JPH09110835A - 6-azaspiro(4,5)decane derivative - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a new 6-azaspiro[4,5]decane derivative, having inhibiting activities against cytoplasmic phospholipase A2 and useful as an antiinflammatory agent. SOLUTION: This 6-azaspiro[4,5]decane derivative is represented by formula I [R<1> is H, a lower alkyl, a tri-lower alkylsilyl, an acyl or a sulfo-lower alkyl; R<2> to R<4> are each H, a lower alkyl, a tri-lower alkylsilyl or an acyl; X is a halogen; Y is O or NR<5> (R<5> is H, a lower alkyl, a tri-lower alkylsilyl or an acyl)], e.g. a compound represented by formula II. The compound represented by formula I is obtained by pulverizing and extracting a shucked shellfish which is shellfishes of the genus Pinna such as Pinna muricate Linnaeus in an organic solvent and further separating the extracted substance according to chromatography.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a novel 6-azaspiro [4,5] decane derivative. This compound has cytoplasmic phospholipase A2 inhibitory activity and is expected to be used as an anti-inflammatory agent.

[0002]

2. Description of the Related Art An inflammatory reaction is a kind of host defense effect that is activated when a harmful stimulus enters a living body, but as a result, it is fatal with swelling, pain, and impairment of organ functions. Not a few. Specifically, the causes, developmental processes, and symptoms thereof are extremely wide and complicated, from acute inflammation associated with type I allergy to an external action to chronic inflammation due to nephritis or rheumatic disease. What is used as this symptomatic treatment agent is called an anti-inflammatory agent, which is roughly classified into a steroid anti-inflammatory agent and a non-steroidal anti-inflammatory agent, and various glucocorticoids, indomethacin and the like are representative. However, steroidal anti-inflammatory drugs
It has an inhibitory effect on protein synthesis including various proteinaceous mediators, has a wide range of pharmacological effects and a large curative effect, but is known to cause serious side effects, and it is almost impossible to separate pharmacological effects and side effects. Was also recently revealed. On the other hand, nonsteroidal anti-inflammatory drugs have limited effects because cyclooxygenase inhibitory (prostaglandin production inhibition) action is the main pharmacological action. Therefore, development of anti-inflammatory agents based on new pharmacological action is required,
A compound having cytoplasmic phospholipase A ₂ inhibitory activity is considered to be a substance that fulfills its purpose. Cytoplasmic phospholipase A ₂ is an enzyme whose existence has been revealed very recently, and it is known that its activation commonly initiates the enzymatic production of prostaglandins, leukotrienes, and PAFs, which are the major mediators of inflammation. Has been.

[0003]

The object of the present invention is therefore to discover an anti-inflammatory substance having a novel structure.

[0004]

Means for Solving the Problems As a result of searching the components contained in the extract of marine organisms, the present inventors have found that a novel 6-azaspiro isolated from the extract of the peeled sardine of the pinna genus Pinna We have found that the [4,5] decane derivative has a cytoplasmic phospholipase A2 inhibitory activity, and completed the present invention.

That is, the present invention has the general formula (I)

[0006]

Embedded image

(Wherein R ¹ represents a hydrogen atom, a substituted or unsubstituted lower alkyl group, a tri-lower alkylsilyl group, an acyl group or a sulfo lower alkyl group, and R ² , R ³ and R ⁴ are independently hydrogen. Atom, a substituted or unsubstituted lower alkyl group, a tri-lower alkylsilyl group, or an acyl group, X represents a halogen atom, Y represents an oxygen atom or -NR ⁵
-Represents. R ⁵ represents a hydrogen atom, a substituted or unsubstituted lower alkyl group, a tri-lower alkylsilyl group or an acyl group. The present invention provides a 6-azaspiro [4,5] decane derivative represented by (4).

[0008]

BEST MODE FOR CARRYING OUT THE INVENTION In the above general formula (I), the lower alkyl group means a linear or branched alkyl group having 1 to 6 carbon atoms, and specific examples thereof include a methyl group and an ethyl group. , Propyl group, isopropyl group, butyl group, isobutyl group, pentyl group, hexyl group and the like. Examples of the substituent of the lower alkyl group include a halogen atom, a lower alkoxy group such as a methoxy group and an ethoxy group, an aryl group such as a phenyl group, a p-tolyl group and a furyl group.

