N-formyl peptide 類化合物的藥理作用之一是促使嗜中性白血球去顆

參、抗發炎活性試驗

將前述合成出及經結構判定正確之化合物

21-33、41-48、50-53、

61-68、70-73、81-93、101-109、111-114、116-118 及 120-123 提供抗

發炎活性試驗，測試之方法分別採用化合物對於 fMLP 誘導的嗜中性 白血球去顆粒作用 (neutrophil degranulation)抑制試驗、化合物對於 fMLP 誘 導 的 嗜 中 性 白 血 球 超 氧 自 由 基 生 成 作 用 (neutrophil superoxide formation)抑制試驗及化合物對於 PMA 誘導的嗜中性白血 球超氧自由基生成作用 (neutrophil superoxide formation)抑制試驗，依 其抑制百分率來判定其活性強度，篩選結果分別如：Table 13 至 Table 18、Table 19 至 Table 24 及 Table 25 至 Table 30 所示。

fMLP 為 一 種 趨 化 性 (chemotactic peptide)

N-formyl-

methionylleucylphenylalanine (CHO-Met-Leu-Phe-OH) 之 簡 稱 ⁽⁸⁷⁾ 。

N-formyl peptide 類化合物的藥理作用之一是促使嗜中性白血球去顆

粒作用，故 fMLP 可作為化合物測定抗發炎的藥理活性試驗時之誘導 劑。fMLP 除可促使嗜中性白血球去顆粒作用外，亦可促使嗜中性白 血球超氧自由基生成作用。

PMA 之 全 名 為 phorbol 12-myristate 13-acetate diester ( 亦 稱 12-o-tetradecanoylphorbol-13-acetate；故亦簡稱 TPA)，其結構如下所 示：

O H

₃

C

HO H

OH H

CH

₃

CH

₃

H

O C

O CH

₃

HO H

₃

C

O C O CH

₂

H

₃

C ( )

12

PMA

PMA 與 fMLP 兩者皆能促使嗜中性白血球超氧自由基生成作 用，彼此差異在於作用位置不相同，fMLP 會與嗜中性白血球細胞膜 上接受器結合產生超氧自由基之生成⁽⁸⁸⁾，而 PMA 則直接進入嗜中性 白血球細胞內與細胞內 protein kinase C (PKC)結合產生超氧自由基生 成作用⁽⁸⁹⁾，根據彼此的差異點可知，抗發炎化合物其產生藥理活性的

作用位置。

選用 trifluoperazine⁽⁹⁰⁾ (TFP)當作 positive control 的原因是它可以 抑制嗜中性白血球 (neutrophils)釋放出溶菌酵素 (lysozyme)，同時減 少 O2. –的形成，而它的作用是 calmodulin antagonist 和 protein kinase C inhibitor。

trifluoperazine

S N N

N H

₃

C

CF

₃

由測試的結果發現：

(一)對於fMLP誘導的嗜中性白血球去顆粒作用抑制試驗

從化合物21-33、41-48、50-53、61-68、70-73、81-93、101-109、

111-114、116-118及120-123對以fMLP誘導的嗜中性白血球去顆粒作用

之體外試驗中，由

β

-glucuronidase或lysozyme的抑制百分率 (見Table 13至Table 18)看來，在濃度30

µ

M時，化合物26、27、30、33、41、

42、50、53、61-63、65、68、71、81、83-85、87、88、90、92、93

及113分別呈現弱的抑制活性 (具有約20-42%的抑制百分率)，但是發 現化合物25、28及32呈現明顯的抑制活性，其抑制

β

-glucuronidase的 IC50值分別為10.5

±

0.8

µ

M 、35.7

±

3.8

µ

M 及12.7

±

4.4

µ

M ，而抑制 lysozyme的IC50值分別為15.5±2.8

µM、50.6±4.6 µM及18.5±5.3 µM。

其他化合物則無明顯的抑制活性。

綜 合 上 述 ， 發 現 ethyl 5-(2'-alkoxycarbonyl substituted phenoxy)-furan-2-carboxylates (21-33)類衍生物的活性較明顯。在ethyl 5-(2'-alkoxycarbonyl substituted phenoxy)furan-2-carboxylates (21-33)類 衍生物中將甲基、甲氧基或溴原子導入苯環時，具有較高的活性，而

化 合 物 ethyl

5-(2'-methoxycarbonyl-4'-bromophenoxy)furan-2-carboxylate (32)的IC50

值 (抑制

β

-glucuronidase的IC50 = 12.7

±

4.4

µ

M及抑制lysozyme的IC50 = 18.5±5.3

µM)約為trifluoperazine之IC

50值 (抑制β-glucuronidase的IC50

= 24.4

±

0.5

µ

M及抑制lysozyme的IC50 = 22.8

±

0.5

µ

M)的二分之一倍，化 合 物 ethyl 5-(2'-ethoxycarbonyl-3'-methyl-phenoxy)furan-2-carboxylate (25) 的 IC50值 ( 抑 制

β

-glucuronidase 的 IC₅₀ = 10.5

±

0.8

µ

M 及 抑 制 lysozyme 的 IC50 = 15.5

±

2.8

µ

M) 約 與 trifluoperazine 的 IC50值 (抑 制

β

-glucuronidase的IC₅₀ = 14.2

±

0.7

µ

M及抑制lysozyme的IC₅₀ = 16.0

±

0.9

µ

M) 相 當 ， 化 合 物 ethyl

5-(2'-methoxy-carbonyl-4'-methoxyphenoxy)furan-2-carboxylate (28) 的 IC50值 (抑制β-glucuronidase的IC50 = 35.7± 3.8

µM及抑制lysozyme的

IC50 = 50.6±4.6

µM)約為trifluoperazine之IC

50值 (抑制β-glucuronidase 的IC50 = 24.4

±

0.5

µ

M及抑制lysozyme的IC50 = 22.8

±

0.5

µ

M)的二倍。

與甲基、甲氧基或溴原子相較之下，若將碘原子導入苯環，則其活性 降低，此外，若將氯原子導入苯環，則其活性降得更低。

Table 13. The inhibitory effect of ethyl 5-(2'-alkoxycarbonyl substituted phenoxy)- furan-2-carboxylates on rat neutrophil degranulation (in vitro)

21: R=CH

3

, R

1

=R

2

=R

3

=R

4

=H 28: R=CH

3

, R

1

=R

3

=R

4

=H, R

2

=OCH

3

22: R=CH

3

, R

1

=R

2

=R

3

=H, R

4

=CH

3

29: R=CH

3

, R

2

=R

3

=R

4

=H, R

1

=OCH

3

23: R=CH

3

, R

1

=R

2

=R

4

=H, R

3

=CH

3

30: R=CH

3

, R

1

=R

2

=R

4

=H, R

3

=Cl 24: R=CH

3

, R

1

=R

3

=R

4

=H, R

2

=CH

3

31: R=CH

3

, R

1

=R

3

=R

4

=H, R

2

=Cl 25: R=C

2

H

5

, R

2

=R

3

=R

4

=H, R

1

=CH

3

32: R=CH

3

, R

1

=R

3

=R

4

=H, R

2

=Br 26: R=CH

3

, R

1

=R

2

=R

3

=H, R

4

=OCH

3

33: R=CH

3

, R

1

=R

3

=R

4

=H, R

2

=I 27: R=CH

3

, R

1

=R

2

=R

4

=H, R

3

=OCH

3

Percent Release

Compound conc.--- (µM) β-Glucuronidase (%inh) Lysozyme (%inh)

Control 15.1±0.9 -- 30.4±1.2 -- 21 (30) 13.8±0.3 9.0±4.5 22.9±1.0* 28.0±5.1

(100) 13.5±2.3 7.2±10.7 24.8±3.3 24.7±6.6 22 (30) 12.8±1.8 18.8±9.0 28.0±1.8 11.2±6.3 (100) 12.7±1.8 14.7±8.3 31.7±2.5 3.0±3.9 23 (30) 13.5±1.5 8.3±8.0 28.1±0.3 17.1±3.9

(100) 16.4±0.9 -7.7±5.0 32.5±3.3 6.4±8.3 TFP (3) 13.9±2.0 9.6±4.8 30.5±2.9 8.7±4.1 (10) 8.1±0.8* 52.1±7.1 15.1±1.2** 52.7±3.7 (30) 2.2±1.3** 87.8±4.5 3.2±1.7** 89.7±4.8

IC

50

14.3±3.9 12.5±3.7

Control 22.6±1.7 -- 46.4±2.1 -- 24 (30) 22.6±2.5 -0.4±10.0 49.3±6.2 -5.5±9.2

(100) 21.5±0.7 3.7±8.2 52.5±6.9 -12.5±10.9 Control 14.8±1.0 -- 16.7±1.5 -- TFP (3) 13.4±1.2 9.6±1.8 14.9±0.9 10.5±2.2

(10) 8.5±1.3** 43.1±5.3 9.7±0.9* 39.7±9.8 (30) 5.1±0.4** 64.7±3.1 6.6±0.4** 59.5±5.2

COOR

O R

₄

R

₃

R

₂

R

₁

O COOCH

₂

CH

₃

IC

50

19.3±1.3 21.9±2.5 Control 42.5±0.8 -- 61.5±3.3 -- 25 (3) 33.9±2.1** 20.2±3.6 54.4±7.0 12.2±6.8

(10) 14.8±1.6** 65.1±3.3 29.9±6.3** 51.9±8.0 (30) 0.7±0.5** 98.1±1.4 14.3±4.5** 77.4±6.3

IC

₅₀

10.5±0.8 15.5±2.8

TFP (3) 35.2±0.8 16.8±2.2 55.7±5.3 8.9±1.9 (10) 22.7±1.6** 44.9±8.6 44.0±6.8 28.0±6.1 (30) 4.8±0.7** 88.6±2.7 2.0±2.2** 96.9±4.1

