JPWO2001036420A1 - Tricyclic azaindolizine derivatives with sPLA2 inhibitory activity - Google Patents

Abstract

(57) [Abstract] A compound having _sPLA2 inhibitory activity is provided. The compound has the general formula (I): [wherein E is N or C- ^R4 ; G is N or C- ^R25 ; ^R1 is a carbocyclic group, a heterocyclic group, or the like; one of ^R3 and ^R4 is -( ^L2 )- (an acidic group), the other is a hydrogen atom, and ^L2 is a linking group to the acidic group; ring A is an optionally substituted 5- to 8-membered cyclohexane ring or cyclohexene ring; ^R24 and ^R25 are hydrogen atoms, or the like], a prodrug thereof, or a pharmaceutically acceptable salt thereof, or a hydrate thereof.

Description

Description: TECHNICAL FIELD The present invention relates to tricyclic azaindolizine derivatives that are effective in inhibiting _sPLA2- mediated fatty acid release.

BACKGROUND ART _sPLA2 (secretory phospholipase _A2 ) is an enzyme that hydrolyzes membrane phospholipids and is considered to be the rate-limiting enzyme controlling the so-called arachidonic acid cascade, which uses arachidonic acid produced during the hydrolysis as a starting material. Furthermore, lysophospholipids, which are by-produced during phospholipid hydrolysis, are known to be important mediators of cardiovascular diseases. Therefore, in order to normalize the excessive activity of the arachidonic acid cascade or lysophospholipids, it is important to develop compounds that inhibit _sPLA2 -mediated fatty acid (e.g., arachidonic acid) release, i.e., compounds that inhibit the activity or production of _sPLA2 . Such compounds are effective in preventing sPLA2-mediated fatty acid (e.g., arachidonic acid) release in various diseases, such as septic shock, adult respiratory distress syndrome, pancreatitis, trauma, bronchial asthma, allergic rhinitis, rheumatoid arthritis, arteriosclerosis, stroke, cerebral infarction, inflammatory bowel disease, psoriasis, heart failure, and myocardial infarction.
These compounds are useful in the general treatment of conditions caused and/or sustained by the overproduction of sPLA _2. The involvement of sPLA ₂ in pathological conditions is thought to be extremely diverse, and its effects are potent.

Examples of _sPLA2 inhibitors include indole derivatives [EP-620214 (JP-A-7-010838), EP-620215 (JP-A-7-025850), EP-
675110 (JP-A-7-285933, and WO96/03376), indene derivatives (WO96/03120), indolizine derivatives (WO96/03
383), naphthyl derivatives (WO97/21664 and WO9721716
), carbazole derivatives (WO98/18464), pyrazine derivatives (W
O99/51605) are known.

DISCLOSURE OF THE INVENTION The present invention provides tricyclic azaindolizine derivatives which have _sPLA2 inhibitory activity and are useful as therapeutic agents for septic shock, adult respiratory distress syndrome, pancreatitis, trauma, bronchial asthma, allergic rhinitis, rheumatoid arthritis, arteriosclerosis, stroke, cerebral infarction, inflammatory bowel disease, psoriasis, heart failure, and myocardial infarction.

The present invention relates to a compound of formula I) (I): wherein E is N or C-R ⁴ ; when E is N, G is C-R ²⁵ , and when E is C-R ⁴ , G is N; R ¹ is (a) a C1-C20 alkyl, a C1-C20 alkenyl, a C1-C20 alkynyl, a carbocyclic group, or a heterocyclic group; (b) the group represented by (a) substituted with one or more groups, each independently selected from non-interfering substituents;
or (c) -(L ¹ )-R ⁵ (wherein L ¹ is a hydrogen atom, a nitrogen atom, a carbon atom,
a divalent linking group of 1 to 18 atoms selected from an oxygen atom and a sulfur atom, ^R5 being a group selected from (a) or (b); one of ^R3 and ^R4 being -( ^L2 )- (acidic group) (wherein ^L2 is a linking group to the acidic group, and the length of the linking group to the acidic group is 1 to 5), and the other being a hydrogen atom; ring A being a group represented by the formula: (wherein ^R2 is _CONH2 , _CONHNH2 , or _CSNH2 ; ^R18 , ^R19 , ^R20 , ^R21 , ^R22 , ^R23 , ^R26 , and ^R27 are each independently a hydrogen atom or lower alkyl; and ^R24 and ^R25 are each independently a hydrogen atom, C1-C6 alkyl, aryl, halogen, or aralkyl), a prodrug thereof, or a pharmaceutically acceptable salt or hydrate thereof.

More specifically, the following items II) to XVI) are described below.

II) General formula (II): wherein ^R24 and ring A are as defined above; J is N or C- ^R8 , and when J is N, G is C- ^R25 ( ^R25 is as defined above), and when J is C- ^R8 , G is N; ^R6 is -( _CH2 )m- ^R9 (m is an integer of 1 to 6, ^R9 is a group represented by the formula (d): (In the formula, a, c, e, n, q, t, and v each independently represent 0, 1, or 2.
R ¹⁰ and R ¹¹ are each independently halogen, C1-C10 alkyl, C
a is an oxygen atom or a sulfur atom; β is —CH ₂ — or —(CH 2 ) ₂ —; γ is an oxygen atom or a sulfur atom; b is an integer of 0 to 3; d is an integer of 0 to 4; f, p, and w are each independently an integer of 0 to 5; r is an integer of 0 to 7; and u is an integer of 0 to 4), or ( _e ) a group represented by C1-C6 alkyl, C1-C6 alkyloxy, C1-C
a group selected from (d) substituted with one or more substituents selected from the group consisting of C1-C6 haloalkyloxy, C1-C6 haloalkyl, phenyl, and halogen); one of ^R7 and ^R8 is a hydrogen atom, and the other is -( ^L3 ) ^-R12 (wherein ^L3 is a group represented by the formula: (wherein M is —CH ₂ —, —O—, —N(R ¹⁵ )—, or —S—; R ¹³ and R ¹⁴ each independently represent a hydrogen atom, a C1-C10 alkyl, an aryl, an aralkyl, a carboxy, or a halogen; and R ¹⁵ represents a hydrogen atom or a C1-C6 alkyl; and R ¹² represents a group of the formula: (wherein R ¹⁶ is a hydrogen atom, a metal, or a C1-C10 alkyl; R ¹⁷ is each independently a hydrogen atom or a C1-C10 alkyl; and h is an integer of 1 to 8), a prodrug thereof, or a pharmaceutically acceptable salt thereof, or a hydrate thereof.

When b, d, f, p, r, u, and/or w are 2 or more, a plurality of R¹⁰
and a plurality of R¹¹may be different from each other.¹⁰The naphthyl group is
When the substituent is a substituted group, it may be substituted at any position on the naphthyl group.
CH₂- and -(CH₂)₂- is R¹⁰may be substituted with.

III) R ¹ and R ⁶ are of the formula: (wherein R ¹⁰ , R ¹¹ , b, d, f, p, r, u, w, α, β, and γ are as defined above), a prodrug thereof, or a pharmaceutically acceptable salt thereof, or a hydrate thereof, according to either I) or II).

When b, d, f, p, r, u, and/or w are 2 or more, a plurality of R¹⁰
and a plurality of R¹¹may be different from each other.¹⁰The naphthyl group is
When it is a substituent, it may be substituted at any position on the naphthyl group.
CH₂- and -(CH₂)₂- is R¹⁰may be substituted with.

IV) R ¹ and R ⁶ are of the formula: (wherein R ¹⁰ , R ¹¹ , d, p, u, and w are as defined above)
1. A compound according to any one of (1) to (3), a prodrug thereof, a pharmaceutically acceptable salt thereof, or a hydrate thereof.

When p, u, and/or w are 2 or more, a plurality of R ¹⁰ and a plurality of R
¹¹ may be different from each other.

V) A compound according to any one of I) to IV), a prodrug thereof, or a pharmaceutically acceptable salt or hydrate thereof, wherein ^R3 and ^R7 are -O-( _CH2 )m-COOH (m is as defined above) or -O-CH( ^-R30 )-COOH ( ^R30 is a hydrogen atom or a C1-C3 alkyl).

VI) General formula (III): (wherein G, J, R ¹⁰ , R ³⁰ , ring A, and m are as defined above; ring B is a benzene ring or a cyclohexane ring; and R ³⁸ is a hydrogen atom or a C1-C3 alkyl), a prodrug thereof, or a pharmaceutically acceptable salt thereof, or a hydrate thereof.

VII) A compound according to any one of I) to VI), a prodrug thereof, a pharmaceutically acceptable salt thereof, or a hydrate thereof, wherein R ² is —CONH ₂ .

VIII) A compound according to any one of I) to VII), a prodrug thereof, a pharmaceutically acceptable salt thereof, or a hydrate thereof, wherein R ¹⁸ , R ¹⁹ , R ²⁰ , R ²¹ , R ²² , and R ²³ are all hydrogen atoms.

IX) A pharmaceutical composition comprising, as an active ingredient, a compound according to any one of I) to VIII).

X) The pharmaceutical composition according to IX) which is an sPLA ₂ inhibitor.

XI) The pharmaceutical composition according to IX), which is a type I sPLA ₂ inhibitor.

XII) The pharmaceutical composition according to IX), which is a type II sPLA ₂ inhibitor.

XIII) The pharmaceutical composition according to IX), which is a type V sPLA ₂ inhibitor.

XIV) The pharmaceutical composition according to IX), which is a type X sPLA ₂ inhibitor.

XV) Use of a compound according to any one of claims I) to VIII) for the manufacture of a medicament for treating a disease caused by _sPLA2 .

XVI) A method for treating a disease caused by _sPLA2 in a mammal, including a human, which comprises administering to the mammal a therapeutically effective amount of a compound according to any one of I) to VIII).

