A novel dual antagonist of thromboxane A2 and leukotriene D 4 receptors: Synthesis and structure-activity relationships of chloroquinolylvinyl derivatives

Article abstract of DOI:10.1248/cpb.54.603

To discover an orally active thromboxane A₂ (TXA₂) and leukotriene D₄ (LTD₄) dual antagonist, we designed and synthesized chloroquinolylvinyl derivatives based on the structures of the TXA₂ antagonist daltroban and the LTD₄ antagonist montelukast. Among these derivatives, 4-{[(2-(4-chlorophenylsulfonylamino)-1-{3- [(E)-2-(7-chloro-2-quinolyl)vinyl]phenyl}ethyl)thio]methyl}benzoic acid (18d) showed potent inhibitory activity against U46619-induced aggregation of guinea pig platelets and LTD₄-induced contraction in the guinea pig ileum, with IC₅₀ values of 340 nm and 0.40 nm, respectively. Oral administration of 18 d also inhibited both the LTD₄-induced acceleration of plasma leakage to skin in guinea pig and the U46619-induced increase in airway resistance in guinea pig with ED₅₀ values of 0.47 mg/kg and 3.3 mg/kg, respectively.

Full text of DOI:10.1248/cpb.54.603

May 2006
Chem. Pharm. Bull. 54(5) 603—610 (2006)
603
A Novel Dual Antagonist of Thromboxane A2 and Leukotriene D4
Receptors: Synthesis and Structure–Activity Relationships of
Chloroquinolylvinyl Derivatives
Yoshinori OKAMOTO,*,a Masaki YOKOTA,a Soichiro KAWAZOE,b Hirokazu KUBOTA,a Hitoshi NAGAOKA,1)
a
Yasuhito ARAKIDA,c and Makoto TAKEUCHI
a Chemistry Research Laboratories, Astellas Pharmaceutical Inc.; 2–1–6 Kashima, Yodogawa-ku, Osaka 532–8514, Japan:
b Process Chemistry Laboratories, Astellas Pharmaceutical Inc.; 160–2 Matsukubo, Akahama, Takahagi, Ibaraki
318–0001, Japan: and c Planning & Administration Dept., Astellas Pharmaceutical Inc.; 3–17–1 Hasune, Itabashi-ku,
Tokyo 174–8612, Japan. Received October 7, 2005; accepted January 27, 2006
To discover an orally active thromboxane A2 (TXA2) and leukotriene D4 (LTD4) dual antagonist, we designed
and synthesized chloroquinolylvinyl derivatives based on the structures of the TXA2 antagonist daltroban and
the LTD4 antagonist montelukast. Among these derivatives, 4-{[(2-(4-chlorophenylsulfonylamino)-1-{3-[(E)-2-(7-
chloro-2-quinolyl)vinyl]phenyl}ethyl)thio]methyl}benzoic acid (18d) showed potent inhibitory activity against
U46619-induced aggregation of guinea pig platelets and LTD4-induced contraction in the guinea pig ileum, with
IC50 values of 340 nm and 0.40 nm, respectively. Oral administration of 18 d also inhibited both the LTD4-induced
acceleration of plasma leakage to skin in guinea pig and the U46619-induced increase in airway resistance in
guinea pig with ED50 values of 0.47 mg/kg and 3.3 mg/kg, respectively.
Key words leukotriene D4; thromboxane A2; dual antagonist; structure–activity relationship
Asthma is regarded as an inflammatory disease of the res- (Fig. 1). In this report, we describe the structure–activity re-
piratory tract. Arachidonic acid released from membrane lationships of TXA2 and LTD4 receptor dual antagonists and
phospholipids by phospholipase A2 is converted to various the pharmacological profiles of selected inhibitors.
metabolites that play important roles in asthma2): thrombox-
Chemistry A series of the 4-chlorobenzensulfonamide
ane A2 (TXA2) is produced from arachidonic acid through propyl derivatives 7a—h was synthesized using the proce-
the cyclooxygenase pathway and leukotrienes (LTs) are syn- dure shown in Chart 1. The allylic alcohol 125) was oxidized
thesized through the 5-lipoxygenase pathway. TXA2 and by manganese dioxide to afford the a,b-unsaturated ketone
LTD4 are considered to be aggravating factors in asthma with 2. Michael addition of chlorobenzenesulfonamide to com-
TXA2 inducing bronchial hyperreactivity and bronchocon- pound 2 gave the sulfonamide 3, and the carbonyl group of
striction,3,4) and LTD4 inducing potent bronchoconstriction, compound 3 was reduced by sodium borohydride to afford
enhanced vascular permeability and mucus secretion.5,6) In the alcohol derivative 4, which was treated with thionylchlo-
addition, the levels of TXA2 and LTs in the plasma, bron- ride to give the benzyl chloride 5. Thioalkylation of com-
choalveolar lavage fluid (BALF) and urine from patients with pound 5 with various thiols gave the esters 6a—h, which
bronchial asthma have been shown to be elevated.7) Based on were hydrolyzed to afford compounds 7a—h.
these observations, TXA2 and LTD4 are thought to be poten-
A series of benzenesulfonamide ethyl derivatives 18a—j
tial targets for anti-asthmatic drugs: in fact, highly potent and was synthesized using the procedure shown in Chart 2. Con-
selective antagonists for these mediators have been used for densation of the 7-chloro-2-methylquinoline 826) and methyl
treatment of asthma.8—10) However, TXA2 and LTD4 play dif- 3-formylbenzoate in the presence of acetic anhydride gave
ferent roles in the symptoms of asthma, and clinical trial re- the ester 9, which was hydrolyzed to afford the benzoic acid
sults of TXA2 receptor antagonists11—13) and LTD4 receptor derivative 10. Compound 10 was reacted with ethyl iso-
antagonists14,15) suggest that the predominant mediator varies cyanoacetate in the presence of diphenylphosphoryl azide
from patient to patient. These results suggest that an antago- (DPPA) to afford the oxazole derivative 11, which was
nist for both TXA2 and LTD4 receptors would be more effec- treated with hydrochloric acid to give the aminoketone deriv-
tive in the treatment of asthma, compared to the selective an- ative 12, followed by the reduction with sodium borohydride
tagonists. Some TXA2 and LTD4 dual receptor antagonists,
such as RS-60116) and YM-158,17—22) have been reported to
have potent efficacy in various antiasthmatic models.
Based on the above, we planned to design a novel dual
antagonist for the TXA2 and LTD4 receptors. The potent
and selective LTD4 antagonist montelukast23) contains a
lipophilic chloroquinolylvinyl group and a carboxyl group,
whereas the potent and selective TXA2 antagonist dal-
troban24) is a chlorobenzenesulfonamide that also contains a
carboxyl group. Since both montelukast and daltroban have a
carboxyl group, the TXA2 and LTD4 receptor dual antago-
nists were designed by introducing the chlorobenzenesulfon-
Fig. 1
amide group of daltroban into the structure of montelukast
∗ To whom correspondence should be addressed. e-mail: yoshinori.okamoto@jp.astellas.com
© 2006 Pharmaceutical Society of Japan

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Vol. 54, No. 5
Chart 1
Chart 2
to afford the aminoalcohol 13. The amino group of com- sulting esters 17a—j were hydrolyzed to give compounds
pound 13 was protected with phthalic anhydride, and the re- 18a—j.
sulting phthalimide derivative was treated with methanesul-
fonyl chloride to give the mesylate 15, which was reacted Results and Discussion
with various thiols to give the sulfides 16a—f. Compounds
The inhibitory activities of compounds 7a—h and 18a—j
16a—f were treated with methylhydrazine, and the resulting against U46619-induced aggregation in guinea pig platelets
amines were reacted with various sulfonyl chlorides. The re- and against LTD4-induced contraction in guinea pig ileum

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605
Table 1. TXA2 and LTD4 Antagonist Activities of Chloroquinolylvinyl
Derivatives
were evaluated, and the results are listed in Table 1.
Among the 4-chlorobenzenesulfonamide propyl deriva-
tives, the compound possessing an acetic acid group on the
sulfur atom (7a) showed no TXA2 antagonist activity. Re-
placement of the acetic acid group (7a) with propanoic acid
group (7b) or butanoic acid group (7c) enhanced the TXA2
antagonistic activity, and derivative 7c showed particularly
potent TXA2 antagonist activity with an IC50 value of 150 nM.
The introduction of 3,3-dimethyl (7d) or dimethylene (7e)
groups on the butanoic acid reduced the TXA2 antagonist ac-
tivity, with IC50 values of 1100 nM and 500 nM, respectively.
Replacement of butanoic acid (7c) on the sulfur atom with
4-carboxyphenylmethyl (7f), 4-carboxymethylphenylmethyl
(7g) or 2-(4-carboxyphenyl)ethyl (7h) groups reduced the
TXA2 inhibitory activity compared with compound 7c. Intro-
duction of a phenyl ring or bulky substituent between the sul-
fur atom and the carboxylic acid group were unfavorable for
TXA2 antagonist activity.
Among the 4-chlorobenzenesulfonamide ethyl derivatives
18a—f, the compounds possessing propanoic acid (18a),
3,3-dimethylbutanoic acid (18b) and 3,3-dimethylenebu-
tanoic acid (18c) on the sulfur atom showed less potent
TXA2 antagonist activities than that of daltroban. On the
contrary, the compounds with 4-carboxyphenylmethyl (18d)
and 2-(4-carboxyphenyl)ethyl (18f) groups on the sulfur
atom showed potent TXA2 antagonistic activities with IC50
values of 340 nM and 510 nM, respectively. The compound
with a 4-carbonylmethylphenylmethyl group (18e) on the
sulfur atom showed reduced TXA2 antagonistic activity,
compared with compound 18d. In the 4-chlorobenzenesul-
fonamide ethyl derivatives, introduction of a phenyl ring on
the sulfur atom was favorable for TXA2 antagonist activity.
