/mL).
The metabolic stability and in vivo pharmacokinetic (PK) prop-
erties of hybrid 3b and 3d were evaluated in mice after oral (po)
administration 50 mg/kg, respectively [37]. As shown in Table 4,
after oral dosing, both hybrids reached a maximum concentration
in plasma within 1.5 h; their elimination half-life was favorable (4.1
and 3.3 h, respectively); and the area under curve (AUC) was 4988
Fig. 1. Illustration of design strategy for propylene-tethered isatin-ciprofloxacin and 2,865, respectively. In general, the metabolic stability and
hybrids. in vivo PK profiles were less favorable than the parent CPFX, need to
582 R. Wang et al. / European Journal of Medicinal Chemistry 156 (2018) 580e586

Scheme 1. Synthetic route for mono-/bis-isatin-CPFX hybrids 3a-f and 4a-f.

be further optimized. column with formic acid as additive to provide the desired targets
In summary, a series of novel propylene-tethered isatin-CPFX 3a,b and bis-isatin-CPFX hybrids 4a,b.
hybrids were synthesized and evaluated for their in vitro antibac-
terial and anti-mycobacterial activities as well as cytotoxicity. All [Link]. 1-Cyclopropyl-7-(4-(3-(2,3-dioxoindolin-1-yl)propyl)piper-
hybrids exhibited considerable inhibitory activity against the tested azin-1-yl)-6-fluoro-4-oxo-1,4-dihydroquinoline-3-carboxylic acid
strains, and the most active conjugate 3b with low cytotoxicity was (3a). Brown solid, yield: 16%. Mp: 174e176  C. 1H NMR (400 MHz,
more potent than the parent CPFX and RIF against MTB H37Rv and DMSO‑d6) d 1.12e1.27 (4H, m, 2  cyclopropyl-CH2), 2.04 (2H, t,
MDR-TB, warrant further investigations. -CH2-), 2.92 (4H, s, piperazine-4H), 3.19 (4H, s, piperazine-4H),
3.66e3.68 (1H, m, cyclopropyl-CH), 3.86 (2H, t, -CH2-), 4.25 (2H,
3. Experimental section t, -CH2-), 7.08 (1H, t, Ar-H), 7.34 (1H, d, Ar-H), 7.43e7.49 (2H, m, Ar-
H), 7.60 (1H, t, Ar-H), 7.76 (1H, d, Ar-H), 8.44 (1H, s, C2-H). 13C NMR
3.1. Synthesis (DMSO‑d6, 100 MHz) dC: 183.886, 172.078, 165.005, 163.985,
158.728, 154.171, 151.786, 151.232, 148.658, 138.480, 124.707,
3.1.1. General procedure for the preparation of 3a,b and 4a,b 123.445, 122.811, 118.101, 112.122, 111.108, 109.511, 106.930, 62.133,
N-(3-bromopropyl)isatins 2a,b were obtained via literature re- 60.222, 48.571, 44.072, 37.350, 35.272, 26.515, 21.229, 14.557, 8.024.
ported method [34,35]. To a mixture of N-(3-bromopropyl)isatins ESI-MS m/z: 519 [MþH]þ. HRMS-ESI: m/z Calcd for C28H28FN4O5
2a,b (5 mmol) and CPFX (3 mmol) in DMF (50 mL), K2CO3 [MþH]þ: 519.2043; Found: 519.2033.
(20 mmol) was added. The mixture was stirred at room tempera-
ture for 3 days. After filtration, the filtrate was concentrated under [Link]. 1-Cyclopropyl-6-fluoro-7-(4-(3-(5-methyl-2,3-dioxoindolin-
reduced pressure. The residue was purified by reverse phase 1-yl)propyl)piperazin-1-yl)-4-oxo-1,4-dihydroquinoline-3-carboxylic
 R. Wang et al. / European Journal of Medicinal Chemistry 156 (2018) 580e586 583

Table 1 Table 3
In vitro antibacterial activity of hybrids 3a-f and 4a-f against Gram-positive strains. Anti-mycobacterial activity and cytotoxicity of hybrids 3a-f and 4a-f.

Compd. MIC (mg/mL) Compd. R1 R2 MIC (mg/mL) CC50b (mg/mL)