As used herein, the term "alkyl" as used alone or in combination with other terms
The term "alkyl" refers to a straight or branched chain monovalent hydrocarbon radical having a number of carbon atoms in the specified range, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, n-nonanyl, n-decanyl, n-undecanyl, n-dodecanyl, n-tridecanyl, n-tetradecanyl, n-pentadecanyl, n-hexadecanyl, n-heptadecanyl, n-octadecanyl, n-nonadecanyl, n-icosanyl, and the like.

As used herein, the term "alkenyl," alone or in combination with other terms, refers to a straight- or branched-chain monovalent hydrocarbon radical having a number of carbon atoms in the specified range and one or more double bonds, such as, for example, vinyl, allyl, propenyl, crotonyl, isopentenyl, the various butenyl isomers, and the like.

As used herein, the term "alkynyl" refers to a straight- or branched-chain monovalent hydrocarbon group having a number of carbon atoms in the specified range and one or more triple bonds, and optionally further having double bonds. Examples include ethynyl, propynyl,
Examples include 6-heptynyl, 7-octynyl, and 8-nonyl.

In this specification, the term "carbocyclic group" refers to a group derived from a saturated or unsaturated, substituted or unsubstituted organic skeleton of a 5- to 14-membered ring, preferably a 5- to 10-membered ring, more preferably a 5- to 7-membered ring, in which the atoms forming the ring, other than hydrogen atoms, are carbon atoms only. It also includes groups in which two or three of the above carbocyclic rings are connected in succession. Representative carbocyclic groups include (f) cycloalkyl (e.g., cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl), cycloalkenyl (cyclobutylenyl, cyclopentenyl, cyclohexenyl, cycloheptenyl, and cyclooptenyl), phenyl, naphthyl, norbornyl, bicycloheptadienyl, indenyl, stilbenyl, terphenylyl, phenylcyclohexenyl, acenaphthyl, anthryl, biphenylyl, bibenzylyl, and the group represented by formula (IV): (wherein x is an integer of 1 to 8).

As used herein, the term "heterocyclic group" refers to a group derived from a monocyclic or polycyclic, saturated or unsaturated heterocyclic skeleton having 5 to 14 ring atoms including 1 to 3 heteroatoms selected from the group consisting of nitrogen atoms, oxygen atoms, and sulfur atoms. Examples thereof include pyridyl, pyrrolyl, furanyl, benzofuranyl, thienyl, benzothienyl, pyrazolyl, imidazolyl, phenylimidazolyl, triazolyl, isoxazolyl, oxazolyl, thiazolyl, thiadiazolyl, indolyl, carbazolyl, norharmanyl, azaindolyl, benzofuranyl, dibenzofuranyl, dibenzothiophenyl, indazolyl, imidazo[1,2-a]pyridinyl, benzotriazolyl, anthranilyl, 1,2-benzisoxazolyl, benzoxazolyl, benzothiazolyl, purinyl, pridinyl, dipyridinyl, phenylpyridinyl, benzylpyridinyl, pyrimidinyl, phenylpyrimidinyl, pyrazinyl, 1,3,5-triazinyl, quinolyl, phthalazinyl, quinazolinyl, and quinoxalinyl.

The carbocyclic group and heterocyclic ring in R ¹ include (g) formula: (wherein v is an integer of 0 to 2, R ¹⁰ and R ¹¹ each independently represent a halogen atom,
a group independently selected from C1-C10 alkyl, C1-C10 alkyloxy, C1-C10 alkylthio, optionally substituted phenyl, and C1-C10 haloalkyl; α is an oxygen atom or a sulfur atom; β is —CH ₂ — or —(CH
₂ ) _2- , γ is an oxygen atom or a sulfur atom, b is an integer of 0 to 3, d is an integer of 0 to 4, f, p and w are integers of 0 to 5, r is an integer of 0 to 7, and u is an integer of 0 to 4. When b, d, f, p, r, u, and/or w are 2 or more, multiple ^R10s and multiple ^R11s may be different from each other. When ^R10 is a substituent of a naphthyl group, it may be substituted at any position on the naphthyl group. _-CH2- and -( _CH2 ) _2- in β may be substituted with ^R10 .

More preferably, (h) formula: (wherein R ¹⁰ , R ¹¹ , α, β, and γ are as defined above, and each y is independently 0 or 1). When ^{R 10} is a substituent of a naphthyl group, it may be substituted at any position on the naphthyl group. -CH ₂ - and -(CH
₂ ) _2- may be substituted with ^R10 .

In the present specification, the term "non-interfering substituent" refers to a group of the tricyclic compound represented by formula (I),
The groups listed in (a) in R ¹ (e.g., "alkyl", "alkenyl",
"Carbocyclic group" and "heterocyclic group" refer to groups suitable for substitution on C1
-C10 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C7-C1
2aralkyl (e.g., benzyl and phenethyl), C7-C12 alkaryl, C3-C8 cycloalkyl, C3-C8 cycloalkenyl, phenyl, tolyl, xylyl, biphenylyl, C1-C10 alkyloxy, C1-C6 alkyloxyC1-C6 alkyl (e.g., methyloxymethyl, ethyloxymethyl, methyloxyethyl, and ethyloxyethyl), C1-C6 alkyloxyC1-C6 alkyloxy (e.g., methyloxymethyloxy, and methyloxyethyloxy), C1-C6 alkylcarbonyl (e.g., methylcarbonyl and ethylcarbonyl), C1-C6 alkylcarbonylamino (e.g., methylcarbonylamino and ethylcarbonylamino), C1-C6 alkyloxyamino (e.g., methyloxyamino and ethyloxyamino),
C1-C6 alkyloxyaminocarbonyl (e.g., methyloxyaminocarbonyl and ethyloxyaminocarbonyl), mono- or di-C1-C6 alkylamino (e.g., methylamino, ethylamino, dimethylamino, and ethylmethylamino), C1-C10 alkylthio, C1-C6 alkylthiocarbonyl (e.g., methylthiocarbonyl and ethylthiocarbonyl), C1-C
C1-C6 alkylsulfinyl (e.g., methylsulfinyl and ethylsulfinyl), C1-C6 alkylsulfonyl (e.g., methylsulfonyl and ethylsulfonyl), C2-C6 haloalkyloxy (e.g., 2-chloroethyloxy and 2-bromoethyloxy), C1-C6 haloalkylsulfonyl (e.g., chloromethylsulfonyl and bromomethylsulfonyl), C1-C10 haloalkyl, C1-C6 hydroxyalkyl (e.g., hydroxymethyl and hydroxyethyl), C1-C6 alkyloxycarbonyl (e.g., methyloxycarbonyl and ethyloxycarbonyl), —(CH ₂ )z—O—(C1
-C6 alkyl), benzyloxy, aryloxy (e.g., phenyloxy), arylthio (e.g., phenylthio), _-CONHSO2R20 , formyl, amino, amidino, halogen, carbamyl, carboxyl, carbalkyloxy, -( _CH2 )z-COOH (e.g., carboxymethyl, carboxyethyl, and carboxypropyl) ^, cyano, cyanoguanidino, guanidino, hydrazide, hydrazino, hydroxy, hydroxyamino, nitro, phosphono, -
Examples thereof include SO ₃ H, thioacetal, thiocarbonyl, a carbocyclic group, and a heterocyclic group (where z is an integer of 1 to 8, and R ²⁰ is a C1-C6 alkyl or aryl).

Halogen, C1-C6 alkyl, C1-C6 alkyloxy, C1-C6 alkylthio, and C1-C6 haloalkyl are preferred. More preferred are halogen,
C1-C3 alkyl, C1-C3 alkyloxy, C1-C3 alkylthio, C
1-C3 haloalkyl is exemplified.

In this specification, the term "halogen" means fluorine, chlorine, bromine, or iodine.

As used herein, the term "cycloalkyl" refers to a cyclic monovalent hydrocarbon group having a number of carbon atoms in the specified range, including, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, and the like.

As used herein, the term "cycloalkenyl" refers to a cyclic monovalent hydrocarbon group having a number of carbon atoms in the specified range and one or more double bonds, such as 1-cyclopropenyl, 2-cyclopropenyl, 1-cyclobutenyl, and 2-cyclobutenyl.

In the present specification, examples of "alkyloxy" include methyloxy, ethyloxy, n-propyloxy, isopropyloxy, n-butyloxy, n-pentyloxy, n-hexyloxy and the like.

In the present specification, examples of "alkylthio" include methylthio, ethylthio,
n-Propylthio, isopropylthio, n-butylthio, n-pentylthio, n
-hexylthio and the like.

As used herein, the term "acidic group" refers to an organic group that acts as a proton donor to enable hydrogen bonding when attached to a tricyclic skeleton via a suitable linking atom (defined below as a "linking group to an acidic group"). For example, (k) a group of formula: (wherein R ¹⁶ is a hydrogen atom, a metal, or a C1-C10 alkyl, R ¹⁷ are each independently a hydrogen atom or a C1-C10 alkyl, and h is an integer of 1 to ₈ ).
(O)(OH) ₂ is preferred. (m)-COOH is more preferred. Esters and prodrugs thereof are also preferred.

In the present specification, the term "a linking group to an acidic group" refers to a divalent linking group represented by the symbol -(L ² )-, which normally serves to link a tricyclic skeleton to an "acidic group". For example, a group represented by the formula (n): (wherein M is —CH ₂ —, —O—, —N(R ¹⁵ )—, or —S—; R ¹³ and R ¹⁴ each independently represent a hydrogen atom, a C1-C10 alkyl, an aryl, an aralkyl, a carboxy, or a halogen; (wherein R ¹⁵ is a hydrogen atom or a C1-
C alkyl). Preferably, (o)-O-CH
_2- , -S- _CH2- , -N( ^R15 ) _-CH2- , _-CH2 - _CH2- , -O
-CH( _CH3 )-, or -O-CH(( _CH2 ) _2Ph )- (wherein ^R15
is a hydrogen atom or _C1 - _C6 alkyl, and Ph is phenyl). More preferred are (p) -O- _CH2- , -O-CH( _CH3 )-, or -S-C
H ₂ − is an example.

