Acta Pharmaceutica Sinica 2010, 45 (7): 860868

860

Synthesis and in vitro antibacterial activity of

7-(4-alkoxyimino-3-methyl-3-methylaminopiperidin-1-yl)quinolones
WAN Zhi-long, CHAI Yun, LIU Ming-liang*, GUO Hui-yuan, SUN Lan-ying
(Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100050, China)

Abstract: To explore new agents of quinolone derivatives with high antibacterial activity, 7-(4-alkoxyimino3-methyl-3-methylaminopiperidin-1-yl)quinolones were designed and synthesized, and their activity against
gram-positive and gram-negative strains was tested in vitro. Sixteen target compounds were obtained. Their
structures were established by 1H NMR, HRMS and X-ray crystallographic analysis. Compounds 14k and
14m14o show good antibacterial activity against the tested five gram-positive strains and five gram-negative
strains (MIC: 0.2516 gmL1), of which the most active compound 14o is 8-fold more potent than levofloxacin
against S. pneumoniae (MIC: 4 gmL1), and comparable to levofloxacin against S. aureus, S. epidermidis,
E. faecalis and E. coli (MIC: 0.251 gmL1), but generally less potent than gemifloxacin.
Key words : fluoroquinolone; chemical synthesis; in vitro antibacterial activity
CLC number: R916
Document code: A
Article ID: 0513-4870 (2010) 07-0860-09

7-(3--3--4--1-)

, *, ,

(, 100050)
: , 16 7-(3--3--4--1-)
, 1H NMR HRMS , X-
14k 14m14o 5 5
(MIC: 0.2516 gmL1), 14o (MIC:
0.251 gmL1) , (MIC: 4 gmL1) 8 ,

: ; ;

The quinolones were evolved from agents used

only for the treatment of urinary tract infections (UTIs)
to the latest fluoroquinolones with a remarkably broad
spectrum of activity and excellent pharmacokinetics
allowing once-daily dosing and thus improving patient

compliance[1]. New quinolones, such as gatifloxacin,

moxifloxacin and gemifloxacin (GMFX), have been
prescribed extensively for the treatment of respiratory
tract infections, UTIs, sexually transmitted diseases
and infections of the skin and soft tissue. However, as
use of fluoroquinolones has increased, the continuous

Received 2009-10-30.
Project supported by the Center Commonweal Basic Scientific Research
Operation Foundation (No. IMBF-20060404).
*
Corresponding author Tel / Fax: 86-10-63036965,
E-mail: lmllyx@yahoo.com.cn

increase in the resistance among bacteria is generally

recognized[2]. Thus, recent efforts have been directed
toward the synthesis of new quinolones that can

WAN Zhi-long, et al: Synthesis and in vitro antibacterial activity of 7-(4-alkoxyimino-3-methyl-3methylaminopiperidin-1-yl)quinolones

provide improved antibacterial activity against grampositive cocci and anaerobes and against fluoroquinolone-resistant strains, while retaining the good gramnegative activity of early fluoroquinolones, such as
ciprofloxacin and ofloxacin.
The structure-activity relationship (SAR) study of
quinolone antibacterial agents showed that the basic
group at the C-7 position is the most adaptable site for
chemical change and an area that greatly influences
their potency, spectrum and safety[3]. In general, 5and 6-membered nitrogen heterocycles including
piperazinyl, pyrrolidinyl and piperidinyl type side
chains have been proven to be the optimal substituents.
However, quinolone antibacterial agents possessing
piperidine substitution reported in the literature were
significantly fewer than that of piperazinyl- and
pyrrolidinyl-based derivatives.
As part of an ongoing program to find potent new
quinolones displaying strong gram-positive activity,
we have focused our attention on introducing new
functional groups to the piperidine ring[46]. Methyl
group was introduced into 3-position of pyrrolidine
ring for the purpose of increasing gram-positive
antibacterial activity[7]. For example, as an analogue
of GMFX, DW286 (Figure 1) possessing an additional
methyl group at 3-position of pyrrolidine ring displays
far more potent antibacterial activity than GMFX
against important gram-positive organisms, methicillinresistant staphylococcus aureus (MRSA) and ofloxacin
resistant organisms, and with excellent pharmacokinetic
profiles[8].
In this study, structural modifications
were made on the basis of 7-(4-alkoxyimino-3-methylaminopiperidin-1-yl)-1, 8-naphthyridones (DZH, Figure
1) exhibiting significant in vitro antibacterial activity
against gram-positive organisms, which is more
potent than GMFX, linezolid and vancomycin[9]. New
piperidine derivatives and a series of fluoroquinolone
compounds derived from these amines at the C-7 position
were designed and synthesized. These piperidine

Figure 1

Chemical structures of GMFX, DW286 and DZH

861

derivatives are structurally novel, having a methylamino

and a methyl group at 3-position and an alkoxyimino
group at 4-position of the piperidine ring.
Our
primary objective was to optimize the potency of these
compounds against gram-positive and gram-negative
organisms.