MSSE MRSE MSSA MRSA [Link].1 [Link].2 [Link].1 [Link].2 MTB H37Rv MDR-TBa

3a 0.25 4 0.25 0.25 1 1 8 64 3a H O 0.2 0.5 128

3b 0.125 2 0.125 0.125 0.5 0.5 4 32 3b Me O 0.1 0.5 64
3c 0.125 2 0.125 0.25 0.5 1 8 32 3c H NOMe 0.39 2.0 32
3d 0.06 1 0.06 0.06 0.5 0.5 2 8 3d Me NOMe 0.78 1.0 256
3e 0.25 2 0.25 0.25 1 1 8 32 3e H NOEt 0.78 1.0 64
3f 0.125 2 0.125 0.125 0.5 1 4 32 3f Me NOEt 3.12 4.0 128
4a 64 >128 128 >128 64 >128 >128 >128 4a H O 12.5 32 32
4b 32 128 64 >128 64 >128 128 >128 4b Me O 50 32 64
4c 32 128 128 >128 64 128 128 >128 4c H NOMe 12.5 64 64
4d 16 64 32 32 32 128 128 >128 4d Me NOMe 25 32 16
4e 64 128 128 >128 64 >128 >128 >128 4e H NOEt 25 64 32
4f 32 64 64 128 64 128 128 >128 4f Me NOEt 50 64 64
CFFX 0.125 64 0.25 0.5 0.5 0.5 8 128 CPFX 0.78 2.0 128
levofloxacin 0.125 4 0.125 0.125 1 1 16 64 INH 0.05 >128 128
RIF 0.39 32 512
Abbreviations: MSSE, methicillin-sensitive Staphylococcus epidermidis ATCC
a
12228; MRSE, methicillin-resistant Staphylococcus epidermidis13-3; MSSA, MDR-TB: resistant to INH, RIF and EMB.
b
methicillin-sensitive Staphylococcus aureus ATCC 29213; MRSA, methicillin- CC50: The 50% cytotoxic concentration in a mammalian VERO cell line.
resistant Staphylococcus aureus ATCC 33591; [Link].1, Enterococcus faecalis ATCC
29212; [Link].2, Enterococcus faecalis ATCC 51299; [Link].1, Enterococcus faecium ATCC
700221; [Link].2, Enterococcus faecium 13-7. 4H), 3.66e3.67 (1H, m, cyclopropyl-CH), 3.77 (2H, t, -CH2-), 3.87
(2H, t, -CH2-), 4.24 (2H, t, -CH2-), 7.07 (1H, t, Ar-H), 7.13 (1H, t, Ar-H),
7.26 (1H, d, Ar-H), 7.34 (1H, d, Ar-H), 7.36 (1H, d, Ar-H), 7.47 (1H, d,
acid (3b). Brown solid, yield: 21%. Mp: 158e160  C. 1H NMR
Ar-H), 7.56 (1H, d, Ar-H), 7.61 (1H, t, Ar-H), 7.68 (1H, t, Ar-H), 7.74
(400 MHz, DMSO‑d6) d 1.10e1.26 (4H, m, 2  cyclopropyl-CH2), 2.02
(1H, d, Ar-H), 8.44 (1H, s, C2-H). 13C NMR (DMSO‑d6, 100 MHz) dC:
(2H, t, -CH2-), 2.22 (3H, s, CH3), 2.98 (4H, s, piperazine-4H), 3.23
184.211, 183.875, 172.076, 165.042, 163.620, 158.742, 154.273,
(4H, s, piperazine-4H), 3.66e3.68 (1H, m, cyclopropyl-CH), 3.85
151.502, 151.225, 148.644, 144.320, 144.218, 138.662, 138.480,
(2H, t, -CH2-), 4.22 (2H, t, -CH2-), 7.23 (1H, d, Ar-H), 7.29 (1H, s, Ar-
124.874, 124.707, 123.496, 122.381, 118.079, 118.006, 112.151,
H), 7.43 (2H, t, Ar-H), 7.76 (1H, d, Ar-H), 8.41 (1H, s, C2-H). 13C NMR
111.925, 111.312, 111.108, 109.409, 106.566, 62.082, 55.396, 52.844,
(DMSO‑d6, 100 MHz) dC: 184.101, 172.056, 164.932, 158.742, 154.236,
49.862, 38.473, 37.342, 35.206, 26.537, 23.824, 8.010. ESI-MS m/z:
151.799, 149.089, 148.629, 144.174, 138.764, 138.456, 132.800,
706 [MþH]þ. HRMS-ESI: m/z Calcd for C39H37FN5O7 [MþH]þ:
124.947, 117.991, 112.224, 110.984, 109.424, 103.746, 62.104, 49.205,
706.2677; Found: 706.2692.
37.364, 36.038, 36.023, 35.235, 26.537, 26.536, 20.427, 20.425,
7.995. ESI-MS m/z: 533 [MþH]þ. HRMS-ESI: m/z Calcd for
C29H30FN4O5 [MþH]þ: 533.2200; Found: 533.2175. [Link]. 3-(5-methyl-2,3-dioxoindolin-1-yl)propyl 1-cyclopropyl-6-
fluoro-7- (4-(3-(5-methyl-2,3-dioxoindolin-1-yl)propyl)piperazin-1-
[Link]. 3-(2,3-Dioxoindolin-1-yl)propyl 1-cyclopropyl-7-(4-(3-(2,3- yl)-4-oxo-1,4-dihydroquinoline-3-carboxylate (4b). Brown solid,
dioxoindolin-1- yl)propyl)piperazin-1-yl)-6-fluoro-4-oxo-1,4- yield: 23%. Mp: 144e145  C. 1H NMR (400 MHz, DMSO‑d6)
dihydroquinoline-3-carboxylate (4a). Brown solid, yield: 26%. Mp: d 1.19e1.39 (4H, m, 2  cyclopropyl-CH2), 2.02 (2H, t, -CH2-), 2.20
152e153  C. 1H NMR (400 MHz, DMSO‑d6) d 1.10e1.30 (4H, m, (2H, t, -CH2-), 2.30 (3H, s, CH3), 2.35 (3H, s, CH3), 2.68 (2H, t, -CH2-),
2  cyclopropyl-CH2), 1.84 (2H, t, -CH2-), 2.04 (2H, t, -CH2-), 2.45 2.78 (4H, s, piperazine-4H), 3.30 (4H, s, piperazine-4H), 3.31e3.47
(2H, t, -CH2-), 3.17 (4H, s, piperazine-4H), 3.38 (4H, s, piperazine- (1H, m, cyclopropyl-CH), 3.86 (2H, t, -CH2-), 4.00 (2H, t, -CH2-), 4.38

Table 2
In vitro antibacterial activity of hybrids 3a-f and 4a-f against Gram-negative strains.