As used herein, the term "length of the linking group to the acidic group" refers to the number of atoms (excluding hydrogen atoms) in the shortest chain of the linking group -( ^L2 )- connecting the tricyclic skeleton to the "acidic group." When -( ^L2 )- contains a carbon ring, the number of atoms is counted as approximately equal to the calculated diameter of the carbon ring. Therefore, for benzene rings and cyclohexane rings in the linking group to the acidic group, the length of -( ^L2 )- is counted as 2 atoms. The preferred length is 2 to 3.

As used herein, the term "haloalkyl" refers to the above-mentioned "alkyl" substituted with the above-mentioned "halogen" at any position. Examples thereof include chloromethyl, trifluoromethyl, 2-chloromethyl, 2-bromomethyl, etc.

As used herein, "hydroxyalkyl" refers to the above-mentioned "alkyl" substituted with hydroxy at any position. Examples include hydroxymethyl, 2-hydroxyethyl, 3-hydroxypropyl, etc. Hydroxymethyl is preferred.

In the present specification, the "haloalkyl" in "haloalkyloxy" has the same meaning as defined above. Examples thereof include 2-chloroethyloxy, 2-trifluoroethyloxy, 2-chloroethyloxy, etc.

As used herein, the term "aryl" refers to a monocyclic or fused-ring aromatic hydrocarbon. Examples include phenyl, 1-naphthyl, 2-naphthyl, anthryl, etc. Phenyl and 1-naphthyl are particularly preferred.

As used herein, "aralkyl" refers to a C1-C8 alkyl substituted with the above-mentioned "aryl", which may be substituted at any possible position. For example, benzyl,
Examples include phenethyl, phenylpropyl (for example, 3-phenylpropyl), naphthylmethyl (for example, 1-naphthylmethyl), and the like.

As used herein, the term "alkyloxycarbonyl" includes C1-C6 alkyloxycarbonyl, such as methyloxycarbonyl, ethyloxycarbonyl, and n-propyloxycarbonyl.

In the present specification, the term "acyl" refers to a C1-
C6 alkylcarbonyl or arylcarbonyl, for example, acetyl, trifluoroacetyl, propionyl, benzoyl, etc.

^{In the compounds} represented by general formula (I), preferred groups ^of substituents for ^{R 1 to} R ³ are shown as (A) to (O ⁾ .
^{R 3} , R ²⁵ , R ²⁶ , and R ²⁷ are preferably hydrogen atoms.
) are defined as the same as the groups of substituents described above.

In R ¹ , (A): —(L ¹ )—R ⁵ , (B): —(CH ₂ ) _1-2 —
(f), (C):-( _CH2 ) _1-2- (g),(D):-( _CH2 ) _1-2-
(h) is preferred.

In ^R2 , (E): _CONH2 or _CONHNH2 , (F): CON
_H2 is preferred.

In ^R3 , (G): -(n)-(k), (H): -(n)-(l), (
I):-(n)-(m), (J):-(o)-(k), (K):-(o)-(l
), (L):-(o)-(m), (M):-(p)-(k), (N):-(p)
-(l), (O):-(p)-(m) are preferred.

A preferred group of compounds represented by general formula (I) is shown below:
¹ , R ² , R ³ ) = (A, E, G), (A, E, H), (A, E, I), (A,
E, J), (A, E, K), (A, E, L), (A, E, M), (A, E, N)
, (A, E, O), (A, F, G), (A, F, H), (A, F, I), (A,
F, J), (A, F, K), (A, F, L), (A, F, M), (A, F, N)
, (A, F, O), (B, E, G), (B, E, H), (B, E, I), (B,
E, J), (B, E, K), (B, E, L), (B, E, M), (B, E, N)
, (B, E, O), (B, F, G), (B, F, H), (B, F, I), (B,
F, J), (B, F, K), (B, F, L), (B, F, M), (B, F, N)
, (B, F, O), (C, E, G), (C, E, H), (C, E, I), (C,
E, J), (C, E, K), (C, E, L), (C, E, M), (C, E, N)
, (C, E, O), (C, F, G), (C, F, H), (C, F, I), (C,
F, J), (C, F, K), (C, F, L), (C, F, M), (C, F, N)
, (C, F, O), (D, E, G), (D, E, H), (D, E, I), (D,
E, J), (D, E, K), (D, E, L), (D, E, M), (D, E, N)
, (D, E, O), (D, F, G), (D, F, H), (D, F, I), (D,
F, J), (D, F, K), (D, F, L), (D, F, M), (D, F, N)
, (D, F, O) are exemplified.

For each, R ⁴ is preferably a hydrogen atom, C 1 -C 3 alkyl, or aryl.

BEST MODE FOR CARRYING OUT THE INVENTION The compounds of the present invention represented by general formula (I) can be synthesized by known methods described in chemical literature. The compounds of the present invention represented by general formula (I) can be synthesized by the following methods A and B. Representative examples are shown below.
The ring-expanded compounds can be synthesized in the same manner.

(Method A) wherein R ¹ , R ⁴ , and R ²⁴ are as defined above, R ²⁸ , R ²⁹ , and R
³¹ is a C1-C3 alkyl, R ³⁶ is a protecting group for a hydroxyl group (e.g., tetrahydropyranyl, etc.), R ³⁰ is a hydrogen atom or a C1-C3 alkyl, and each Hal is independently a halogen. (Step 1) Compound (VI) can be obtained by dissolving compound (V) in a solvent such as dimethylformamide, adding an alkyl halide derivative and a base (e.g., potassium carbonate, potassium t-butoxide, sodium hydride, etc.), and reacting at room temperature to 180°C, preferably 20 to 150°C, for 3 to 80 hours, preferably 5 to 70 hours.

(Step 2) Compound (VI) is dissolved in a solvent such as dimethyl sulfoxide, and a reagent such as potassium acetate or sodium acetate is added. The mixture is heated at 20°C to 200°C, preferably 100°C to 180°C.
The compound (VII) can be obtained by reacting at 0°C for 3 to 17 hours.

(Step 3) Compound (VI) is added to a solution of a Grignard reagent (R ¹ MgHal, Hal is a halogen) or R ¹ Li in ether, tetrahydrofuran, or dimethoxyethane.
I) in ether, tetrahydrofuran, or dimethoxyethane at −20° C.
The compound (VIII) can be obtained by adding the compound (VIII) at 0°C to 70°C, preferably 20°C to 60°C, for 1 to 48 hours, preferably 2 to 24 hours.

(Step 4) Compound (VIII) can be dissolved in a solvent such as ethanol, methanol, dioxane, or tetrahydrofuran, and N-aminophthalimide and an acid such as trifluoroacetic acid, hydrochloric acid, or sulfuric acid can be added thereto, followed by reaction at 20°C to 120°C, preferably 50°C to 100°C, for 5 minutes to 2 hours, preferably 10 minutes to 1 hour, to obtain compound (IX).

(Step 5) This step is a step of deprotecting the phthalimide of compound (IX).
Synthesis, Theodora W Green (John Wilk
For example, compound (IX)
is dissolved in an alcohol solvent such as ethanol, hydrazine is added, and the mixture is heated at 50°C to 100°C.
The reaction is carried out at 0° C. for 0.5 to 3 hours to give the amino compound (X).

(Step 6) Compound (X) is dissolved in a solvent such as chloroform, dichloroethane, tetrahydrofuran, or toluene, and a β-ketoester (e.g., methyl acetoacetate) and an acid catalyst (e.g., p-toluenesulfonic acid, methanesulfonic acid, hydrochloric acid, sulfuric acid, or trifluoroacetic acid) are added. The mixture is heated at 20°C to 150°C, preferably 30°C to 100°C.
C. for 1 to 20 hours, preferably 3 to 16 hours, to give Compound (XI).
It is preferable to remove water generated in the reaction system by a method using a Dean-Stark filter filled with molecular sieves 4A or the like.

(Step 7) Compound (XI) is dissolved in a solvent such as tetrahydrofuran or dimethylformamide, and a base such as potassium carbonate or sodium hydride and an alkyl halide derivative are added thereto, followed by reaction at 0°C to 100°C, preferably 20 to 40°C, for 1 to 15 hours to obtain compound (XII).

(Step 8) This step is a step of converting a hydroxyl group into a halogen. Compound (XII) is dissolved in a solvent such as methylene chloride, and a reagent such as triphenylphosphine, carbon tetrachloride, or N-bromosuccinimide is added. The mixture is stirred at 0° C. to 50° C., preferably 0° C. to 30° C. for 1 hour.
The compound (XIII) can be obtained by reacting for 1 to 10 hours, preferably 1 to 5 hours.

(Step 9) This step is a step of introducing a substituent into the 5-position. Compound (XIII) is dissolved in a solvent such as methylene chloride or tetrahydrofuran, and Hal-C(═X)—C(═X) is added.
)-Hal (Hal is halogen) (e.g., oxalyl chloride) and a base such as N-methylmorpholine or triethylamine are added to the reaction mixture, and the mixture is stirred at −30° C. to 70° C., preferably −20° C. to 40° C., for 10 minutes to 10 hours, preferably 10 minutes to 2 hours. The reaction mixture is poured into aqueous ammonia, and the resulting mixture is subjected to a conventional post-treatment to obtain compound (XI).
V) can be obtained.

(Step 10) This step is a step of preparing a phosphonium salt. Compound (XIV) and triphenylphosphine or the like are heated at 60° C. to 150° C. in a solvent such as acetonitrile or toluene.
C., preferably 80.degree. C. to 120.degree. C., for 5 to 100 hours, preferably 10 to 70 hours, to give compound (XV).