Further, as the tri-lower alkylsilyl group, trimethylsilyl group, ethyldimethylsilyl group, t-
Examples thereof include a butyldimethylsilyl group and the like, and examples of the acyl group include a formyl group, an acetyl group, a propionyl group, a butyryl group, a benzoyl group, a p-chlorobenzoyl group and the like. Further, examples of the sulfo lower alkyl group include a 2-sulfoethyl group, a 2-sulfopropyl group, a 3-sulfopropyl group and a 4-sulfobutyl group.

6- represented by the general formula (I) of the present invention
Among the azaspiro [4,5] decane derivatives, compounds represented by the following formulas (1) and (2) are pinna shells such as, for example, scalloped shellfish (Pinna genus) and the like in organic solvents. It is obtained by a usual method by crushing, extracting in, and separating by chromatography.

[0011]

Embedded image

[0012]

Embedded image

Examples of the solvent used for crushing and extraction include ethanol, methanol, acetone, and the like. Column chromatography, high performance liquid and thin layer chromatography are used, and column chromatography is TSK-.
In addition to G3000S, Diaion HP-20, Sephadex LH-20, DEAE Sephadex, reverse phase RP-18, etc. are used, and reverse phase RP-18 etc. are used for high performance liquid chromatography, and thin layer For chromatography, RP-18 is used in addition to silica gel.

The other compounds represented by the above general formula (I) of the present invention include, for example, the methyl iodide and the like obtained by subjecting the 6-azaspiro [4,5] decane derivative represented by the above formula (1) to a conventional method. Can be obtained by alkylation with an alkylating agent, a silylation with a silylating agent such as trimethylsilyl chloride, or an acylation with acetyl chloride.
In addition, 6-azaspiro [4 represented by the above formula (2)
5] Esterification or amidation of a decane derivative in the presence of a suitable condensing agent, or alkylation similar to the above,
It can also be obtained by combining silylation, acylation and the like, if desired.

Hereinafter, the details will be described with reference to Examples and Test Examples.

[0016]

【Example】

Example 1 Approximately 10 pieces of peeled sardine mussels (10 kg) were used.
It was mixed with L ethanol and pulverized with a blender. The obtained immersion liquid was cold-soaked for 10 days, and then suction-filtered with a Buchner funnel having an inner diameter of 30 cm. The dark brown filtrate was concentrated to dryness under reduced pressure with a rotary evaporator (about 40 ° C.), and this concentrate was partitioned with 80% methanol and n-hexane. 80%
The methanol layer was concentrated, partitioned with water and ethyl acetate, and the aqueous layer was concentrated.

T suspended in a glass column with ethanol
After filling SK-G3000S and replacing the solvent with water,
The concentrated aqueous layer was passed through. Sequentially on this column,
25% ethanol-water, 50% ethanol-water, 75%
Ethanol-water and 99% ethanol were poured, and the fractions eluted with 50% ethanol were collected and concentrated to give a small amount of methanol solution. This was placed on a Sephadex LH-20 column swollen with methanol and eluted with methanol, and active fractions were collected and concentrated to give a small amount of an aqueous solution. Place this on a DEAE Sephadex column and apply 0.02M
Elution with a phosphate buffer (pH 6.9), placing the active fraction on TSK-G3000S (water) as it is, and eluting with an ethanol-water mixed solvent in which the ratio of ethanol was sequentially changed to 25, 50, 75, 99%. The 50% ethanol eluate was collected and concentrated. Put on a reverse phase column, 60% methanol-
Elution with water was performed, and active fractions were collected, concentrated, and loaded onto a normal phase column, and eluted with 15% methanol-chloroform to collect and concentrate active fraction 1. This was adsorbed on a preparative silica gel thin layer plate (thickness 0.5 mm, 20 × 20 cm), developed with methanol: chloroform (3: 7) for 1 hour, and R
Scraping off the part with f value around 0.5, chloroform:
Add methanol (1: 1), shake vigorously, and let stand,
The lower layer was separated and concentrated. The single active fraction was concentrated to dryness and then 6-
Azaspiro [4,5] decane derivative (1) (4 mg) was obtained as a pale yellow solid. The physical properties of this product were as follows.