Compd. MIC (mg/mL)

[Link].1 [Link].2 K.p.1 K.p.2 P.a. A.c. E.c. E.a. S.m.1 M.m. P.r. P.v. P.m. S.m.2 C.f.

3a
0.03
0.03 1 0.125 1 1 0.03
0.03 0.25
0.03 0.03
0.03
0.03 0.125
0.03
3b
0.03
0.03 1 0.06 2 0.25
0.03 0.06 0.25
0.03
0.03
0.03
0.03 0.06
0.03
3c
0.03
0.03 0.5
0.03 1 0.5
0.03
0.03 0.06
0.03
0.03
0.03
0.03 0.06
0.03
3d
0.03
0.03 0.25
0.03 0.5 0.25
0.03
0.03 0.06
0.03
0.03
0.03
0.03
0.03
0.03
3e
0.03
0.03 0.5 0.03 1 1 0.06
0.03 0.125 0.03
0.03
0.03
0.03 0.125
0.03
3f
0.03
0.03 0.5
0.03 1 0.5
0.03
0.03 0.06
0.03
0.03
0.03
0.03 0.06
0.03
4a 4 16 64 64 >128 128 128 64 32 64 128 64 64 >128 64
4b 8 8 64 64 128 64 32 128 >128 64 128 64 64 128 64
4c 4 4 64 32 64 128 32 64 64 128 64 32 64 128 64
4d 8 16 32 16 32 64 16 32 32 64 32 32 64 64 32
4e 8 32 64 64 64 >128 64 64 >128 64 128 128 64 >128 128
4f 16 16 128 32 128 64 128 64 64 32 64 64 64 128 64
CPFX
0.03
0.03 0.5
0.03 0.25 0.5
0.03
0.03 0.06
0.03
0.03
0.03
0.03 2
0.03
levofloxacin
0.03
0.03 0.5
0.03 2 0.125
0.03 0.06 0.125
0.03
0.03
0.03
0.03 1
0.03

[Link].1, Escherichia coli ATCC 25922 ESBLs(); [Link].2, Escherichia coli ATCC 35218 ESBLs(þ); K.p.1, Klebsiella pneumoniae ATCC 700603 ESBLs(þ); K.p.2, Klebsiella
pneumonia 15-2 ESBLs(); P.a., Pseudomonas aeruginosa ATCC 27853; A.c., Acinetobacter calcoacetious ATCC 19606; E.c., Enterobacter cloacae ATCC 43560; E.a., Enterobacter
aerogenes ATCC 13048; S.m.1, Serratia marcescens ATCC 21074; M.m., Morganella morganii ATCC 25830; P.r., Providentia rettgeri ATCC 31052; P.v., Proteus vulgaris ATCC
29905; P.m., Proteus mirabilis 13-1; S.m.2, Stenotrophomonas maltophilia ATCC 13636; C.f., Citrobacter freundii ATCC 43864. ESBLs(þ): Extended spectrum beta-lactamases
(ESBLs)-producing.
584 R. Wang et al. / European Journal of Medicinal Chemistry 156 (2018) 580e586

Table 4
Metabolic stability and in vivo pharmacokinetic values of hybrids 3c and 3d in mice.

Compd. microsomal stability (t1/2, min) pharmacokinetics (p.o.)

Cmax (ng$mL1) tl/2 (h) Tmax (h) AUC0-inf (ng$h$mL1)