(Step 11) This step is a step of forming a ring by using the Wittig reaction.
Compound (XV) is reacted with 1,8-diazabicyclo[5,4,0]-7-undecene (DBU), t-
A base such as potassium butoxide is added, and the mixture is heated at 20°C to 120°C, preferably 30°C to 10°C.
The compound (X) is obtained by reacting the compound (X) at 0° C. for 3 to 24 hours, preferably 5 to 10 hours.
VI) can be obtained.

(Step 12) This step is a hydrolysis step. Compound (XVI) is dissolved in a solvent such as methanol or tetrahydrofuran, and a base such as sodium hydroxide is added. The mixture is heated at 0° C. to 40° C.
The compound (XVII) can be obtained by reacting the mixture at 10° C., preferably 10° C. to 30° C., for 0.5 to 6 hours, preferably 0.5 to 2 hours.

(Method B)[In the formula, R¹, R²⁴, R²⁵, R³⁰, R³¹and Hal are as defined above.
, R³²and R³³is C1-C3 alkyl, R³⁴is R¹Precursor of R³⁵
represents C1-C3 alkyl, optionally substituted aryl (e.g., tolyl),
, R³⁷is acyl (e.g., acetyl, benzoyl, etc.), and Ms is mesyl). (Step 1) This step constructs the pyrrolo[1,2-a]pyrazine ring.
XVIII) and Hal-CH₂—C(═O)—CH₂−(CH₂)_m-CO₂R
³³the mixture at 40°C to 90°C, preferably 50°C to 70°C, for 3 to 36 hours,
Preferably, the mixture is stirred for 12 to 24 hours to obtain a quaternary salt.
Dissolved in a solvent such as dichloroethane or acetonitrile, 1,8-diazabicyclo[
5,4,0]-7-undecene (DBU), a base such as triethylamine, etc., is added, and the resulting mixture is 4
0°C to 90°C, preferably 60°C to 80°C for 1 to 10 hours, preferably 1 to 5 hours
The mixture is stirred for 1 hour. Compound (XIX) can be obtained by carrying out a usual post-treatment.
can.

(Step 2) This step is a step of introducing a substituent into the 6-position of pyrrolo[1,2-a]pyrazine, and can be carried out using the Friedel-Crafts reaction. Compound (XIX) is dissolved in a solvent such as methylene chloride or chlorobenzene, and the solution is reacted with R34COHal and a Lewis acid (e.g., _AlCl3 , S) at -78°C to ¹⁰ °C, preferably -20°C to 0°C.
_BF5 , _BF3, etc.) is slowly added and the temperature is -10°C to 10°C, preferably 0°C to 1
The mixture is stirred at 0° C. for 30 minutes to 5 hours, preferably 1 hour to 3 hours. Compound (XX) can be obtained by carrying out a usual post-treatment.

(Step 3) This step is a step of reducing esters and ketones to alcohols. Compound (XX) is reacted with a reducing agent such as lithium aluminum hydride or lithium borohydride in a solvent such as ether or tetrahydrofuran at 0° C. to 80° C., preferably 10° C. to 120° C.
By reacting at 40°C for 30 minutes to 10 hours, preferably 1 hour to 5 hours,
Compound (XXI) can be obtained.

(Step 4) This step involves reductively removing the hydroxyl group and converting it to methylene. A Lewis acid (e.g., _AlCl3 , etc.) is dissolved in a solvent such as methylene chloride or tetrahydrofuran, and a reducing agent such as boron-t-butylamine complex or sodium borohydride is added at -20°C to 10°C, preferably under ice-cooling, and the mixture is stirred for 5 to 30 minutes, preferably 10 to 20 minutes. Compound (XXI) is added to this reaction solution at -20°C to 10°C, preferably under ice-cooling, and the mixture is stirred at 15°C to 40°C, preferably 20°C.
Compound (XXII) can be obtained by reacting at 30° C. or below for 30 minutes to 5 hours, preferably 1 hour to 3 hours.

(Step 5) This step is a step of converting the alkyloxy group at position 1 into a ketone.
XII) is added with an acid such as concentrated hydrochloric acid and heated at 80°C to 150°C, preferably 100°C to 120°C.
The mixture is stirred for 1 to 5 hours, preferably 1 to 3 hours, at 0° C. Compound (XXIII) can be obtained by carrying out a usual post-treatment.

(Step 6) This step is a step of protecting the hydroxyl group with acetyl. Compound (XXIII) is dissolved in pyridine, and the solution is heated with acetic anhydride at 0°C to 50°C, preferably 20°C to 40°C for 1 to 3 hours.
The reaction is carried out for 10 hours, preferably 3 to 8 hours, to obtain Compound (XXIV
) can be obtained.

(Step 7) This step is a step of converting the ketone at position 1 into a halogen. Compound (XXIV)
A halogenating agent such as phosphorus oxychloride or phenylphosphonic dichloride is added to the reaction mixture, and the mixture is stirred at 60° C. to 150° C., preferably 80° C. to 110° C., for 10 minutes to 3 hours, preferably 30 minutes to 1 hour.
can be obtained.

(Step 8) This step is a step of converting the halogen at the 1-position into a sulfonyl group.
Compound (XXVI) can be obtained by dissolving compound (XV) in ethanol, adding R ³⁵ SO ₂ Na, and heating under reflux for 10 to 50 hours, preferably 24 to 36 hours.

(Step 9) This step is a step of converting the sulfonyl group at position 1 to an alkyloxy group. HOCH(R ³⁰ )CO ₂ R ³¹ and a reagent such as sodium hydride or potassium t-butoxide are suspended in a solvent such as tetrahydrofuran, and the suspension is stirred at -20°C to 50°C, preferably 0°C to 30°C, for 15 minutes to 2 hours, preferably 30 minutes to 1 hour. Compound (XXVI) is added to this solution, and the reaction is continued at -20°C to 50°C, preferably 0°C to 30°C, for a further 15 minutes to 5 hours, preferably 30 minutes to 2 hours, to obtain compound (XXVII).

(Step 10) This step can be carried out in the same manner as Method A - Step 9.

(Step 11) This step is a step of removing the acetyl group. Compound (XXIX) can be obtained by dissolving compound (XXVIII) in a mixture of tetrahydrofuran and methanol, adding hydrochloric acid, and stirring the mixture at 0°C to 50°C, preferably 20°C to 30°C, for 1 to 10 hours, preferably 3 to 8 hours.

(Step 12) This step is a step of converting the hydroxyl group into _-OSO2Me .
) is dissolved in a solvent such as methylene chloride or tetrahydrofuran, and triethylamine,
Compound (XXX) can be obtained by reacting with methanesulfonyl chloride in the presence of a base such as pyridine at -30°C to 50°C, preferably 0°C to 30°C, for 10 minutes to 5 hours, preferably 30 minutes to 2 hours.

(Step 13) This step is a step of converting _-OSO2Me into a halogen.
) is dissolved in a solvent such as acetonitrile, acetone, or dimethylformamide, and heated at 0°C to 100°C, preferably 30°C, with a reagent such as lithium bromide or lithium chloride.
The compound (
XXXI) can be obtained.

(Step 14) This step can be carried out in the same manner as Method A - Step 10.

(Step 15) This step can be carried out in the same manner as Method A-Step 11.

(Step 16) This step can be carried out in the same manner as Method A - Step 12.

When the compounds of the present invention contain acidic or basic functional groups, they can form a variety of physiologically suitable salts that are more water soluble than the parent compound. Representative pharmaceutically acceptable salts include, but are not limited to, alkali metal and alkaline earth metal salts, such as lithium, sodium, potassium, calcium, magnesium, and aluminum. Salts are conveniently prepared from the free acid by treating the acid in solution with a base or by contacting the acid with an ion exchange resin. Relatively non-toxic inorganic and organic base addition salts of the compounds of the present invention, such as amine cations, ammonium, and quaternary ammonium derived from nitrogenous bases sufficiently basic to form salts with the compounds of the present invention, are included within the definition of pharmaceutically acceptable salts (see, for example, S.M. Berge et al., "Pharmaceuticals of the Invention").
ical Salts,” J. Phar. Sci., 66, 1-19 (1977
Additionally, the basic groups of the compounds of the present invention can be reacted with suitable organic or inorganic acids to give acetates, benzenesulfonates, benzoates, bicarbonates, bisulfates, bitartrates, borates, bromides, camsylates, carbonates,
It forms salts such as chloride, clavulanate, citrate, edetate, edisylate, estolate, esylate, fluoride, fumarate, gluceptate, gluconate, glutamate, glycolisanilate, hexylresorcinate, hydroxynaphthoate, iodide, isothionate, lactate, lactobionate, laurate, malate, marceate, mandelate, mesylate, methyl bromide, methyl nitrate, methyl sulfate, mucate, napsylate, nitrate, oleate, oxalate, palmitate, panthothenate, phosphate, polygalacturonate, salicylate, stearate, subacetate, succinate, thionate, tartrate, tosylate, trifluoroacetate, trifluoromethanesulfonate, and valerate. When forming a hydrate, it may be coordinated with any number of water molecules.

When a compound of the present invention contains one or more chiral centers, it may exist as an optically active form. Similarly, when a compound contains an alkenyl or alkenylene group, cis and trans isomers are possible. Mixtures of R- and S-isomers, including R- and S-isomers, mixtures of cis and trans isomers, and racemic mixtures, are encompassed within the scope of the present invention. Asymmetric carbon atoms may also be present in substituents, such as alkyl groups. All such isomers, as well as mixtures thereof, are encompassed within the present invention. If a specific stereoisomer is desired, it can be prepared by methods known to those skilled in the art, either by subjecting a starting material having a pre-resolved asymmetric center to stereospecific reactions, or by preparing a mixture of stereoisomers and then resolving them by known methods.