Results and discussion

1

Chemistry
The detailed synthetic pathways to 4-alkoxyimino3-methyl-3-methylaminopiperidine dihydrochlorides 11a,
11b starting from 1-tert-butoxycarbonyl-4-piperidone
(1) and novel fluoroquinolones 14a14p were depicted
in Schemes 1 and 2, respectively.
Sixteen target compounds 14a14p were synthesized in our study, and their structures were established
by 1 H NMR and HRMS spectra. Their physical
constants and spectral data were depicted in Table 1.
It is obvious that the target compounds 14a14p
and intermediates 11a, 11b are all racemes. However,
it was necessary to determine the geometry of the
oximino group because it could exist in the E or Z
configuration. Although we were not successful in
preparing X-ray quality single crystals of compounds
14a14p, we were able to obtain X-ray data for the
intermediate 10a. As expected, the six-membered
piperidine ring adopts a chair conformation and the
methyloxime geometry of 10a was confirmed to have
the E-configuration [10].
2 Pharmacology
The synthesized fluoroquinolones 14a14p were
evaluated for their in vitro antibacterial activity against
representative gram-positive and gram-negative strains
using standard techniques. The minimum inhibitory
concentration (MIC) values of the compounds against
the ten strains are presented in Table 2. The antibacterial activity of the reference compounds GMFX and
levofloxacin (LVFX) are also included.

Acta Pharmaceutica Sinica 2010, 45 (7): 860868

862

(a) (CH3O) 2CO, 70% NaH, C6H5CH 3, 80 ; (b) CH3I, K2 CO3, (CH 3) 2CO, 40 ; (c) RONH 2HCl, Et 3N, EtOH; 5560 ; (d) NaOH,
MeOH, 50 , and then HOAc; (e) ClCOOBu-i, Et 3N, 14 to 12 ; (f) NH3 gas, 0 5 ; (g) NaBrO, CH3CN, 5 ; (h) Boc2 O,
EtOH, rt; (i) CH3I, 70% NaH, THF, rt; (j) HCl gas, CH2Cl2, rt.
Scheme 1

Compd.

Synthesis of 4-alkoxyimino-3-methyl-3-methylaminopiperidine dihydrochlorides 11a, 11b

R1

Compd.

CH3

14b

CH3

14d

CH3

14f

COCHF2

R1

14a

14c

14e

COCHF2

14g

CF

C2H4 F

CH3

14h

CF

C2H4 F

C2H5

14i

CF

C2H5

CH3

14j

CF

C2H5

C2H5

14k

CF

CH3

14l

CF

C2H5

14m

COCH3

CH3

14n

COCH3

C2H5

CH3

14p

2,4-F2 -C6 H3

14o

Scheme 2

Synthesis of target compounds 14a14p

C2H5
2,4-F2 -C6 H3

C2H5
C2H5

C2H5

WAN Zhi-long, et al: Synthesis and in vitro antibacterial activity of 7-(4-alkoxyimino-3-methyl-3methylaminopiperidin-1-yl)quinolones

Table 1

863

Physical constants and spectral data of compounds 14a14p

Compd.

Yield
/%

mp/C

14a

76.7

216217

14b

66.3

199200

14c

74.8

203205

14d

72.5

210212

14e

57.6

206207

14f

54.8

186189

14g

65.6

201204

14h

63.2

177179

14i

69.8

204206

14j

62.7

185187

14k

72.1

207208

14l

75.7

194195

14m

52.9

181183

14n

63.3

159161

14o

48.2

159162

14p

76.7

216217

H NMR / (400 MHz, CDCl3 )

HR-FAB-MS
Calcd (Found)