3b 31 1228 4.1 1.3 4988

3d 17 832 3.3 1.2 2865
CPFX >60 3129 5.2 1.8 17,835

(2H, t, -CH2-), 6.87 (1H, d, Ar-H), 7.20 (1H, s, Ar-H), 7.26 (1H, d, Ar- carboxylic acid (3e). Yellow solid, yield: 33%. Mp: 126e128  C. 1H
H), 7.32 (1H, d, Ar-H), 7.40e7.43 (3H, m, Ar-H), 8.06 (1H, d, Ar-H), NMR (400 MHz, DMSO‑d6) d 1.10e1.28 (4H, m, 2  cyclopropyl-
8.58 (1H, s, C2-H). 13C NMR (CDCl3, 100 MHz) dC: 183.785, CH2), 1.35 (3H, t, NOCH2CH3), 2.03 (2H, t, -CH2-), 3.00 (4H, s,
173.026, 165.822, 165.472, 158.574, 154.526, 152.056, 148.775, piperazine-4H), 3.24 (4H, s, piperazine-4H), 3.66e3.69 (1H, m,
148.491, 144.160, 144.050, 139.121, 138.859, 138.050, 133.580, cyclopropyl-CH), 3.90 (2H, t, -CH2-), 4.18 (2H, t, -CH2-), 4.38 (2H, q,
133.420, 125.691, 125.443, 122.942, 117.619, 117.510, 113.135, NOCH2CH3), 7.07 (1H, t, Ar-H), 7.28 (1H, d, Ar-H), 7.41e7.45 (2H, m,
110.612, 110.102, 109.657, 105.180, 61.695, 55.170, 52.435, 48.950, Ar-H), 7.77 (1H, t, Ar-H), 7.85 (1H, d, Ar-H), 8.40 (1H, s, C2-H). 13C
48.906, 38.290, 37.372, 34.790, 26.515, 23.394, 20.638, 20.589, NMR (DMSO‑d6, 100 MHz) dC: 172.071, 165.450, 164.750, 162.884,
8.177. ESI-MS m/z: 734 [MþH]þ. HRMS-ESI: m/z Calcd for 151.794, 158.556, 144.145, 138.465, 133.274, 127.636, 123.051,
C41H41FN5O7 [MþH]þ: 734.2990; Found: 734.2997. 122.461, 115.490, 112.195, 111.969, 109.869, 109.366, 106.631,
72.698, 61.987, 49.584, 44.947, 44.590, 37.124, 35.243, 36.763,
3.1.2. The general procedure for preparing targets 3c-f and 4c-f 36.697, 14.871, 7.986. ESI-MS m/z: 562 [MþH]þ. HRMS-ESI: m/z
To a solution of methylhydroxylamine or ethylhydroxylamine Calcd for C30H33FN5O5 [MþH]þ: 562.2465; Found: 562.2438.
hydrochloride (15 mmol) and sodium bicarbonate (15 mmol) dis-
solved in water (10 mL) and methanol (10 mL) was added 3a,b or [Link]. 1-Cyclopropyl-7-(4-(3-(3-(ethoxyimino)-5-methyl-2-
4a,b (5 mmol). The reaction mixture was stirred at room temper- oxoindolin-1-yl)propyl)piperazin-1-yl)-6-fluoro-4-oxo-1,4-
ature for 24 h. After removal of the solvent, the residue was diluted dihydroquinoline-3-carboxylic acid (3f). Yellow solid, yield: 57%.
with water (20 mL) and stirred for 10 min, and then filtered. The Mp: 118e119  C. 1H NMR (400 MHz, DMSO‑d6) d 1.09e1.26 (4H, m,
solid crude product was purified by column chromatography (silica 2  cyclopropyl-CH2), 1.35 (3H, t, NOCH2CH3), 2.01 (2H, t, -CH2-),
gel) eluted with DCM to v (DCM):v (MeOH) ¼ 10:1 to give the title 2.23 (3H, s, CH3), 3.14 (4H, s, piperazine-4H), 3.34 (4H, s, piperazine-
3c-f and 4c-f. 4H), 3.66e3.68 (1H, m, cyclopropyl-CH), 3.88 (2H, t, -CH2-), 4.17
(2H, t, -CH2-), 4.38 (2H, q, NOCH2CH3), 7.13e7.18 (1H, m, Ar-H), 7.24
[Link]. 1-Cyclopropyl-6-fluoro-7-(4-(3-(3-(methoxyimino)-2- (1H, d, Ar-H), 7.45 (1H, d, Ar-H), 7.67 (1H, s, Ar-H), 7.78 (1H, d, Ar-H),
oxoindolin-1-yl)propyl)piperazin-1-yl)-4-oxo-1,4-dihydroquinoline- 8.40 (1H, s, C2-H). 13C NMR (DMSO‑d6, 100 MHz) dC: 172.093,
3-carboxylic acid (3c). Yellow solid, yield: 53%. Mp: 149e151  C. 1H 164.779, 162.891, 156.786, 151.86, 148.576, 143.817, 141.980,
NMR (400 MHz, DMSO‑d6) d 1.10e1.26 (4H, m, 2  cyclopropyl- 138.509, 133.500, 132.056, 128.119, 122.636, 121.134, 115.519,
CH2), 2.02 (2H, t, -CH2-), 2.92 (4H, s, piperazine-4H), 3.24 (4H, s, 112.289, 109.679, 106.809, 79.770, 79.442, 72.654, 62.002, 49.060,