A prodrug is a derivative of the compound of the present invention having a chemically or metabolically decomposable group, which becomes a pharmaceutically active compound of the present invention in vivo by solvolysis or under physiological conditions. The derivatives of the compound of the present invention are active in both acid and base forms, but acid derivatives are advantageous in terms of solubility, tissue binding, and controlled release in mammalian organisms (Bungard, H.
, Design of Prodrugs, pp. 7-9, 21-24, Els
(Evier, Amsterdam 1985). Prodrugs containing acidic derivatives such as esters prepared by reacting the parent acidic compound with an appropriate alcohol, or amides prepared by reacting the parent acidic compound with an appropriate amine, are well known to those skilled in the art. Simple aliphatic or aromatic esters derived from the acidic group of the compounds of the present invention are preferred prodrugs. More preferred are C1-C6 alkyl esters (e.g., methyl ester, ethyl ester) of the acidic group. In some cases, double ester prodrugs such as (acyloxy)alkyl esters or ((alkoxycarbonyl)oxy)alkyl esters can also be prepared.

The term "inhibit" refers to a preventative or therapeutically significant reduction in _sPLA2 -initiated fatty acid release by a compound of the present invention. The term "pharmaceutically acceptable" refers to a carrier, diluent, or excipient that is compatible with the other ingredients in the formulation and not deleterious to the recipient.

As described in the Experimental Examples below, the compound of the present invention has an sPLA ₂ inhibitory activity.
Therefore, by administering a therapeutically effective amount of the compounds represented by general formulas (I), (II), and (III), their prodrug derivatives, or pharmaceutically acceptable salts thereof, or hydrates thereof to mammals (including humans), they can be used as therapeutic agents for diseases such as septic shock, adult respiratory distress syndrome, pancreatitis, trauma, bronchial asthma, allergic rhinitis, rheumatoid arthritis, arteriosclerosis, stroke, cerebral infarction, inflammatory bowel disease, psoriasis, heart failure, and myocardial infarction.

The compounds of the present invention can be administered by a variety of routes, including oral, aerosol, rectal, transdermal, subcutaneous, intravenous, intramuscular, and intranasal. The formulations of the present invention are prepared by combining (e.g., mixing) a therapeutically effective amount of the compound of the present invention with a pharmaceutically acceptable carrier or diluent. The formulations of the present invention are prepared by known procedures using well-known and readily available ingredients.

In preparing the compositions of the present invention, the active ingredient is mixed with or diluted by a carrier, or placed into a carrier in the form of a capsule, sachet, paper, or other container. When the carrier serves as a diluent, it is a solid, semi-solid, or liquid material that acts as a vehicle and can be in the form of a tablet, pill, powder, mouthwash, elixir, suspension, emulsion, solution, syrup, aerosol (a solid in a liquid medium), or ointment, containing, for example, up to 10% of the active compound. The compounds of the present invention are preferably formulated prior to administration.

Any suitable carrier known to those skilled in the art can be used for this formulation. In such formulations, the carrier can be a solid, liquid, or a mixture of a solid and a liquid. For example,
For intravenous injection, the compound of the present invention is dissolved in 4% dextrose/0.5% sodium citrate aqueous solution to a concentration of 2 mg/ml.
Solid carriers include flavoring agents, lubricants, solubilizers, suspending agents,
One or more substances that also serve as binders, tablet disintegrants, encapsulating materials, etc. Tablets for oral administration contain suitable excipients such as calcium carbonate, sodium carbonate, lactose, calcium phosphate, etc., along with disintegrating agents such as corn starch, alginic acid, and/or binders such as gelatin, acacia, and lubricants such as magnesium stearate, stearic acid, talc, etc.

In powders, the carrier is a finely divided solid mixed with the finely divided active ingredient. In tablets, the active ingredient is mixed with the carrier in suitable proportions having the necessary binding properties and compacted in the shape and size desired. Powders and tablets contain about 1
The active ingredient is a novel compound of the present invention, and the active ingredient is present in an amount of from about 99% by weight. Suitable solid carriers are magnesium carbonate, magnesium stearate, talc, sugar, lactose, pectin, dextrin, starch, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose, a low melting wax, or cocoa butter.

Sterile liquid preparations include suspensions, emulsions, syrups, and elixirs. The active ingredient can be dissolved or suspended in a pharmaceutically acceptable carrier, such as sterile water, sterile organic solvent, or a mixture of both. The active ingredient can often be dissolved in a suitable organic solvent, such as aqueous propylene glycol. Other compositions can also be prepared by dispersing the finely divided active ingredient in aqueous starch, sodium carboxymethylcellulose solution, or a suitable oil.

The dosage varies depending on the state of the disease, the administration route, the age and body weight of the patient, but is usually 0.01 to 50 mg/kg/day when administered orally to an adult.

The present invention will be explained in more detail below with reference to examples and test examples, but the present invention is not limited to these.

The following abbreviations are used in the examples:

Me: Methyl Et: Ethyl Hex: Hexyl Ph: Phenyl Bn: Benzyl Tol: Tolyl Ac: Acetyl Ms: Mesyl DBU: 1,8-diazabicyclo[5.4.0]-7-undecene Example Example 1 (First Step) Compound (1) (30.0 g, 0.313 mol) was dissolved in sodium hydride (3.
The resulting mixture was added to a suspension of 2-(3-chloropropyl)tetrahydro-2H-pyran (25.7 g, 0.144 mol) in dimethylformamide (100 ml) at 20° C. or below under a nitrogen stream, and stirred for 30 minutes at room temperature. 2-(3-chloropropyl)tetrahydro-2H-pyran (25.7 g, 0.144 mol) was added at the same temperature, and the mixture was heated and stirred in an oil bath at 60° C. for 16 hours. Dimethylformamide was removed from the reaction mixture under reduced pressure, and water was added to the residue, which was then extracted with toluene. The toluene layer was washed with water,
After drying over magnesium sulfate and removing the toluene, excess 2-(3-chloropropyl)tetrahydro-2H-pyran was removed under reduced pressure in an oil bath at 150°C to obtain compound (2) (41.5 g, 85.4%). ^1H -NMR ( _CDCl3 ): 1.18 (3H, t, J = 6.9 Hz), 1.2
1 (3H, t, J = 6.9Hz), 1.33 (3H, t, J = 7.2Hz), 1
．． 52-2.04 (12H, m), 2.40 (1H, m), 3.39-3.86
(7H, m), 4.24 (2H, m), 4.57 (1H, m), 4.77 (1H
, m). (Step 2) Compound (2) (41.5 g, 0.112 mol), potassium acetate (12.1 g
A mixture of 3, 0.123 mol) and dimethyl sulfoxide (125 ml) was heated in a nitrogen stream in an oil bath at 160°C for 17 hours. After cooling the reaction mixture, water was added and the mixture was extracted with ether. The ether layer was washed with water and dried over magnesium sulfate, and the ether was distilled off. The residue was distilled under reduced pressure to obtain a colorless liquid (3) (29.14 g, 87.2%) with a boiling point of 153-155°C (0.5 mmHg). ^1H -NMR ( _CDCl3 ): 1.22 (3H, t, J = 6.9 Hz), 1.2
3 (3H, t, J = 6.9Hz), 1.54-1.96 (12H, m), 2.7
9 (1H, m), 3.40-3.86 (8H, m), 4.57 (1H, t, J=
3.5 Hz), 4.68 (1H, dd, J = 7.5, 4.5 Hz). (Step 3) Magnesium (1.12 g, 0.0459 mol), ether (30 ml),
A Grignard reagent prepared from 1,2-dibromoethane (0.15 ml, 0.00186 mol) and benzyl bromide (5.24 ml, 0.044 mol) was added to a solution of compound (3) (11.0 g, 0.0367 mol) in ether (33 ml), and the mixture was stirred at room temperature for 14 hours, and then heated to reflux in an oil bath at 60° C. for 5 hours. The reaction mixture was added to an aqueous solution of ammonium chloride (3.93 g, 0.0734 mol) (40 ml) under ice cooling.
38 ml of 2N sulfuric acid was added, and the mixture was stirred for 30 minutes. The reaction mixture was neutralized with sodium bicarbonate and extracted with ether. The ether layer was dried over magnesium sulfate, and the ether was removed to give a light brown crude product (4) (15 ml).
2 g) was obtained, which was used crude in the fourth step.

(Step 4) Compound (4) (15.2 g) obtained by the above method and N-aminophthalimide (5
The resulting mixture (95 g, 0.0367 mol) was suspended in 95% ethanol (100 ml), 1N hydrochloric acid (18.4 ml, 0.0184 mol) was added, and the mixture was heated under reflux in an oil bath for 30 minutes. The ethanol was removed under reduced pressure, and water was added to the residue, which was then extracted with chloroform.
The organic layer was washed with water, dried over anhydrous magnesium sulfate, and subjected to silica gel column chromatography. 10.7 g of crude product was obtained from the fraction eluted with a 5% acetonitrile-chloroform mixture. The crude product was recrystallized from acetone-isopropyl ether to give pale yellow crystals (5) (7.20 g, 54.6%) having a melting point of 127-129°C.
%) was obtained.