1.071.11 (m, 2H, cyclopropyl), 1.241.30 (m, 5H, cyclopropyl, CH3 ), 2.25 (s, 3H, NCH3 ), 2.522.59 [C20 H25FN5 O4 +H]+
(m, 1H, piperidine), 3.053.12 (m, 1H, piperidine), 3.433.53 (m, 2H, piperidine), 3.613.67 (m, 1H,
418.189 1
cyclopropyl), 3.90 (s, 3H, OCH3 ), 4.284.31 (m, 1H, piperidine), 4.404.45 (m, 1H, piperidine),
(418.187 0)
8.078.10 (d, J = 13.2 Hz, 1H, H-5 quinolone), 8.73 (s, 1H, H-2 quinolone)
1.081.09 (m, 2H, cyclopropyl), 1.261.34 (m, 8H, cyclopropyl, OCH2CH3, CH3 ), 2.28 (s, 3H, [C21 H27FN5 O4 +H]+
NCH3 ), 2.602.61 (m, 1H, piperidine), 3.073.11 (m, 1H, piperidine), 3.513.58 (m, 2H, piperidine),
432.204 7
3.663.68 (m, 1H, cyclopropyl), 4.134.18 (q, J = 7.2 Hz, 2H, OCH2CH3), 4.314.38 (m, 2H,
(432.205 4)
piperidine), 8.078.10 (d, J = 13.2 Hz, 1H, H-5 quinolone), 8.73 (s, 1H, H-2 quinolone)
1.16 (s, 3H, CH3 ), 2.142.16 (d, J = 10.4 Hz, 3H, NCH3 ), 2.332.39 (m, 1H, piperidine), 2.802.90
(m, 1H, piperidine), 3.183.35 (m, 2H, piperidine), 3.86 (s, 3H, OCH3), 3.954.03 (m, 2H, piperidine),
7.077.09 (m, 2H, ArH), 7.397.44 (1H, m, ArH), 8.108.14 (d, J = 13.2 Hz, 1H, H-5 quinolone),
8.67 (s, 1H, H-2 quinolone)
1.20 (s, 3H, CH3), 1.231.26 (t, J = 7.2 Hz, 3H, OCH2CH3), 2.142.17 (d, J = 10.0 Hz, 3H, NCH3),
2.362.37 (m, 1H, piperidine), 2.822.89 (m, 1H, piperidine), 3.193.36 (m, 2H, piperidine), 3.96
4.01 (m, 2H, piperidine), 4.084.13 (q, J = 7.2 Hz, 2H, OCH2CH3), 7.107.12 (m, 2H, ArH), 7.40
7.44 (1H, m, ArH), 8.108.14 (d, J = 13.2 Hz, 1H, H-5 quinolone), 8.67 (s, 1H, H-2 quinolone)
0.870.92 (m, 1H, cyclopropyl), 1.041.11 (m, 1H, cyclopropyl), 1.191.39 (m, 5H, cyclopropyl and
CH3 ), 2.28 (s, 3H, NCH3 ), 2.402.48 (m, 1H, piperidine), 3.173.24 (m, 2H, piperidine), 3.363.42
(m, 2H, piperidine), 3.533.56 (m, 1H, piperidine), 3.91 (s, 3H, OCH3), 4.074.12 (m, 1H, cyclopropyl),
6.576.94 (t, J = 74.8 Hz, 1H, OCHF2 ), 8.038.06 (d, J = 11.6 Hz, 1H, H-5 quinolone), 8.85 (s, 1H,
H-2 quinolone)
0.831.08 (m, 3H, cyclopropyl), 1.251.57 (m, 7H, cyclopropyl, CH3 and OCH2CH3 ), 2.31 (s, 3H,
NCH3 ), 2.402.48 (m, 1H, piperidine), 3.213.24 (m, 2H, piperidine), 3.433.52 (m, 3H, piperidine),
4.074.12(m, 1H, cyclopropyl), 4.084.13 (q, J = 6.8 Hz, 2H, OCH2CH3 ), 6.717.18 (t, J = 74.8 Hz,
1H, OCHF2 ), 8.048.07 (d, J = 12.0 Hz, 1H, H-5 quinolone), 8.85 (s, 1H, H-2 quinolone)
1.27 (s, 3H, CH3 ), 2.27 (s, 3H, NCH3), 2.31-2.34 (m, 1H, piperidine), 3.213.43 (m, 5H, piperidine),
3.91 (s, 3H, OCH3), 4.764.88 (m, 4H, CH2CH2F), 7.998.02 (d, J = 11.2 Hz, 1H, H-5 quinolone),
8.63 (s, 1H, H-2 quinolone)
1.261.29 (t, J = 6.8 Hz, 3H, OCH2CH3 ), 1.39 (s, 3H, CH3 ), 2.30 (s, 3H, NCH3 ), 2.723.00 (m, 1H,
piperidine), 3.243.42 (m, 5H, piperidine), 4.104.17 (q, J = 6.8 Hz, 2H, OCH2CH3 ), 4.714.88 (m,
4H, CH2CH2F), 7.998.02 (d, J = 10.8 Hz, 1H, H-5 quinolone), 8.62 (s, 1H, H-2 quinolone)
1.28 (s, 3H, CH3), 1.541.58 (t, J = 7.0 Hz, 3H, NCH2CH3 ), 2.32 (s, 3H, NCH3 ), 2.352.37 (m, 1H,
piperidine), 3.203.42 (m, 5H, piperidine), 3.91 (s, 3H, OCH3), 4.454.52 (m, 2H, NCH2), 7.988.01
(d, J = 11.2 Hz, 1H, H-5 quinolone), 8.64 (s, 1H, H-2 quinolone)
1.261.30 (m, 6H, CH3 and OCH2CH3 ), 1.541.58 (t, 3H, NCH2CH3), 2.29 (s, 3H, NCH3 ), 2.342.38
(m, 1H, piperidine), 3.233.45 (m, 5H, piperidine), 4.124.18 (q, J = 6.8 Hz, 2H, OCH2CH3), 4.45
4.53 (m, 2H, NCH2 ), 7.998.02 (d, J = 11.6 Hz, 1H, H-5 quinolone), 8.63 (s, 1H, H-2 quinolone)
1.121.22 (m, 2H, cyclopropyl), 1.27 (s, 3H, CH3 ), 1.291.32 (m, 2H, cyclopropyl), 2.30 (s, 3H,
NCH3 ), 2.322.40 (m, 1H, piperidine), 3.203.42 (m, 5H, piperidine), 3.90 (s, 3H, OCH3), 3.994.01
(m, 1H, cyclopropyl), 7.937.96 (d, J = 11.6 Hz, 1H, H-5 quinolone), 8.80 (s, 1H, H-2 quinolone)
1.161.22 (m, 2H, cyclopropyl), 1.261.35 (m, 8H, cyclopropyl, CH3 and OCH2CH3 ), 2.32 (s, 3H,
NCH3), 2.352.43 (m, 1H, piperidine), 3.203.43 (m, 5H, piperidine), 3.994.01 (m, 1H, cyclopropyl),
4.134.18 (q, J = 7.2 Hz, 2H, OCH2CH3 ), 7.937.96 (d, J = 11.6 Hz, 1H, H-5 quinolone), 8.80 (s, 1H,
H-2 quinolone)
1.021.06 (m, 2H, cyclopropyl), 1.221.26 (m, 2H, cyclopropyl), 1.49 (s, 3H, CH3 ), 2.51 (s, 3H,
NCH3 ), 2.702.72 (m, 1H, piperidine), 3.023.06 (m, 1H, piperidine), 3.353.37 (m, 1H, piperidine),
3.483.55 (m, 2H, piperidine), 3.673.69 (m, 1H, piperidine), 3.71 (s, 3H, OCH3 -8 quinolone), 3.94
(s, 3H, OCH3 ), 3.964.02 (m, 1H, cyclopropyl), 7.867.89 (d, J = 11.6 Hz, 1H, H-5 quinolone), 8.83
(s, 1H, H-2 quinolone)
1.011.03 (m, 2H, cyclopropyl), 1.221.31 (m, 5H, cyclopropyl and OCH2CH3 ), 1.49 (s, 3H, CH3 ),
2.50 (s, 3H, NCH3 ), 2.722.74 (m, 1H, piperidine), 3.033.06 (m, 1H, piperidine), 3.353.37 (m, 1H,
piperidine), 3.503.56 (m, 2H, piperidine), 3.703.72 (m, 4H, piperidine and OCH3 ), 3.964.00 (m,
1H, cyclopropyl), 4.024.13 (q, J = 6.8 Hz, 2H, OCH2CH3 ), 7.867.89 (d, J = 11.6 Hz, 1H, H-5
quinolone), 8.83 (s, 1H, H-2 quinolone)