IC

50

14.2±0.7 16.0±0.9

Control 24.5±0.3 -- 46.1±1.1 -- 26 (10) 21.5±1.3 12.0±6.4 40.1±1.2 13.1±0.8

(30) 17.4±0.3** 29.1±1.9 33. 7±1.5** 27.0±1.6 (100) 11.2±2.3** 54.1±9.6 28.0±4.2** 39.5±8.4

IC

50

92.1±12.2

27 (10) 22.9±0.4 6.4±2.9 43.3±3.1 5.9±7.8 (30) 18.6±1.4** 23.6±6.8 36.9±1.1 19.6±3.8 (100) 10.7±2.6** 56.4±10.6 23.2±3.5** 49.6±7.2

IC

50

91.5±12.6

28 (10) 18.5±1.2** 24.6±5.4 33.5±0.4** 27.1±2.6 (30) 10.2±1.0** 58.4±4.6 24.5±2.0** 46.9±3.9 (100) 2.3±0.8** 90.4±3.5 13.1±2.1** 71.7±4.5

IC

₅₀

35.7±3.8 50.6±4.6

TFP (10) 27.2±2.8 -10.9±12.5 49.1±2.7 -7.5±4.8 (20) 13.9±1.8** 43.3±8.2 24.1±5.2** 48.4±4.5 (30) 9.6±1.5** 60.9±7.0 11.7±4.7** 74.9±8.6

IC

50

24.4±0.5 22.8±0.5

Control 31.5±1.5 -- 60.5±3.7 -- 29 (10) 30.4±1.3 3.5±0.7 62.2±1.5 -3.2±4.2

(30) 26.0±1.8 17.7±3.0 56.6±3.6 6.4±1.9 TFP (3) 30.2±0.9 3.4±2.6 58.3±0.8 2.6±5.4 (10) 20.0±0.5** 35.1±5.1 32.9±1.0** 44.7±4.6 (30) 4.5±0.4** 85.9±1.2 9.3±1.1** 84.8±3.7

IC

50

11.3±0.8 11.0±0.2

Control 24.5±0.3 -- 46.1±1.1 -- 30 (30) 21.3±0.6 12.9±3.0 33.4±1.9* 27.7±3.4

(100) 14.2±2.9** 41.9±11.4 23. 2±4.4** 49.6±9.5 31 (30) 20.6±1.1 15.7±5.2 39.3±3.1 14.9±6.0 (100) 20.1±1.5 17.9±7.4 36.2±2.8 21.5±5.8 32 (10) 13.1±1.6** 46.5±7.0 27.9±2.9** 39.7±5.2 (30) 2.7±1.0** 88.6±4.1 12.7±4.0** 72.3±8.9 (100) -0.3±0.5** 101.3±2.3 0.7±1.1** 98.6±2.6

IC

50

12.7±4.4 18.5±5.3

TFP (10) 27.2±2.8 -10.9±12.5 49.1±2.7 -7.5±4.8

(20) 13.9±1.8** 43.3±8.2 24.1±5.2** 48.4±4.5 (30) 9.6±1.5** 60.9±7.0 11.7±4.7** 74.9±8.6

IC

50

24.4±0.5 22.8±0.5

Control 30.4±0.6 -- 64.8±1.6 -- 33 (10) 24.6±3.1* 19.3±8.6 51.7±3.8* 20.4±4.7

(30) 24.1±4.5 20.8±13.9 44.9±4.3** 30.8±5.8 TFP (3) 31.3±0.3 -1.0±2.1 63.9±5.8 2.6±2.1

(10) 23.7±1.4** 23.2±6.5 54.4±9.7 18.1±10.0 (20) 5.5±0.7** 82.0±2.6 2.0±1.3** 97.2±1.9

IC

50

14.0±0.5 12.6±0.6

Neutrophil suspensions were preincubated at 37 ℃ with 0.5 % DMSO or test compounds in the

presence of cytochalasin B (5 μg/ml) for 3 min. Forty-five minutes after the addition of fMLP (1 μ

M ), β-glucuronidase and lysozyme activities in the supernatant were determined. Trifluoperazine

(TFP) is a positive control. Values are presented as mean±S.E., N=3-8. *: P<0.05, **: P<0.01.

Table 14. The inhibitory effect of 5-(2'-carboxyl substituted phenoxy)furan-2- carboxylic acids on rat neutrophil degranulation (in vitro)

41: R

1

=R

2

=R

3

=R

4

=H 47: R

1

=R

2

=R

4

=H, R

3

=OCH

3

42: R

1

=R

2

=R

3

=H, R

4

=CH

3

48: R

1

=R

3

=R

4

=H, R

2

=OCH

3

43: R

1

=R

2

=R

4

=H, R

3

=CH

3

50: R

1

=R

2

=R

4

=H, R

3

=Cl 44: R

1

=R

3

=R

4

=H, R

2

=CH

3

51: R

1

=R

3

=R

4

=H, R

2

=Cl 45: R

2

=R

3

=R

4

=H, R

1

=CH

3

52: R

1

=R

3

=R

4

=H, R

2

=Br 46: R

1

=R

2

=R

3

=H, R

4

=OCH

3

53: R

1

=R

3

=R

4

=H, R

2

=I

Percent Release

Compound conc.--- (µM) β-Glucuronidase (%inh) Lysozyme (%inh)

Control 15.1±0.9 -- 30.4±1.2 -- 41 (30) 10.8±0.3 30.8±5.4 28.3±1.0 9.4±5.9

(100) 12.5±2.0 14.6±10.1 25.2±0.9* 22.0±4.4

42 (10) 13.7±1.7 12.1±6.2 -- --

(30) 11.6±0.2* 25.1±3.3 32.8±1.2 -3.0±3.8 (100) 14.0±0.6 7.0±5.3 35.9±4.0 -11.5±5.3 43 (30) 10.0±0.2** 36.9±4.8 30.7±0.7 -0.3±3.9

(100) 10.7±0.9* 33.9±4.3 32.4±2.1 -0.5±4.8 TFP (3) 13.9±2.0 9.6±4.8 30.5±2.9 8.7±4.1

(10) 8.1±0.8* 52.1±7.1 15.1±1.2** 52.7±3.7 (30) 2.2±1.3** 87.8±4.5 3.2±1.7** 89.7±4.8

IC

₅₀

14.3±3.9 12.5±3.7

Control 22.6±1.7 -- 46.4±2.1 --

44 (30) 21.0±0.7 6.1±4.5 47.5±1.6 -3.2±8.0

(100) 18.6±2.9 18.9±7.3 48.0±6.6 -2.9±10.3 Control 14.8±1.0 -- 16.7±1.5 -- TFP (3) 13.4±1.2 9.6±1.8 14.9±0.9 10.5±2.2

(10) 8.5±1.3** 43.1±5.3 9.7±0.9* 39.7±9.8 (30) 5.1±0.4** 64.7±3.1 6.6±0.4** 59.5±5.2

IC

₅₀

19.3±1.3 21.9±2.5

COOH

O R

₄

R

₃

R

₂

R

₁

O COOH

Control 42.5±0.8 -- 61.5±3.3 -- 45 (10) 35.9±2.5* 15.4±4.5 56.7±5.4 8.1±4.4

(30) 36.7±2.6* 13.5±4.7 59.1±5.0 4.1±3.6 TFP (3) 35.2±0.8 16.8±2.2 55.7±5.3 8.9±1.9 (10) 22.7±1.6** 44.9±8.6 44.0±6.8 28.0±6.1 (30) 4.8±0.7** 88.6±2.7 2.0±2.2** 96.9±4.1

IC

₅₀

14.2±0.7 16.0±0.9

Control 24.5±0.3 -- 46.1±1.1 -- 46 (30) 22.3±0.5 8.8±3.1 45.3±0.8 1.6±3.5

(100) 22.9±1.5 6.1±6.9 39.6±1.5 13.7±4.8 47 (30) 24.8±0.5 -1.4±3.2 43.3±2.6 6.1±5.0

(100) 21.0±0.9 14.5±2.9 41.3±3.2 10.5±6.4 48 (30) 21.8±0.9 10.9±4.5 43.6±3.4 5.6±5.8

(100) 20.7±1.0 15.5±4.2 41.3±3.3 10.9±5.1 50 (10) 22.1±0.8 9.9±3.0 42.0±2.6 8.9±4.5

(30) 14.6±1.4** 40.6±5.8 30.3±3.7** 34.3±7.8 (100) 10.9±1.8** 55.3±8.4 15.8±1.8** 66.0±3.1

IC

50

66.6±4.1 68.0±5.7

51 (30) 20.4±1.8 17.8±7.9 41.0±1.6 11.1±2.1 (100) 17.7±1.7** 27.8±7.0 32.3±2.4** 30.0±5.0 52 (30) 21.1±0.7 13.9±3.1 41.2±2.4 10.7±3.8 (100) 22.3±1.8 9.1±8.1 37.2±2.4 19.3±5.4 TFP (10) 27.2±2.8 -10.9±12.5 49.1±2.7 -7.5±4.8

(30) 13.9±1.8** 43.3±8.2 24.1±5.2** 48.4±4.5 (100) 9.6±1.5** 60.9±7.0 11.7±4.7** 74.9±8.6

IC

₅₀

24.4±0.5 22.8±0.5

Control 30.4±0.6 -- 64.8±1.6 -- 53 (10) 24.2±2.6** 20.6±7.3 54.8±4.3 15.5±5.3

(30) 22.0±3.3** 27.9±9.7 49.5±5.2* 23.7±6.8 TFP (3) 31.3±0.3 -1.0±2.1 63.9±5.8 2.6±2.1

(10) 23.7±1.4** 23.2±6.5 54.4±9.7 18.1±10.0 (20) 5.5±0.7** 82.0±2.6 2.0±1.3** 97.2±1.9

IC

50

14.0±0.5 12.6±0.6

Neutrophil suspensions were preincubated at 37 ℃ with 0.5 % DMSO or test compounds in the presence of cytochalasin B (5 μg/ml) for 3 min. Forty-five minutes after the addition of fMLP (1 μ M ), β-glucuronidase and lysozyme activities in the supernatant were determined. Trifluoperazine (TFP) is a positive control. Values are presented as mean±S.E., -- not determined. N=3-8. *: P<0.05,