Elemental analysis; C₂₂H₂₀N₂O₃ Measured values: C; 73.31, H; 5.52, N; 7.89 Calculated values: C; 73.31, H; 5.59, N; 7.77¹ H-NMR (CDCl₃); 1.85 (2H, m), 2.58 (2H, t, J
= 7.2Hz), 3.68 (2H, t, J = 6.2Hz), 3.68 (2H, s
), 6.22 (1H, d, J = 3.0Hz), 6.61 (1H, d, J = 3.0
Hz), 6.95-7.04 (7H, m). (Step 5) Compound (5) (1.80 g, 5 mmol), hydrazine hydrate (626
mg, 12.5 mmol) and 18 ml of 99.5% ethanol were mixed to obtain an oil
The mixture was heated to reflux for 45 minutes in a bath, the ethanol was removed under reduced pressure, and the residue was treated with toluene.
The toluene layer was washed with water, dried over anhydrous magnesium sulfate, and then
The mixture was subjected to lumina column chromatography and eluted with a 20% ethyl acetate-toluene mixture.
The resulting fractions gave a colorless oil (6) (1.20 g, 100%).¹ H-NMR (CDCl₃); 1.80 (2H, m), 2.50 (2H, t, J
= 7.4Hz), 3.65 (2H, t, J = 6.2Hz), 4.01 (2H, s
), 5.93 (1H, d, J = 3.0Hz), 6.66 (1H, J = 3.0Hz
), 7.07-7.28 (5H, m). (Step 6) Compound (6) (1.20 g, 5 mmol), methyl acetoacetate (581 mg,
5 mmol), paratoluenesulfonic acid hydrate (48 mg, 0.25 mmol
A mixture of 2,4-dimethylaminobenzoate (2,4-dimethylaminobenzoate) and chloroform (40 ml) was heated to reflux in an oil bath for 16 hours.
The resulting water was dehydrated using a Dean-Stark filter packed with molecular sieves 4A.
After cooling the reaction mixture, it was subjected to silica gel column chromatography.
From the fraction eluted with nitrile-chloroform, 1.497 g of crude crystals was obtained.
The crude crystals were recrystallized from acetone-isopropyl ether, melting at 148-149°C.
Crystals of (7) (1.21 g, 81.6%) were obtained.¹ H-NMR (CDCl₃); 1.68 (2H, m), 2.28 (3H, s),
2.60 (2H, t, J = 7.2Hz), 3.41 (2H, q, J = 5.2Hz
), 4.25 (2H, s), 4.42 (1H, t, J=5.2Hz), 5.84
(1H, s), 6.39 (1H, s), 7.12-7.21 (5H, m), 11
. 00 (1H, s). (Step 7) Compound (7) (1.08 g, 3.63 mmol), methyl bromoacetate (0.4
5 ml, 4.72 mmol), potassium carbonate (652 mg, 4.72 mmol)
A mixture of dimethylformamide (10 ml) was stirred at room temperature for 1 hour and 45 minutes.
Water was added to the reaction mixture, and the mixture was extracted with toluene. The organic layer was washed with water and extracted with magnesium sulfate.
After drying, the mixture was subjected to silica gel column chromatography and eluted with toluene.
The eluate (1.18 g) was recrystallized from acetone-isopropyl ether to give a melting point of 102
Compound (8) (1.07 g, 80.1%) was obtained as white crystals at −103° C.

Elemental analysis; C₂₁H₂₄N₂O₄ Measured values: C; 68.48, H; 6.51, N; 7.58. Calculated values: C; 68.46, H; 6.57, N; 7.60.¹ H-NMR (CDCl₃); 1.85 (2H, m), 2.40 (3H, s),
2.72 (2H, t, J = 7.2Hz), 3.61 (2H, t, J = 6.2Hz
), 3.84 (3H, s), 4.38 (2H, s), 4.77 (2H, s), 5
．． 63 (1H, s), 6.53 (1H, br.s), 7.12-7.22 (5H
, m). (Step 8) Compound (8) (942 mg, 2.56 mmol), triphenylphosphine (
805 mg, 3.07 mmol), carbon tetrachloride (5 ml), and methylene chloride
(5 ml) was stirred at room temperature for 4.5 hours.
The oily compound was extracted from the fraction eluted with chloroform.
Compound (9) (808 mg, 81.6%) was obtained.¹ H-NMR (CDCl₃); 2.02 (2H, m), 2.39 (3H, s),
2.78 (2H, t, J = 7.4Hz), 3.49 (2H, t, J = 6.6Hz
), 3.83 (3H, s), 4.37 (2H, s), 4.77 (2H, s), 5
．． 62 (1H, s), 6.51 (1H, s), 7.12-7.23 (5H, m)
(Step 9) Oxalyl chloride (0.91 ml) cooled in an ice-methanol bath (-14°C)
To a solution of compound (9) (80, 10.5 mmol) in methylene chloride (10 ml),
8 mg, 2.09 mmol) and N,N-diisopropylethylamine (0.4
A solution of 4 ml of 2.51 mmol of methyl methyl chloride (5 ml) was added, and the mixture was stirred at the same temperature for 1
The reaction mixture was stirred for 5 minutes. The reaction mixture was added to concentrated aqueous ammonia (3 ml) and chloroform (20 ml) under ice cooling.
The organic layer was poured into a mixture of 10 ml of ethanol, and the insoluble matter was removed by filtration, followed by extraction with chloroform.
The mixture was washed with water, dried over magnesium sulfate, and then purified by silica gel column chromatography.
The eluate with 50% acetonitrile-chloroform was subjected to acetone-
Recrystallization from ethyl acetate gave compound (10) (854 mg, 82.9%) with a melting point of 1
It was obtained as pale yellow crystals at 94-196°C.

Elemental analysis: C₂₃H₂₄ClN₃O₅ Measured values: C; 60.27, H; 5.21, N; 9.21, Cl; 7.89. Calculated values: C; 60.33, H; 5.28, N; 9.18, Cl; 7.74.¹ H-NMR (CDCl₃); 2.01 (2H, m), 2,46 (3H, s),
2.96 (2H, t, J = 7.7Hz), 3.54 (2H, t, J = 6.4Hz
), 3.82 (3H, s), 4.38 (2H, s), 4.75 (2H, s), 5
．． 52 (1H, br.s), 5.97 (1H, s), 6.63 (1H, br.s
), 7.24 (5H, m). (Step 10) Compound (10) (750 mg, 1.64 mmol), triphenylphosphine
(516 mg, 1.97 mmol), potassium iodide (327 mg, 1.97 mmol),
A mixture of 100 ml of acetonitrile (30 ml) and 100 ml of acetonitrile was heated in an oil bath for 70 hours.
The acetonitrile was removed under reduced pressure, and the residue was treated with chloroform to remove the insoluble matter.
The product was filtered off, and subjected to silica gel column chromatography.
From the fraction eluted with 1-chloroform, a pale yellow powder of compound (11) (1.32 g) was obtained.
, 99.2%) was obtained.¹ H-NMR (CDCl₃); 1.82 (2H, m), 2.46 (3H, s),
3.14 (2H, t, J = 6.4Hz), 3.31 (2H, m), 3.82 (3
H, s), 4.40 (2H, s), 4.84 (2H, s), 5.78 (1H, b
r. s), 6.02 (1H, s), 6.86 (1H, br.s), 7.04-7
．． 78 (20H, m). (11th step) Compound (11) (1.32g, 1.63mmol), DBU (0.61ml,
A mixture of 4.08 mmol) and methylene chloride (15 ml) was heated in an oil bath for 6 hours.
Water was added to the reaction mixture, which was then acidified with 2N hydrochloric acid and then diluted with chloroform.
The organic layer was washed with water, dried over anhydrous magnesium sulfate, and then filtered through a silica gel column.
The mixture was subjected to column chromatography. Elution was with 40% acetonitrile-chloroform.
The fractions containing the compound (12) were collected and recrystallized from acetone-isopropyl ether.
(375 mg, 56.9%) was obtained as pale yellow crystals, melting point 181-183°C.
Elemental analysis: C₂₃H₂₃N₃O₄ Measured values: C; 68.27, H; 5.64, N; 10.41. Calculated values: C; 68.13, H; 5.72, N; 10.37.¹ H-NMR (d₆-DMSO); 2.33 (2H, m), 2.38 (3H, s
), 2.63 (2H, t, J=7.2Hz), 3.81 (3H, s), 4.21
(2H, s), 4.74 (2H, s), 5.62 (1H, s), 5.63 (2H
, br), 6.42 (1H, t, J = 5.1Hz), 7.14-7.25 (5H
, m). (Step 12) Compound (12) (37 mg, 0.091 mmol) was dissolved in methanol (3 ml).
Dissolved, and 1N sodium hydroxide solution (0.18 ml, 0.18 mmol) was added to the chamber.
The mixture was stirred at room temperature for 45 minutes, and 1N hydrochloric acid (0.23 ml, 0.23 mmol) was added.
The mixture was concentrated under reduced pressure, and the precipitated crystals were collected by filtration.
The compound (I-1) (34 mg, 94.4%) was obtained by recrystallization from ethyl acetate, melting at 25°C.
Obtained as pale yellow crystals at 6-258°C (decomposition).

Elemental analysis: C₂₂H₂₁N₃O₄ Measured values: C; 67.33, H; 5.46, N; 10.69. Calculated values: C; 67.50, H; 5.41, N; 10.74.¹ H-NMR (d₆-DMSO); 2.25 (2H, m), 2.32 (3H, s
), 2.58 (2H, t, J=7.6Hz), 4.26 (2H, s), 4.75
(2H, s), 6.01 (1H, s), 6.08 (1H, t, J=4.8Hz)
, 6.91 (1H, br.s), 7.14-7.29 (5H, m), 7.32 (
1H, br. s). Compounds (I-2) to (I-10) were synthesized by carrying out the same reaction as in Example 1.
The physical parameters are shown in Tables 1 and 2.

