Transcript 김미금

Synthesis and Structure-Activity Relationships of
5-Amino-6-fluoro-1-[(1R,2S)-2-fluorocyclopropan-1-yl]8-methylquinolonecarboxylic Acid Antibacterials Having Fluorinated
7-[(3R)-3-(1-Aminocyclopropan-1-yl)pyrrolidin-1-yl] Substituents
Hiroaki Inagaki etc.
J. Med. Chem. 2003, 46, 1005-1015
분자생명과학부
김미금
2003-06-03
1. Introduction
(1)
Multidrug-resistant Gram-positive pathogens
a. Acqusition of resistance to vancomycin(VRE and vancomycinintermediate-resistant staphylococcal strains; VISA), an antibacterial agents generally regarded as the agent of last resort
for serious infection
b. Quinolone antibacterial agents의 영역에서는 내성을 가진 Grampositive 박테리아에 효능이 있는 약품을 얻기 위한 다양한 시도 및
travafloxacin(TVFX), moxifloxacin(MFLX), gemifloxacin(GMFX),
gatifloxacin(GFLX) 등의 계발
c. Driving force for the development of new antibacterial agents
- Antibacterial agents에 대한 Gram-positive bacteria의 내성
범위의 증가
- 임상에서 사용되고 있는 Quinolone이 아닌 몇몇의 물질이 보이는
내성 돌연변이와 부작용
2/19
(2) Target molecule
a. 선행연구
R3
R2
O
F
COOH
H2N
N
N
CH 3
R1
- C-7 위치에 3-(aminocycleopropal-1-yl)pyrrolidin-1-yl 치환기(R2= c-C3H5)를
포함하는 3-(1-amino-1-substituted-methyl)pyrrolidin-1-yl
groups를 갖는 여러 quinolone 유도체의 Gram-positive bacteria에
대한 antibacterial activity
- 7-{(3R)-3-(1-amino-1-substituted-methyl)pyrrolidin-1-yl}
quinolone 유도체 : 좋은 활성을 보였으나 높은 genotoxicity 가짐
- Genotoxicity를 줄이기 위한 N-1 위치에 cyclopropyl 치환기(R1= H) 대신에
(1R, 2S)-2-fluorocyclopropan-1-yl 치환기((R1= F)의 도입의 유용함
- 5-amino-8-methylquinolone 유도체(R3= NH2)의 Gram-positive bacteria에
대해 antibacterial activity 및 8-methylquinolone 유도체(R3= H)에 비해 적은
chromosomal toxicity의 보고
3/19
b. Target molecule
NH2
O
F
COOH
4: R1 = F, R2 = F
H
N
N
CH3
H2N
R1
1: R1 = H, R2 = H
2: R1 = H, R2 = F
3 (= DQ113): R1 = F, R2 = H
F
R2
Figure 1.
- Genotoxicity를 줄이면서 Gram-positive bacteria에 대해 높은 효능을
보이는 화합물
- C-7 위치에 (3R)-3-(1-aminocyclo-propan-1-yl)pyrrolidin-1-yl
치환기를 갖는 5-amino-1-[(1R,2S)-2-fluorocyclopropan-1-yl]8-methylquinolone(1, Figure 1)
- 화합물 1 : 높은 antibacterial 활성을 보였지만, micronucleus test에서
양성 반응을 보이고 chromosome injuring test에서 독성
- C-7 pyrrolidine 치환기로의 fluorine atom의 도입의 genotoxicity 감소에
효과를 화합물 1에 도입
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2. Chemistry
(1) Retrosynthesis
Scheme 1
NH2
O
NH2
F
COOH
H
O
F
COOH
NH
+
H
Boc
N
N
NH
R1
CH3
H2N
R1
F
5: R1 = H, R2 = H
6: R1 = H, R2 = F
7: R1 = F, R2 = H
R2
1-4
8: R1 = F, R2 = F
F
N
R2
CH3
F
9
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(2) Synthesis of compound 9
Scheme 2a
NO2
O
NO2
F
COOEt
NH2
O
F
F
COOEt
F
COOR3
d
a, b, c
F
O
F
F
N
CH3
CH3
10
N
CH3
F
11
F
12: R3 = Et
e
9: R3 = H
a
Reagents: (a) Ac2O, CH(OEt)3, reflux; (b) (1R, 2S)2-2-fluoro-1-cyclopropylamine p-toluenesulfonic acid salt, Et3N/CH2CL2; (c) NaH/1,4-dioxane;
(d) H2, Raney Ni (W-6)/MeOH, 1,4-dioxane; (e) concentrated aqueous HCl, AcOH, reflux
----------------------------------------------Mechanism(1)
NO2
O
NO2
O
H O+Et
OEt
F
F
COEt
H
H
F
+
C
H
EtO
O-
O
COEt
EtO
+
OEt
F
F
C
H
OEt
F
CH3
CH3
10
NO2
O
F
OCOEt
NO2
O
H
F
NO2