**: P<0.01.

Table 15. The inhibitory effect of substituted furo[2,3-b]chromone-2-carboxylic acid ethyl esters on rat neutrophil degranulation (in vitro)

O O

O

COOCH

₂

CH

₃

R

₁

R

₂

R

₃

R

₄

61: R

1

=R

2

=R

3

=R

4

=H 67: R

1

=R

2

=R

4

=H, R

3

=OCH

3

62: R

1

=R

2

=R

3

=H, R

4

=CH

3

68: R

1

=R

3

=R

4

=H, R

2

=OCH

3

63: R

1

=R

2

=R

4

=H, R

3

=CH

3

70: R

1

=R

2

=R

4

=H, R

3

=Cl 64: R

1

=R

3

=R

4

=H, R

2

=CH

3

71: R

1

=R

3

=R

4

=H, R

2

=Cl 65: R

2

=R

3

=R

4

=H, R

1

=CH

3

72: R

1

=R

3

=R

4

=H, R

2

=Br 66: R

1

=R

2

=R

3

=H, R

4

=OCH

3

73: R

1

=R

3

=R

4

=H, R

2

=I

Percent Release

Compound conc.--- (µM) β-Glucuronidase (%inh) Lysozyme (%inh)

Control 15.1±0.9 -- 30.4±1.2 -- 61 (10) 9.8±0.7** 37.5±3.9 -- --

(30) 11.9±1.4* 24.8±7.1 23.8±2.1* 7.3±7.0 (100) 7.4±1.1** 56.0±5.0 16.4±1.1** 34.1±5.9

IC

50

70.5±7.8

62 (10) 13.4±2.1* 12.5±10.2 -- -- (30) 9.9±0.3** 38.3±2.9 31.2±4.6 10.9±11.5 (100) 7.7±0.5** 52.1±2.8 24.2±2.7 29.7±5.8

IC

50

77.2±8.1

TFP (3) 13.9±2.0 9.6±4.8 30.5±2.9 8.7±4.1 (10) 8.1±0.8* 52.1±7.1 15.1±1.2** 52.7±3.7 (30) 2.2±1.3** 87.8±4.5 3.2±1.7** 89.7±4.8

IC

₅₀

14.3±3.9 12.5±3.7

Control 22.6±1.7 -- 46.4±2.1 -- 63 (30) 17.9±1.1 20.6±1.1 44.8±1.1 3.2±2.7

(100) 17.3±1.8 23.7±3.5 49.2±6.0 -5.5±8.3 64 (30) 16.1±2.0 29.0±4.7 50.5±6.0 -8.8±10.8

(100) 14.9±1.6 34.4±3.0 29.4±3.4 36.3±8.0

Control 14.8±1.0 -- 16.7±1.5 --

TFP (3) 13.4±1.2 9.6±1.8 14.9±0.9 10.5±2.2

(10) 8.5±1.3** 43.1±5.3 9.7±0.9* 39.7±9.8 (30) 5.1±0.4** 64.7±3.1 6.6±0.4** 59.5±5.2

IC

50

19.3±1.3 21.9±2.5

Control 21.6±0.6 -- 54.7±1.6 -- 65 (10) 22.1±0.4 -2.7±4.7 46.0±7.5 16.4±11.8

(30) 16.7±1.4** 22.1±8.4 40.7±4.3** 25.8±5.8 66 (10) 18.3±0.5** 15.0±5.2 43.9±5.6* 20.0±8.1 (30) 19.6±0.1* 8.9±3.2 47.9±4.5 12.6±5.9 67 (10) 19.4±0.6* 10.0±2.7 49.3±2.5 9.6±5.3

(30) 18.1±0.7** 15.9±3.0 48.3±3.3 11.6±5.1 68 (10) 17.9±0.3** 16.9±3.0 45.9±3.7 16.2±4.5 (30) 17.4±0.2** 17.7±2.1 38.7±6.5** 27.8±10.1 TFP (3) 24.8±2.0 -14.6±12.7 64.9±6.7 -17.6±11.5

(10) 17.0±2.0** 21.3±11.4 45.6±3.2 17.5±3.1 (30) 2.6±0.7** 87.8±3.4 1.3±2.7** 97.3±4.9

IC

₅₀

18.9±2.1 18.3±0.9

Control 32.5±0.4 -- 48.2±3.0 -- 70 (10) 29.2±1.1 10.1±4.7 44.9±3.4 6.7±1.5

(30) 27.3±1.1 15.9±4.5 45.8±2.8 4.9±3.1 71 (10) 29.2±0.7 10.1±3.1 47.1±3.5 2.4±2.4 (30) 22.6±1.3** 30.6±3.6 38.3±5.1 20.9±7.0 72 (10) 30.5±0.5 6.1±0.8 51.2±3.0 -6.4±2.0

(30) 32.3±0.7 0.9±1.2 51.4±4.2 -6.4±3.1 73 (10) 32.7±2.3 -0.7±8.2 53.1±2.0 -10.7±5.1

(30) 31.3±0.5 3.8±1.4 48.4±3.3 -0.4±2.4 TFP (3) 37.8±1.3 -14.6±12.7 56.7±4.0 -17.6±11.5

(10) 24.9±1.3** 21.3±11.4 39.4±1.9 17.5±3.1 (30) 3.7±0.4** 87.8±3.4 1.1±1.6** 97.3±4.9

IC

50

18.9±2.1 18.3±0.9

Neutrophil suspensions were preincubated at 37 ℃ with 0.5 % DMSO or test compounds in the presence of cytochalasin B (5 μg/ml) for 3 min. Forty-five minutes after the addition of fMLP (1 μ M ), β-glucuronidase and lysozyme activities in the supernatant were determined. Trifluoperazine (TFP) is a positive control. Values are presented as mean±S.E., -- not determined. N=3-8. *: P<0.05,

**: P<0.01.

Table 16. The inhibitory effect of 5-(2'-alkoxycarbonyl substituted phenoxy)furfurals on rat neutrophil degranulation (in vitro)

81: R=CH

3

, R

1

=R

2

=R

3

=R

4

=H 88: R=CH

3

, R

1

=R

3

=R

4

=H, R

2

=OCH

3

82: R=CH

3

, R

1

=R

2

=R

3

=H, R

4

=CH

3

89: R=CH

3

, R

2

=R

3

=R

4

=H, R

1

=OCH

3

83: R=CH

3

, R

1

=R

2

=R

4

=H, R

3

=CH

3

90: R=CH

3

, R

1

=R

2

=R

4

=H, R

3

=Cl 84: R=CH

3

, R

1

=R

3

=R

4

=H, R

2

=CH

3

91: R=CH

3

, R

1

=R

3

=R

4

=H, R

2

=Cl 85: R=C

2

H

5

, R

2

=R

3

=R

4

=H, R

1

=CH

3

92: R=CH

3

, R

1

=R

3

=R

4

=H, R

2

=Br 86: R=CH

3

, R

1

=R

2

=R

3

=H, R

4

=OCH

3

93: R=CH

3

, R

1

=R

3

=R

4

=H, R

2

=I 87: R=CH

3

, R

1

=R

2

=R

4

=H, R

3

=OCH

3

Percent Release

Compound conc.--- (µM) β-Glucuronidase (%inh) Lysozyme (%inh) Control 20.2±0.6 -- 25.6±0.7 --

81 (10) 18.3±0.6 9.4±4.1 21.9±0.5 14.3±0.8

(30) 16.5±0.6** 18.1±4.5 14.8±0.1** 41.9±1.9 82 (10) 19.8±0.6 1.8±2.8 33.7±3.3 -30.8±9.3

(30) 16.8±0.8** 17.6±4.7 20.8±1.2 18.1±7.7 83 (10) 17.1±0.6* 15.3±3.6 24.5±1.3 4.3±2.4

(30) 15.9±0.8** 21.2±6.0 16.1±2.2** 37.6±6.9 84 (10) 18.0±0.4 10.7±2.8 37.3±8.6 -43.6±8.0

(30) 15.1±0.4** 25.3±1.1 17.5±2.3** 32.1±7.0 85 (10) 18.4±1.2 9.0±4.5 32.9±1.3 -28.4±1.2

(30) 15.8±1.1* 21.6±7.8 20.7±1.4 18.5±8.5 TFP (3) 20.4±0.6 -0.8±0.2 27.5±0.8 -7.6±5.7

(10) 15.1±0.6** 25.4±0.8 16.8±0.4** 34.4±0.3 (30) 2.4±0.4** 87.7±2.1 1.8±0.8** 92.5±3.4

IC

50

12.9±0.2 12.0±0.5

Control 31.5±1.5 -- 60.5±3.7 -- 86 (10) 25.3±1.0 19.6±1.7 60.7±5.1 -0.06±2.2

COOR

O R

₄

R

₃

R

₂

R

₁

O CHO

(30) 26.4±2.3 16.5±3.3 59.5±7.5 2.4±6.6 87 (10) 27.1±1.2 13.8±0.5 60.5±4.1 0.09±1.0

(30) 24.2±1.9* 23.2±3.2 54.7±6.5 10.3±5.2 88 (10) 28.6±2.4 9.4±3.4 59.6±4.2 1.6±2.7

(30) 22.6±0.8* 28.2±2.0 50.8±6.0 16.7±4.5 89 (10) 27.4±3.1 13.3±7.4 60.1±3.0 0.3±3.8

(30) 25.6±2.5 19.1±4.1 58.2±4.2 3.9±1.8 90 (10) 26.4±3.8 17.0±9.0 56.3±10.0 8.0±12.5

(30) 23.6±1.3* 25.0±1.1 53.2±5.7 12.4±4.4 91 (10) 31.4±1.9 0.3±2.1 61.1±3.3 -1.0±1.6

(30) 29.2±2.3 7.5±3.1 56.4±4.4 6.9±2.0 92 (10) 24.5±2.9 22.5±6.5 -1.4±4.9 -1.4±4.9

(30) 22.8±1.7* 27.6±2.1 54.9±3.4 9.1±1.7 93 (10) 27.3±1.8 13.4±1.5 59.7±2.7 1.1±2.3 (30) 20.8±1.5** 34.1±2.4 49.5±2.3 18.1±1.3 TFP (3) 30.2±0.9 3.4±2.6 58.3±0.8 2.6±5.4

(10) 20.0±0.5** 35.1±5.1 32.9±1.0** 44.7±4.6 (30) 4.5±0.4** 85.9±1.2 9.3±1.1** 84.8±3.7

IC

50

11.3±0.8 11.0±0.2

Neutrophil suspensions were preincubated at 37 ℃ with 0.5 % DMSO or test compounds in the

presence of cytochalasin B (5 μg/ml) for 3 min. Forty-five minutes after the addition of fMLP (1 μ

M ), β-glucuronidase and lysozyme activities in the supernatant were determined. Trifluoperazine

(TFP) is a positive control. Values are presented as mean±S.E., N=3. *: P<0.05, **: P<0.01.