Next, we investigated the influence of the substituent on
the 4-position of the benzene sulfonamide group on TXA2
antagonistic activity. Removal of the chloro group (18g)
caused loss of activity. The bromo derivative 18h was almost
as potent as the chloro derivative 18d, but the nitro (18i) and
fluoro (18j) derivatives showed decreased TXA2 antagonist
activity. From these results, it was concluded that the nature
of the substituents on the 4-position of the benzene sulfon-
amide group are important for TXA2 antagonistic activity.
As shown in Table 1, montelukast has potent LTD4 antago-
nistic activity with an IC50 value of 0.085 nM. In the series of
4-chlorobenzenesulfonamide propyl derivatives, the influ-
ence of the length of the methylene chain on the sulfur atom
IC50 (nM)
No.
R
n
X
TXA2
LTD4
7a
7b
7c
2
2
2
Cl
Cl
Cl
51000
1000
150
1.2
0.41
0.85
7d
7e
2
2
Cl
Cl
1100
500
0.41
0.21
7f
2
Cl
980
3.5
7g
7h
2
2
Cl
Cl
4400
1000
11
8.4
18a
18b
1
1
Cl
Cl
9100
1.7
10000
0.24
18c
18d
1
1
Cl
Cl
1400
340
0.23
0.40
18e
18f
1
1
Cl
Cl
1100
510
2.1
3.0
18g
18h
18i
1
1
1
1
H
Br
680
500
0.041
0.077
0.23
NO2
F
1000
18j
750
370
0.12
Daltroban
N.D.a)
0.085
Montelukast
N.D.a)
a) Not determined.
(7a—c) on the LTD4 antagonistic activity differed from that in LTD4 antagonist activity suggesting that a phenyl group in
for TXA2 antagonistic activity. The compound with a this position causes a loss of this activity.
propanoic acid group on the sulfur atom (7b) showed potent
Among the 4-chlorobenzenesulfonamide ethyl derivatives,
LTD4 antagonist activity, with an IC50 value of 0.41 nM. Re- the compound with a 2-propanoic acid group (18a) on the
placement of the propanoic acid group in compound 7b with sulfur atom showed LTD4 antagonist activity, with an IC50
acetic acid (7a) or butanoic acid (7c) resulted in a decrease in value of 1.7 nM. Replacement of the carboxyethyl group in
the LTD4 antagonistic activity. Compounds with dimethyl compound 18a with 3,3-dimethylbuanoic acid (18b) or 3,3-
(7d) or dimethylene (7e) groups at the 3 position of the bu- dimethylenebutanoic acid (18c) groups on the sulfur atom in-
tanoic acid group in compound 7c retained potent LTD4 an- creased the LTD4 antagonist activity, with IC50 values of
tagonist activities, with IC50 values of 0.41 and 0.21 nm, re- 0.24 nM and 0.23 nM, respectively. These results suggest that
spectively. These results suggest that a bulky substituent at bulky substituents on the butanoic acid in compound 18a
the 3 position of the butanoic acid group in compound 7c is are favorable for LTD4 antagonist activity. Replacement of
tolerated for LTD4 antagonistic activity. Replacement of the propanoic acid group in compound 18a with a car-
the propanoic acid group of compound 7b with 4-car- boxyphenylmethyl group (18d) resulted in an increase in this
boxyphenylmethyl (7f), 4-carboxymethylphenylmethyl (7g) activity, with an IC50 value of 0.40 nM. The compounds with
or 4-carboxyphenylethyl (7h) groups resulted in a decrease carboxymethylphenylmethyl (18e) or carboxyphenylethyl

606
Vol. 54, No. 5
n-hexaneꢀ3 : 30 : 70) gave 2 (10.6 g, 41%) as a colorless solid. 1H-NMR
(CDCl3): 5.97 (1H, dd, Jꢀ10.8, 1.6 Hz), 6.48 (1H, dd, Jꢀ17.2, 1.6 Hz), 7.18
(1H, dd, Jꢀ17.2, 10.8 Hz), 7.37—7.44 (2H, m), 7.50 (1H, t, Jꢀ7.6 Hz), 7.59
(1H, d, Jꢀ8.4 Hz), 7.69 (1H, d, Jꢀ8.4 Hz), 7.73 (1H, d, Jꢀ16.8 Hz), 7.80
(1H, d, Jꢀ7.6 Hz), 7.88 (1H, d, Jꢀ7.6 Hz), 8.05—8.09 (2H, m), 8.18 (1H,
s). FAB-MS m/z: 320 [MꢁH]ꢁ.
4-Chloro-N-(3-{3-[(E)-2-(7-chloro-2-quinolyl)vinyl]phenyl}-3-oxo-
propyl)benzenesulfonamide (3) A mixture of 2 (8.70 g, 27.2 mmol), 4-
chlorobenzenesulfonamide (5.73 g, 29.9 mmol), potassium tert-butoxide
(0.10 g, 0.89 mmol), 1,4-dioxane (80 ml) and benzene (80 ml) was stirred at
80 °C for 10 h. The reaction mixture was evaporated in vacuo. Column chro-
matography of the residue on silica gel (0.2% MeOH–CHCl3) gave 3
(8.10 g, 58%) as a colorless solid. 1H-NMR (DMSO-d6): 3.15—3.21 (2H,
m), 3.29 (2H, br t, Jꢀ6.2 Hz), 7.57—7.63 (3H, m), 7.69 (2H, br d,
Jꢀ8.8 Hz), 7.83—7.89 (4H, m), 7.95 (1H, br s), 7.98 (1H, d, Jꢀ6.4 Hz),
8.01—8.04 (3H, m), 8.23 (1H, br s), 8.44 (1H, d, Jꢀ8.8 Hz). FAB-MS m/z:
511 [MꢁH]ꢁ.
Table 2. Inhibitory Activities of U46619-Induced Increase in Airway Re-
sistance in Guinea Pig and LTD4-Induced Acceleration of Plasma Leakage
in Guinea Pig Skin
U46619-induced
LTD4-induced
plasma leakage
airway
resistance
No.
ED50 (mg/kg)
ED50 (mg/kg)
p.o.
p.o.
7c
7e
18d
18g
18h
18j
N.D.a)
38%b)
3.3
54%c)
0.12
0.47
N.D.a)
N.D.a)
N.D.a)
63%c)
24%c)
44%c)
(ꢀ)-4-Chloro-N-(3-{3-[(E)-2-(7-chloro-2-quinolyl)vinyl]phenyl}-3-hy-
a) Not determined. b) % inhibition at 10 mg/kg. c) % inhibition at 1 mg/kg.
droxypropyl)benzenesulfonamide (4)
A mixture of 3 (8.40 g, 17.7
mmol), tetrahydrofuran (40 ml) and ethanol (40 ml) was added to sodium
borohydride (0.33 g, 8.8 mmol) at 5 °C. The reaction mixture was stirred at
room temperature for 1 h, and then acidified with a 10% aqueous solution of
citric acid. The mixture was extracted with CHCl3, washed with brine, dried
over Na2SO4, and evaporated in vacuo to give 4 (8.50 g, 90%) as a yellow
(18f) groups showed less potent LTD4 antagonist activity
compared to compound 18d, with IC50 values of 2.1 nM and
3.0 nM, respectively. These results suggest that the distance
between the sulfur atom and the carboxylic acid group is im-
portant for potent LTD4 antagonist activity.
Removal (18g) or replacement of the chloro group at the
4-position of the benzenesulfonamide group in compound
18d with bromo (18h), nitro (18i) or fluoro (18j) groups in-
creased the LTD4 antagonistic activity, with IC50 values of
0.041 nM, 0.077 nM, 0.23 nM and 0.12 nM, respectively. These
results suggest that substituents on the benzenesulfonamide
group do not significantly influence the activity.
Selected compounds (7c, 7e, 18d, 18g, 18h, 18j) which
possessed potent TXA2 and LTD4 antagonist activities were
tested for their ability to inhibit the LTD4-induced accelera-
tion of plasma leakage to skin and U46619-induced increase
in airway resistance in guinea pig after oral administration.
The results are shown in Table 2. Compounds 7e and 18d
showed potent inhibition of LTD4-induced acceleration of
plasma leakage, with ED50 values of 0.12 and 0.47 mg/kg, re-
spectively. Compound 18d also showed inhibitory activity
against the U46619-induced increase in airway resistance
with an ED50 value of 3.3 mg/kg. Compound 7e was less po-
tent than compound 18d in this respect.
In conclusion, in order to find an orally active TXA2 and
LTD4 dual antagonist, we designed and synthesized chloro-
quinolylvinylphenyl derivatives based on the molecular
structures of the TXA2 antagonist daltroban and the LTD4 an-
tagonist montelukast. Among these compounds, 18d showed
potent TXA2 and LTD4 antagonist activity both in vitro and
in vivo.
1
amorphous solid. H-NMR (CDCl3): 1.88—1.93 (2H, m), 3.07—3.25 (2H,
m), 4.86 (1H, br t, Jꢀ6.2 Hz), 5.65 (1H, br t, Jꢀ5.8 Hz), 7.16 (1H, br d,
Jꢀ8.0 Hz), 7.26—7.31 (2H, m), 7.44 (1H, dd, Jꢀ8.8, 1.8 Hz), 7.46 (d,
Jꢀ8.4 Hz), 7.52 (1H, br s), 7.57—7.62 (2H, m), 7.68 (2H, d, Jꢀ8.8 Hz),
8.79 (2H, d, Jꢀ8.0 Hz), 7.99 (1H, br d, Jꢀ1.6 Hz), 8.07 (1H, d, Jꢀ8.8 Hz).
FAB-MS m/z: 513 [MꢁH]ꢁ.