[C23 H23F3 N5 O4 +H]+

490.170 2
(490.169 6)
[C24 H25F3 N5 O4 +H]+
504.185 9
(504.187 7)
[C22 H26F3 N4 O5 +H]+
483.185 5
(483.182 5)

[C23 H28F3 N4 O5 +H]+

497.201 2
(497.201 7)
[C20 H24F3 N4 O4 +H]+
441.175 0
(441.175 6)
[C21 H26F3 N4 O4 +H]+
455.190 6
(455.190 2)
[C20 H25F2 N4 O4 +H]+
423.184 4
(423.183 6)
[C21 H27F2 N4 O4 +H]+
437.200 0
(437.200 5)
[C21 H25F2 N4 O4 +H]+
435.184 4
(435.185 6)
[C22 H27F2 N4 O4 +H]+
449.200 0
(449.201 5)
[C22 H28FN4 O5 +H]+
447.204 4
(447.205 3)

[C23 H30FN4 O5 +H]+

461.220 0
(461.224 1)

1.451.49 (m, 6H, 2CH3), 2.49 (s, 3H, NCH3 ), 2.822.88 (m, 2H, piperidine), 3.353.56 (m, 4H, [C21 H26FN4 O5 +H]+
piperidine), 3.90 (s, 3H, OCH3), 4.364.44 (m, 1H, OCH2), 4.624.71 (m, 1H, OCH2 ), 4.934.95 (m,
433.188 7
1H, NCH), 7.647.66 (d, J = 11.6 Hz, 1H, H-5 quinolone), 8.94 (s, 1H, H-2 quinolone)
(433.188 8)
1.141.29 (m, 9H, 3CH3), 2.122.22 (m, 2H, piperidine), 2.26 (s, 3H, NCH3 ), 3.353.56 (m, 4H, [C22 H28FN4 O5 +H]+
piperidine), 4.114.16 (q, J = 6.8 Hz, 2H, OCH2CH3 ), 4.354.49 (m, 3H, OCH2CH), 7.737.76 (d,
447.204 4
J = 11.2 Hz, 1H, H-5 quinolone), 8.62 (s, 1H, H-2 quinolone)
(447.202 8)

Acta Pharmaceutica Sinica 2010, 45 (7): 860868

864

Table 2 In vitro antibacterial activity of compounds 14a14p against selected strains. S. a.: Staphylococcus aureus ATCC29213;
S. e.: Staphylococcus epidermidis ATCC12228; S. p.: Streptococcus pneumoniae 97100; S. py.: Streptococcus pyogenes 9619; E. f.:
Enterococcus faecalis ATCC29212; E. c.: Escherichia coli ATCC26; K. p.: Klebsiella pneumoniae 7; P. a.: Pseudomonas aeruginosa
17; S. s.: Shigella sonnei 51592; E. c.: Enterobacter cloacae 45301
MIC / gmL 1

Compd.
S. a.

S. e.

S. p.

S. py.

E. f.

E. c.

K. p.

P. a.

S. s.

E.c.

14a

32

128

64

16

64

16

16

14b

128

>128

16

32

32

>128

128

32

14c

32

>128

32

32

32

>128

32

32

14d

>128

>128

128

>128

>128

>128

>128

>128

14e

0.5

64

32

14f

64

64

32

16

128

32

32

14g

14h

64

14i

14j

64

14k

0.25

14l

14m

0.25

14n

0.25

32

64

16

128

16

16

>128

>128

>128

>128

>128

128

64

16

32

64

128

16

16

>128

>128

>128

>128

>128

>128

128

32

16

32

>128

>128

>128

>128

>128

16

0.5

16

0.5

16

16

14o

0.25

0.5

14p

0.25

16

128

128

64

64

64

16

GMFX

<0.03

<0.03

0.06

0.06

0.5

0.125

0.06

0.06

LVFX

0.25

0.5

32

0.5

0.125

The novel fluoroquinolones 14a14p have generally

potent antibacterial activity against the ten strains.
Compounds 14k and 14m14o showed good antibacterial
activity against the tested five gram-positive strains and
five gram-negative strains (MIC: 0.2516 gmL1 ), of
which the most active compound 14o was found to be
8-fold more potent than LVFX against S. pneumoniae
(MIC: 4 gmL1), and comparable to LVFX against
S. aureus, S. epidermidis, E. faecalis and E. coli (MIC:
0.251 gmL1), but generally less potent than GMFX.
3 Conclusion and discussion
In summary, a series of novel 7-(4-alkoxyimino3-methyl-3-methylaminopiperidin-1-yl) fluoroquinolone
derivatives were designed, synthesized and evaluated for
their in vitro antibacterial activity against representative
gram-positive and gram-negative strains. All of the
novel fluoroquinolones demonstrate potent antibacterial
activity, of which compounds 14k and 14m14o show
good antibacterial activity (MIC: 0.2516 gmL1),
and the most active compound 14o is 8-fold more potent
than LVFX against S. pneumoniae, and comparable
to LVFX against S. aureus, S. epidermidis, E. faecalis
and E. coli, but they are generally less potent than
GMFX against the tested gram-positive and gram-