piperazine-4H), 3.67e3.68 (1H, m, cyclopropyl-CH), 3.90 (2H, t, 44.240, 37.138, 35.257, 26.726, 20.930, 20.799, 14.973, 8.002. ESI-MS
-CH2-), 4.15e4.21 (5H, m, -CH2- and NOCH3), 7.05 (1H, t, Ar-H), 7.29 m/z: 576 [MþH]þ. HRMS-ESI: m/z Calcd for C31H35FN5O5 [MþH]þ:
(1H, d, Ar-H), 7.41e7.46 (2H, m, Ar-H), 7.78 (1H, t, Ar-H), 7.92 (1H, d, 576.2622; Found: 576.2637.
Ar-H), 8.40 (1H, s, C2-H). 13C NMR (CDCl3, 100 MHz) dC: 173.237,
165.536, 163.642, 157.415, 148.571, 143.875, 138.057, 132.800, [Link]. 3-(2-(methoxyimino)-3-oxoindolin-1-yl)propyl 1-
131.582, 127.871, 122.577, 120.857, 115.724, 113.565, 109.494, cyclopropyl-7- (4-(3-(3-(methoxyimino)-2-oxoindolin-1-yl)propyl)
105.013, 64.707, 61.659, 58.458, 52.953, 49.845, 18.163, 44.036, piperazin-1-yl)-6-fluoro-4-oxo-1,4-dihydroquinoline-3-carboxylate
36.949, 34.783, 26.814, 8.265. ESI-MS m/z: 548 [MþH]þ. HRMS-ESI: (4c). Yellow solid, yield: 36%. Mp: 162e164  C. 1H NMR (400 MHz,
m/z Calcd for C29H31FN5O5 [MþH]þ: 548.2309; Found: 548.2289. DMSO‑d6) d 1.11e1.28 (4H, m, 2  cyclopropyl-CH2), 1.86 (2H, t,
-CH2-), 2.00 (2H, t, -CH2-), 2.68 (2H, t, -CH2-), 3.23 (4H, s,
[Link]. 1-Cyclopropyl-6-fluoro-7-(4-(3-(3-(methoxyimino)-5- piperazine-4H), 3.34 (4H, s, piperazine-4H), 3.65e3.67 (1H, m,
methyl-2-oxoindolin-1-yl)propyl)piperazin-1-yl)-4-oxo-1,4- cyclopropyl-CH), 3.77 (2H, t, -CH2-), 3.87 (2H, t, -CH2-), 4.10e4.21
dihydroquinoline-3-carboxylic acid (3d). Yellow solid, yield: 37%. (8H, m, -CH2- and 2  NOCH3), 7.05e7.10 (2H, m, Ar-H), 7.20 (1H, d,
Mp: 141e143  C. 1H NMR (400 MHz, DMSO‑d6) d 1.09e1.26 (4H, m, Ar-H), 7.28 (1H, d, Ar-H), 7.42e7.50 (3H, m, Ar-H), 7.76e7.89 (3H, m,
2  cyclopropyl-CH2), 2.00 (2H, t, -CH2-), 2.22 (3H, s, CH3), 3.10 (4H, Ar-H), 8.40 (1H, s, C2-H). 13C NMR (CDCl3, 100 MHz) dC: 173.201,
s, piperazine-4H), 3.37 (4H, s, piperazine-4H), 3.60e3.66 (1H, m, 165.355, 163.853, 163.635, 148.520, 143.676, 143.416, 138.042,
cyclopropyl-CH), 3.86 (2H, t, -CH2-), 4.15e4.20 (5H, m, -CH2- and 132.771, 132.654, 128.066, 127.871, 123.394, 123.212, 122.913,
NOCH3), 7.12e7.15 (1H, m, Ar-H), 7.23 (1H, d, Ar-H), 7.44 (1H, d, Ar- 120.857, 115.804, 115.702, 113.361, 113.150, 109.854, 109.132,
H), 7.64 (1H, d, Ar-H), 7.77 (1H, d, Ar-H), 8.38 (1H, s, C2-H). 13C NMR 108.709, 105.385, 65.049, 64.852, 64.707, 61.710, 54.907, 52.122,
(CDCl3, 100 MHz) dC: 172.034, 164.634, 162.752, 154.156, 151.706, 48.090, 37.671, 36.963, 34.827, 26.755, 23.343, 8.210. ESI-MS m/z:
148.556, 143.897, 141.965, 138.385, 133.558, 132.093, 128.177, 764 [MþH]þ. HRMS-ESI: m/z Calcd for C41H43FN7O7 [MþH]þ:
122.745, 115.417, 112.275, 109.657, 109.344, 106.879, 64.765, 61.965, 764.3208; Found: 764.3230.
48.112, 47.988, 37.109, 35.279, 26.683, 20.842, 7.988. ESI-MS m/z:
562 [MþH]þ. HRMS-ESI: m/z Calcd for C30H33FN5O5 [MþH]þ: [Link]. 3-(2-(methoxyimino)-5-methyl-3-oxoindolin-1-yl)propyl 1-
562.2465; Found: 562.2454. cyclopropyl-6- fluoro-7-(4-(3-(3-(methoxyimino)-5-methyl-2-
oxoindolin-1-yl)propyl)piperazin-1-yl)-4-oxo-1,4-dihydroquinoline-
[Link]. 1-Cyclopropyl-7-(4-(3-(3-(ethoxyimino)-2-oxoindolin-1-yl) 3-carboxylate (4d). Yellow solid, yield: 27%. Mp: 136e138  C. 1H
propyl)piperazin-1-yl)-6-fluoro-4-oxo-1,4-dihydroquinoline-3- NMR (400 MHz, DMSO‑d6) d 1.10e1.26 (4H, m, 2  cyclopropyl-
 R. Wang et al. / European Journal of Medicinal Chemistry 156 (2018) 580e586 585