Table 17. The inhibitory effect of 5-(2'-alkoxycarbonyl substituted phenoxy)-2- furanacrylic acids on rat neutrophil degranulation (in vitro)

101: R=CH

3

, R

1

=R

2

=R

3

=R

4

=H 106: R=CH

3

, R

1

=R

2

=R

3

=H, R

4

=OCH

3

102: R=CH

3

, R

1

=R

2

=R

3

=H, R

4

=CH

3

107: R=CH

3

, R

1

=R

2

=R

4

=H, R

3

=OCH

3

103: R=CH

3

, R

1

=R

2

=R

4

=H, R

3

=CH

3

108: R=CH

3

, R

1

=R

3

=R

4

=H, R

2

=OCH

3

104: R=CH

3

, R

1

=R

3

=R

4

=H, R

2

=CH

3

109: R=CH

3

, R

2

=R

3

=R

4

=H, R

1

=OCH

3

105: R=C

2

H

5

, R

2

=R

3

=R

4

=H, R

1

=CH

3

Percent Release

Compound conc.--- (µM) β-Glucuronidase (%inh) Lysozyme (%inh) Control 20.2±0.6 -- 25.6±0.7 -- 101 (10) 19.9±0.8 1.5±5.7 47.9±4.8** -85.9±13.6

(30) 18.4±0.9 8.8±5.6 41.1±5.1* -59.1±15.6 102 (10) 19.6±0.5 2.9±1.3 40.1±5.7* -55.2±12.3 (30) 21.5±0.8 -5.9±1.6 40.2±8.1 -55.2±17.8 103 (10) 16.5±0.5** 18.4±2.0 41.3±4.2** -60.4±11.9 (30) 15.9±0.5** 21.3±0.2 33.9±7.1 -30.6±15.7 104 (10) 18.4±0.5 9.0±1.3 40.8±4.6* -58.4±13.8 (30) 18.0±0.4 10.7±2.4 37.6±5.6 -45.5±18.0 105 (10) 19.7±0.2 2.3±1.9 37.3±3.4* -45.0±8.9

(30) 21.3±0.7 -5.2±1.1 33.6±0.5 -31.4±6.1 TFP (3) 20.4±0.6 -0.8±0.2 27.5±0.8 -7.6±5.7

(10) 15.1±0.6** 25.4±0.8 16.8±0.4** 34.4±0.3 (30) 2.4±0.4** 87.7±2.1 1.8±0.8** 92.5±3.4

IC

50

12.9±0.2 12.0±0.5

Control 31.5±1.5 -- 60.5±3.7 -- 106 (10) 31.2±2.0 1.1±2.1 63.2±2.5 -4.6±3.1

(30) 28.4±4.1 10.6±8.9 58.9±6.3 3.0±7.1 107 (10) 32.6±2.7 -3.0±4.1 63.7±2.8 -5.5±3.9

O COOR R

₁

R

₂

R

₃

R

₄

O CH CH COOH

(30) 29.8±1.9 5.6±1.8 61.5±2.1 -1.9±2.8 108 (10) 33.6±2.6 -6.3±4.0 62.4±2.8 -3.4±2.8 (30) 31.8±2.0 -0.7±1.8 63.5±2.7 -5.2±2.7 109 (10) 35.8±1.9 -13.6±1.7 64.2±2.4 -6.3±2.4 (30) 35.3±2.6 -11.8±3.5 63.1±2.8 -4.5±2.3 TFP (3) 30.2±0.9 3.4±2.6 58.3±0.8 2.6±5.4

(10) 20.0±0.5** 35.1±5.1 32.9±1.0** 44.7±4.6 (30) 4.5±0.4** 85.9±1.2 9.3±1.1** 84.8±3.7

IC

50

11.3±0.8 11.0±0.2

Neutrophil suspensions were preincubated at 37 ℃ with 0.5 % DMSO or test compounds in the

presence of cytochalasin B (5 μg/ml) for 3 min. Forty-five minutes after the addition of fMLP (1 μ

M ), β-glucuronidase and lysozyme activities in the supernatant were determined. Trifluoperazine

(TFP) is a positive control. Values are presented as mean±S.E., N=3. *: P<0.05, **: P<0.01.

Table 18. The inhibitory effect of 5-(2'-carboxyl substituted phenoxy)-2-furanacrylic acids on rat neutrophil degranulation (in vitro)

111: R

1

=R

2

=R

3

=R

4

=H 118: R

1

=R

3

=R

4

=H, R

2

=OCH

3

112: R

1

=R

2

=R

3

=H, R

4

=CH

3

120: R

1

=R

2

=R

4

=H, R

3

=Cl 113: R

1

=R

2

=R

4

=H, R

3

=CH

3

121: R

1

=R

3

=R

4

=H, R

2

=Cl 114: R

1

=R

3

=R

4

=H, R

2

=CH

3

122: R

1

=R

3

=R

4

=H, R

2

=Br 116: R

1

=R

2

=R

3

=H, R

4

=OCH

3

123: R

1

=R

3

=R

4

=H, R

2

=I 117: R

1

=R

2

=R

4

=H, R

3

=OCH

3

Percent Release

Compound conc.--- (µM) β-Glucuronidase (%inh) Lysozyme (%inh)

Control 20.2±0.6 -- 25.6±0.7 -- 111 (10) 19.7±1.2 2.7±4.0 41.0±6.9* -58.6±14.8

(30) 20.4±0.8 -0.6±1.8 37.2±7.8 -43.5±16.8 112 (10) 16.2±1.8 19.2±11.0 45.0±3.7** -74.8±9.4

(30) 19.7±0.7 2.5±4.6 43.0±6.5** -66.5±14.9 113 (10) 21.1±0.6 -4.2±2.7 37.4±4.2 -45.2±12.5 (30) 15.7±1.9* 21.7±11.1 35.7±4.6 -38.4±14.1 114 (10) 20.0±0.9 0.7±5.8 34.1±1.9 -33.7±11.9 (30) 20.7±1.7 -3.0±10.9 35.1±0.9 -37.5±8.0 TFP (3) 20.4±0.6 -0.8±0.2 27.5±0.8 -7.6±5.7

(10) 15.1±0.6** 25.4±0.8 16.8±0.4** 34.4±0.3 (30) 2.4±0.4** 87.7±2.1 1.8±0.8** 92.5±3.4

IC

50

12.9±0.2 12.0±0.5

Control 31.5±1.5 -- 60.5±3.7 -- 116 (10) 28.6±2.2 9.3±2.9 65.8±1.7 -9.3±5.0

(30) 28.3±2.0 10.4±2.8 63.1±1.7 -4.7±3.4 117 (10) 31.3±1.8 0.7±1.7 64.3±0.9 -6.8±5.1 (30) 29.6±1.6 6.2±0.7 64.6±1.0 -7.4±5.1

COOH

O R

₁

R

₂

R

₃

O CH

R

₄

CH COOH

118 (10) 30.6±1.3 2.8±1.1 64.4±1.5 -6.9±4.3 (30) 30.1±2.3 4.8±2.9 62.7±2.5 -3.9±2.5 120 (10) 32.9±2.8 -4.1±3.9 60.3±1.1 -0.1±4.5 (30) 33.2±3.1 -4.9±5.3 62.9±1.6 -4.3±3.6 121 (10) 35.3±2.3 -11.7±2.5 63.4±2.3 -5.1±2.8 (30) 32.4±2.6 -2.5±3.6 59.9±2.7 0.8±2.1 122 (10) 37.0±2.4 -17.2±2.4 64.1±2.8 -6.2±3.1

(30) 35.2±3.4 -11.3±5.9 61.6±2.3 -2.2±4.8 123 (10) 28.7±3.0 9.2±5.8 61.9±0.8 -2.9±4.9 (30) 28.5±2.1 9.8±2.5 61.4±2.0 -1.7±3.2 TFP (3) 30.2±0.9 3.4±2.6 58.3±0.8 2.6±5.4

(10) 20.0±0.5** 35.1±5.1 32.9±1.0** 44.7±4.6 (30) 4.5±0.4** 85.9±1.2 9.3±1.1** 84.8±3.7

IC

50

11.3±0.8 11.0±0.2

Neutrophil suspensions were preincubated at 37 ℃ with 0.5 % DMSO or test compounds in the

presence of cytochalasin B (5 μg/ml) for 3 min. Forty-five minutes after the addition of fMLP (1 μ

M ), β-glucuronidase and lysozyme activities in the supernatant were determined. Trifluoperazine

(TFP) is a positive control. Values are presented as mean±S.E., N=3. *: P<0.05, **: P<0.01.