Example 11 (Step 1) 3-Methoxy-2,5-dimethylpyrazine (13) (13.71 g, 99.2
mmol) and ethyl 5-bromo-4-oxopentanoate (24.34 g, 1
A mixture of 100 ml of acetonitrile (23 ml, 153.8 mmol) and 100 ml of DBU (23 ml, 153.8 mmol) was stirred at 60°C for 16 hours. The reaction mixture was dissolved in acetonitrile (143 ml), and DBU (23 ml, 153.8 mmol) was added, followed by heating under reflux for 2 hours. The reaction mixture was concentrated, poured into water, and extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over magnesium sulfate, and the solvent was distilled off. The residue was purified by silica gel column chromatography (hexane:ethyl acetate=9
The target compound (14) (10.57 g, 40.6%) was obtained as a pale yellow oil. ¹ H-NMR (CDCl ₃ ): 1.24 (3H, t, J = 7.2 Hz), 2.2
6 (3H, s), 2.63 (2H, t, J=7.5Hz), 2.97 (2H, t
, J=7.5Hz), 4.03 (3H, s), 4.13 (2H, q, J=7.2
Hz), 6.53 (1H, s), 7.07 (1H, m), 7.22 (1H, s)
(Step 2) To a suspension of aluminum chloride (11.24 g, 84.3 mmol) in chlorobenzene (27 ml) was added benzoyl chloride (9.78 ml, 84.3 mmol) under ice cooling.
The mixture was stirred for 10 minutes under the same conditions.
8.1 mmol) was gradually added, and the mixture was stirred under the same conditions for another 3 hours. The reaction mixture was poured into a mixture of ice, 28% aqueous ammonia, and ethyl acetate, and the insoluble matter was filtered off. The mixture was extracted with ethyl acetate, and the organic layer was washed with saturated brine and dried over magnesium sulfate. The solvent was evaporated, and the residue was purified by silica gel column chromatography (hexane:ethyl acetate = 9:1). The target product (15) (3.49 g, 33.9%) was obtained as yellow crystals. ¹ H-NMR (CDCl ₃ ): 1.16 (3H, t, J = 7.2 Hz), 2.3
5 (3H, s), 2.40-2.48 (2H, m), 2.58-2.66 (2H
, m), 4.03 (2H, q, J=7.2Hz), 4.08 (3H, s), 6.
61 (1H, s), 7.44-7.60 (3H, m), 7.64-7.70 (2
(1H, m), 8.50 (1H, s). (Step 3) Compound (15) (5.49 g, 15.0 mmol) was dissolved in diethyl ether (10
The reaction mixture was dissolved in a mixed solvent of 2.5 ml of water and tetrahydrofuran (50 ml), and lithium aluminum hydride (1 g, 26.4 mmol) was gradually added at room temperature, followed by stirring for 3 hours. The reaction mixture was diluted with tetrahydrofuran (30 ml), cooled on ice, and then diluted with water (2.5
mL) was gradually added, and the mixture was stirred at room temperature for 1 hour. The precipitated inorganic matter was filtered off and washed with tetrahydrofuran. The solvent was evaporated, the residue was dissolved in ethyl acetate, and dried over magnesium sulfate. The solvent was evaporated to obtain the target product (16) (5.0 g, 100%). This was used in the next reaction without purification.

(Step 4) A suspension of aluminum chloride (4.0 g, 30 mmol) in tetrahydrofuran (
95 ml) under ice-cooling, borane-t-butylamine complex (5.21 g, 60 mm
ol) was added and stirred for 10 minutes. Then, under the same conditions, compound (16) (5.0 g,
30 ml of a tetrahydrofuran solution of 15.0 mmol) of toluene was added, and the mixture was stirred at room temperature for 1 hour. The reaction solution was poured into 2N hydrochloric acid and extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over magnesium sulfate, and the solvent was distilled off. The residue was purified by silica gel chromatography (toluene:ethyl acetate=5:1) to obtain the target compound (17).
(4.71 g, 100%) was obtained as a pale yellow oil. ¹ H-NMR (CDCl ₃ ): 1.23 (1 H, br.s), 1.82-1.9
4 (2H, m), 2.20 (3H, s), 2.73 (2H, t, J = 7.5Hz
), 3.60-3.70 (2H, m), 4.04 (3H, s), 4.20 (2H
, s), 6.65 (1H, s), 6.96-7.03 (3H, m), 7.15-
7.29 (3H, m). (Step 5) A mixture of compound (17) (4.71 g, 15.0 mmol) and concentrated hydrochloric acid (88 ml) was heated under reflux for 70 minutes. The reaction mixture was diluted with water and extracted with a mixture of chloroform and methanol (9:1). The organic layer was washed with saturated brine, dried over magnesium sulfate, and the solvent was distilled off. The residue was washed with ethyl acetate-diethyl ether to obtain the target product (18) (3.60 g, 81%) as a colorless powder. ^1H -NMR ( _d6 -DMSO): 1.61-1.72 (2H, m), 1.97
(3H, s), 2.54 (2H, t, J=7.8Hz), 3.40 (2H, q,
J = 6.6Hz), 4.14 (2H, s), 4.42 (1H, t, J = 5.1H
z), 6.72 (1H, s), 6.87 (1H, s), 7.04-7.31 (5
(1H, m), 10.37 (1H, br.s). (Step 6) Compound (18) (3.53 g, 11.9 mmol) was dissolved in pyridine (71 ml), and acetic anhydride (3.37 ml, 35.7 mmol) was added. The mixture was stirred at room temperature for 6.5 minutes.
The mixture was stirred for 1 hour. The reaction mixture was diluted with water, acidified with 1N hydrochloric acid under ice cooling, and extracted with ethyl acetate. The organic layer was washed with water and then saturated brine, dried over magnesium sulfate, and the solvent was distilled off. The residue was recrystallized from methylene chloride-diethyl ether. The target product (19) (3.86 g, 95.6%) was obtained as colorless crystals.

Melting point: 160-161°C ¹ H-NMR (CDCl ₃ ); 1.87-1.92 (2H, m), 2.01 (3
H, s), 2.11 (3H, s), 2.65 (2H, t, J=7.5Hz), 4
．． 06 (2H, t, J = 6.3Hz), 4.13 (3H, s), 6.46 (1H
, s), 6.98-7.04 (3H, m), 7.17-7.32 (3H, m),
10.04 (1H, br.s). (Step 7) Compound (19) (3.94 g, 11.64 mmol) and phosphorus oxychloride (
A mixture of 31 ml of toluene and 100 ml of hexane (20%) was stirred at 80° C. for 45 minutes. Phosphorus oxychloride was evaporated under reduced pressure, and the residue was dissolved in ethyl acetate. The organic layer was washed with saturated aqueous sodium bicarbonate and saturated brine, dried over magnesium sulfate, and the solvent was evaporated. The residue was purified by silica gel column chromatography (toluene:ethyl acetate=7:1) to obtain the target product (20) (4.25 g, 100%) as a pale yellow oil. ¹ H-NMR (CDCl ₃ ): 1.93-2.05 (2H, m), 2.02 (3
H, s), 2.29 (3H, s), 2.77 (2H, t, J=7.5Hz), 4
．． 09 (2H, t, J = 6.3Hz), 4.23 (2H, s), 6.80 (1H
, s), 6.95-7.01 (2H, m), 7.18-7.32 (4H, m). (Step 8) Compound (20) (4.25 g, 11.64 mmol) in ethanol (62
ml) and sodium p-toluenesulfinate (4.15 g, 23.3 mmol)
) was added, and the mixture was heated under reflux for 36 hours. The reaction mixture was poured into water and extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over magnesium sulfate, and the solvent was distilled off. The residue was purified by silica gel column chromatography (ethyl acetate) to obtain the target product (21) (4.86 g, 87.5%) as yellow crystals.

Melting point: 141-142°C ¹ H-NMR (CDCl ₃ ); 1.97-2.09 (2H, m), 2.04 (3
H, s), 2.31 (3H, s), 2.41 (3H, s), 2.82 (2H, t
, J=7.5Hz), 4.10 (2H, t, J=6.3Hz), 4.24 (2H
, s), 6.92-6.98 (2H, m), 7.17-7.35 (6H, m),
7.46 (1H, s), 8.01 (2H, d, J = 8.1 Hz). (Step 9) Potassium t-butoxide (3.36 g, 29.9 mmol) was added to a solution of methyl glycolate (3.85 ml, 49.9 mmol) in tetrahydrofuran (120 ml), and the mixture was stirred at room temperature for 30 minutes. Then, the reaction mixture was cooled on ice to obtain compound (21) (4
.75 g, 9.97 mmol) was added and stirred for an additional 40 minutes. The reaction mixture was poured into water and extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over magnesium sulfate, and the solvent was distilled off. The residue was purified by silica gel column chromatography (hexane:ethyl acetate=1:1) to obtain the target compound (22) (2.71 g, 66.2%).
) was obtained as a yellow oil. ¹ H-NMR (CDCl ₃ ): 1.90-2.02 (2H, m), 2.01 (3
H, s), 2.14 (3H, s), 2.72 (2H, t, J=7.5Hz), 3
．． 77 (3H, s), 4.07 (2H, t, J=6.6Hz), 4.18 (2H
, s), 5.01 (2H, s), 6.74 (1H, s), 6.96-7.03 (
3H,m), 7.16-7.30 (3H,m). (Step 10) Compound (22) (2.60 g, 6.34 mmol) in methylene chloride (78
ml), N-methylmorpholine (1.39 ml, 12.6 mmol) was added under ice cooling.
Then, oxalyl chloride (1.1 ml, 12.6 mmol) was added, and the mixture was stirred at room temperature for 2.
The reaction mixture was stirred for 5 hours. Then, the reaction mixture was poured into 28% aqueous ammonia (78 ml) and
The mixture was stirred for 1 minute and extracted with methylene chloride. The organic layer was washed with saturated brine, dried over magnesium sulfate, and the solvent was evaporated. The residue was purified by silica gel column chromatography (ethyl acetate) to obtain the target product (23) (2.42 g, 79.3%) as pale yellow crystals.