CH2), 1.78 (2H, t, -CH2-), 2.00 (2H, t, -CH2-), 2.21 (3H, s, CH3), 2.27 0.06, 0.03, 0.015 and 0.008 mg/mL. Petri dishes were incubated
(3H, s, CH3), 2.41 (2H, t, -CH2-), 2.52 (4H, s, piperazine-4H), 3.19 (4H, with 104 colony-forming units (cfu) and incubated at 35  C for
s, piperazine-4H), 3.62e3.64 (1H, m, cyclopropyl-CH), 3.74 (2H, t, 18e24 h.
-CH2-), 3.86 (2H, t, -CH2-), 4.14e4.20 (8H, m, -CH2- and 2  NOCH3),
7.05e7.40 (5H, m, Ar-H), 7.64e7.74 (3H, m, Ar-H), 8.37 (1H, s, C2-H).
13 3.3. Antimycibacterial MIC determination
C NMR (CDCl3, 100 MHz) dC: 173.215, 165.581, 165.144, 163.897,
163.657, 151.998, 148.513, 143.853, 143.591, 141.506, 140.981,
Conjugates 3a-f, 4a-f along with CPFX, RIF and INH were dis-
138.035, 132.953, 132.873, 132.501, 128.695, 128.491, 115.680,
solved in DMSO and evaluated in vitro activity against MTB H37Rv
113.303, 113.070, 109.723, 108.848, 108.440, 105.472, 62.129, 64.954,
and MDR-TB via rapid direct susceptibility test technique [34]. The
64.787, 64.634, 61.717, 58.473, 54.805, 52.020, 47.973, 37.590,
wells of a sterile 48-well plate were filled with 100 mL two-fold
36.956, 34.871, 26.734, 23.227, 20.952, 8.192. ESI-MS m/z: 792
diluted tested compounds and 100 mL MTB H37Rv or MDR-TB
[MþH]þ. HRMS-ESI: m/z Calcd for C43H47FN7O7 [MþH]þ: 792.3521;
suspension containing 4  103 mg cells. Pure medium replaced
Found: 792.3537.
the diluted compounds in two wells as the positive control of
growth, and deionized water instead of the culture in other two
[Link]. 3-(2-(ethoxyimino)-3-oxoindolin-1-yl)propyl 1-cyclopropyl-
wells as the negative control of growth in the plates. The plates
7- (4-(3-(3-(ethoxyimino)-2-oxoindolin-1-yl)propyl)piperazin-1-yl)-
were covered and sealed, then incubated at 37  C in a wet box. The
6-fluoro-4-oxo-1,4-dihydroquinoline-3-carboxylate (4e). Yellow
positive and negative control wells should show obvious difference
solid, yield: 47%. Mp: 157e159  C. 1H NMR (400 MHz, DMSO‑d6)
after 3 days. The MIC was determined by observing the quantity
d 1.10e1.26 (4H, m, 2  cyclopropyl-CH2), 1.34 (6H, t,
and state of the cells in each test well by a continuous visual high
2  NOCH2CH3), 1.86 (2H, t, -CH2-), 2.02 (2H, t, -CH2-), 2.48 (2H, t,
magnification system, and re-determined 7 days later. The MIC is
-CH2-), 3.51 (4H, s, piperazine-4H), 3.16 (4H, s, piperazine-4H),
defined as the concentration of the compound required to give
3.65e3.67 (1H, m, cyclopropyl-CH), 3.79 (2H, t, -CH2-), 3.88 (2H,
complete inhibition of bacterial growth.
t, -CH2-), 4.18 (2H, t, -CH2-), 4.42 (4H, q, 2  NOCH2CH3), 7.05e7.10
(2H, m, Ar-H), 7.20 (1H, d, Ar-H), 7.28 (1H, d, Ar-H), 7.42e7.51 (3H,
m, Ar-H), 7.74 (1H, d, Ar-H), 7.82 (1H, d, Ar-H), 7.86 (1H, d, Ar-H), 3.4. Cytotoxicity
8.39 (1H, s, C2-H). 13C NMR (CDCl3, 100 MHz) dC: 173.084, 165.618,
165.129, 163.868, 163.751, 154.572, 152.100, 148.425, 143.839, The synthesized conjugates 3a-f and 4a-f were dissolved in
143.598, 143.401, 138.050, 132.616, 132.333, 127.944, 127.732, DMSO and examined for toxicity (CC50) in a mammalian VERO cell
122.957, 122.774, 115.964, 115.797, 113.390, 113.157, 110.029, 109.176, line at concentrations from 1000 to 4 mg/mL [35]. The VERO cells
108.615, 104.998, 73.310, 73.062, 72.924, 61.681, 55.191, 52.567, were maintained in culture medium (Minimum Essential Medium
49.169, 49.125, 37.882, 36.941, 34.652, 26.901, 24.080, 24.021, with Earle's salt, supplemented with 10% fetal bovine serum) at
14.703, 8.206. ESI-MS m/z: 792 [MþH]þ. HRMS-ESI: m/z Calcd for 37  C under 5% CO2. Cells were seeded in 96-well plates at the