negative strains.
These results show that introduction of another
methyl group into 3-position of piperidine ring causes
reduced antibacterial activity, which is contrary to the
activity profiles of pyrrolidine-containing fluoroquinolones. In addition, the fluoroquinolones featuring
methyloxime-incorporated piperidino-substitution at C-7
position are more active than those of the corresponding
analogues containing ethyloxime.

Experimental
1

General experimental procedures

Melting points were determined on X6 precision
melting-point apparatus and uncorrected. 1H NMR
spectra were determined on a Varian Mercury-400 MHz
spectrometer using tetramethylsilane as an internal
standard. Electrospray ionization (ESI) mass spectra
were obtained on a MDSSCIEX Q-Tap mass spectrometer.
Fast Atom Bombardment (FAB) mass spectra and
HR-MS were obtained on an AutoSpec Ultima-TOF
mass spectrometer. The reference drugs GMFX and
LVFX were synthesized in our lab according to the
references[1113].

WAN Zhi-long, et al: Synthesis and in vitro antibacterial activity of 7-(4-alkoxyimino-3-methyl-3methylaminopiperidin-1-yl)quinolones

2 Chemistry
2.1 Methyl 1-tert-butoxycarbonyl-4-oxopiperidine3-carboxylate (2) To a suspension of 70% sodium
hydride (30.20 g, 0.88 mol) in dry toluene (500 mL),
dimethyl carbonate (43.20 g, 0.48 mol) was added
dropwise in 0.5 h at room temperature under an
atmosphere of nitrogen. After addition of a few drops
of methanol, a solution of 1-tert-butoxycarbonyl-4piperidone (1, 48.00 g, 0.24 mol) dissolved in dry
toluene (200 mL) was added dropwise to the reaction
mixture while stirring at 80 in 1 h. The reaction
mixture was stirred for 3 h at the same temperature and
then cooled to 0 (ice bath), adjusted to pH 66.5
with acetic acid. The resulting mixture was diluted
with cold distilled water (100 mL) and adjusted to pH 8
with 5% sodium hydroxide solution. The toluene
layer was separated and the aqueous layer was extracted
with toluene (100 mL). The combined toluenes were
dried over anhydrous sodium sulfate, and concentrated
under reduced pressure. The solid obtained was dried
in vacuo to give the title compound 2 (54.50 g, 88.0%)
as an off-white solid, mp 3234 . 1H NMR (400
MHz, CDCl3) : 1.47 (s, 9H, Boc), 2.352.38 (t, J = 6.0
Hz, 2H, H-5), 3.543.57 (t, J = 6.0 Hz, 2H, H-6), 3.76
(s, 3H, OCH3), 4.05 (s, 2H, H-2). ESI-MS m/z: 258
[M+H]+.
2.2 Methyl 1-tert-butoxycarbonyl-3-methyl-4-oxopiperidine-3-carboxylate (3)
To a suspension of
compound 2 (53.68 g, 0.21 mol) and anhydrous
potassium carbonate (100.93 g, 0.73 mol) in dry acetone
(500 mL), a solution of methyl iodide (23.38 mL, 0.38
mol) dissolved in dry acetone (250 mL) was added at
room temperature under an atmosphere of nitrogen.
The reaction mixture was heated to 40 and stirred
for 6 h, cooled to room temperature and filtered. The
filtrate was concentrated under reduced pressure. The
residue was diluted with dichloromethane (300 mL)
and washed with distilled water and saturated saline
separately, dried over anhydrous sodium sulfate, filtered
and concentrated. The yellow oily residue was treated
with petroleum ether (300 mL) and filtered. The solid
obtained was washed twice with petroleum ether and
dried in vacuo to give the title compound 3 (45.36 g,
80.1%) as a white solid, mp 4243 . 1 H NMR (400
MHz, CDCl3) : 1.31 (s, 3H, CH3), 1.49 (s, 9H, Boc),
2.452.51 (m, 1H, H-5), 2.76 (s, 1H, H-2), 3.063.10
(m, 1H, H-6), 3.303.37 (m, 1H, H-5), 3.72 (s, 3H,

865

OCH3), 4.12 (s, 1H, H-2), 4.484.51 (m, 1H, H-6).