(ꢀ)-4-Chloro-N-(3-chloro-3-{3-[(E)-2-(7-chloro-2-quinolyl)vinyl]-
phenyl}propyl)benzenesulfonamide (5) A solution of 4 (8.40 g, 16.4
mmol) in 1,2-dichloroethane was combined with thionyl chloride (1.43 ml,
19.6 mmol) at 5 °C. The reaction mixture was stirred at room temperature
for 5 h, and then ice and saturated aqueous sodium bicarbonate solution
were added. The reaction mixture was extracted with CHCl3, washed with
brine, dried over Na2SO4, and evaporated in vacuo. Column chromatography
of the residue on silica gel (AcOEt : n-hexaneꢀ1 : 4→3 : 7) gave 5 (8.52 g,
quant.) as a yellow amorphous solid. 1H-NMR (CDCl3): 2.24—2.31 (2H, m),
3.10—3.25 (2H, m), 4.98 (1H, dd, Jꢀ8.4, 6.0 Hz), 5.03 (1H, br t, Jꢀ6.0 Hz),
7.26 (1H, d, Jꢀ8.0 Hz), 7.34—7.39 (2H, m), 7.44 (1H, dd, Jꢀ8.8, 2.0 Hz),
7.49 (2H, br d, Jꢀ8.8 Hz), 7.56 (1H, d, Jꢀ8.0 Hz), 7.58—7.66 (2H, m), 7.71
(2H, d, Jꢀ8.8 Hz), 7.81 (2H, br d, Jꢀ8.4 Hz), 8.05 (1H, br d, Jꢀ2.0 Hz),
8.10 (1H, d, Jꢀ8.4 Hz). FAB-MS m/z: 531 [MꢁH]ꢁ.
(ꢀ)
Methyl
4-[(3-{4-Chlorophenylsulfonylamino}-1-{3-[(E)-2-(7-
chloro-2-quinolyl)vinyl]phenyl}propyl)thio]propionate (6b) A solution
of 5 (0.50 g, 0.9 mmol) in N,Nꢂ-dimethylimidazolinone was combined with
cesium carbonate (1.61 g, 1.9 mmol) and methyl 3-mercaptopropionate
(0.17 g, 1.4 mmol) at 10 °C. The reaction mixture was stirred at 40 °C for
12 h, water was added, the reaction mixture was extracted with ethyl acetate,
washed with water and brine, dried over MgSO4, and evaporated in
vacuo. Column chromatography of the residue on silica gel (AcOEt : n-
1
hexaneꢀ1 : 3) gave 6b (0.32 g) as a yellow amorphous solid. H-NMR
(CDCl3): 2.00—2.06 (2H, m), 2.47—2.64 (4H, m), 3.03—3.08 (2H, m),
3.68 (3H, s), 3.90 (1H, t, Jꢀ6.6 Hz), 4.91 (1H, t, Jꢀ6.4 Hz), 7.19—7.80
(13H, m), 8.08—8.13 (2H, m), FAB-MS m/z: 615 [MꢁH]ꢁ.
(ꢀ) Ethyl 4-[(3-{4-Chlorophenylsulfonylamino}-1-{3-[(E)-2-(7-chloro-
2-quinolyl)vinyl]phenyl}propyl)thio]acetate (6a) The title compound
was prepared from ethyl mercaptoacetate in the same manner as described
above, and obtained as a pale yellow solid. (82%). 1H-NMR (CDCl3):
1.20—1.30 (3H, m), 2.03—2.08 (2H, m), 2.71—3.13 (4H, m), 4.03—4.19
(3H, m), 7.00—8.13 (15H, m). FAB-MS m/z: 615 [MꢁH]ꢁ.
Experimental
1H-NMR spectra were obtained on a JEOL JNM-EX90 or JNM-A500
spectrometer and chemical shifts are expressed as d (ppm) values with
tetramethylsilane as the internal standard. Abbreviations of the 1H-NMR sig-
nal patterns are as follows: s, singlet; d, doublet; dd, double doublet; t,
triplet; q, quartet; m, multiplet: br, broad. Mass spectra were obtained on a
JEOL JMS-DX300 or Hitachi M-80 spectrometer. Column chromatography
on silica gel was performed with Wakogel C-200. Preparative thin-layer
(ꢀ)
Methyl
4-[(3-{4-Chlorophenylsulfonylamino}-1-{3-[(E)-2-(7-
chloro-2-quinolyl)vinyl]phenyl}propyl)thio]butylate (6c) The title com-
pound was prepared from methyl 4-mercaptobutylate in the same manner as
described above, and obtained as a pale yellow solid. (44%). 1H-NMR
(CDCl3): 1.21—1.28 (3H, m), 1.71—1.85 (2H, m), 2.01—2.06 (2H, m),
2.26—2.39 (4H, m), 3.02—3.08 (2H, m), 3.84—3.88 (1H, m), 4.08—4.15
(2H, m), 4.79 (1H, t, Jꢀ6.4 Hz), 7.18—7.75 (11H, m), 8.08—8.13 (2H, m).
FAB-MS m/z: 643 [MꢁH]ꢁ.
chromatography was performed with Merck PLC plate Silica gel 60 F254
.
1-{3-[(E)-2-(7-Chloro-2-quinolyl)vinyl]phenyl}propenone (2) A mix-
ture of 1-{3-[(E)-2-(7-chloro-2-quinolyl)vinyl]phenyl}propenol (26.2 g,
81.4 mmol), manganese dioxide (131 g, 1.51 mol) and toluene (500 ml) was
refluxed for 3 h. The reaction mixture was cooled to room temperature
and then filtered through celite. The filtrate was evaporated in vacuo.
Column chromatography of the residue on silica gel (AcOEt : CHCl3 :
(ꢀ)
Methyl
4-[(3-{4-Chlorophenylsulfonylamino}-1-{3-[(E)-2-(7-
chloro-2-quinolyl)vinyl]phenyl}propyl)thio]-3,3-dimethylbutylate (6d)
The title compound was prepared from methyl 3,3-dimethyl-4-mercapto-
butylate in the same manner as described above, and obtained as a pale yel-

May 2006
607
low solid. (94%). 1H-NMR (CDCl3): 0.96 (3H, s), 0.98 (3H, s), 2.03 (2H, q,
Jꢀ6.8 Hz), 2.23, 2.36 (each 1H, each d, Jꢀ14.0 Hz), 2.38, 2.43 (each 1H,
each d, Jꢀ12.4 Hz), 3.07 (2H, m), 3.64 (3H, s), 3.81 (1H, t, Jꢀ7.2 Hz), 4.90
(1H, t, Jꢀ6.0 Hz), 7.17—7.87 (13H, m), 8.07—8.13 (2H, m). FAB-MS m/z:
657 [MꢁH]ꢁ.
(DMSO-d6) d: 2.52 (2H, br d, Jꢀ5.9 Hz), 7.72 (1H, t, Jꢀ7.6 Hz), 7.78 (1H,
dd, Jꢀ8.8, 2.0 Hz), 7.87 (1H, d, Jꢀ16.6 Hz), 8.06—8.10 (2H, m), 8.19 (1H,
d, Jꢀ8.8 Hz), 8.25 (1H, d, Jꢀ8.8 Hz), 8.30—8.38 (3H, m), 8.59 (3H, br s),
8.80 (1H, d, Jꢀ8.8 Hz). FAB-MS m/z: 323 [MꢁH]ꢁ.
(ꢀ)-2-Amino-1-{3-[(E)-2-(7-chloro-2-quinolyl)vinyl]phenyl}ethanol
(13) An ice-cold mixture of potassium carbonate (25.3 g, 198 mmol),
sodium borohydride (5.54 g, 147 mmol) and ethanol (580 ml) was added 12
(38.7 g, 98 mmol) in portions with the temperature maintained below 15 °C.
The reaction mixture was stirred at room temperature for 1 h and then
poured into ice-water. The resulting precipitate was collected by filtration to
give 13 (21.20 g, 55%) as a colorless solid. 1H-NMR (DMSO-d6) d: 2.65
(1H, dd, Jꢀ13.0, 7.8 Hz), 2.74 (1H, t, Jꢀ13.0, 4.4 Hz), 4.51 (1H, br dd,
Jꢀ7.2, 4.4 Hz) 7.33 (1H, d, Jꢀ8.0 Hz), 7.39 (1H, t, Jꢀ7.6 Hz), 7.47 (1H, d,
Jꢀ16.4 Hz), 7.56—7.63 (2H, m), 7.70 (1H, br s), 7.88 (1H, d, Jꢀ16.4 Hz),
7.93 (1H, d, Jꢀ8.4 Hz), 7.99—8.03 (2H, m), 8.40 (1H, d, Jꢀ8.4 Hz). FAB-
MS m/z: 325 [MꢁH]ꢁ.
(ꢀ) Benzyl 4-{[(3-{4-Chlorophenylsulfonylamino}-1-{3-[(E)-2-(7-
chloro-2-quinolyl)vinyl]phenyl}propyl)thio]methyl}-1-cyclopropanac-
etate (6e) The title compound was prepared from benzyl 1-mercapto-
methylcyclopropanacetate in the same manner as described above, and ob-
1
tained as a pale yellow solid. (quant.). H-NMR (CDCl3): 0.31—0.62 (4H,
m), 1.92—2.02 (2H, m), 2.31—2.52 (4H, m), 2.99—3.10 (2H, m), 3.86 (1H,
t, Jꢀ7.6 Hz), 4.99 (1H, t, Jꢀ6.4 Hz), 5.11 (2H, s), 7.11—8.12 (20H, m).
FAB-MS m/z: 731 [MꢁH]ꢁ.
(ꢀ) Methyl 4-{[(3-{4-Chlorophenylsulfonylamino}-1-{3-[(E)-2-(7-
chloro-2-quinolyl)vinyl]phenyl}propyl)thio]methyl}benzoate (6f) The
title compound was prepared from methyl 4-mercaptomethylbenzoate in the
same manner as described above, and obtained as a pale yellow solid. (93%).
1H-NMR (CDCl3): 1.97—2.03 (2H, m), 2.97 (2H, q, Jꢀ6.8 Hz), 3.45 (1H, d,
Jꢀ13.6 Hz), 3.57 (1H, d, Jꢀ13.6 Hz), 3.65 (1H, t, Jꢀ8.0 Hz), 3.90 (3H, s),
4.38 (1H, t, Jꢀ6.4 Hz), 7.12—8.14 (19H, m). FAB-MS m/z: 677 [MꢁH]ꢁ.