C43H47FN7O7 [MþH]þ: 792.3521; Found: 792.3539. plating density of 1  104 cells per well and allowed to recover for
24 h. Culture medium was replaced by assay medium containing
[Link]. 3-(2-(ethoxyimino)-5-methyl-3-oxoindolin-1-yl)propyl 1- the compound to be tested or drug-free. After 72 h of exposure,
cyclopropyl-7- (4-(3-(3-(ethoxyimino)-5-methyl-2-oxoindolin-1-yl) cells were harvested and cell viability was assessed by MTT assay.
propyl)piperazin-1-yl)-6-fluoro-4-oxo-1,4-dihydroquinoline-3- The CC50 values were calculated by Bliss analyses.
carboxylate (4f). Yellow solid, yield: 14%. Mp: 144e146  C. 1H NMR
(400 MHz, DMSO‑d6) d 1.09e1.26 (4H, m, 2  cyclopropyl-CH2), 1.36
3.5. Pharmacokinetic Profiles
(6H, t, 2  NOCH2CH3), 1.79 (2H, t, -CH2-), 2.00 (2H, t, -CH2-), 2.22
(3H, s, CH3), 2.29 (3H, s, CH3), 2.40 (2H, t, -CH2-), 2.52 (4H, s,
Pharmacokinetic Profiles determination SPF female ICR mice
piperazine-4H), 3.19 (4H, s, piperazine-4H), 3.62e3.64 (1H, m,
weighing 20e25 g were used in the pharmacokinetic study. The
cyclopropyl-CH), 3.75 (2H, t, -CH2-), 3.86 (2H, t, -CH2-), 4.18 (2H, t,
rats were fasted overnight before dosing. Every treatment group
-CH2-), 4.43 (4H, q, 2  NOCH2CH3), 7.07 (1H, d, Ar-H), 7.16 (1H, d,
contained 3 mice. Mice were dosed with the tested compounds
Ar-H), 7.21 (1H, d, Ar-H), 7.28 (1H, d, Ar-H), 7.41 (1H, d, Ar-H),
along with CPFX suspension at 50 mg/kg (p.o.). Compounds were
7.66e7.77 (3H, m, Ar-H), 8.42 (1H, s, C2-H). 13C NMR (CDCl3,
suspended in 0.5% CMC for oral administration. Blood was collected
100 MHz) dC: 173.230, 165.406, 164.036, 163.824, 148.527, 143.802,
from the jugular vein of each animal at the following times after
143.708, 141.513, 140.986, 138.064, 132.909, 132.727, 132.370,
administration of drugs: 0.25, 0.5, 1, 2, 4, 6, 8 and 24 h after a single
128.615, 128.367, 115.950, 115.804, 113.485, 113.259, 109.993,
oral dosing. All blood samples were centrifuged at 3000 r/min for
108.855, 108.352, 105.304, 73.303, 73.092, 72.895, 61.768, 54.929,
10 min to obtain serum which was then stored at 20  C. 150 mL of
52.166, 48.345, 48.301, 37.678, 36.956, 34.769, 26.836, 23.489,
the serum was added to 500 mL of acetonitrile and the mixture was
21.025, 20.966, 14.776, 14.725, 8.214. ESI-MS m/z: 820 [MþH]þ.
centrifuged at 13000 r/min for 10 min to remove protein. The su-
HRMS-ESI: m/z Calcd for C45H51FN7O7 [MþH]þ: 820.3834; Found:
pernatant was dried and dissolved in 100 mL of acetonitrile, the
820.3860.
solution was centrifuged at 13000 r/min for 10 min. The superna-
tant was moved to a sample bottle for HPLC analysis. Total area
3.2. Antibacterial MIC determination
under the concentration time curve (AUC), the elimination half-
time (t1/2), the peak concentration (Cmax) and the time to reach
All hybrids were screened for their in vitro antibacterial activity
peak concentration (Tmax) of samples were determined directly
against representative Gram-positive and Gram-negative strains
from the experimental data using WinNonlin V6.2.1.
(from Chinese Center for Disease Control and Prevention), by
means of standard two-fold serial dilution method using agar
media [38]. Hybrids (10.0 mg) were dissolved in 0.1 N NaOH solu- Appendix A. Supplementary data
tion and water (10 mL). Further progressive two folds serial dilution
with melted Mueller-Hinton agar was performed to obtain the Supplementary data related to this article can be found at
required concentrations of 128, 64, 32, 16, 8, 4, 2, 1, 0.5, 0.25, 0.125, [Link]
586 R. Wang et al. / European Journal of Medicinal Chemistry 156 (2018) 580e586