(二)對於fMLP誘導的嗜中性白血球超氧自由基生成作用抑制試驗 從化合物

21-33、41-48、50-53、61-68、70-73、81-93、101-109、

111-114、116-118及120-123對以fMLP誘導的嗜中性白血球超氧自由基

生成作用之體外試驗中，由superoxide formation的抑制百分率 (見 Table 19至Table 24)看來，在濃度30

µ

M時，化合物23、26、27、30、

31、44-46、48、51-53、61、62、65-68、70、72、73、81、83-91、

101、108、117、118及120分別呈現弱的抑制活性 (具有約20-47%的

抑制百分率)，但是發現化合物25、28、32、50、64、71、92及93呈 現 明 顯 的 抑 制 活 性 ， 其 抑 制 superoxide formation的 IC50值 分 別 為 15.0

±

1.9

µ

M 、39.9

±

2.8

µ

M、24.3

±

5.1

µ

M 、35.4

±

8.0

µ

M、45.0

±

3.5

µ

M、24.4

±

1.5

µ

M、13.4

±

2.9

µ

M及19.6

±

5.6

µ

M。其他化合物則無明 顯的抑制活性。

綜 合 上 述 ， 發 現 ethyl 5-(2'-alkoxycarbonyl substituted phenoxy)-furan-2-carboxylates (21-33) 類 、 5-(2'-carboxyl substituted phenoxy)-furan-2-carboxylic acids (41-48 及 50-53) 類 、 substituted furo[2,3-b]-chromone-2-carboxylic acid ethyl esters (61-68及70-73)類及 5-(2'-alkoxycarbonyl substituted phenoxy)furfurals (81-93)類衍生物的 活 性 較 明 顯 。 在 ethyl 5-(2'-alkoxycarbonyl substituted phenoxy)furan-2-carboxylates (21-33)類衍生物中將甲基、甲氧基或溴 原 子 導 入 苯 環 時 ， 具 有 較 高 的 活 性 ， 而 化 合 物 ethyl 5-(2'-ethoxycarbonyl-3'-methyl-phenoxy)furan-2-carboxylate (25)的IC50

= 15.0±1.9

µM約與trifluoperazine的IC

50 = 14.7± 0.4

µM相當，化合物

ethyl 5-(2'-methoxy-carbonyl-4'-methoxyphenoxy)furan-2-carboxylate (28)的IC50 = 39.9

±

2.8

µ

M約為trifluoperazine之IC50 = 20.1

±

1.5

µ

M的二

倍 ， 化 合 物 ethyl

5-(2'-methoxycarbonyl-4'-bromophenoxy)furan-2-carboxylate (32)的IC50

= 24.3

±

5.1

µ

M約與trifluoperazine的IC₅₀ = 20.1

±

1.5

µ

M相當。與甲基、

甲氧基或溴原子相較之下，若將氯原子導入苯環，則其活性降低，此 外，若將碘原子導入苯環，則其活性降得更低。在5-(2'-carboxyl substituted phenoxy)furan-2-carboxylic acids (41-48及50-53)類衍生物 中 將 氯 原 子 導 入 苯 環 時 ， 具 有 較 高 的 活 性 ， 而 化 合 物 5-(2'-carboxyl-5'-chlorophenoxy)furan-2-carboxylic acid (50) 的 IC50 = 35.4±8.0 µM約為trifluoperazine之IC50 = 20.1±1.5 µM的二倍。與氯原子 相較之下，若將溴原子或碘原子導入苯環，則其活性降低，此外，若 將 甲 基 或 甲 氧 基 導 入 苯 環 ， 則 其 活 性 降 得 更 低 。 在 substituted furo[2,3-b]chromone- 2-carboxylic acid ethyl esters (61-68及70-73)類衍 生 物 中 將 氯 原 子 導 入 環 上 時 ， 具 有 較 高 的 活 性 ， 而 化 合 物 ethyl

6-chlorofuro[2,3-b]chromone- 2-carboxylate (71)的IC50 = 24.4

±

1.5

µ

M 約為trifluoperazine之IC50 = 12.9

±

1.0

µ

M的二倍。與氯原子相較之下，

若將甲基、甲氧基或碘原子導入環上，則其活性降低，此外，若將溴 原子導入環上，則其活性降得更低。另外，在5-(2'-alkoxycarbonyl substituted phenoxy)furfurals (81-93)類衍生物中將溴原子或碘原子導 入 苯 環 時 ， 具 有 較 高 的 活 性 ， 而 化 合 物 5-(2'-methoxycarbonyl-4'-bromophenoxy)furfural (92)的IC50 = 13.4±2.9

µ

M 約 為 trifluoperazine 之 IC50 = 6.2

±

0.3

µ

M 的 二 倍 ， 化 合 物 5-(2'-methoxycarbonyl-4'-iodophenoxy)furfural (93) 的 IC50 = 19.6

±

5.6

µM約為trifluoperazine之IC

50 = 6.2±0.3 µM的三倍。與溴原子或碘原子 相較之下，若將甲氧基或氯原子導入苯環，則其活性降低，此外，若 將甲基導入苯環，則其活性降得更低。

Table 19. The inhibitory effect of ethyl 5-(2'-alkoxycarbonyl substituted phenoxy)- furan-2-carboxylates on rat neutrophil superoxide formation (in vitro)

21: R=CH

3

, R

1

=R

2

=R

3

=R

4

=H 28: R=CH

3

, R

1

=R

3

=R

4

=H, R

2

=OCH

3

22: R=CH

3

, R

1

=R

2

=R

3

=H, R

4

=CH

3

29: R=CH

3

, R

2

=R

3

=R

4

=H, R

1

=OCH

3

23: R=CH

3

, R

1

=R

2

=R

4

=H, R

3

=CH

3

30: R=CH

3

, R

1

=R

2

=R

4

=H, R

3

=Cl 24: R=CH

3

, R

1

=R

3

=R

4

=H, R

2

=CH

3

31: R=CH

3

, R

1

=R

3

=R

4

=H, R

2

=Cl 25: R=C

2

H

5

, R

2

=R

3

=R

4

=H, R

1

=CH

3

32: R=CH

3

, R

1

=R

3

=R

4

=H, R

2

=Br 26: R=CH

3

, R

1

=R

2

=R

3

=H, R

4

=OCH

3

33: R=CH

3

, R

1

=R

3

=R

4

=H, R

2

=I 27: R=CH

3

, R

1

=R

2

=R

4

=H, R

3

=OCH

3

Superoxide Formation

Compound conc.--- (µM) nmol/10

⁶

cells/30 min (%inh)

Control 0.9±0.1 --

21 (30) 0.9±0.0 -1.2±2.3

(100) 0.6±0.0 6.6±6.6

22 (30) 0.8±0.1 14.9±2.4

(100) 0.7±0.0* 23.3±6.3

23 (30) 0.8±0.1 20.0±2.1

(100) 0.6±0.1* 36.4±1.4

TFP (3) 0.5±0.1** 35.7±2.2

(5) 0.2±0.0** 71.4±3.4

(10) 0.1±0.0** 85.7±1.0

IC

₅₀

4.3±0.3

Control 1.25±0.21 --

24 (30) 1.02±0.21 19.8±3.9

(100) 0.80±0.11 34.7±4.7

Control 1.30±0.17 --

TFP (3) 1.10±0.27 17.8±1.2

(10) 0.20±0.03** 84.5±1.2

(30) 0.05±0.04** 96.1±3.2

COOR

O R

₄

R

₃

R

₂

R

₁

O COOCH

₂

CH

₃

IC

50

9.4±2.2

Control 2.15±0.23 --

25 (3) 1.61±0.26 24.8±7.9

(10) 1.31±0.24** 40.5±8.1

(30) 0.52±0.09** 76.5±1.8

IC

₅₀

15.0±1.9

TFP (1) 2.59±0.15 29.4±12.0

(10) 0.47±0.08** 77.1±7.2

(30) 0.12±0.03** 93.0±3.0

IC

50

14.7±0.4

Control 1.68±0.20 --

26 (30) 1.31±0.25 23.3±6.7

(100) 1.13±0.22* 33.2±7.5

27 (30) 1.04±0.15* 38.4±2.9

(100) 0.94±0.25** 47.7±10.1

28 (3) 1.26±0.08 14.6±8.5

(10) 0.88±0.03** 48.8±2.8

(30) 0.85±0.08** 49.7±2.5

(100) 0.53±0.16** 71.2±9.7

IC

₅₀

39.9±2.8

TFP (3) 1.52±0.04 3.1±5.3

(10) 1.05±0.04* 33.0±5.2

(30) 0.52±0.04** 69.5±10.4

IC

50

20.1±1.5

Control 5.47±0.57 --

29 (10) 4.93±0.67 9.7±7.6

(30) 5.12±0.96 15.6±12.2

TFP (3) 4.12±0.04 24.3±1.9

(10) 1.16±0.08** 78.1±5.4

(30) 0.02±0.05** 99.1±3.3

IC

50

6.2±0.3

Control 1.68±0.20 --

30 (30) 1.26±0.22 25.3±7.6

(100) 1.38±0.24 18.9±7.7

31 (30) 1.05±0.20** 39.0±5.6

(100) 1.13±0.21* 33.9±7.8

32 (1) 1.23±0.14 15.5±7.3

(3) 0.89±0.12** 44.0±5.8

(10) 0.80±0.12** 54.1±5.0

(30) 0.64±0.09** 61.2±5.8

IC

50

24.3±5.1

TFP (3) 1.52±0.04 3.1±5.3

(10) 1.05±0.04* 33.0±5.2

IC

50

20.1±1.5

Control 1.59±0.05 --

33 (10) 1.23±0.18* 22.5±13.1

(30) 2.08±0.03** -30.7±3.5

TFP (3) 1.27±0.22 20.5±5.3

(10) 0.91±0.16** 42.9±2.8

(30) 0.03±0.02** 98.2±0.9

IC

50

13.0±0.3

Neutrophil suspensions were preincubated at 37 °C with 0.5 % DMSO or test compounds in the

presence of cytochalasin B (5 µg/ml) for 3 min. Fifteen minutes after the addition of fMLP (0.3 µM),

the absorbance was determined at 550 nm. Trifluoperazine (TFP) is a positive control. Values are

presented as mean± S.E., N=3-7. *: P<0.05, **: P<0.01.