Molecular weight: 532 Molecular formula: C ₂₅ H ₄₁ ClN ₂ O ₆ S ¹ H-NMR (CD ₃ OD, 400MHz): δ 6.28 (1H, t), 5.82 (1H, dd),
5.72 (1H, d), 5.64 (1H, dd), 5.03 (1H, dd), 3.78 (2H, t),
3.68 (2H, t), 3.40 (1H, m), 3.00 (2H, t), 2.56 (2H, br.d
d), 2.55 (2H, t), 2.40 (1H, m), 2.15 (1H, dd), 1.98 (1H),
1.92 (1H), 1.88 (3H, s), 1.86 (1H), 1.84 (1H), 1.82 (2
H), 1.79 (1H), 1.68 (1H), 1.64 (1H), 1.56 (1H), 1.54 (1
H), 1.40 (1H, dddd), 1.08 (3H, d).

¹³ C-NMR (CD ₃ OD, 125 MHz): δ169.04 (s), 1
36.98 (d), 134.51 (s), 132.14 (s), 130.69 (d), 128.29
(d), 127.71 (d), 68.80 (d), 68.74 (s), 58.06 (t), 54.2
1 (d), 53.46 (d), 49.05 (t), 41.65 (t), 36.17 (d), 35.0
5 (t), 34.11 (t), 33.74 (t), 31.24 (t), 28.37 (t), 27.2
1 (t), 21.55 (t), 19.38 (t), 18.96 (q), 11.30 (q).

HRFAB-MS: m / z 533.2452 (△ -0.5mmu) (M + H)
⁺

The 15% methanol-chloroform elution active fraction 2 of the normal phase column was adsorbed on a preparative silica gel thin layer plate (thickness 0.5 mm, 20 × 20 cm), and methanol / chloroform (3: 7) was used. Unfold for 1 hour, R
Scraping off the part with f value around 0.2, chloroform:
Add methanol (1: 1), shake vigorously, and let stand,
The lower layer was separated and concentrated. The single active fraction was concentrated to dryness and then 6-
Azaspiro [4,5] decane derivative (2) (1 mg) was obtained as a pale yellow solid. The physical properties of this product were as follows.

Molecular weight: 425 Molecular formula: C ₂₃ H ₃₈ ClNO ₄ ¹ H-NMR (CD ₃ OD, 400 MHz): δ6.45 (1H, t), 5.82 (1H, dd),
5.72 (1H, d), 5.64 (1H, dd), 5.03 (1H, dd), 3.78 (2H, t),
3.40 (1H, m), 2.56 (2H, br.dd), 2.55 (2H, t), 2.40 (1H,
m), 2.15 (1H, dd), 1.98 (1H), 1.92 (1H), 1.88 (3H, s),
1.86 (1H), 1.84 (1H), 1.82 (2H), 1.79 (1H), 1.68 (1H),
1.64 (1H), 1.56 (1H), 1.54 (1H), 1.40 (1H, dddd), 1.08
(3H, d).

¹³ C-NMR (CD ₃ OD, 125 MHz): δ 170.14 (s), 1
36.97 (d), 134.51 (s), 132.01 (s), 130.75 (d), 128.22
(d), 127.71 (d), 68.84 (d), 68.48 (s), 58.06 (t), 54.2
1 (d), 53.46 (d), 41.65 (t), 36.37 (d), 34.17 (t), 33.7
0 (t), 31.53 (t), 28.54 (t), 27.20 (t), 21.66 (t), 19.4
6 (t), 19.03 (q), 11.86 (q).