References Chem. 13 (2005) 5774e5778.

[19] D. Sriram, A. Aubry, P. Yogeeswaria, Bioorg. Med. Chem. Lett 16 (2006)
2982e2985.
[1] Y.Q. Hu, S. Zhang, Z. Xu, Z.S. Lv, M.L. Liu, L.S. Feng, Eur. J. Med. Chem. 141
[20] S.N. Pandeya, D. Sriram, P. Yogeeswari, S. Ananthan, Chemotherapy 47 (2001)
(2017) 335e345.
266e269.
[2] G.F. Zhang, S. Zhang, B.F. Pan, M.L. Liu, L.S. Feng, Eur. J. Med. Chem. 143 (2018)
[21] S. Talath, B.A. Bhongade, Am. J. PharmTech Res. 3 (2013) 570e581.
710e723.
[22] T. Aboul-Fadl, F.A.S. Bin-Jubair, O. Aboul-Wafa, Eur. J. Med. Chem. 45 (2010)
[3] World Health Organization, Global Tuberculosis Report, 2017.
4578e4586.
[4] GBD 2013 mortality and causes of death collaborators, Lancet 385 (2015)
[23] T.R. Bal, B. Anand, P. Yogeeswari, D. Sriram, Bioorg. Med. Chem. Lett 15 (2005)
117e171.
4451e4455.
[5] R.J. Kerns, M.J. Rybak, G.W. Kaatz, F. Vaka, R. Cha, R.G. Grucz, V.U. Diwadkar,
[24] D. Sriram, T.R. Bal, P. Yogeeswari, Med. Chem. Res. 14 (2005) 211e228.
T.D. Ward, Bioorg. Med. Chem. Lett 13 (2003) 1745e1749.
[25] D. Sriram, T.R. Bal, P. Yogeeswari, J. Pharm. Pharmaceut. Sci. 8 (2005)
[6] R. Mather, L.M. Karenchak, E.G. Romanowski, R.P. Kowalski, Am. J. Oph-
565e577.
thalmol. 133 (2002) 463e466.
[26] D. Banerjee, P. Yogeeswari, P. Bhat, A. Thomas, M. Srividya, D. Sriram, Eur. J.
[7] Y.L. Fan, J.B. Wu, X.W. Cheng, F.Z. Zhang, L.S. Feng, Eur. J. Med. Chem. 146
Med. Chem. 46 (2011) 106e121.
(2018) 554e563.
[27] D. Sriram, P. Yogeeswari, G. Gopal, Eur. J. Med. Chem. 40 (2005) 1373e1376.
[8] G.F. Zhang, X.F. Liu, S. Zhang, B.F. Pan, M.L. Liu, Eur. J. Med. Chem. 146 (2018)
[28] D. Sriram, P. Yogeeswari, K. Meena, Pharmazie 61 (2006) 274e277.
599e612.
[29] Z. Xu, X.F. Song, M. Qiang, Z.S. Lv, J. Heterocycl. Chem. 54 (2017) 3735e3741.
[9] Y.L. Fan, X.W. Cheng, J.B. Wu, M. Liu, F.Z. Zhang, Z. Xu, L.S. Feng, Eur. J. Med.
[30] L.S. Feng, M.L. Liu, B. Wang, Y. Chai, X.Q. Hao, S. Meng, H.Y. Guo, Eur. J. Med.
Chem. 146 (2018) 1e14.
Chem. 45 (2010) 3407e3412.
[10] Y.Q. Hu, C. Gao, S. Zhang, L. Xu, Z. Xu, L.S. Feng, X. Wu, F. Zhao, Eur. J. Med.
[31] L.S. Feng, M.L. Liu, Y.B. Zhang, Chem. Res. Chin. Univ. 28 (2012) 61e66.
Chem. 139 (2017) 22e47.
[32] Y.Q. Hu, L.D. Meng, M. Qiang, X.F. Song, J. Heterocycl. Chem. 54 (2017)
[11] X.D. Jia, S. Wang, M.H. Wang, M.L. Liu, G.M. Xia, X.J. Liu, Y. Chai, H.W. He, Chin.
3725e3729.
Chem. Lett. 28 (2017) 235e239.
[33] Z. Xu, M. Qiang, Z.S. Lv, Asian J. Chem. 29 (2017) 1039e1041.
[12] Z. Xu, S. Zhang, C. Gao, F. Zhao, Z.S. Lv, L.S. Feng, Chin. Chem. Lett. 28 (2017)
[34] Z. Xu, S. Zhang, X.F. Song, M. Qiang, Z.S. Lv, Bioorg. Med. Chem. Lett 27 (2017)
159e167.
3643e3646.
[13] S. Zhang, Z. Xu, C. Gao, Q.C. Ren, C. Le, Z.S. Lv, L.S. Feng, Eur. J. Med. Chem. 138
[35] Z. Xu, X.F. Song, Y.Q. Hu, M. Qiang, Z.S. Lv, Eur. J. Med. Chem. 138 (2017)
(2017) 501e513.
66e71.
[14] Y.Q. Hu, S. Zhang, F. Zhao, C. Gao, L.S. Feng, Z.S. Lv, Z. Xu, X. Wu, Eur. J. Med.
[36] C.X. Zhi, Z.Y. Long, A. Manikowski, J. Comstock, W.C. Xu, N.C. Brown,
Chem. 133 (2017) 255e267.
P.M. Tarantino, K.A. Holm, E.J. Dix, G.E. Wright, M.H. Barnes, M.M. Butler,
[15] Y.Q. Hu, Z. Xu, S. Zhang, X. Wu, J.W. Ding, Z.S. Lv, L.S. Feng, Eur. J. Med. Chem.
K.A. Foster, W.A. Lamarr, B. Bachand, R. Bethell, C. Cadilhac, S. Charron,
136 (2017) 122e130.
S. Lamothe, I. Motorina, R. Storer, J. Med. Chem. 49 (2006) 1455e1465.
[16] M.T. Gabr, N.S. El-Gohary, E.R. Elbendary, M.M. El-Kerdawy, N. Ni, Eur. J. Med.
[37] X. Yan, Z.S. Lv, J. Wen, S.J. Zhao, Z. Xu, Eur. J. Med. Chem. 143 (2018) 899e904.
Chem. 128 (2017) 36e44.
[38] X.H. Jin, Y. Xu, X.H. Yang, X.L. Chen, M.H. Wu, J.G. Guan, L.S. Feng, Curr. Top.
[17] T.L. Devale, J. Parikh, P. Miniyar, P. Sharma, B. Shrivastava, P. Murumlkar,
Med. Chem. 17 (2018) 3213e3218.
Bioorg. Chem. 70 (2017) 256e266.
[18] D. Sriram, P. Yogeeswaria, J.S. Basha, D.R. Radha, V. Nagaraja, Bioorg. Med.