(ꢀ) Methyl 4-{[(3-{4-Chlorophenylsulfonylamino}-1-{3-[(E)-2-(7-
(ꢀ)-N-(2-{3-[(E)-2-(7-Chloro-2-quinolyl)vinyl]phenyl}-2-hydroxy-
ethyl)phthalimide (14) A mixture of 13 (16.10 g, 49.6 mmol), phthalic an-
hydride (7.34 g, 49.6 mmol) and toluene (20 ml) was stirred and heated to
160 °C, and then stirred under reduced pressure (3 mmHg) for 3 h. The reac-
tion mixture was cooled to room temperature, and then acetonitrile was
added. The resulting precipitate was collected by filtration to give 14
chloro-2-quinolyl)vinyl]phenyl}propyl)thio]methyl}phenylacetate
(6g)
1
The title compound was prepared from methyl 4-mercaptometylphenylac-
etate in the same manner as described above, and obtained as a pale yellow
(18.8 g) as a colorless solid. H-NMR (DMSO-d6) d: 3.72 (1H, dd, Jꢀ13.2,
4.6 Hz), 3.83 (1H, dd, Jꢀ13.2, 8.8 Hz), 4.96—5.01 (1H, m), 5.74 (1H, d,
Jꢀ4.4 Hz) 7.33—7.48 (3H, m), 7.59 (1H, dd, Jꢀ8.0, 2.0 Hz), 7.76 (1H, br d,
Jꢀ7.6 Hz), 7.75 (1H, br s), 7.82—7.95 (6H, m), 8.00 (1H, d, Jꢀ8.0 Hz),
8.03 (1H, d, Jꢀ2.0 Hz), 8.40 (1H, d, Jꢀ8.8 Hz). FAB-MS m/z: 455
[MꢁH]ꢁ.
1
solid. (39%). H-NMR (CDCl3): 2.00—2.06 (2H, m), 2.95—3.00 (2H, m),
3.27—3.72 (4H, m), 3.68 (3H, s), 4.76—4.80 (1H, m), 7.04—7.89 (17H,
m), 8.25—8.40 (2H, m). FAB-MS m/z: 691 [MꢁH]ꢁ.
(ꢀ) Methyl 4-{2-[(3-{4-Chlorophenylsulfonylamino}-1-{3-[(E)-2-(7-
chloro-2-quinolyl)vinyl]phenyl}propyl)thio]ethyl}benzoate
(6h) The
(ꢀ)-1-{3-[(E)-2-(7-Chloro-2-quinolyl)vinyl]phenyl}-2-phthalimido-
ethylmethanesulfonate (15) An ice-cold mixture of 14 (17.8 g,
39.1 mmol), pyridine (53 ml) and 1,2-dichloroethane was combined with
methanesulfonylchloride (3.36 ml, 47.0 mmol), and the mixture was stirred
at room temperature for 12 h. The reaction mixture was acidified with 10%
aqueous solution of citric acid, and then the mixture was extracted with
CHCl3. The organic layer was washed with a 10% aqueous solution of citric
acid, a 5% aqueous solution of potassium carbonate and brine, dried over
MgSO4, and evaporated in vacuo. The residue was combined with acetoni-
trile, and the resulting precipitate was collected by filtration to give 15
(19.0 g) as a colorless solid. 1H-NMR (DMSO-d6) d: 3.19 (3H, s), 3.95 (1H,
dd, Jꢀ14.8, 4.0 Hz), 4.27 (1H, dd, Jꢀ14.8, 8.8 Hz), 5.89 (1H, dd, Jꢀ8.8,
4.0 Hz), 7.51—7.56 (2H, m), 7.59—7.63 (2H, m), 7.79—7.83 (1H, m),
7.84—7.89 (2H, m), 7.90—7.96 (5H, m), 8.00—8.04 (2H, m), 8.43 (1H, d,
Jꢀ8.8 Hz). FAB-MS m/z: 533 [MꢁH]ꢁ.
title compound was prepared from methyl 4-(2-mercaptoethyl)benzoate in
the same manner as described above, and obtained as a pale yellow amor-
phous solid. (49%). 1H-NMR (CDCl3) d: 2.00—2.06 (2H, m), 2.50—2.60
(2H, m), 2.70—2.82 (2H, m), 2.95—3.02 (2H, m), 3.82 (1H, t, Jꢀ7.4 Hz),
3.87 (3H, s), 4.54 (1H, t, Jꢀ6.4 Hz), 7.20 (1H, d, Jꢀ7.6 Hz), 7.33—7.41
(2H, m), 7.44—7.50 (3H, m), 7.53 (1H, br s), 7.66 (2H, d, Jꢀ8.8 Hz), 7.72
(2H, d, Jꢀ8.8 Hz), 8.08 (1H, br d, Jꢀ1.6 Hz), 8.12 (1H, d, Jꢀ8.4 Hz). FAB-
MS m/z: 691 [MꢁH]ꢁ.
Methyl 3-[(E)-2-(7-Chloro-2-quinolyl)vinyl]benzoate (9) A mixture
of 7-chloroquinaldine (8, 5.40 g, 30.4 mmol), methyl isophthalaldehyde
(5.99 g, 36.5 mmol) and acetic anhydride (8.61 ml, 91.2 mmol) in xylene
(54 ml) was stirred under reflux for 8 h. The reaction mixture was cooled to
room temperature, and the solvent was evaporated under reduced pressure.
Diethylether was added to the residue and the resulting precipitate was col-
1
lected by filtration to give 9 (4.65 g) as a colorless solid. H-NMR (DMSO-
(ꢀ)-Methyl 4-{[(1-{3-[(E)-2-(7-Chloro-2-quinolyl)vinyl]phenyl}-2-
d6) d: 3.91 (3H, s), 7.50—7.65 (3H, m), 7.90—8.08 (6H, m), 8.27 (1H, br s),
phthalimidoethyl)thio]methyl}benzoate (16d) A mixture of 15 (3.00 g, 5.6
mmol), methyl 4-mercaptomethylbenzoate (1.54 g, 8.4 mmol) and dimeth-
ylsulfoxide (90 ml) was combined with cesium carbonate (3.96 g, 11.3
mmol) at 20 °C. The reaction mixture was stirred at room temperature for
1 h, and then benzene, ice and water were added. The reaction mixture was
filtered, and the organic layer of the filtrate was washed with water and brine,
dried over MgSO4, and evaporated in vacuo. Column chromatography of the
residue on silica gel (benzene→5% AcOEt-benzene) gave 16d (1.54 g) as a
8.42 (1H, d, Jꢀ8.8 Hz). FAB-MS m/z: 324 [MꢁH]ꢁ.
3-[(E)-2-(7-Chloro-2-quinolyl)vinyl]benzoic Acid (10) A mixture of 9
(4.60 g, 14.2 mmol), tetrahydrofurane (46 ml) and methanol (23 ml) was
combined with aqueous 1 M NaOH solution (21 ml) and stirred at 40 °C. for
3 h. The reaction mixture was acidified with an aqueous 10% citric acid so-
lution and the resulting precipitate was collected by filtration to give 10 as a
colorless solid. 1H-NMR (DMSO-d6) d: 7.53—7.62 (3H, m), 7.93—8.04
(6H, m), 8.28 (1H, br s), 8.42 (1H, d, Jꢀ8.8 Hz), 12.6—13.8 (1H, br s).
FAB-MS m/z: 308 [MꢃH]ꢁ.
Ethyl 5-{3-[(E)-2-(7-Chloro-2-quinolyl)vinyl]phenyl}oxazol-4-carbox-
ylate (11) A solution of 10 (4.70 g, 15.2 mmol) in dimethylformamide
(70 ml) was combined with potassium carbonate, 1.5 hydrate (5.01 g,
30.3 mmol) and ethyl isocyanoacetate (2.23 g, 19.7 mmol), and then stirred
for 5 min. at room temperature. Diphenylphosphoryl azide (5.01 g,
18.2 mmol) was added to the reaction mixture at 5 °C, and the mixture was
stirred for 20 h at room temperature. Ice-water was added to the reaction
mixture, which was then extracted with ethyl acetate. The organic layer was
washed with water and brine, dried over MgSO4, and evaporated in vacuo.
Column chromatography of the residue on silica gel (5% ethyl
acetate–CHCl3) gave 11 (5.01 g, 93%) as a colorless solid. 1H-NMR
(DMSO-d6) d: 1.29 (3H, t, Jꢀ7.2 Hz), 4.33 (2H, q, Jꢀ7.2 Hz), 7.54—7.65
(3H, m), 7.89—7.96 (4H, m), 8.01 (2H, d, Jꢀ8.8 Hz), 8.04 (1H, br d,
Jꢀ2.0 Hz), 8.32 (1H, br s), 8.42 (1H, d, Jꢀ8.4 Hz), 8.63 (1H, s). EI-MS m/z:
404 [Mꢁ].
1
colorless solid. H-NMR (DMSO-d6) d: 3.58 (1H, d, Jꢀ13.2 Hz), 3.70 (1H,
d, Jꢀ13.2 Hz), 3.88 (3H, s), 4.00—4.16 (3H, m), 4.28 (1H, t, Jꢀ8.0 Hz),
7.24—7.93 (17H, m), 8.08—8.13 (2H, m). FAB-MS m/z: 619 [MꢁH]ꢁ.
(ꢀ)-Methyl 3-[(1-{3-[(E)-2-(7-Chloro-2-quinolyl)vinyl]phenyl}-2-ph-
thalimidoethyl)thio]propanoate (16a) The title compound was prepared
from methyl 3,3-dimethyl-4-mercapto-butylate in the same manner as de-
scribed above, and obtained as a pale yellow solid. (15%). 1H-NMR (CDCl3)
d: 2.40—2.43 (2H, m), 2.64—2.76 (2H, m), 3.61 (3H, s), 4.11—4.20 (2H,
m), 4.46 (1H, t, Jꢀ8.0 Hz), 7.33—7.91 (13H, m), 8.07—8.12 (2H, m). FAB-
MS m/z: 557 [MꢁH]ꢁ.