HRFAB-MS: m / z 426.2412 (△ + 0.1mmu) (M + H)
⁺

Test Example 1 Measurement of Phospholipase A2 Inhibitory Activity As phospholipase A2 (PLA2), 85 kDa cytoplasmic PLA2 (cPLA2) purified from rabbit platelets based on a previous report (FEBS Lett., 282, 326-330, 1991) was used. The inhibitory activity against was measured as follows.
The 6-azaspiro [4,5] decane derivative (1) or (2) was dissolved in methanol and used as a test solution. 1
25 μL of M Tris-hydrochloric acid (pH 9.0) and 20 μL of 50 mM calcium chloride solution were mixed with a test solution and cP.
LA2 solution was added to make 200 μL, and the mixture was reacted at 37 ° C. for 20 minutes. Then the substrate 1-palmitoyl-2
-[ ^14C ] arachidonoyl-glycerophosphoethanolamine (0.5 nmol / 50000 dpm / 50μ)
L) was added, and the mixture was further reacted at 37 ° C. for 20 minutes. Dole reagent (isopropanol: heptane: 1N sulfuric acid = 10:
The reaction was stopped by adding 1.25 mL of 40: 1), the [ ¹⁴ C] arachidonic acid fraction was collected by the method of Dole, and the radioactivity was measured by a liquid scintillation counter to measure the enzyme activity. Table 1 shows the results.

[0026]

[Table 1]

[0027]

INDUSTRIAL APPLICABILITY Since the compound of the present invention has cytoplasmic phospholipase A2 inhibitory activity, cytoplasmic phospholipase A2
It has a use as an inhibitor and is expected to be used as, for example, an anti-inflammatory agent or a prodrug thereof.

────────────────────────────────────────────────── ───

[Procedure amendment]

[Submission date] August 9, 1996

[Procedure amendment 2]

[Document name to be amended] Statement

[Correction target item name] Full text

[Correction method] Change

[Correction contents]

[Document Name] Statement

Title of the invention 6-azaspiro [4 . 5] Decane derivative

[Claims]

Embedded image (In the formula, R ¹ represents a hydrogen atom, a substituted or unsubstituted lower alkyl group, a tri-lower alkylsilyl group, an acyl group or a sulfo lower alkyl group, and R ² , R ³ and R ⁴ are independently a hydrogen atom or a substituted group. Or, it represents an unsubstituted lower alkyl group, a tri-lower alkylsilyl group, or an acyl group, X represents a halogen atom, Y represents an oxygen atom or -NR ^5- , R ⁵ represents a hydrogen atom, a substituted or unsubstituted 6-azaspiro represented by a lower alkyl group, a tri-lower alkylsilyl group or an acyl group) [4 . 5] Decane derivative.

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a novel 6-azaspiro [4 . 5] A decane derivative. This compound has cytoplasmic phospholipase A ₂ inhibitory activity and is expected to be used as an anti-inflammatory agent.

[0002]

2. Description of the Related Art An inflammatory reaction is a kind of host defense effect that is activated when a harmful stimulus enters a living body, but as a result, it is fatal with swelling, pain, and impairment of organ functions. Not a few. Specifically, the causes, developmental processes, and symptoms thereof are extremely wide and complicated, from acute inflammation associated with type I allergy to an external action to chronic inflammation due to nephritis or rheumatic disease. What is used as this symptomatic treatment agent is called an anti-inflammatory agent, which is roughly classified into a steroid anti-inflammatory agent and a non-steroidal anti-inflammatory agent, and various glucocorticoids, indomethacin and the like are representative. However, steroidal anti-inflammatory drugs
It has an inhibitory effect on protein synthesis including various proteinaceous mediators, has a wide range of pharmacological effects and a large curative effect, but is known to cause serious side effects, and it is almost impossible to separate pharmacological effects and side effects. Was also recently revealed. On the other hand, nonsteroidal anti-inflammatory drugs have limited effects because cyclooxygenase inhibitory (prostaglandin production inhibition) action is the main pharmacological action. Therefore, development of anti-inflammatory agents based on new pharmacological action is required,
A compound having cytoplasmic phospholipase A ₂ inhibitory activity is considered to be a substance that fulfills its purpose. Cytoplasmic phospholipase A ₂ is an enzyme whose existence has been revealed very recently, and it is known that its activation commonly initiates the enzymatic production of prostaglandins, leukotrienes, and PAFs, which are the major mediators of inflammation. Has been.