Table 20. The inhibitory effect of 5-(2'-carboxyl substituted phenoxy)furan-2- carboxylic acids on rat neutrophil superoxide formation (in vitro)

41: R

1

=R

2

=R

3

=R

4

=H 47: R

1

=R

2

=R

4

=H, R

3

=OCH

3

42: R

1

=R

2

=R

3

=H, R

4

=CH

3

48: R

1

=R

3

=R

4

=H, R

2

=OCH

3

43: R

1

=R

2

=R

4

=H, R

3

=CH

3

50: R

1

=R

2

=R

4

=H, R

3

=Cl 44: R

1

=R

3

=R

4

=H, R

2

=CH

3

51: R

1

=R

3

=R

4

=H, R

2

=Cl 45: R

2

=R

3

=R

4

=H, R

1

=CH

3

52: R

1

=R

3

=R

4

=H, R

2

=Br 46: R

1

=R

2

=R

3

=H, R

4

=OCH

3

53: R

1

=R

3

=R

4

=H, R

2

=I

Superoxide Formation

Compound conc.--- (µM) nmol/10

⁶

cells/30 min (%inh)

Control 0.9±0.1 --

41 (30) 0.9±0.0 -4.8±2.3

(100) 1.0±0.1 -9.1±8.3

42 (30) 0.9±0.1 5.9±2.9

(100) 0.7±0.0 21.9±2.5

43 (30) 1.0±0.2 0.0±6.3

(100) 0.7±0.0 23.1±7.1

TFP (3) 0.5±0.1** 35.7±2.2

(5) 0.2±0.0** 71.4±3.4

(10) 0.1±0.0** 85.7±1.0

IC

₅₀

4.3±0.3

Control 1.29±0.22 --

44 (30) 0.83±0.06 31.5±8.4

(100) 0.72±0.12 44.0±1.7

Control 1.30±0.17 --

TFP (3) 1.10±0.27 17.8±1.2

(10) 0.20±0.03** 84.5±1.2

(30) 0.05±0.04** 96.1±3.2

COOH

O R

₄

R

₃

R

₂

R

₁

O COOH

Control 2.15±0.23 --

45 (10) 1.36±0.17** 36.6±3.7

(30) 1.40±0.20** 35.2±3.8

TFP (1) 2.59±0.15 29.4±12.0

(10) 0.47±0.08** 77.1±7.2

(30) 0.12±0.03** 93.0±3.0

IC

50

14.7±0.4

Control 1.68±0.20 --

46 (30) 1.14±0.24* 28.2±6.0

(100) 1.11±0.25* 36.8±9.8

47 (30) 1.31±0.04 11.6±7.5

(100) 1.05±0.19* 38.4±5.0

48 (30) 1.20±0.09 26.3±8.0

(100) 1.00±0.12* 37.9±9.1

50 (10) 1.19±0.03 26.7±9.7

(30) 0.81±0.04** 49.9±4.3

(100) 0.46±0.09** 75.2±5.0

IC

50

35.4±8.0

51 (10) 1.18±0.05 20.3±8.2

(30) 0.96±0.10** 41.3±6.5

(100) 0.77±0.06** 57.4±6.3

IC

50

72.6±8.9

52 (30) 1.11±0.04* 38.6±6.4

(100) 0.98±0.09** 40.1±6.1

TFP (3) 1.52±0.04 3.1±5.3

(10) 1.05±0.04* 33.0±5.2

(30) 0.52±0.04** 69.5±10.4

IC

50

20.1±1.5

Control 1.59±0.05 --

53 (10) 1.27±0.10* 19.7±8.6

(30) 1.06±0.07** 33.6±5.1

TFP (3) 1.27±0.22 20.5±5.3

(10) 0.91±0.16** 42.9±2.8

(30) 0.03±0.02** 98.2±0.9

IC

₅₀

13.0±0.3

Neutrophil suspensions were preincubated at 37 °C with 0.5 % DMSO or test compounds in the

presence of cytochalasin B (5 µg/ml) for 3 min. Fifteen minutes after the addition of fMLP (0.3 µM),

the absorbance was determined at 550 nm. Trifluoperazine (TFP) is a positive control. Values are

presented as mean± S.E., N=3-7. *: P<0.05, **: P<0.01.

Table 21. The inhibitory effect of substituted furo[2,3-b]chromone-2-carboxylic acid ethyl esters on rat neutrophil superoxide formation (in vitro)

O O

O

COOCH

₂

CH

₃

R

₁

R

₂

R

₃

R

₄

61: R

1

=R

2

=R

3

=R

4

=H 67: R

1

=R

2

=R

4

=H, R

3

=OCH

3

62: R

1

=R

2

=R

3

=H, R

4

=CH

3

68: R

1

=R

3

=R

4

=H, R

2

=OCH

3

63: R

1

=R

2

=R

4

=H, R

3

=CH

3

70: R

1

=R

2

=R

4

=H, R

3

=Cl 64: R

1

=R

3

=R

4

=H, R

2

=CH

3

71: R

1

=R

3

=R

4

=H, R

2

=Cl 65: R

2

=R

3

=R

4

=H, R

1

=CH

3

72: R

1

=R

3

=R

4

=H, R

2

=Br 66: R

1

=R

2

=R

3

=H, R

4

=OCH

3

73: R

1

=R

3

=R

4

=H, R

2

=I

Superoxide Formation

Compound conc.--- (µM) nmol/10

⁶

cells/30 min (%inh)

Control 0.9±0.1 --

61 (10) 1.0±0.0 -6.1±3.1

(30) 0.6±0.0** 34.3±3.5

(100) 0.4±0.1** 54.9±1.9

IC

50

75.9±3.9

62 (10) 0.8±0.1 2.5±3.1

(30) 0.5±0.0** 42.2±3.5

(100) 0.3±0.1** 65.5±7.9

IC

₅₀

57.6±4.1

TFP (3) 0.5±0.1** 35.7±2.2

(5) 0.2±0.0** 71.4±3.4

(10) 0.1±0.0** 85.7±1.0

IC

50

4.3±0.3

Control 1.25±0.21 --

63 (30) 1.29±0.24 -3.2±9.6

(100) 0.82±0.16 34.8±5.0

Control 1.63±0.05 --

64 (10) 1.24±0.12* 24.5±5.1

(30) 0.73±0.06** 54.9±5.9

(100) 0.42±0.07** 73.7±4.3

IC

₅₀

45.0±3.5

Control 1.30±0.17 --

TFP (3) 1.10±0.27 17.8±1.2

(10) 0.20±0.03** 84.5±1.2

(30) 0.05±0.04** 96.1±3.2

IC

50

9.4±2.2

Control 2.02±0.34 --

65 (10) 1.54±0.44 24.8±14.1

(30) 1.50±0.48 28.1±12.3

66 (10) 1.46±0.26 28.1±1.2

(30) 1.27±0.37* 39.9±8.6

67 (10) 1.32±0.38* 37.5±8.6

(30) 1.41±0.43 32.5±9.8

68 (10) 1.37±0.30* 33.9±4.9

(30) 1.07±0.11** 45.9±3.0

TFP (3) 2.42±0.10 -29.4±12.0

(10) 0.44±0.05** 77.1±7.2

(30) 0.10±0.08** 93.0±3.0

IC

50

14.7±0.4

Control 1.83±0.05 --

70 (10) 1.55±0.06 15.3±1.7

(30) 1.16±0.02** 36.7±1.5

71 (10) 1.49±0.05 18.6±0.4

(20) 1.01±0.08** 44.3±5.8

(30) 0.73±0.03** 60.1±2.8

IC

50

24.4±1.5

72 (10) 1.78±0.13 3.0±4.2

(30) 1.34±0.12* 24.7±7.4

73 (10) 1.42±0.15 21.7±10.4

(30) 1.08±0.04** 41.2±0.7

TFP (3) 2.50±0.42 -30.2±11.7

(10) 1.09±0.28 40.5±11.1

(30) 0.11±0.06** 93.6±3.3

IC

50

12.9±1.0

Neutrophil suspensions were preincubated at 37 °C with 0.5 % DMSO or test compounds in the

presence of cytochalasin B (5 µg/ml) for 3 min. Fifteen minutes after the addition of fMLP (0.3 µM),

the absorbance was determined at 550 nm. Trifluoperazine (TFP) is a positive control. Values are

presented as mean± S.E., N=3-6. *: P<0.05, **: P<0.01.

Table 22. The inhibitory effect of 5-(2'-alkoxycarbonyl substituted phenoxy)furfurals on rat neutrophil superoxide formation (in vitro)

81: R=CH

3

, R

1

=R

2

=R

3

=R

4

=H 88: R=CH

3

, R

1

=R

3

=R

4

=H, R

2

=OCH

3

82: R=CH

3

, R

1

=R

2

=R

3

=H, R

4

=CH

3

89: R=CH

3

, R

2

=R

3

=R

4

=H, R

1

=OCH

3

83: R=CH

3

, R

1

=R

2

=R

4

=H, R

3

=CH

3

90: R=CH

3

, R

1

=R

2

=R

4

=H, R

3

=Cl 84: R=CH

3

, R

1

=R

3

=R

4

=H, R

2

=CH

3

91: R=CH

3

, R

1

=R

3

=R

4

=H, R

2

=Cl 85: R=C

2

H

5

, R

2

=R

3

=R

4

=H, R

1

=CH

3

92: R=CH

3

, R

1

=R

3

=R

4

=H, R

2

=Br 86: R=CH

3

, R

1

=R

2

=R

3

=H, R

4

=OCH

3

93: R=CH

3

, R

1

=R

3

=R

4

=H, R

2

=I 87: R=CH

3

, R

1

=R

2

=R

4

=H, R

3

=OCH

3

Superoxide Formation

Compound conc.--- ( µ M) nmol/10

⁶

cells/30 min (%inh)