(ꢀ)-Methyl 4-[(1-{3-[(E)-2-(7-Chloro-2-quinolyl)vinyl]phenyl}-2-ph-
thalimidoethyl)thio]-3,3-dimethylbutylate (16b) The title compound was
prepared from methyl 3,3-dimethyl-4-mercapto-butylate in the same manner
as described above, and obtained as a pale yellow solid. (41%). 1H-NMR
(CDCl3) d: 0.99 (6H, s), 2.18—2.27 (2H, m), 2.51—2.60 (2H, m), 3.60 (3H,
s), 4.10—4.13 (2H, m), 4.40 (1H, t, Jꢀ8.0 Hz), 7.31—7.83 (13H, m),
8.07—8.12 (2H, m). FAB-MS m/z: 599 [MꢁH]ꢁ.
2-Amino-1-{3-[(E)-2-(7-chloro-2-quinolyl)vinyl]phenyl}ethanone Di-
hydrochloride (12) A mixture of 11 (5.00 g, 12.4 mmol), ethanol (25 ml)
and concentrated HCl (25 ml) was stirred under reflux for 6 h. The reaction
mixture was cooled to room temperature, and the resulting precipitate was
collected by filtration to give 12 (4.80 g, 98%) as a colorless solid. 1H-NMR
(ꢀ)-Benzyl {[(1-{3-[(E)-2-(7-Chloro-2-quinolyl)vinyl]phenyl}-2-ph-
thalimidoethyl)thio]methyl}cyclopropanacetate (16c) The title com-
pound was prepared from benzyl (1-mercaptomethyl)cyclopropanacetate in
the same manner as described above, and obtained as a pale yellow solid.
(46%). 1H-NMR (CDCl3) d: 0.43—0.51 (4H, m), 2.30—2.45 (2H, m), 2.56—

608
Vol. 54, No. 5
8.14 (2H, m). FAB-MS m/z: 677 [MꢁH]ꢁ.
2.60 (2H, m), 4.07—4.10 (2H, m), 4.43—4.47 (1H, m), 5.05 (2H, s), 7.27—
7.81 (18H, m), 8.07—8.11 (2H, m). FAB-MS m/z: 673 [MꢁH]ꢁ.
(ꢀ)-Methyl 4-{[(2-{Phenylsulfonylamino}-1-{3-[(E)-2-(7-chloro-2-
quinolyl)vinyl]phenyl}ethyl)thio]methyl}benzoate (17g) The title com-
pound was prepared from 16d and benzenesulfonyl chloride in the same
manner as described above, and obtained as a pale yellow amorphous solid.
(ꢀ)-Methyl 4-{[(1-{3-[(E)-2-(7-Chloro-2-quinolyl)vinyl]phenyl}-2-
phthalimidoethyl)thio]methyl}phenylacetate (16e) The title compound
was prepared from methyl 3,3-dimethyl-4-mercapto-butylate in the same
1
1
manner as described above, and obtained as a pale yellow solid. (34%). H-
(69%). H-NMR (CDCl3) d: 3.30—3.49 (2H, m), 3.50—3.75 (3H, m), 3.91
NMR (CDCl3) d: 3.70 (1H, d, Jꢀ14.0 Hz), 3.82 (3H, s), 3.84 (1H, d,
Jꢀ14.0 Hz), 3.98 (1H, dd, Jꢀ14.4, 8.0 Hz), 4.09 (1H, dd, Jꢀ14.4, 8.0 Hz),
4.24 (1H, t, Jꢀ8.0 Hz), 7.29—7.40 (4H, m), 7.45 (1H, d, Jꢀ16.8 Hz),
7.59—7.68 (3H, m), 7.77—7.86 (7H, m), 7.93 (1H, d, Jꢀ8.8 Hz), 8.00—
8.04 (2H, m), 8.41 (1H, d, Jꢀ8.4 Hz). FAB-MS m/z: 619 [MꢁH]ꢁ.
(3H, s), 4.66 (1H, t, Jꢀ6.4 Hz), 6.91—7.95 (18H, m), 8.08—8.15 (2H, m).
FAB-MS m/z: 627 [MꢃH]ꢁ.
(ꢀ)-Methyl 4-{[(2-{4-Bromophenylsulfonylamino}-1-{3-[(E)-2-(7-chloro-
2-quinolyl)vinyl]phenyl}ethyl)thio]methyl}benzoate (17h) The title com-
pound was prepared from 16d and 4-bromobenzenesulfonyl chloride in the
same manner as described above, and obtained as a pale yellow amorphous
solid. (67%). 1H-NMR (CDCl3) d: 3.25—3.40 (2H, m), 3.52—3.72 (3H, m),
3.91 (3H, s), 4.72 (1H, t, Jꢀ6.4 Hz), 7.03—7.75 (15H, m), 7.92—8.02 (4H,
m). FAB-MS m/z: 706 [MꢃH]ꢁ.
(ꢀ)-Methyl 4-{[(1-{3-[(E)-2-(7-Chloro-2-quinolyl)vinyl]phenyl}-2-
phthalimidoethyl)thio]-2-ethyl}benzoate (16f) The title compound was
prepared from methyl 4-(2-mercaptoethyl)benzoate in the same manner as
described above, and obtained as a pale yellow solid. (26%). 1H-NMR
(CDCl3) d: 2.64—2.73 (2H, m), 2.80—2.84 (2H, m), 3.85 (3H, s), 4.10—
4.15 (2H, m), 4.44 (1H, t, Jꢀ8.4 Hz), 7.11—7.94 (17H, m), 8.07—8.12 (2H,
m). FAB-MS m/z: 633 [MꢁH]ꢁ.
(ꢀ)-Methyl 4-{[(2-{4-Nitrophenylsulfonylamino}-1-{3-[(E)-2-(7-chloro-
2-quinolyl)vinyl]phenyl}ethyl)thio]methyl}benzoate (17i) The title com-
pound was prepared from 16d and 4-nitrobenzenesulfonyl chloride in the
same manner as described above, and obtained as a pale yellow amorphous
solid. (68%). 1H-NMR (CDCl3) d: 3.30—3.95 (2H, m), 3.54—3.75 (3H, m),
3.92 (3H, s), 4.96 (1H, t, Jꢀ6.4 Hz), 6.90—8.30 (19H, m). FAB-MS m/z:
672 [MꢁH]ꢁ.
(ꢀ)-Methyl 4-{[(2-{4-Chlorophenylsulfonylamino}-1-{3-[(E)-2-(7-
chloro-2-quinolyl)vinyl]phenyl}ethyl)thio]methyl}benzoate (17d) A mix-
ture of 16d (1.50 g, 2.4 mmol), 1,4-dioxane (10 ml) and ethanol (20 ml) was
com-bined with methylhydrazine (1.55 ml, 29.0 mmol), and stirred under re-
flux for 7 h. The reaction mixture was cooled to room temperature, and then
ice and a 5% aqueous solution of potassium carbonate were added. The re-
action mixture was extracted with 1,2-dichloroethane, washed with a 5%
aqueous solution of potassium carbonate and brine, dried over Na2SO4 and
evaporated in vacuo. The residue was dissolved in 1,2-dichloromethane,
cooled to 5 °C and combined with triethylamine (0.51 ml, 3.6 mmol) and 4-
chlorobenzenesulfonyl chloride (0.61 g, 2.9 mmol). The reaction mixture
was stirred for 12 h at 5 °C, and then acidified with a 10% aqueous solution
of citric acid. The organic layer was washed with brine, dried over Na2SO4,
and evaporated in vacuo. Column chromatography of the residue on silica
gel (3% AcOEt–benzene→5% AcOEt–benzene) gave 17d (1.03 g, 65%) as a
colorless solid. 1H-NMR (DMSO-d6) d: 3.30—3.45 (2H, m), 3.55 (1H, d,
Jꢀ13.6 Hz), 3.67 (1H, d, Jꢀ13.6 Hz), 3.71 (1H, t, Jꢀ6.4 Hz), 3.91 (3H, s),
4.69 (1H, t, Jꢀ6.0 Hz), 7.06—7.75 (13H, m), 7.97 (2H, d, Jꢀ8.0 Hz),
8.08—8.15 (2H, m). FAB-MS m/z: 663 [MꢁH]ꢁ.
(ꢀ)-Methyl 4-{[(2-{4-Fluorophenylsulfonylamino}-1-{3-[(E)-2-(7-chloro-
2-quinolyl)vinyl]phenyl}ethyl)thio]methyl}benzoate (17j) The title com-
pound was prepared from 16d and benzenesulfonyl chloride in the same
manner as described above, and obtained as a pale yellow amorphous solid.
1
(70%). H-NMR (CDCl3) d: 3.34—3.45 (2H, m), 3.54—3.75 (3H, m), 3.92
(3H, s), 4.90 (1H, t, Jꢀ6.4 Hz), 7.06—8.32 (19H, m). FAB-MS m/z: 672
[MꢃH]ꢁ.
(ꢀ)-4-[(3-{4-Chlorophenylsulfonylamino}-1-{3-[(E)-2-(7-chloro-2-
quinolyl)vinyl]phenyl}propyl)thio]propionic Acid (7b) A mixture of 6b
(0.30 g, 0.49 mmol), tetrahydrofuran (6 ml) and methanol (3 ml) was com-
bined with a 1 M aqueous solution of NaOH (1.2 ml, 1.2 mmol). The reaction
mixture was stirred at room temperature for 12 h, and then acidified with
10% aqueous solution of citric acid. The reaction mixture was extracted with
chloroform, washed with brine, dried over MgSO4, and evaporated in vacuo.
Following column chromatography of the residue on silica gel (AcOEt : n-
1
(ꢀ)-Methyl 3-[(2-{4-Chlorophenylsulfonylamino}-1-{3-[(E)-2-(7-chloro-
hexaneꢀ3 : 7) gave 7b (0.08 g, 27%) as a pale yellow amorphous. H-NMR
2-quinolyl)vinyl]phenyl}ethyl)thio]propanoate (17a) The title compound
was prepared from 16a in the same manner as described above, and obtained
(DMSO-d6) d: 1.89—2.00 (2H, m), 2.31—2.50 (2H, m), 2.71—2.76 (2H,
m), 3.33—3.39 (2H, m), 3.82—4.01 (1H, m), 7.21—8.16 (14H, m), 8.42
(1H, d, Jꢀ8.8 Hz), 12.23 (1H, s). FAB-MS m/z: 601 [MꢁH]ꢁ. Anal. Calcd
for C29H26N2O4S2Cl2: C, 57.90; H, 4.36; N, 4.66; S, 10.66; Cl, 11.79. Found:
C, 57.78; H, 4.45; N, 4.58; S, 10.50; Cl, 11.89.