O
COEt
O-
O
F
COEt
H
EtO+
C
OEt
+
CH
EtO
F
F
CH3
F
F
CH3
EtO+
H
F
OEt
F
CH
OEt
CH3
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Mechanism(2)
NO2
NO2
O
F
O
F
H N+H
2
COEt
COEt
Et3N
+
HC
OEt
F
O
O
F
HC
F
N-H
+
+OHEt
F
F
F
CH3
CH3
NO2
O
NO2
O
F
O
O
F
COEt
NO2
COEt
O
F
COOEt
NaH
F
CH
CH
N H
NF
F
CH3
F
11
F
CH3
F
F
F
N
CH3
F
7/19
(3) Synthesis of compound 17a
a
b
OH
EtOOC
EtOOC
EtOOC
O
COOEt
COOEt
14
13
15
Reagents: (a) Zn, BrCH2CO2Et, cat. I2/THF, reflux; (b) (1) SOCl2/pyridine, (2) DBU/CH2Cl2
---------------------------------------------Mechanism (13 → 14 ; The Reformatsky reaction)
O- Zn2+
Zn + BrCH2CO2Et
H2C
C
0Et
O- Zn2+
H
+
H2C
EtOOC
C
N
O
0Et
EtOOC
O
14
13
COOEt
O
S
Cl
O-
O
Cl
S
Cl
O
EtOOC
Cl-
DBU
Cl
EtOOC
EtOOC
EtOOC
H
COOEt
COOEt
H
COOEt
COOEt
15
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H
EtOOC
EtOOC
c
d
N
COOEt
EtOOC
N
Ph
Ph
15
16
O
O
17a(3S) : 17b(3r) = 3.5 : 1
H
H
e
EtOOC
f
HOOC
N
N
Ph
Ph
O
O
18
17a
a
Reagents: (c) (1) NBS, AIBN/CCl4, (2 ) (S)-1-phenylethylamine, NaHCO3 /EtOH, reflux; (d) H2, PtO2 /MeOH; (e) silica gel column chromatography;
(f) aqueous NaOH/EtOH.
---------------------------------------------Mechanism
AIBN, NBS
EtOOC
Br
EtOOC
H2N
Ph , NaHCO3
EtOOC
N
COOEt
COOEt
Ph
15
N
AIBN(2,2'-Azobisisobutyronitrile) :
16
N
NC
O
CN
NBS(N-bromosuccinimide) : O
O
N
Br
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(4) Incorporation of fluorine atom(s) into the pyrrolidinone ring
H
H
EtOOC
a
EtOOC
N
N
Ph
R1
O
Ph
R2
Boc
O
17a
a
b
19: R1 = H, R2 = F
20: R1 = F, R2 = H
21: R1 = F, R2 = F
H
N
H
N
c,d
F
Ph
H
O
22
a
Reagents: (a) LDA, then (PhSO2)2NF/THF; (b) LDA, then 2,6-di-tert-butylphenol/THF; (c) aqueous NaOH/EtOH;
(d) diphenylphosphoryl azide, Et3N/toluene, then tert-BuOH.
----------------------------------------------LDA(Lithium diisopropylamide) : hindered non-nucleophilic strong
[(CH3)2CH]2NLi
(PhSO2)2NF(N-fluorobenzenesulfonimide) : transfer F+ to enolates
and carbanions
10/19
Mechanism (20 → 22 : subsequent Curtius rearrangement reaction)
O
Et3N/toluene
C
R
O
O
(PhO) 2P
N3
+
C
O-
R
OH
DPPA
O
O
C
P(OPh) 2
R
R
N
O
+
N3-
+
-
R
C
N
N
N
O
C
O
+
+
H
N
H+
OC(CH3)3
R
Boc
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(5) Synthesis of compound 27a-d
H
H
EtOOC
H
b
EtOOC
a
N
R1
N
Ph
R2
R1
R1
Ph
R2
S
23a: R1 = H, R2 = H
23b: R1 = H, R2 = F
23c: R1 = F, R2 = H
23d: R1 = F, R2 = F
17a: R1 = H, R2 = H
19: R1 = H, R2 = F
20: R1 = F, R2 = H
21: R1 = F, R2 = F
H
d
EtOOC
H
25a: R1 = H, R2 = H
25b: R1 = H, R2 = F
25c: R1 = F, R2 = H
25d: R1 = F, R2 = F
e
N Cbz
R1
R2
24a: R1 = H, R2 = H
24b: R1 = H, R2 = F
24c: R1 = F, R2 = H
24d: R1 = F, R2 = F
HOOC
N Cbz
R1
N
Ph
R2
O
c
EtOOC
Boc
H
N
H
R1
R2
26a: R1 = H, R2 = H
26b: R1 = H, R2 = F
26c: R1 = F, R2 = H
26d: R1 = F, R2 = F
N Cbz
R2
27a: R1 = H, R2 = H
27b: R1 = H, R2 = F
27c: R1 = F, R2 = H
27d: R1 = F, R2 = F
Reagents: (a) Lawesson's reagent/benzene; (b) Raney Ni (W-6)/EtOH; (c) Cbz-Cl/CH2Cl2; (d) aqueous NaOH/EtOH;
(e) diphenylphosphoryl azide, Et3N/tert-BuOH.