ESI-MS m/z: 272 [M+H]+.
2.3 Methyl 1-tert-butoxycarbonyl-4-methoxyimino3-methylpiperidine-3-carboxylate (4a) To a stirring
solution of compound 3 (15.00 g, 55.3 mmol) dissolved
in ethanol (50 mL), a solution of methoxylamine
hydrochloride (5.05 g, 60.5 mmol) and triethylamine
(8.37 mL, 60.5 mmol) in 80% ethanol (25 mL) was
added dropwise at 5560 in 15 min. The reaction
mixture was stirred at the same temperature for 1.5 h
and concentrated under reduced pressure. The residue
was diluted with distilled water (30 mL) and extracted
with ethyl acetate (350 mL). The combined extracts
were washed with 1 molL1 HCl (210 mL) and
saturated brine separately, dried over anhydrous sodium
sulfate and concentrated under reduced pressure. The
crude product was recrystallized from petroleum ether
to give the title compound 4a (14.71 g, 88.6%) as a
white solid, mp 7576 . 1H NMR (400 MHz,
CDCl3) : 1.36 (s, 3H, CH3 ), 1.46 (s, 9H, Boc), 2.39
2.40 (m, 1H, piperidine), 2.923.16 (m, 3H, piperidine),
3.70 (s, 3H, COOCH3), 3.86 (s, 3H, NOCH3), 3.82
3.83 (m, 1H, piperidine), 4.324.36 (m, 1H, piperidine).
ESI-MS m/z: 301 [M+H]+.
2.4 Methyl 1-tert-butoxycarbonyl-4-ethoxyimino-3methylpiperidine-3-carboxylate (4b)
The title
compound was obtained in a similar manner as for the
preparation of compound 4a. White solid (90.3%),
mp 5153 . 1 H NMR (400 MHz, CDCl3) : 1.22
1.26 (t, J = 7.2 Hz, 3H, OCH2 CH3), 1.36 (s, 3H, CH3),
1.46 (s, 9H, Boc), 2.422.44 (m, 1H, piperidine), 2.92
3.19 (m, 3H, piperidine), 3.70 (s, 3H, OCH3 ), 3.79
3.80 (m, 1H, piperidine), 4.074.12 (q, J = 7.2 Hz, 2H,
OCH2CH3), 4.304.33 (m, 1H, piperidine). ESI-MS
m/z: 315 [M+H]+.
2.5 1-tert-Butoxycarbonyl-3-carbamyl-4-methoxyimino-3-methylpiperidine (7a) To a stirring solution
of compound 4a (17.00 g, 56.6 mmol) in methanol
(100 mL), a solution of 18% sodium hydroxide solution
(25 mL) was added dropwise at room temperature.
The reaction mixture was heated to 50 and stirred
for 2 h at the same temperature. After removal of the
methanol under reduced pressure, the reaction mixture
was diluted with distilled water (30 mL), adjusted to
pH 66.5 with acetic acid and filtered to give the
acid 5a as a white solid. The solid 5a was dissolved
in dichloromethane (150 mL), and to this solution

866

Acta Pharmaceutica Sinica 2010, 45 (7): 860868

triethylamine (8.8 mL, 63.6 mmol) was added. The

reaction mixture was cooled to 14 to 12 using an
ice-salt bath, isobutyl chloroformate (9.0 mL, 69.2 mmol)
was added and stirred for 0.5 h at the same temperature
to give the ester 6a. To the reaction mixture containing
the ester 6a, ammonia gas was pumped cautiously at
05 for 0.5 h, washed with distilled water and
saturated brine, separately, dried over anhydrous sodium
sulfate and concentrated under reduced pressure. The
resulting yellow residue was recrystallized from ethyl
acetate to give the title compound 7a (13.50 g, 83.6%)
as a white solid, mp 126127 .

H NMR (400 MHz,

CDCl3) : 1.37 (s, 3H, CH3), 1.47 (s, 9H, Boc),

2.142.31 (m, 1H, piperidine), 2.953.10 (m, 3H,
piperidine), 3.723.80 (m, 1H, piperidine), 3.90 (s, 3H,
OCH3), 4.354.37 (m, 1H, piperidine), 5.37 (br, 1H,
CONH), 6.19, 6.78 (2s, 1H, CONH). ESI-MS m/z:
286 [M+H]+.
2.6 1-tert-Butoxycarbonyl-3-carbamyl-4-ethoxyimino3-methylpiperidine (7b) The title compound was
obtained in a similar manner as for the preparation of
compound 7a. White solid (78.6%), mp 108109 .
1

H NMR (400 MHz, CDCl3) : 1.241.27 (t, J = 7.2

Hz, 3H, OCH2CH3), 1.38 (s, 3H, CH3), 1.52 (s, 9H,
Boc), 2.202.35 (m, 1H, piperidine), 2.993.15 (m,
3H, piperidine), 3.753.78 (m, 1H, piperidine), 4.11
4.16 (q, J = 7.2 Hz, 2H, OCH2CH3), 4.334.35 (m, H,
piperidine), 5.46 (br, 1H, CONH), 6.19, 6.66 (2s, 1H,
CONH). ESI-MS m/z: 300 [M+H]+.
2.7 1-tert-Butoxycarbonyl-3-amino-4-methoxyimino3-methylpiperidine (8a) To a solution of compound
7a (2.00 g, 7.0 mmol) in acetonitrile (80 mL), freshly
prepared sodium hypobromite solution[14] (14 mL)
was added dropwise at 05 in 0.5 h. The reaction

mixture was stirred at the room temperature for 2 h, the

organic layer was separated and concentrated under
reduced pressure. The residue was dissolved in distilled
water (30 mL), adjusted to pH 2.5 with 2 molL1 HCl,
washed with ethyl acetate (330 mL). The water layer
was adjusted to pH 12 with 15% sodium hydroxide
solution, and then extracted with ethyl acetate (350
mL). The combined organic extracts were washed
with saturated brine (230 mL), dried over anhydrous
sodium sulfate, and then concentrated under reduced
pressure to give the title compound 8a (0.91 g, 50.6%)
as a colorless liquid. 1 H NMR (400 MHz, CDCl3)