[0003]

The object of the present invention is therefore to discover an anti-inflammatory substance having a novel structure.

[0004]

Means for Solving the Problems As a result of searching the components contained in the extract of marine organisms, the present inventors have found that a novel 6-azaspiro isolated from the extract of the peeled sardine of the pinna genus Pinna [4 . 5] It was found that the decane derivative has cytoplasmic phospholipase A ₂ inhibitory activity, and completed the present invention.

That is, the present invention has the general formula (I)

[0006]

Embedded image

(Wherein R ¹ represents a hydrogen atom, a substituted or unsubstituted lower alkyl group, a tri-lower alkylsilyl group, an acyl group or a sulfo lower alkyl group, and R ² , R ³ and R ⁴ are independently hydrogen. Atom, a substituted or unsubstituted lower alkyl group, a tri-lower alkylsilyl group, or an acyl group, X represents a halogen atom, Y represents an oxygen atom or -NR ⁵
-Represents. R ⁵ represents a hydrogen atom, a substituted or unsubstituted lower alkyl group, a tri-lower alkylsilyl group or an acyl group. 6-azaspiro [4 . 5] A decane derivative is provided.

[0008]

BEST MODE FOR CARRYING OUT THE INVENTION In the above general formula (I), the lower alkyl group means a linear or branched alkyl group having 1 to 6 carbon atoms, and specific examples thereof include a methyl group and an ethyl group. , Propyl group, isopropyl group, butyl group, isobutyl group, pentyl group, hexyl group and the like. Examples of the substituent of the lower alkyl group include a halogen atom, a lower alkoxy group such as a methoxy group and an ethoxy group, an aryl group such as a phenyl group, a p-tolyl group and a furyl group.

Further, as the tri-lower alkylsilyl group, trimethylsilyl group, ethyldimethylsilyl group, t-
Examples thereof include a butyldimethylsilyl group and the like, and examples of the acyl group include a formyl group, an acetyl group, a propionyl group, a butyryl group, a benzoyl group, a p-chlorobenzoyl group and the like. Further, examples of the sulfo lower alkyl group include a 2-sulfoethyl group, a 2-sulfopropyl group, a 3-sulfopropyl group and a 4-sulfobutyl group.

6- represented by the general formula (I) of the present invention
Azaspiro [4 . 5] Among the decane derivatives, the compounds represented by the following formulas (1) and (2) are prepared by crushing a shell of a pinna genus such as Iwakawahagoromoga (Pinna genus) in an organic solvent, It is obtained by a usual method by extracting and further separating by chromatography.

[0011]

Embedded image

[0012]

Embedded image

Examples of the solvent used for crushing and extraction include ethanol, methanol, acetone, and the like. Column chromatography, high performance liquid and thin layer chromatography are used, and column chromatography is TSK-.
In addition to G3000S, Diaion HP-20, Sephadex LH-20, DEAE Sephadex, reverse phase RP-18, etc. are used, and reverse phase RP-18 etc. are used for high performance liquid chromatography, and thin layer For chromatography, RP-18 is used in addition to silica gel.

The compound represented by the general formula (I) of the present invention.
The other compound is, for example, 6-aza represented by the above formula (1).
Spiro [4.5] Iodination of the decane derivative according to a conventional method
Alkylation with alkyl agents such as methyl, trimethyl
Silylation with silylating agents such as lyl chloride, acetyl
It can be obtained by acylation with chloride or the like.
In addition, 6-azaspiro [4 represented by the above formula (2).
5] Esterification of decane derivative in the presence of a suitable condensing agent
Or amidation, or the same alkylation as above,
If desired, combine silylation, acylation, etc.
Therefore, it can also be obtained.

Hereinafter, the details will be described with reference to Examples and Test Examples.