Control 2.40±0.23 --

81 (10) 1.85±1.52 24.29±6.8

(30) 1.52±0.37* 38.5±8.9

82 (10) 2.18±0.34 9.8±8.5

(30) 1.97±0.33 17.9±10.0

83 (10) 1.42±0.14* 40.7±0.7

(30) 1.43±0.12* 38.2±10.3

84 (10) 1.51±0.10* 36.6±2.7

(30) 1.60±0.10* 32.6±2.6

85 (10) 1.81±0.05 23.2±7.6

(30) 1.70±0.01 27.9±5.9

TFP (3) 1.81±0.04* 24.3±1.9

(10) 0.51±0.08** 78.1±5.4

(30) 0.01±0.05** 99.1±3.3

IC

50

6.2±0.3

COOR

O R

₄

R

₃

R

₂

R

₁

O CHO

86 (10) 3.93±0.43* 27.6±6.1

(30) 3.40±0.44** 36.7±9.3

87 (10) 3.61±0.57** 33.8±7.5

(30) 2.94±0.37** 45.9±5.0

88 (10) 3.72±0.62** 31.6±9.0

(30) 2.90±0.35** 46.5±7.7

89 (10) 3.80±0.60** 30.4±7.6

(30) 3.02±0.29** 43.6±8.0

90 (10) 3.80±0.51** 30.1±6.6

(30) 2.99±0.35** 44.8±6.3

91 (10) 4.33±0.59 20.8±6.5

(30) 3.83±0.37* 29.3±6.4

92 (3) 3.95±0.29* 27.3±2.4

(10) 2.96±0.16** 45.2±3.0

(30) 1.88±0.12** 64.8±4.6

IC

50

13.4±2.9

93 (3) 3.81±0.27* 29.3±6.0

(10) 3.28±0.21** 39.1±4.4

(30) 2.34±0.12** 56.7±2.9

IC

50

19.6±5.6

TFP (3) 4.12±0.04 24.3±1.9

(10) 1.16±0.08** 78.1±5.4

(30) 0.02±0.05** 99.1±3.3

IC

50

6.2±0.3

Neutrophil suspensions were preincubated at 37 °C with 0.5 % DMSO or test compounds in the

presence of cytochalasin B (5 µg/ml) for 3 min. Fifteen minutes after the addition of fMLP (0.3 µM),

the absorbance was determined at 550 nm. Trifluoperazine (TFP) is a positive control. Values are

presented as mean± S.E., N=3-5. *: P<0.05, **: P<0.01.

Table 23. The inhibitory effect of 5-(2'-alkoxycarbonyl substituted phenoxy)-2- furanacrylic acids on rat neutrophil superoxide formation (in vitro)

101: R=CH

3

, R

1

=R

2

=R

3

=R

4

=H 106: R=CH

3

, R

1

=R

2

=R

3

=H, R

4

=OCH

3

102: R=CH

3

, R

1

=R

2

=R

3

=H, R

4

=CH

3

107: R=CH

3

, R

1

=R

2

=R

4

=H, R

3

=OCH

3

103: R=CH

3

, R

1

=R

2

=R

4

=H, R

3

=CH

3

108: R=CH

3

, R

1

=R

3

=R

4

=H, R

2

=OCH

3

104: R=CH

3

, R

1

=R

3

=R

4

=H, R

2

=CH

3

109: R=CH

3

, R

2

=R

3

=R

4

=H, R

1

=OCH

3

105: R=C

2

H

5

, R

2

=R

3

=R

4

=H, R

1

=CH

3

Superoxide Formation

Compound conc.--- ( µ M) nmol/10

⁶

cells/30 min (%inh)

Control 2.40±0.23 --

101 (10) 2.11±0.22 12.0±1.3

(30) 1.82±0.14 23.7±2.0

102 (10) 2.12±0.05 9.6±10.3

(30) 1.93±0.06 17.6±8.4

103 (10) 2.42±0.13 -1.8±4.2

(30) 2.33±0.18 2.5±2.2

104 (10) 2.53±0.11 -6.5±5.2

(30) 2.09±0.14 12.1±3.7

105 (10) 2.34±0.41 3.9±7.5

(30) 2.43±0.09 -2.4±5.6

TFP (3) 1.81±0.04* 24.3±1.9

(10) 0.51±0.08** 78.1±5.4

(30) 0.01±0.05** 99.1±3.3

IC

50

6.2±0.3

Control 5.47±0.57 --

106 (10) 4.42±0.42 17.8±9.5

O COOR R

₁

R

₂

R

₃

R

₄

O CH CH COOH

107 (10) 4.40±0.43 18.4±8.8

(30) 4.69±0.35 12.5±10.5

108 (10) 4.61±0.56 15.4±7.0

(30) 4.22±0.43 22.2±6.3

109 (10) 4.41±0.62 19.2±7.5

(30) 4.44±0.59 17.9±10.4

TFP (3) 4.12±0.04 24.3±1.9

(10) 1.16±0.08** 78.1±5.4

(30) 0.02±0.05** 99.1±3.3

IC

50

6.2±0.3

Neutrophil suspensions were preincubated at 37 °C with 0.5 % DMSO or test compounds in the

presence of cytochalasin B (5 µg/ml) for 3 min. Fifteen minutes after the addition of fMLP (0.3 µM),

the absorbance was determined at 550 nm. Trifluoperazine (TFP) is a positive control. Values are

presented as mean± S.E., N=3-5. *: P<0.05, **: P<0.01.

Table 24. The inhibitory effect of 5-(2'-carboxyl substituted phenoxy)-2-furanacrylic acids on rat neutrophil superoxide formation (in vitro)

111: R

1

=R

2

=R

3

=R

4

=H 118: R

1

=R

3

=R

4

=H, R

2

=OCH

3

112: R

1

=R

2

=R

3

=H, R

4

=CH

3

120: R

1

=R

2

=R

4

=H, R

3

=Cl 113: R

1

=R

2

=R

4

=H, R

3

=CH

3

121: R

1

=R

3

=R

4

=H, R

2

=Cl 114: R

1

=R

3

=R

4

=H, R

2

=CH

3

122: R

1

=R

3

=R

4

=H, R

2

=Br 116: R

1

=R

2

=R

3

=H, R

4

=OCH

3

123: R

1

=R

3

=R

4

=H, R

2

=I 117: R

1

=R

2

=R

4

=H, R

3

=OCH

3

Superoxide Formation

Compound conc.--- ( µ M) nmol/10

⁶

cells/30 min (%inh)

Control 2.40±0.23 --

111 (10) 2.03±0.15 14.5±6.9

(30) 2.37±0.14 0.3±4.3

112 (10) 2.28±0.23 5.1±2.4

(30) 2.16±0.10 7.6±12.6

113 (10) 2.18±0.16 7.7±9.8

(30) 2.07±0.19 13.2±3.7

114 (10) 2.49±0.13 -4.7±6.1

(30) 2.35±0.06 0.8±6.6

TFP (3) 1.81±0.04* 24.3±1.9

(10) 0.51±0.08** 78.1±5.4

(30) 0.01±0.05** 99.1±3.3

IC

50

6.2±0.3

Control 5.47±0.57 --

116 (10) 4.66±0.75 14.6±10.5

(30) 4.57±0.63 15.8±8.9

COOH

O R

₁

R

₂

R

₃

O CH

R

₄

CH COOH

(30) 3.76±0.03* 29.4±8.7

118 (10) 3.96±0.86* 29.5±8.9

(30) 3.68±0.75** 34.1±7.3

120 (10) 4.50±0.65 17.2±9.2

(30) 4.07±0.54 24.5±10.0

121 (10) 5.01±0.48 7.8±4.8

(30) 4.65±0.50 14.2±7.6

122 (10) 4.89±0.57 9.9±8.3

(30) 4.52±0.49 16.5±8.5

123 (10) 4.91±0.68 10.0±7.9

(30) 4.57±0.65 15.9±9.4

TFP (3) 4.12±0.04 24.3±1.9

(10) 1.16±0.08** 78.1±5.4

(30) 0.02±0.05** 99.1±3.3

IC

50

6.2±0.3

Neutrophil suspensions were preincubated at 37 °C with 0.5 % DMSO or test compounds in the

presence of cytochalasin B (5 µg/ml) for 3 min. Fifteen minutes after the addition of fMLP (0.3 µM),

the absorbance was determined at 550 nm. Trifluoperazine (TFP) is a positive control. Values are

presented as mean± S.E., N=3-5. *: P<0.05, **: P<0.01.

(三)對於PMA誘導的嗜中性白血球超氧自由基生成作用抑制試驗 從化合物

21-33、41-48、50-53、61-68、70-73、81-93、101-109、

111-114、116-118 及 120-123 對以 PMA 誘導的嗜中性白血球超氧自由

基生成作用之體外試驗中，由 superoxide formation 的抑制百分率(見 Table 25 至 Table 30)看來，在濃度 30

µ

M 時，化合物

26、28、45、

65、66、70、81 及 83 分別呈現弱的抑制活性 (具有約 20-34%的抑制

百分率)。其他化合物則無明顯的抑制活性。

Table 25. The inhibitory effect of ethyl 5-(2'-alkoxycarbonyl substituted phenoxy)- furan-2-carboxylates on rat neutrophil superoxide formation (in vitro)

21: R=CH

3

, R

1

=R

2

=R

3

=R

4

=H 28: R=CH

3

, R

1

=R

3

=R

4

=H, R

2

=OCH

3

22: R=CH

3

, R

1

=R

2

=R

3

=H, R

4

=CH

3

29: R=CH

3

, R

2

=R

3

=R

4

=H, R

1

=OCH

3

23: R=CH

3

, R

1

=R

2

=R

4

=H, R

3

=CH

3

30: R=CH

3

, R

1

=R

2

=R

4

=H, R

3

=Cl 24: R=CH

3

, R

1

=R

3

=R

4

=H, R

2

=CH

3

31: R=CH

3

, R

1

=R

3

=R

4

=H, R

2

=Cl 25: R=C

2

H

5

, R

2

=R

3

=R

4

=H, R

1

=CH

3

32: R=CH

3

, R

1

=R

3

=R

4

=H, R

2

=Br 26: R=CH

3

, R

1

=R

2

=R

3

=H, R

4

=OCH

3

33: R=CH

3

, R

1

=R

3

=R

4

=H, R

2

=I 27: R=CH

3

, R

1

=R

2

=R

4

=H, R

3

=OCH

3

Superoxide Formation

Compound conc.--- (µM) nmol/10

⁶

cells/30 min (%inh)