: 1.27 (s, 3H, CH3), 1.43 (s, 9H, Boc), 1.75 (br, 2H,
NH2), 2.302.33 (m, 1H, piperidine), 2.903.00 (m,
3H, piperidine), 3.673.70 (m, 1H, piperidine), 3.82 (s,
3H, OCH3), 3.853.88 (m, 1H, piperidine). ESI-MS
m/z: 258 [M+H]+.
2.8 1-tert-Butoxycarbonyl-3-amino-4-ethoxyimino3-methylpiperidine (8b) The title compound was
obtained in a similar manner as for the preparation of
compound 8a. Colorless liquid (66.0%). 1 H NMR
(400 MHz, CDCl3) : 1.221.26 (t, J = 7.2 Hz, 3H,
OCH2CH3), 1.28 (s, 3H, CH3), 1.46 (s, 9H, Boc), 1.79
(br, 2H, NH2), 2.282.36 (m, 1H, piperidine), 2.97
3.07 (m, 3H, piperidine), 3.693.72 (m, 1H, piperidine),
3.903.93 (m, 1H, piperidine), 4.054.10 (q, J = 7.2
Hz, 2H, OCH2CH3 ). ESI-MS m/z: 272 [M+H]+.
2.9 1-tert-Butoxycarbonyl-3-(N-tert-butoxycarbonyl)
amino-4-methoxyimino-3-methylpiperidine (9a) To
a stirring solution of compound 8a (1.73 g, 6.7 mmol)
in ethanol (30 mL), di-tert-butyl dicarbonate (1.62 g,
7.4 mmol) was added portionwise. The reaction
mixture was stirred for 2 h at the room temperature
and then concentrated under reduced pressure to give
the title compound 9a (2.40 g, 100%) as a colorless oil.
1
H NMR (400 MHz, CDCl3) : 1.43 (s, 9H, Boc), 1.48
(s, 9H, Boc), 1.52 (s, 3H, CH3 ), 2.352.38 (m, 1H,
piperidine), 2.893.18 (m, 3H, piperidine), 3.663.68
(m, 1H, piperidine), 3.86 (s, 3H, OCH3), 4.294.30 (m,
1H, piperidine), 5.67 (s, 1H, NH). ESI-MS m/z: 358
[M+H]+.
2.10 1-tert-Butoxycarbonyl-3-(N-tert-butoxycarbonyl)
amino-4-ethoxyimino-3- methylpiperidine (9b) The
title compound was obtained in a similar manner as for
the preparation of compound 9a. Colorless oil (98.5%).
1
H NMR (400 MHz, CDCl3) : 1.231.27 (t, J = 7.2
Hz, 3H, OCH2CH3), 1.44 (s, 9H, Boc), 1.49 (s, 9H,
Boc), 1.52 (s, 3H, CH3), 2.312.33 (m, 1H, piperidine),
2.983.17 (m, 3H, piperidine), 3.953.96 (m, 1H,
piperidine), 4.084.13 (q, J = 7.2 Hz, 2H, OCH2CH3),
4.354.38 (m, 1H, piperidine), 5.73 (s, 1H, NH).
ESI-MS m/z: 372 [M+H]+.
2.11 1-tert-Butoxycarbonyl-3-(N-tert-butoxycarbonyl)
methylamino-3-methyl-4-methoxyiminopiperidine (10a)
To a stirring solution of compound 9a (2.4 g, 6.7 mmol)
in dry tetrahydrofuran (40 mL), 70% sodium hydride
(0.46 g, 13.4 mmol) was added at 0 in 0.5 h using
an ice bath, and then stirred for 0.5 h at the room
temperature. After addition of methyl iodide (0.84

WAN Zhi-long, et al: Synthesis and in vitro antibacterial activity of 7-(4-alkoxyimino-3-methyl-3methylaminopiperidin-1-yl)quinolones

mL, 13.4 mol), the reaction mixture was stirred at

40 for 5 h and cooled to room temperature, adjusted
to pH 7 with 1 molL1 HCl and then concentrated
under reduced pressure. The residue was diluted with
ethyl acetate (50 mL), washed with distilled water (25
mL), dried over anhydrous sodium sulfate, and filtered.
The filtrate was concentrated under reduced pressure,
dried in vacuo to give the title compound 10a (2.37 g,
95.0%) as a white solid, mp 107109 . 1H NMR
(400 MHz, CDCl3) : 1.34 (s, 3H, CH3), 1.41 (s, 9H,
Boc), 1.46 (s, 9H, Boc), 2.242.25 (m, 1H, piperidine),
2.872.88 (m, 1H, piperidine), 2.91 (s, 3H, NCH3),
2.953.08 (m, 2H, piperidine), 3.82 (s, 3H, OCH3),
3.843.86 (m, 1H, piperidine), 4.304.31 (m, 1H,
piperidine). ESI-MS m/z: 372 [M+H]+.
2.12 1-tert-Butoxycarbonyl-3-(N-tert-butoxycarbonyl)
methylamino-4-ethoxyimino-3-methylpiperidine (10b)
The title compound was obtained in a similar manner as
for the preparation of compound 10a. White solid
(93.6%), mp 5658 . 1H NMR (400 MHz, CDCl3)
: 1.201.24 (t, J = 7.2 Hz, 3H, OCH2CH3), 1.34 (s, 3H,
CH3), 1.41 (s, 9H, Boc), 1.46 (s, 9H, Boc), 2.282.30
(m, 1H, piperidine), 2.91 (s, 3H, NCH3 ), 2.95-3.03 (m,
3H, piperidine), 3.783.82 (m, 1H, piperidine), 4.03
4.07 (q, J = 7.2 Hz, 2H, OCH2CH3), 4.304.32 (m, 1H,
piperidine). ESI-MS m/z: 386 [M+H]+.
2.13
4-Methoxyimino-3-methyl-3-methylaminopiperidine dihydrochloride (11a) To a stirring solution
of compound 10a (2.30 g, 6.2 mmol) in dichloromethane (50 mL), dried hydrochloride gas was pumped
at 05 using an ice bath for 0.5 h. The reaction
mixture was allowed to stir for another 0.5 h at room
temperature, the resulting solid was collected by
suction, and dried in vacuo to afford the title compound
11a (1.04 g, 97.8%) as a white solid, mp 224226 .
1