[0016]

[Examples] Example 1 About 10 kg of peeled sardine mussels (10 kg) were used.
It was mixed with L ethanol and pulverized with a blender. The obtained immersion liquid was cold-soaked for 10 days, and then suction-filtered with a Buchner funnel having an inner diameter of 30 cm. The dark brown filtrate was concentrated to dryness under reduced pressure with a rotary evaporator (about 40 ° C.), and this concentrate was partitioned with 80% methanol and n-hexane. 80%
The methanol layer was concentrated, partitioned with water and ethyl acetate, and the aqueous layer was concentrated.

T suspended in a glass column with ethanol
After filling SK-G3000S and replacing the solvent with water,
The concentrated aqueous layer was passed through. Sequentially on this column,
25% ethanol-water, 50% ethanol-water, 75%
Ethanol-water and 99% ethanol were poured, and the fractions eluted with 50% ethanol were collected and concentrated to give a small amount of methanol solution. This was placed on a Sephadex LH-20 column swollen with methanol and eluted with methanol, and active fractions were collected and concentrated to give a small amount of an aqueous solution. Place this on a DEAE Sephadex column and apply 0.02M
Elution with a phosphate buffer (pH 6.9), placing the active fraction on TSK-G3000S (water) as it is, and eluting with an ethanol-water mixed solvent in which the ratio of ethanol was sequentially changed to 25, 50, 75, 99%. The 50% ethanol eluate was collected and concentrated. Put on a reverse phase column, 60% methanol-
Elution with water was performed, and active fractions were collected, concentrated, and loaded onto a normal phase column, and eluted with 15% methanol-chloroform to collect and concentrate active fraction 1. This was adsorbed on a preparative silica gel thin layer plate (thickness 0.5 mm, 20 × 20 cm), developed with methanol: chloroform (3: 7) for 1 hour, and R
Scraping off the part with f value around 0.5, chloroform:
Add methanol (1: 1), shake vigorously, and let stand,
The lower layer was separated and concentrated. The single active fraction was concentrated to dryness and then 6-
Azaspiro [4 . 5] Decane derivative (1) (4 mg) was obtained as a pale yellow solid. The physical properties of this product were as follows.

Molecular weight: 532 Molecular formula: C ₂₅ H ₄₁ ClN ₂ O ₆ S ¹ H-NMR (CD ₃ OD, 400MHz): δ 6.28 (1H, t), 5.82 (1H, dd),
5.72 (1H, d), 5.64 (1H, dd), 5.03 (1H, dd), 3.78 (2H, t),
3.68 (2H, t), 3.40 (1H, m), 3.00 (2H, t), 2.56 (2H, br.d
d), 2.55 (2H, t), 2.40 (1H, m), 2.15 (1H, dd), 1.98 (1H),
1.92 (1H), 1.88 (3H, s), 1.86 (1H), 1.84 (1H), 1.82 (2
H), 1.79 (1H), 1.68 (1H), 1.64 (1H), 1.56 (1H), 1.54 (1
H), 1.40 (1H, dddd), 1.08 (3H, d).

¹³ C-NMR (CD ₃ OD, 125 MHz): δ169.04 (s), 1
36.98 (d), 134.51 (s), 132.14 (s), 130.69 (d), 128.29
(d), 127.71 (d), 68.80 (d), 68.74 (s), 58.06 (t), 54.2
1 (d), 53.46 (d), 49.05 (t), 41.65 (t), 36.17 (d), 35.0
5 (t), 34.11 (t), 33.74 (t), 31.24 (t), 28.37 (t), 27.2
1 (t), 21.55 (t), 19.38 (t), 18.96 (q), 11.30 (q).

HRFAB-MS: m / z 533.2452 (△ -0.5mmu) (M + H)
⁺

The 15% methanol-chloroform elution active fraction 2 of the normal phase column was adsorbed on a preparative silica gel thin layer plate (thickness 0.5 mm, 20 × 20 cm), and methanol / chloroform (3: 7) was used. Unfold for 1 hour, R
Scraping off the part with f value around 0.2, chloroform:
Add methanol (1: 1), shake vigorously, and let stand,
The lower layer was separated and concentrated. The single active fraction was concentrated to dryness and then 6-
Azaspiro [4 . 5] Decane derivative (2) (1 mg) was obtained as a pale yellow solid. The physical properties of this product were as follows.