1
as a pale yellow amorphous solid. (49%). H-NMR (CDCl3) d: 2.40—2.71
(4H, m), 3.36—3.39 (2H, m), 3.66 (3H, s), 3.96 (1H, t, Jꢀ7.4 Hz), 7.12—
7
.82 (13H, m), 8.09—8.15 (2H, m). FAB-MS m/z: 601 [MꢁH]ꢁ.
(
ꢀ
)-Methyl 4-[(2-{4-Chlorophenylsulfonylamino}-1-{3-[(E)-2-(7-chloro-
(ꢀ)-4-[(3-{4-Chlorophenylsulfonylamino}-1-{3-[(E)-2-(7-chloro-2-
quinolyl)vinyl]phenyl}propyl)thio]acetic Acid (7a) The title compound
was prepared from 6a in the same manner as described above, and obtained
2-quinolyl)vinyl]phenyl}ethyl)thio]-3,3-dimethylbutylate (17b) The titl
e
compound was prepared from 16b in the same manner as described above,
and obtained as a pale yellow amorphous solid. (60%). 1H-NMR (CDCl3) d:
0.99 (6H, s), 2.22—2.34 (2H, m), 2.48 (1H, d, Jꢀ12.0 Hz), 2.55 (1H, d,
Jꢀ12.0 Hz), 3.38 (2H, t, Jꢀ6.8 Hz), 3.75 (3H, s), 3.88 (1H, t, Jꢀ7.2 Hz),
4.91 (1H, m), 7.17—7.78 (13H, m), 8.08—8.14 (2H, m). FAB-MS m/z: 643
[MꢁH]ꢁ.
1
as a pale yellow amorphous solid. (33%). H-NMR (DMSO-d6) d: 1.91—
2.04 (2H, m), 2.70—2.78 (2H, m), 2.94 (1H, d, Jꢀ14.2 Hz), 3.10 (1H, d,
Jꢀ14.2 Hz), 4.06 (1H, t, Jꢀ7.6 Hz), 6.86—8.16 (14H, m), 8.42 (1H, d,
Jꢀ8.8 Hz). FAB-MS m/z: 587 [MꢁH]ꢃ. Anal. Calcd for C28H24N2O4S2Cl2:
C, 57.24; H, 4.12; N, 4.77; S, 10.92; Cl, 12.07. Found: C, 56.86; H, 4.12; N,
4.56; S, 11.06; Cl, 12.04.
(ꢀ
)-Benzyl 1-{[(2-{4-Chlorophenylsulfonylamino}-1-{3-[(E)-2-(7-chloro-
2-quinolyl)vinyl]phenyl}ethyl)thio]methyl}cyclopropylacetate (17c) The
(ꢀ)-4-[(3-{4-Chlorophenylsulfonylamino}-1-{3-[(E)-2-(7-chloro-2-
quinolyl)vinyl]phenyl}propyl)thio]butylic Acid (7c) The title compound
was prepared from 6c in the same manner as described above, and obtained
as a pale yellow solid. (44%). mp 76—78 °C. 1H-NMR (DMSO-d6) d:
1.83—1.92 (1H, m), 2.10—2.12 (2H, m), 2.35—2.56 (4H, m), 3.01—3.18
(2H, m), 3.88 (1H, t, Jꢀ7.2 Hz), 5.05 (1H, t, Jꢀ6.4 Hz), 7.18—7.79 (13H,
m), 8.05—8.15 (2H, m). FAB-MS m/z: 615 [MꢁH]ꢁ. Anal. Calcd for
C30H28N2O4S2Cl2· 0.3C2H3N·0.3H2O: C, 58.03; H, 4.69; N, 5.09; S, 10.13;
Cl, 11.20. Found: C, 57.98; H, 4.50; N, 5.04; S, 10.19; Cl, 11.33.
t
itle compound was prepared from 16c in the same manner as described
above, and obtained as a pale yellow amorphous solid. (55%). 1H-NMR
(CDCl3) d: 0.42—0.55 (4H, m), 2.40—2.42 (2H, m), 2.48—2.56 (2H, m),
3.33 (2H, t, Jꢀ6.4 Hz), 3.93 (1H, t, Jꢀ7.2 Hz), 5.12 (2H, s), 7.13—7.78 (18H,
m), 8.11—8.40 (2H, m). FAB-MS m/z: 717 [MꢁH]ꢁ.
(ꢀ)-Methyl 4-{[(2-{4-Chlorophenylsulfonylamino}-1-{3-[(E)-2-(7-
chloro-2-quinolyl)vinyl]phenyl}ethyl)thio]methyl}phenylacetate (17e)
The title compound was prepared from 16e in the same manner as described
above, and obtained as a pale yellow amorphous solid. (54%). 1H-NMR
(DMSO-d6) d: 3.23—3.35 (2H, m), 3.69 (1H, d, Jꢀ14.0 Hz), 3.81 (1H, d,
Jꢀ14.0 Hz), 3.84 (3H, s), 3.85—3.89 (1H, m), 4.17 (1H, d, Jꢀ8.0 Hz),
7.35—7.39 (3H, m), 7.45 (1H, d, Jꢀ16.4 Hz), 7.49 (1H, br s), 7.57—7.64
(4H, m), 7.59—7.73 (2H, m), 7.82 (1H, d, Jꢀ16.4 Hz), 7.90 (2H, br d,
Jꢀ8.0 Hz), 7.93 (1H, d, Jꢀ9.2 Hz), 8.00—8.04 (3H, m), 8.42 (1H, d,
Jꢀ8.4 Hz). FAB-MS m/z: 619 [MꢁH]ꢁ.
(ꢀ)-4-[(3-{4-Chlorophenylsulfonylamino}-1-{3-[(E)-2-(7-chloro-2-
quinolyl)vinyl]phenyl}propyl)thio]-3,3-dimethylbutylic Acid (7d) The
title compound was prepared from 6d in the same manner as described
1
above, and obtained as a pale yellow solid. (40%). mp 86—89 °C. H-NMR
(DMSO-d6) d: 0.93 (6H, s), 1.89—2.03 (2H, m), 2.09—2.17 (2H, m), 2.34
(1H, d, Jꢀ12.8 Hz), 2.43 (1H, d, Jꢀ12.8 Hz) 2.70—2.79 (2H, m), 3.88 (1H,
t, Jꢀ8.4 Hz), 7.22—8.04 (14H, m), 8.42 (1H, d, Jꢀ8.4 Hz), 12.00 (1H, br s).
FAB-MS m/z: 643 [MꢁH]ꢁ. Anal. Calcd for C32H32N2O4S2Cl2: C, 59.71; H,
5.01; N, 4.35; S, 9.96; Cl, 11.02. Found: C, 59.80; H, 4.94; N, 4.38; S, 9.72;
Cl, 11.30.
(ꢀ)-4-{[(3-{4-Chlorophenylsulfonylamino}-1-{3-[(E)-2-(7-chloro-
2-quinolyl)vinyl]phenyl}propyl)thio]methyl}-1-cyclopropanacetic Acid
(7e) The title compound was prepared from 6e in the same manner as de-
(ꢀ)-Methyl 4-{[(2-{4-Chlorophenylsulfonylamino}-1-{3-[(E)-2-(7-
chloro-2-quinolyl)vinyl]phenyl}ethyl)thio]-2-ethyl}benzoate (17f) The
title compound was prepared from 16f in the same manner as described
above, and obtained as a pale yellow amorphous solid. (79%). 1H-NMR
(CDCl3) d: 2.64—2.68 (2H, m), 2.80—2.84 (2H, m), 3.34 (2H, t,
Jꢀ6.4 Hz), 3.87 (3H, s), 3.87—3.93 (1H, m), 7.14—7.95 (17H, m), 8.08—

May 2006
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NMR (DMSO-d6) d: 3.30—3.33 (2H, m), 3.57—3.60 (3H, m), 3.69 (1H, d,
Jꢀ12.4 Hz), 3.88 (1H, t, Jꢀ7.4 Hz), 7.01—8.04 (18H, m), 8.42 (1H, d,
Jꢀ8.2 Hz). FAB-MS m/z: 663 [MꢁH]ꢁ. Anal. Calcd for C34H28N2O4S2Cl2:
C, 61.54; H, 4.25; N, 4.22; S, 9.66; Cl, 10.68. Found: C, 61.61; H, 4.31; N,
4.46; S, 9.63; Cl, 10.64.
scribed above, and obtained as a pale yellow solid. (40%). mp 97—100 °C.
1H-NMR (DMSO-d6) d: 0.31—0.45 (4H, m), 1.88—1.99 (2H, m), 2.21—
2.29 (2H, m), 2.43—2.51 (2H, m), 2.67—2.73 (2H, m), 3.93 (1H, t,
Jꢀ8.4 Hz), 7.22—8.03 (14H, m), 8.42 (1H, d, Jꢀ8.4 Hz), 12.01 (1H, br s).
FAB-MS m/z: 641 [MꢁH]ꢁ. Anal. Calcd for C32H30N2O4S2Cl2: C, 59.90; H,
4.71; N, 4.37; S, 10.00; Cl, 11.05. Found: C, 59.69; H, 4.56; N, 4.31; S,
10.05; Cl, 11.00.