H NMR (400 MHz, DMSO-d6 ) : 1.59 (s, 3H, CH3),

2.51 (s, 3H, NCH3), 2.762.82 (m, 1H, piperidine), 2.95
3.02 (m, 1H, piperidine), 3.113.22 (m, 2H, piperidine),
3.493.60 (m, 2H, piperidine), 3.90 (s, 3H, OCH3),
9.82 (br, 4H, 2NH2 +). ESI-MS m/z: 172 [M+H]+.
2.14 4-Ethoxyimino-3-methyl-3-methylaminopiperidine dihydrochloride (11b) The title compound was
obtained in a similar manner as for the preparation of
compound 11a. White solid (98.8%), mp 202203 .
1
H NMR (400 MHz, DMSO-d6) : 1.231.25 (t, J = 7.2
Hz, 3H, OCH2CH3), 1.76 (s, 3H, CH3), 2.58 (s, 3H,
NCH3), 2.952.97 (m, 1H, piperidine), 3.193.20 (m, 1H,

867

piperidine), 3.463.50 (m, 2H, piperidine), 3.863.88

(m, 1H, piperidine), 4.194.24 (q, J = 7.2 Hz, 2H,
OCH2CH3), 4.564.60 (m, 1H, piperidine), 9.97 (br, 2H,
NH2 +), 10.53, 10.60 (2br, 2H, NH2+). ESI-MS m/z:
186 [M+H]+ .
2.15 1-Cyclopropyl-6-fluoro-7-(4-methoxyimino-3methyl-3-methylaminopiperidin-1-yl)-1, 4-dihydro-4oxo-1, 8-naphthyridine-3-carboxylic acid (14a) A
mixture of compound 11a (0.59 g, 2.4 mmol), triethylamine (0.5 mL) and dry acetonitrile (10 mL) was stirred
at room temperature for 10 min, 7-chloro-1-cyclopropyl6-fluoro-1, 4-dihydro-4-oxo-1, 8-naphthyridine-3-carboxylic acid 12a (0.56 g, 2.0 mmol) was added to the
solution and stirred for 1.5 h at the same temperature
under an atmosphere of nitrogen. The resulting solid
was collected by suction, washed with distilled water
and ethanol, respectively, dried in vacuo to afford the
title compounds.
The compounds 14b14d, by coupling reaction of
compound 11b with 12a, or compounds 11a, 11b with
12b, were synthesized in a similar manner.
2.16 1-Cyclopropyl-6-fluoro-7-(4-methoxyimino-3methyl-3-methylaminopiperidin-1-yl)-8-difluoromethoxyl-1, 4-dihydro-4-oxo-quinoline-3-carboxylic acid
(14e) A mixture of boric acid (0.25 g, 4.0 mmol) and
acetic anhydride (3 mL, 19.1 mmol) was stirred at 110
for 1.5 h. Then cooled and acetic acid (6 mL) was
added to the solution and stirred at 110 for another
1 h. Then cooled to 5060 , ethyl 1-cyclopropyl6, 7-difluoro-8-difluoromethoxyl-1, 4-dihydro-4-oxoquinoline-3-carboxylate 13a (0.58 g, 1.6 mmol) was
added and stirred at 110 for 1.5 h. Then cooled to
room temperature and poured into cold water (50 mL).
The resulting solid was collected by suction, and dried.
The solid obtained (0.69 g, 1.5 mmol) was added
to a mixture of compound 11a (0.73 g, 3 mmol),
triethylamine (1.2 mL) and dry acetonitrile (15 mL),
which was stirred at 60 for 6 h. After completion of
the condensation, the reaction mixture was concentrated
under reduced pressure. To the residue 6% sodium
hydroxide solution (15 mL) was added, stirred at 40
for 1 h, and then adjusted to pH 7 with 2 molL1 HCl,
the solid product was collected by suction. The crude
product was recrystallized from dichloromethane and
ethanol to give the title compound as a white solid.
The compounds 14f14p, by coupling reaction of
compound 11b with 13a, or compounds 11a, 11b with

Acta Pharmaceutica Sinica 2010, 45 (7): 860868

868

13b13f, were synthesized in a similar manner.

3 MIC determination
All compounds were screened for their in vitro
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Minimum inhibitory concentration (MIC) is defined as
the minimum concentration of the compound required
to give complete inhibition of bacterial growth after
incubation at 35 for 1824 h.

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