Molecular weight: 425 Molecular formula: C ₂₃ H ₃₈ ClNO ₄ ¹ H-NMR (CD ₃ OD, 400 MHz): δ6.45 (1H, t), 5.82 (1H, dd),
5.72 (1H, d), 5.64 (1H, dd), 5.03 (1H, dd), 3.78 (2H, t),
3.40 (1H, m), 2.56 (2H, br.dd), 2.55 (2H, t), 2.40 (1H,
m), 2.15 (1H, dd), 1.98 (1H), 1.92 (1H), 1.88 (3H, s),
1.86 (1H), 1.84 (1H), 1.82 (2H), 1.79 (1H), 1.68 (1H),
1.64 (1H), 1.56 (1H), 1.54 (1H), 1.40 (1H, dddd), 1.08
(3H, d).

¹³ C-NMR (CD ₃ OD, 125 MHz): δ 170.14 (s), 1
36.97 (d), 134.51 (s), 132.01 (s), 130.75 (d), 128.22
(d), 127.71 (d), 68.84 (d), 68.48 (s), 58.06 (t), 54.2
1 (d), 53.46 (d), 41.65 (t), 36.37 (d), 34.17 (t), 33.7
0 (t), 31.53 (t), 28.54 (t), 27.20 (t), 21.66 (t), 19.4
6 (t), 19.03 (q), 11.86 (q).

HRFAB-MS: m / z 426.2412 (△ + 0.1mmu) (M + H) ⁺

Test Example 1 Measurement of phospholipase A ₂ inhibitory activity As phospholipase A ₂ (PLA ₂ ), 85 kDa cytoplasmic PLA ₂ (cPLA ₂ ) purified from rabbit platelets based on a previous report (FEBS Lett., 282, 326-330, 1991). Was used to measure the inhibitory activity against this enzyme as follows.
6-azaspiro [4 . 5] Decane derivative (1) or (2) was dissolved in methanol and used as a test solution. 1
25 μL of M Tris-hydrochloric acid (pH 9.0) and 20 μL of 50 mM calcium chloride solution were mixed with a test solution and cP.
The LA ₂ solution was added to make 200 μL, and the mixture was reacted at 37 ° C. for 20 minutes. Then the substrate 1-palmitoyl-2
-[ ^14C ] arachidonoyl-glycerophosphoethanolamine (0.5 nmol / 50000 dpm / 50μ)
L) was added, and the mixture was further reacted at 37 ° C. for 20 minutes. Dole reagent (isopropanol: heptane: 1N sulfuric acid = 10:
The reaction was stopped by adding 1.25 mL of 40: 1), the [ ¹⁴ C] arachidonic acid fraction was collected by the method of Dole, and the radioactivity was measured by a liquid scintillation counter to measure the enzyme activity. Table 1 shows the results.

[0026]

[Table 1]

[0027]

INDUSTRIAL APPLICABILITY Since the compound of the present invention has cytoplasmic phospholipase A ₂ inhibitory activity, cytoplasmic phospholipase A ₂
It has a use as an inhibitor and is expected to be used as, for example, an anti-inflammatory agent or a prodrug thereof.

─────────────────────────────────────────────────── ─── Continuation of the front page (51) Int.Cl. ⁶ Identification code Internal reference number FI Technical indication C07F 7/18 C07F 7/18 T

Claims (1)

[Claims] 1. A general formula: (In the formula, R ¹ represents a hydrogen atom, a substituted or unsubstituted lower alkyl group, a tri-lower alkylsilyl group, an acyl group or a sulfo lower alkyl group, and R ² , R ³ and R ⁴ are independently a hydrogen atom or a substituted group. Or, it represents an unsubstituted lower alkyl group, a tri-lower alkylsilyl group, or an acyl group, X represents a halogen atom, Y represents an oxygen atom or -NR ^5- , R ⁵ represents a hydrogen atom, a substituted or unsubstituted A 6-azaspiro [4,5] decane derivative represented by a lower alkyl group, a tri-lower alkylsilyl group or an acyl group.