(ꢀ)-4-{[(2-{4-Chlorophenylsulfonylamino}-1-{3-[(E)-2-(7-chloro-2-
quinolyl)vinyl]phenyl}ethyl)thio]2-ethyl}benzoic Acid (18f) The title
compound was prepared from 17f in the same manner as described above,
(ꢀ)-4-{[(3-{4-Chlorophenylsulfonylamino}-1-{3-[(E)-2-(7-chloro-2-
quinolyl)vinyl]phenyl}propyl)thio]methyl}benzoic Acid (7f) The title
compound was prepared from 6f in the same manner as described above,
1
and obtained as a pale yellow solid. (32%). mp 186—187 °C. H-NMR
(DMSO-d6) d: 2.65—2.87 (4H, m), 3.33 (2H, t, Jꢀ7.0 Hz), 3.81 (1H, t,
Jꢀ7.6 Hz), 4.70—4.72 (1H, m), 7.13—7.75 (14H, m), 7.95—8.15 (5H, m).
FAB-MS m/z: 663 [MꢁH]ꢁ. Anal. Calcd for C34H28N2O4S2Cl2: C, 61.54; H,
4.25; N, 4.22; S, 9.66; Cl, 10.68. Found: C, 61.61; H, 4.31; N, 4.46; S, 9.63;
Cl, 10.64.
(ꢀ)-4-{[(2-{Phenylsulfonylamino}-1-{3-[(E)-2-(7-chloro-2-quinolyl)-
vinyl]phenyl}ethyl)thio]methyl}benzoic Acid (18g) The title compound
was prepared from 17g in the same manner as described above, and ob-
tained as a pale yellow solid. (58%). mp 212—214 °C. 1H-NMR (DMSO-
d6) d: 3.19—3.36 (2H, m), 3.67 (1H, d, Jꢀ13.6 Hz), 3.79 (1H, d, Jꢀ
13.6 Hz), 3.85—3.89 (1H, m), 6.85—8.12 (19H, m), 8.42 (1H, d, Jꢀ
8.4 Hz), 12.90 (1H, br s). FAB-MS m/z: 615 [MꢁH]ꢁ. Anal. Calcd for
C33H27N2O4S2Cl·0.1H2O: C, 64.24; H, 4.44; N, 4.54; S, 10.39; Cl, 5.75.
Found: C, 64.07; H, 4.26; N, 4.63; S, 10.25; Cl, 5.71.
(ꢀ)-4-{[(2-{4-Bromophenylsulfonylamino}-1-{3-[(E)-2-(7-chloro-2-
quinolyl)vinyl]phenyl}ethyl)thio]methyl}benzoic Acid (18h) The title
compound was prepared from 17h in the same manner as described above,
and obtained as a pale yellow solid. (69%). mp 203—205 °C. 1H-NMR
(DMSO-d6) d: 3.23—3.36 (2H, m), 3.78 (1H, d, Jꢀ13.6 Hz), 3.80 (1H, d,
Jꢀ13.6 Hz), 3.86 (1H, t, Jꢀ7.6 Hz), 7.15—8.04 (18H, m), 8.42 (1H, d,
Jꢀ8.8 Hz), 12.92 (1H, br s). FAB-MS m/z: 693 [MꢁH]ꢁ. Anal. Calcd for
C33H26N2O4S2ClBr: C, 57.11; H, 3.78; N, 4.04; S, 9.24; Br, 11.51; Cl, 5.11.
Found: C, 57.13; H, 3.68; N, 4.16; S, 9.14; Br, 11.45; Cl, 5.10.
(ꢀ)-4-{[(2-{4-Nitrophenylsulfonylamino}-1-{3-[(E)-2-(7-chloro-2-
quinolyl)vinyl]phenyl}ethyl)thio]methyl}benzoic Acid (18i) The title
compound was prepared from 17i in the same manner as described above,
and obtained as a pale yellow solid. (63%). mp 233—235 °C. 1H-NMR
(DMSO-d6) d: 3.32—3.42 (2H, m), 3.68 (1H, d, Jꢀ13.6 Hz), 3.81 (1H, d,
Jꢀ13.6 Hz), 3.86 (1H, t, Jꢀ6.4 Hz), 7.15—8.04 (14H, m), 8.28—8.43 (5H,
m), 12.91 (1H, br s). FAB-MS m/z: 660 [MꢁH]ꢁ. Anal. Calcd for
C33H26N3O6S2Cl: C, 60.04; H, 3.97; N, 6.37; S, 9.71; Cl, 5.37. Found: C,
59.91; H, 3.98; N, 6.33; S, 9.65; Cl, 5.40.
(ꢀ)-4-{[(2-{4-Fluorophenylsulfonylamino}-1-{3-[(E)-2-(7-chloro-2-
quinolyl)vinyl]phenyl}ethyl)thio]methyl}benzoic Acid (18j) The title
compound was prepared from 17j in the same manner as described above,
and obtained as a pale yellow solid. (74%). mp 192—193 °C. 1H-NMR
(DMSO-d6) d: 3.21—3.29 (2H, m), 3.68 (1H, d, Jꢀ13.6 Hz), 3.80 (1H, d,
Jꢀ13.6 Hz), 3.88 (1H, t, Jꢀ7.2 Hz), 7.12—8.04 (18H, m), 8.42 (1H, d,
Jꢀ8.4 Hz), 12.91 (1H, br s). FAB-MS m/z: 633 [MꢁH]ꢁ. Anal. Calcd for
C33H26N2O4S2ClF: C, 62.60; H, 4.14; N, 4.42; S, 10.13; Cl, 5.60; F, 3.00.
Found: C, 62.52; H, 4.05; N, 4.36; S, 10.12; Cl, 5.63; F, 3.06.
Biological Methods. Agonist-Induced Contraction of the Guinea Pig
Ileum Guinea pigs weighing 370 to 740 g were sacrificed by exsanguina-
tion. The terminal ileum was removed and suspended in Tyrode’s solution,
which had the following composition: 136.8 mM NaCl, 2.7 mM KCl, 1.8 mM
CaCl2, 1.1 mM MgCl2, 0.42 mM NaH2PO4, 11.9 mM NaHCO3 and 5.6 mM
glucose (pH 7.4). The ileum was divided into segments of approximately
40 mm in length and set in a Magnus vessel containing 10 ml of Tyrode’s so-
lution aerated with a 95% O2–5% CO2 gas mixture. The tissue was placed
under tension using a 1-g mass. The force generated by the tissue was iso-
metrically measured. The ileum contractile response against 1ꢄ10ꢃ9 M LTD4
was first measured in the absence of the test compound and then in the pres-
ence of the compound at various concentrations. An IC50 value was calcu-
lated by linear regression analysis (maximum-likelihood method) using
SAS.
U46619-Induced Platelet Aggregation Using a syringe containing 1
volume of 3.8% sodium citrate aqueous solution, 9 volumes of blood were
collected. Guinea pig and human PRP was obtained by centrifuging the
blood for 10 min at 270 g. The remaining blood was further centrifuged at
110ꢄg for 10 min to yield PPP. The PRP was diluted with PPP to adjust
control of the platelet count to 500000 cells/ml. Platelet aggregation was in-
duced by a stable analog of TXA2, 1ꢄ10ꢃ6 M U46619, and was measured
using a NBS Hema Tracer VI (Nikoh Bioscience, Tokyo, Japan). Various
concentrations of the compounds were added to the PRP 2 min before the
addition of U46619, and an IC50 value (50% inhibition concentration) was
1
and obtained as a pale yellow amorphous solid. (65%). H-NMR (DMSO-
d6) d: 1.93—2.01 (2H, m), 2.68—2.71 (2H, m), 3.56 (1H, d, Jꢀ13.2 Hz),
3.66 (1H, d, Jꢀ13.2 Hz), 3.87 (1H, t, Jꢀ7.2 Hz), 7.20—8.04 (18H, m), 8.42
(1H, d, Jꢀ8.8 Hz), 12.87 (1H, br s). FAB-MS m/z: 663 [MꢁH]ꢁ. Anal.
Calcd for C34H28N2O4S2Cl2: C, 61.54; H, 4.25; N, 4.22; S, 9.66; Cl, 10.68.
Found: C, 61.29; H, 4.29; N, 4.14; S, 9.43; Cl, 10.38.
(ꢀ)-4-{[(3-{4-Chlorophenylsulfonylamino}-1-{3-[(E)-2-(7-chloro-2-
quinolyl)vinyl]phenyl}propyl)thio]methyl}phenylacetic Acid (7g) The
title compound was prepared from 6g in the same manner as described
1
above, and obtained as a pale yellow solid. (20%). mp 76—78 °C. H-NMR
(DMSO-d6) d: 1.94—2.03 (2H, m), 2.65—2.75 (2H, m), 3.36—3.57 (4H,
m), 3.74—3.91 (1H, m), 7.08—8.00 (18H, m), 8.42 (1H, d, Jꢀ8.0 Hz),
12.31 (1H, br s). FAB-MS m/z: 677 [MꢁH]ꢁ. Anal. Calcd for
C35H30N2O4S2Cl2: C, 62.03; H, 4.46; N, 4.13; S, 9.46; Cl, 10.46. Found: C,
62.19; H, 4.60; N, 4.19; S, 9.34; Cl, 10.75.
(ꢀ)-4-{2-[(3-{4-Chlorophenylsulfonylamino}-1-{3-[(E)-2-(7-chloro-2-
quinolyl)vinyl]phenyl}propyl)thio]ethyl}benzoic Acid (7h) The title
compound was prepared from 6h in the same manner as described above,
and obtained as a pale yellow amorphous solid. (70%). 1H-NMR (CDCl3) d:
1.95—2.09 (2H, m), 2.51—2.65 (2H, m), 2.73—2.87 (2H, m), 2.92—3.05
(2H, m), 3.84 (1H, t, Jꢀ7.6 Hz), 4.93 (1H, t, Jꢀ6.4 Hz), 7.14—7.20 (3H,
m), 7.34 (1H, t, Jꢀ7.6 Hz), 7.42—7.51 (6H, m), 7.62—7.76 (5H, m), 7.96—
7.99 (2H, m), 8.10—8.13 (2H, m). FAB-MS m/z: 677 [MꢁH]ꢁ. Anal. Calcd
for C35H30N2O4S2Cl2·0.1H2O: C, 61.87; H, 4.48; N, 4.12; S, 9.44; Cl, 10.44.
Found: C, 61.57; H, 4.52; N, 4.11; S, 9.55; Cl, 10.53.