OCH3
S
P
S
S
P
S
H3CO
Lawesson's reagent
12/19
(6) Synthesis of final product 1-4
NH 2
O
F
H
NH
a
H
b, c
Boc
27a-d
COOH
N
N
NH
R2
R1
CH 3
H2N
R1
5: R1 = H, R2 = H
6: R1 = H, R2 = F
7: R1 = F, R2 = H
8: R1 = F, R2 = F
F
R2
1: R1 = H, R2 = H
2: R1 = H, R2 = F (HCl salt)
3 (= DQ113): R1 = F, R2 = H
4:
R1 = F, R2 = F (HCl salt)
Reagents: (a) H2, Pd-C/EtOH; (b) 9, Et3N/DMSO, heat; (c) concentrted aqueous HCl.
(7) Synthesis of Reference compounds 30, 31
NH 2
R4
R5
NH 2
O
F
NH
O
F
COOH
COOH
+
Boc
H2N
R4
NH
F
28: R4 = H, R5 = H
29: R4 = Me, R5 = H
N
CH 3
F
9
R5
N
N
H
CH 3
F
30: R4 = H, R5 = H
31: R4 = Me, R5 = H
Reagents: (a) Et3N/DMSO, heat; (b) concentrted aqueous HCl.
13/19
3. Results and Discussion
(1) 대표적인 Gram-positive와 Gram-negative bacteria에 대한 1-4,
30, 31의 minimum inhibitory concentrations(MICs) (Table 1)
-화합물 1-4, 30, 31은 Gram-positive bacteria에 대해
reference quinolones TVFX, MFLX, GFLX, CPFX보다 2-128배 활성
-C-7 위치에 fluorinated pyrrolidine을 갖고있는 화합물 2-4와
non-fluorinated 화합물 1은 거의 동일한 antibacterial 활성
→pyrrolidine substituents에 fluorine atom(s)를 도입하는 것은
antibacterial 활성에 크게 영향을 미치지 않음.
14/19
(2) Result of intravenous single-does toxicity study and micronucleus test (Table 2)
a. Intravenous single-does toxicity study
- 1 ((3R)-3-(1-aminocyclopropan-1-yl)pyrrolidin-1-yl substituent) < 30-31
- 2 (trans-oriented) > 1 (non-fluorinated) = 4 (difluorinated) > 3 (cis-oriented)
b. Micronucleus test
- Non-fluorinated 화합물 1, 31 : 양성반응
- Fluorinated 화합물 2-4 : 음성반응
15/19
(3) Solubility in water (Table 3)
: 3 (fluorinated 화합물) > 1 (non-fluorinated 화합물)
→ fluorine atom이 물에서의 용해도를 향상시키는데 기여
16/19
(4) Antibacterial activities of 3 (DQ- 113) (Table 4)
a.
clinically 분리된 내성을 갖는 Gram-positive bacteria에 대한 MICs가 결정
b.
다른 quinoline antibacterial agents, VCM, TEIC, LNZ보다 더 효과적
c.
levofloxacin-resistant MRSA에 대항하는 QPR/DPR과 비슷한 활성
d.
PRSP에 대항해서는, 3이 열거된 화합물들 중에서 가장 효능을 보였다.
e.
VRE strains 모두에서 가장 효능을 보였다.
17/19
(5) pharmacokinetic profiles of 3 (DQ- 113) (Table 5)
a.
time-concentration curve에서 높은 plasma concentration과 area
b.
간, 신장, 폐에서 좋은 분포 경향
18/19
4. Conclusion
(1) C-7 위치에 (3R)-3-(1-aminocyclopropan-1-yl)pyrrolidin-1-yl 치환기를 갖고있는
5-amino-6-fluoro-1-[(1R,2S)-2-fluorocyclopropan-1-yl]-8-methylquinolones
시리즈 합성
(2) Fluorinated 화합물 2-4는 non-fluorinated 화합물 1과 비교해서 효능을 보였고,
Gram-positive bacteria에 대해 reference quinolone보다는 적어도 4배의 효능을
보였다.
(3) Fluorinated 화합물들 중에서는 cis-oriented 유도체 3(DQ-113)이 1에 비해서
줄어든 정맥내로의 single-does toxicity를 보였고, 쥐에서 좋은 pharmacokinetic
profiles를 보였으며, 시험된 다른 quinolines와 비교했을 때 clinically 분리된
내성을 갖는 Gram-positive bacteria에 대해 더 좋은 antibacterial 활성을 보였다.
(4) 화합물 3은 이 논문에서 언급한 것들에 더불어 다른 clinically 분리된 bacteria에
대한 완벽한 antibacterial 활성을 보였고, further preclinical evaluation을
위해 선택되었다.
19/19