Control 4.1±0.3 --

21 (30) 3.9±0.1 -11.4±2.2

(100) 3.6±0.3 -3.0±7.2

22 (30) 3.7±0.2 -3.0±3.8

(100) 3.7±0.1 -5.2±3.0

23 (30) 3.5±0.2 -1.1±4.0

(100) 3.4±0.2 2.2±3.8

TFP (3) 3.5±0.4 19.2±11.2

(5) 0.8±0.0** 81.4±2.1

(10) 0.2±0.1** 96.1±2.0

IC

₅₀

4.7±0.5

Control 1.75±0.29 --

24 (30) 1.71±0.31 3.1±1.7

(100) 1.36±0.24 23.1±3.3

Control 1.92±0.09 --

TFP (3) 1.34±0.07** 29.8±0.3

(10) 0.48±0.02** 74.5±0.2

(30) 0.22±0.01** 88.2±1.3

IC

50

8.8±0.1

COOR

O R

₄

R

₃

R

₂

R

₁

O COOCH

₂

CH

₃

Control 3.14±0.12 --

25 (10) 2.48±0.26* 21.1±6.7

(30) 3.10±0.06 1.2±2.2

TFP (1) 2.55±0.23 16.6±10.1

(3) 1.59±0.43** 49.5±7.4

(10) 0.39±0.19** 87.1±4.9

IC

₅₀

4.5±0.3

Control 2.16±0.18 --

26 (30) 1.44±0.19** 33.9±5.3

(100) 1.58±0.13** 32.8±5.1

27 (30) 1.89±0.27 14.1±6.1

(100) 1.72±0.35* 21.9±7.8

28 (30) 1.56±0.15* 27.1±7.2

(100) 2.17±0.11 7.3±5.3

TFP (3) 1.85±0.53* 21.9±7.1

(5) 1.05±0.08** 53.6±4.0

(10) 0.53±0.11** 76.4±5.1

IC

50

6.0±0.1

Control 6.34±0.25 --

29 (10) 5.99±0.82 5.6±12.1

(30) 6.23±0.82 1.8±12.2

TFP (3) 5.63±0.10 8.6±2.0

(10) 1.18±0.03** 80.2±2.0

(30) 0.47±0.01** 90.9±0.4

IC

₅₀

7.6±0.3

Control 2.16±0.18 --

30 (30) 1.90±0.17 12.2±0.9

(100) 3.05±0.20* -29.7±5.7

31 (30) 1.97±0.14 16.1±5.9

(100) 2.16±0.13 7.3±8.5

32 (30) 2.19±0.35 0.1±11.6

(100) 2.91±0.38** -48.8±4.1

TFP (3) 1.85±0.53* 21.9±7.1

(5) 1.05±0.08** 53.6±4.0

(10) 0.53±0.11** 76.4±5.1

IC

50

6.0±0.1

Control 3.50±0.03 --

33 (10) 3.24±0.15 7.5±3.7

(30) 3.14±0.25 10.4±6.7

TFP (3) 2.33±0.19** 32.5±8.6

(10) 0.86±0.15** 74.9±5.7

(30) 0.08±0.03** 97.6±0.7

IC

50

6.8±2.3

° %

presence of cytochalasin B (5 µg/ml) for 3 min. Fifteen minutes after the addition of PMA (3 nM), the absorbance was determined at 550 nm. Trifluoperazine (TFP) is a positive control. Values are presented as mean± S.E., N=3-5. *: P<0.05, **: P<0.01.

Table 26. The inhibitory effect of 5-(2'-carboxyl substituted phenoxy)furan-2- carboxylic acids on rat neutrophil superoxide formation (in vitro)

41: R

1

=R

2

=R

3

=R

4

=H 47: R

1

=R

2

=R

4

=H, R

3

=OCH

3

42: R

1

=R

2

=R

3

=H, R

4

=CH

3

48: R

1

=R

3

=R

4

=H, R

2

=OCH

3

43: R

1

=R

2

=R

4

=H, R

3

=CH

3

50: R

1

=R

2

=R

4

=H, R

3

=Cl 44: R

1

=R

3

=R

4

=H, R

2

=CH

3

51: R

1

=R

3

=R

4

=H, R

2

=Cl 45: R

2

=R

3

=R

4

=H, R

1

=CH

3

52: R

1

=R

3

=R

4

=H, R

2

=Br 46: R

1

=R

2

=R

3

=H, R

4

=OCH

3

53: R

1

=R

3

=R

4

=H, R

2

=I

Superoxide Formation

Compound conc.--- (µM) nmol/10

⁶

cells/30 min (%inh)

Control 4.1±0.3 --

41 (30) 3.9±0.1 -10.4±2.8

(100) 4.0±0.0 -13.4±0.8

42 (30) 4.1±0.1 -17.0±1.8

(100) 4.0±0.2 -13.2±4.8

43 (30) 3.9±0.3

(100) 3.9±0.1 -14.0±0.5

TFP (3) 3.5±0.4 19.2±11.2

(5) 0.8±0.0** 81.4±2.1

(10) 0.2±0.1** 96.1±2.0

IC

₅₀

4.7±0.5

Control 1.75±0.29 --

44 (30) 1.87±0.23 -9.5±6.2

(100) 1.76±0.35 1.9±8.2

Control 1.92±0.09 --

TFP (3) 1.34±0.07** 29.8±0.3

(10) 0.48±0.02** 74.5±0.2

(30) 0.22±0.01** 88.2±1.3

IC

50

8.8±0.1

Control 3.14±0.12 --

COOH

O R

₄

R

₃

R

₂

R

₁

O COOH

45 (10) 2.61±0.24 17.3±4.6

(30) 2.48±0.13 20.7±7.4

TFP (1) 2.55±0.23 16.6±10.1

(3) 1.59±0.43** 49.5±7.4

(10) 0.39±0.19** 87.1±4.9

IC

50

4.5±0.3

Control 2.16±0.18 --

46 (30) 1.78±0.17 17.4±5.6

(100) 2.01±0.37 4.8±9.9

47 (30) 2.19±0.07 6.2±7.4

(100) 1.77±0.14 17.9±2.5

48 (30) 2.00±0.08 6.1±5.2

(100) 1.65±0.27* 30.0±9.8

50 (30) 1.89±0.25 13.1±7.3

(100) 1.69±0.24* 28.2±8.8

51 (30) 1.99±0.10 6.2±7.3

(100) 1.96±0.14 9.0±3.0

52 (30) 2.14±0.16 8.7±7.2

(100) 1.93±0.31 12.6±9.1

TFP (3) 1.85±0.53* 21.9±7.1

(5) 1.05±0.08** 53.6±4.0

(10) 0.53±0.11** 76.4±5.1

IC

50

6.0±0.1

Control 3.50±0.03 --

53 (10) 3.06±0.30 12.9±7.7

(30) 3.16±0.27 10.0±8.5

TFP (3) 2.33±0.19** 32.5±8.6

(10) 0.86±0.15** 74.9±5.7

(30) 0.08±0.03** 97.6±0.7

IC

₅₀

6.8±2.3

Neutrophil suspensions were preincubated at 37 °C with 0.5 % DMSO or test compounds in the

presence of cytochalasin B (5 µg/ml) for 3 min. Fifteen minutes after the addition of PMA (3 nM), the

absorbance was determined at 550 nm. Trifluoperazine (TFP) is a positive control. Values are presented

as mean± S.E., N=3-4. *: P<0.05, **: P<0.01.

Table 27. The inhibitory effect of substituted furo[2,3-b]chromone-2-carboxylic acid ethyl esters on rat neutrophil superoxide formation (in vitro)

O O

O

COOCH

₂

CH

₃

R

₁

R

₂

R

₃

R

₄

61: R

1

=R

2

=R

3

=R

4

=H 67: R

1

=R

2

=R

4

=H, R

3

=OCH

3

62: R

1

=R

2

=R

3

=H, R

4

=CH

3

68: R

1

=R

3

=R

4

=H, R

2

=OCH

3

63: R

1

=R

2

=R

4

=H, R

3

=CH

3

70: R

1

=R

2

=R

4

=H, R

3

=Cl 64: R

1

=R

3

=R

4

=H, R

2

=CH

3

71: R

1

=R

3

=R

4

=H, R

2

=Cl 65: R

2

=R

3

=R

4

=H, R

1

=CH

3

72: R

1

=R

3

=R

4

=H, R

2

=Br 66: R

1

=R

2

=R

3

=H, R

4

=OCH

3

73: R

1

=R

3

=R

4

=H, R

2

=I

Superoxide Formation

Compound conc.--- (µM) nmol/10

⁶

cells/30 min (%inh)

Control 4.1±0.3 --

61 (30) 4.0±0.4 0.4±4.4

(100) 4.3±0.3 -8.8±1.5

62 (30) 3.8±0.2 -7.9±4.8

(100) 3.6±0.2 -3.0±3.8

TFP (3) 3.5±0.4 19.2±11.2

(5) 0.8±0.0** 81.4±2.1

(10) 0.2±0.1** 96.1±2.0

IC

50

4.7±0.5

Control 1.75±0.29 --

63 (30) 1.97±0.28 -14.9±8.0

(100) 2.17±0.24 -29.1±10.9

64 (30) 1.81±0.34 -1.7±6.6

(100) 1.71±0.31 3.6±6.3

Control 1.92±0.09 --

TFP (3) 1.34±0.07** 29.8±0.3

(10) 0.48±0.02** 74.5±0.2

(30) 0.22±0.01** 88.2±1.3

IC

₅₀

8.8±0.1