(ꢀ)-3-[(2-{4-Chlorophenylsulfonylamino}-1-{3-[(E)-2-(7-chloro-2-
quinolyl)vinyl]phenyl}ethyl)thio]propanoic Acid (18a) The title com-
pound was prepared from 17a in the same manner as described above, and
obtained as a pale yellow amorphous solid. (59%). 1H-NMR (CDCl3) d:
2.38—2.71 (4H, m), 3.31—3.38 (2H, m), 4.02 (1H, t, Jꢀ7.6 Hz), 6.99—
7.78 (13H, m), 8.04—8.17 (2H, m). FAB-MS m/z: 587 [MꢁH]ꢁ. Anal.
Calcd for C28H24N2O4S2Cl2·0.3H2O: C, 56.72; H, 4.18; N, 4.72; S, 10.82;
Cl, 11.96. Found: C, 57.60; H, 4.25; N, 5.01; S, 10.85; Cl, 12.04.
(ꢀ)-4-[(2-{4-Chlorophenylsulfonylamino}-1-{3-[(E)-2-(7-chloro-2-
quinolyl)vinyl]phenyl}ethyl)thio]3,3-dimethylbutylic Acid (18b) The
title compound was prepared from 17b in the same manner as described
above, and obtained as a pale yellow solid. (31%). mp 177—179 °C. 1H-
NMR (DMSO-d6) d: 0.98 (6H, s), 2.11—2.19 (2H, m), 2.43—2.55 (2H, m),
3.26—3.35 (2H, m), 3.90 (1H, t, Jꢀ7.6 Hz), 7.22—8.03 (13H, m), 8.42 (1H,
d, Jꢀ8.4 Hz), 12.02 (1H, br s). FAB-MS m/z: 629 [MꢁH]ꢁ. Anal. Calcd for
C31H30N2O4S2Cl2: C, 59.14; H, 4.80; N, 4.45; S, 10.19; Cl, 11.26. Found: C,
58.75; H, 4.65; N, 4.38; S, 10.29; Cl, 11.37.
(ꢀ)-1-{[(2-{4-Chlorophenylsulfonylamino}-1-{3-[(E)-2-(7-chloro-2-
quinolyl)vinyl]phenyl}ethyl)thio]methyl}cyclopropylacetic Acid (18c)
The title compound was prepared from 17c in the same manner as described
above, and obtained as a pale yellow solid. (45%). mp 173—176ºC. 1H-
NMR (DMSO-d6) d: 0.36—0.44 (4H, m), 2.21—2.33 (2H, m), 2.50—2.58
(2H, m), 3.25—3.35 (2H, m), 3.97 (1H, t, Jꢀ7.2 Hz), 7.21—8.03 (14H, m),
8.42 (1H, d, Jꢀ8.4 Hz), 12.02 (1h, br s). FAB-MS m/z: 627 [MꢁH]ꢁ. Anal.
Calcd for C31H28N2O4S2Cl2: C, 59.33; H, 4.50; N, 4.46; S, 10.22; Cl, 11.30.
Found: C, 58.94; H, 4.45; N, 4.45; S, 10.06; Cl, 11.22.
(ꢀ)-4-{[(2-{4-Chlorophenylsulfonylamino}-1-{3-[(E)-2-(7-chloro-2-
quinolyl)vinyl]phenyl}ethyl)thio]methyl}benzoic Acid (18d) The title
compound was prepared from 17d in the same manner as described above,
and obtained as a pale yellow solid. (55%). mp 188—190 °C. 1H-NMR
(DMSO-d6) d: 3.23—3.29 (2H, m), 3.68 (1H, d, Jꢀ13.6 Hz), 3.80 (1H, d,
Jꢀ13.6 Hz), 3.87 (1H, t, Jꢀ7.6 Hz), 7.15—8.04 (18H, m), 8.42 (1H, d,
Jꢀ8.4 Hz), 12.91 (1H, br s). FAB-MS m/z: 649 [MꢁH]ꢁ. Anal. Calcd for
C33H26N2O4S2Cl2: C, 61.01; H, 4.03; N, 4.31; S, 9.87; Cl, 10.91. Found: C,
60.92; H, 3.97; N, 4.49; S, 9.83; Cl, 10.88.
(ꢀ)-4-{[(2-{4-Chlorophenylsulfonylamino}-1-{3-[(E)-2-(7-chloro-2-
quinolyl)vinyl]phenyl}ethyl)thio]methyl}phenylacetic Acid (18e) The
title compound was prepared from 17e in the same manner as described
above, and obtained as a pale yellow solid. (32%). mp 138—140 °C. 1H-

610
Vol. 54, No. 5
calculated from the inhibition ratio on the basis of the maximum light trans-
mittance. All experiments were carried out within 4 h after blood collection
to avoid a decrease in platelet sensitivity to U46619.
LTD4-Induced Acceleration of Plasma Leakage in Guinea Pig Skin
Male Hartley guinea pigs whose back fur had been shaved with an electric
clipper on the day before the experiment were given an intravenous adminis-
tration of saline (1 ml per animal) containing 1% Evans blue. Two minutes
later, 5 ng LTD4 and the vehicle solution were administered intracutaneously
on the back of the guinea pig (at 2 points for LTD4 and 2 points for vehicle).
5) Rinkema L. E., Bemis K. G., Fleisch J. H., J. Pharmacol. Exp. Ther.,
230, 550—557 (1984).
6) Ueno A., Tanaka K., Katori M., Prostaglandins, 23, 865—880 (1982).
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8) Fujimura M., Sakamoto S., Saito. M., Miyake Y., Matsuda T., Allergy
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T., Aishita H., Jpn. J. Pharmacol., 60, 217—225 (1992).
The guinea pig was sacrificed by decapitation 30 min later. The skin was re- 10) Krell G. D., Aharony D., Buckner C. K., Keith R. A., Kusner E. J.,
moved, and the visible blood in the isolated skin was also removed as much
as possible. The pigment retained within the skin was then extracted by the
addition of extraction buffer ([7 : 3] acetone : 0.5% Na2SO4 solution) and the
amount of LTD4-induced pigment leakage was measured using the 620 nm
absorbance of the extract (UV-visible spectrophotometer, model UV-160A;
Snyder D. W., Bernstein P. R., Matassa V. G., Yee Y. K., Brown F. J.,
Hesp B., Giles R. E., Am. Rev. Respir. Dis., 141, 978—987 (1990).
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rato K., Takishima T., J. Allergy Clin. Immunol., 92, 507—512 (1993).
Shimadzu, Kyoto). LTD4-induced dye leakage was defined by subtracting 13) Adkins J. C., Brogden R. N., Drugs, 55, 121—144 (1998).
the dye content in the vehicle-injected site from that in the LTD4-induced
site, so these calculated dye contents were collected for Evans blue dye re-
14) Samara E., Cao G., Locke C., Grannerman G. R., Dean R., Killian A.,
Clin. Pharmacol. Ther., 62, 426—435 (1997).
maining within the vasculature. This dye amount was used as an index of 15) Kurashima K., Ogawa H., Ohka T., Fujimura M., Matsuda T., Ann. Al-
plasma leakage, although there was a potential uncontrolled hydrostatic lergy, 68, 53—56 (1992).
pressure effect in this system. Test compounds were orally administered 1 h 16) Yamada T., Takahashi Y., Ishizaki M., Musoh K., Ohashi T., Tanaka
before intracutaneous administration of LTD4.
U46619-Induced Increase in Airway Resistance Increase Male Hart-
ley guinea pigs were anesthetized by intraperitoneal injection of 1.2 g/kg
H., Inagaki N., Nagai H., Pharmacology, 69, 51—58 (2003).
17) Arakida Y., Suwa K., Ohga K., Yokota M., Miyata K., Yamada T.,
Honda K., J. Pharmacol. Exp Ther., 287, 633—639 (1998).
urethane and a tracheal cannula was inserted. Spontaneous respiration was 18) Arakida Y., Ohga K., Kobayashi S., Yokota M., Miyata K., Yamada T.,
stopped with gallamine (1 mg/kg i.v.), and artificial respiration was carried Honda K., Eur. J. Pharm., 362, 229—233 (1998).
out at a rate of 60 strokes/min and a volume of 1 ml/100 g body weight per 19) Arakida Y., Ohga K., Suwa K., Yokota M., Miyata K., Yamada T.,
cycle. After intravenous administration of U46619 (3 mg/kg), airway resist-
ance was measured using a respiratory function measuring apparatus (Model
6; Buxco Electronics, Inc., Sharon, CT, U.S.A.). The airway resistance was
Honda K., J. Pharmacol. Exp Ther., 290, 1285—1291 (1999).
20) Arakida Y., Ohga K., Okada Y., Morio H., Suwa K., Yokota M., Eur. J.
Pharm., 403, 169—179 (2000).
measured as the mean of every 5 s and expressed as the percentage change 21) Arakida Y., Ohga K., Suwa K., Okada Y., Morio H., Yokota M., Mi-
compared with the basal resistance level. The test compound was orally ad-
ministered 1 h before intravenous injection of agonists. The effects of each
yata K., Yamada T., Honda K., Jpn. J. Pharmacol., 82, 287—294
(2000).
compound were evaluated using the peak percentage change in lung resist- 22) Arakida Y., Ohga K., Suwa K., Okada Y., Morio H., Yokota M.,
ance.
Mi–yata K., Yamada T., Honda K., Jpn. J. Pharmacol., 83, 63—72
(2000).
Acknowledgements The authors are grateful to Ms. Keiko Ohga and 23) Jones T. R., Labelle M., Belly M., Champion E., Charette L., Evans J.,
Ms. Kiyomi Suwa for biological evaluation and to the staff of the Analysis
& Pharmacokinetics Research Laboratories for elemental analysis and spec-
tral measurements.
Ford-Hutchinson A. W., Gauthier J. Y., Lord A., Masson P., Mcauliffe
M., Mcfarlane C. S., Matters K. M., Pickett C., Piechuta H., Rochette
C., Rodger I. W., Sawyer N., Young R. N., Zamboni R., Abraham W.
M., Can. J. Pharmacol., 73, 191—201 (1995).
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