Melting point: 157-158°C ^1H -NMR ( _CDCl3 ); 1.86-1.97 (2H, m), 2.02 (3
H, s), 2.18 (3H, s), 2.84-2.92 (2H, m), 3.76
(3H, s), 4.07 (2H, t, J=6.3Hz), 4.21 (2H, s)
, 4.97 (2H, s), 5.65 (1H, br.s), 6.64 (1H, br.
. s), 7.02-7.08 (3H, m), 7.19-7.33 (3H, m). (Step 11) Compound (23) (2.66 g, 5.53 mmol) in methanol (67 ml)
To a solution of 27.5 ml of 5N hydrochloric acid and 13 ml of tetrahydrofuran was added 5N hydrochloric acid (27.5 ml), and the mixture was stirred at room temperature for 7 hours. The reaction mixture was diluted with water and extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over magnesium sulfate, and the solvent was removed by distillation. The residue was purified by silica gel column chromatography (ethyl acetate:methanol=19:1) to obtain the target compound (24) (2.0 g, 82.3%) as a yellow powder. ¹ H-NMR (CDCl ₃ ): 1.75-1.86 (2H, m), 2.18 (3H, m).
H, s), 2.93 (2H, t, J = 7.2Hz), 3.57 (2H, t, J =
5.7Hz), 3.76 (3H, s), 4.23 (2H, s), 4.98 (2H
, s), 5.75 (1H, br.s), 6.67 (1H, br.s), 6.99
-7.09 (3H, m), 7.19-7.32 (3H, m). (Step 12) Compound (24) (1.88 g, 4.29 mmol) in methylene chloride (54
To the resulting solution (25) (2.09 g, 94%) was added triethylamine (0.90 ml, 6.43 mmol) and methanesulfonyl chloride (0.50 ml, 6.43 mmol) under ice-cooling, and the mixture was stirred for 30 minutes. The reaction mixture was poured into ice water, acidified with 1N hydrochloric acid, and extracted with methylene chloride. The organic layer was washed with saturated brine, dried over magnesium sulfate, and the solvent was evaporated to obtain the target product (25) (2.09 g, 94%) as a yellow powder. This was used in the next reaction without further purification.

(Step 13) Compound (25) (2.09 g, 4.03 mmol) was dissolved in dimethylformamide (
The reaction mixture was dissolved in 10 ml of ethanol (38 ml), lithium bromide (1.4 g, 16.1 mmol) was added, and the mixture was stirred at 50° C. for 2 hours and 45 minutes. The reaction mixture was poured into water and extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over magnesium sulfate, and the solvent was evaporated.
The residue was purified by silica gel column chromatography (toluene:ethyl acetate=1:1).
The target compound (26) (1.62 g, 75.8%) was obtained.

Melting point: 157-158°C ^1H -NMR ( _CDCl3 ); 2.08-2.19 (2H, m), 2.18 (3
H, s), 2.97 (2H, t, J = 7.5Hz), 3.43 (2H, t, J =
6.6Hz), 3.76 (3H, s), 4.25 (2H, s), 4.97 (2H
, s), 5.64 (1H, br.s), 6.62 (1H, br.s), 7.03
-7.11 (3H, m), 7.20-7.33 (3H, m). (Step 14) A suspension of compound (26) (1.96 g, 3.90 mmol) in acetonitrile (
Triphenylphosphine (1.55 g, 5.91 mmol) was added to the resulting solution (37 ml), and the mixture was heated under reflux for 18 hours. The acetonitrile was removed by distillation, and the residue was washed with ethyl acetate-ether to obtain the target compound (27) (2.71 g, 90.9%) as a yellow powder. This was used in the next reaction without further purification.

(Step 15) Compound (27) (2.71 g, 3.54 mmol) was dissolved in acetonitrile (70 m
1), DBU (1.32 ml, 8.83 mmol) was added, and the mixture was stirred at room temperature for 3
The mixture was stirred for 40 minutes. The reaction mixture was poured into 1N hydrochloric acid and extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over magnesium sulfate, and the solvent was evaporated. The residue was purified by silica gel column chromatography (ethyl acetate:methanol=19:1) to obtain the target product (28) (696 mg, 48.4%) as colorless crystals.

Melting point: 178-180°C ^1H -NMR ( _CDCl3 ); 2.13 (3H, d, J = 0.9Hz), 2.3
2-2.42 (2H, m), 2.68 (2H, t, J=7.5Hz), 3.75
(3H, s), 4.16 (2H, s), 4.97 (2H, s), 5.40-5.
95 (2H, br), 6.55 (1H, t, J=4.8Hz), 6.99 (1H
, d, J=0.9Hz), 7.05-7.10 (2H, m), 7.18-7.3
2 (3H, m). (Step 16) To a solution of compound (28) (395 mg, 0.974 mmol) in tetrahydrofuran (15 ml) was added 1N sodium hydroxide (3.88 ml), and the mixture was stirred at room temperature for 35 minutes.
The mixture was stirred for 10 minutes. 4 ml of 1N hydrochloric acid was added to the reaction mixture, and the mixture was extracted with a mixture of chloroform and methanol (9:1). The organic layer was washed with saturated brine, dried over magnesium sulfate, and the solvent was distilled off. The residue was recrystallized from methylene chloride-methanol-ether to obtain the target product (I-11) (331 mg, 86.8%) as colorless crystals.

Melting point: 204-206°C ^1H -NMR ( _d6 -DMSO); 2.11 (1H, s), 2.23-2.34
(2H, m), 2.64 (2H, t, J=7.5Hz), 4.24 (2H, s)
, 4.76 (2H, s), 6.19 (1H, t, J=4.8Hz), 7.01 (
1H, br. s), 7.13-7.22 (3H, m), 7.24-7.32 (2
H, m), 7.41 (1H, br.s), 7.51 (1H, s), 12.72 (
1H, br). Compounds (I-12) to (I-14) were synthesized by carrying out the same reaction as in Example 11. Their physical properties are shown in Table 3.

Test Example Human Secretory Phospholipase _A2 Inhibition Assay The following chromogenic assay is used to identify and evaluate inhibitors of recombinant human secretory phospholipase _A2 . The assay considered here is 96
This assay has been adapted for high-volume screening using well microtiter plates. A general description of this assay is given in Laure. J. Reynolds, Lo
An article by L. Hughes and Edward A. Dennis,
lysis of Human Synovial Fluid Phosp
holipase A ₂ on Short Chain Phosphati
dylcholine-Mixed Micelles: Developmen
of a Spectrophotometric Assay Suit
"able for a Microtiterplate Reader" (A
analytic biochemistry, 204, pp 190-19
7, 1992).

Reagents (reaction buffer) CaCl _{2.6H 2} O (2.19 g/L), KCl (7.455 g/L), bovine serum albumin (fatty acid free) (1 g/L) (Sigma A-7030), Tris-HCl (3.94 g/L), pH 7.5 (adjusted with NaOH) (Enzyme buffer) 0.05 M AcONa, 0.2 M NaCl, pH 4.5 (adjusted with acetic acid) (DTNB) 198 mg of 5,5'-dithiobis-2-benzoic acid (Wako Pure Chemical Industries, Ltd.) was dissolved in 10 mL of H ₂ O.
Dissolve 100 mg of racemic 1,2-bis(heptanoylthio)-1,2-dideoxy-sn-glycero-3-phosphorylcholine in 1 ml of chloroform.

(Triton-X 100) 624.9 mg of Triton-X 100 was dissolved in 100 ml of reaction buffer.

(Enzyme solution) Type I enzyme: sPLA ₂ solution (330 ng/μl) (A. Kanda et al., Bioc
himica et Biophysica Acta, 1171 (1992)
1-10) is diluted for use in the assay (27 μl of enzyme solution is diluted with 1973 μl of reaction buffer).

Type II enzyme: Dissolve 1 mg of enzyme in 1 ml of enzyme buffer. Store at 4°C. For assay, add 1995 μl of reaction buffer to 5 μl of this solution to dilute it.

Type V and type X enzymes: cDNA sequences encoding human type V and type X _sPLA2 (C
Hen et al., J. Biol. Chem, 1994, 269, 2365-2368 and Cupillard et al., J. Biol. Chem, 1997, 272, 1
5745-15752) was transformed into an expression vector for animal cells, pSVL SV40
Late Promoter Expression Vector (Amer
The resulting expression vector was inserted in the forward direction into the promoter downstream region of a LipofectAMINE reagent (Gi
Using a transfection kit (BCO BRL) according to the manufacturer's instructions, CHO host cells were transfected to obtain CHO cells stably expressing human type V and type X _sPLA2 . Each expressing cell was cultured for 3 days in α-MEM medium containing 10% fetal bovine serum, and the culture supernatant was used to measure the activity of each enzyme.

Enzyme reaction: for one microtiter plate 1) Place 0.106 ml of the substrate solution in a centrifuge tube and spray with nitrogen gas to evaporate the solvent. Add 0.54 ml of Triton-X 100 to this.
After adding and stirring, sonify in a bath type sonication.
To this, add 17.8 ml of reaction buffer and 0.46 ml of DTNB.
and dispense 0.18 ml into each well of a 96-well microtiter plate.

2) Add 10 μl of test compound (or solvent blank) according to the pre-set plate arrangement.

3) Incubate at 40°C for 15 minutes.

4) 90 ng/well for human type I enzyme and 50 ng for human type II enzyme
/well, 40 μl/well for human type V enzyme, 15 μl/well for human type X enzyme
μl/well was reacted.

5) The change in absorbance for human type I, type II, and type X enzymes over 30 minutes, and for human type V enzyme over 45 minutes, was measured using a plate reader, and the inhibitory activity was calculated (OD: 405 nm).
6) _IC50 was determined by plotting the log concentration against inhibition values ranging from 10% to 90% inhibition.

The results are shown in Table 4.

Formulation Examples The following Formulation Examples 1 to 9 are merely illustrative and are not intended to limit the scope of the invention in any way. The term "active ingredient" means a compound of formula (I), a prodrug thereof, or a pharmaceutically acceptable salt thereof, or a solvate thereof.

Formulation Example 1 Hard gelatin capsules are made using the following ingredients: Dose (mg/capsule) Active ingredient 250 Starch (dry) 200 Magnesium stearate 10 Total 460 mg Formulation Example 2 Tablets are made using the following ingredients: Dose (mg/tablet) Active ingredient 250 Cellulose (microcrystalline) 400 Silicon dioxide (fumed) 10 Stearic acid 5 Total 665 mg The ingredients are mixed and compressed into tablets each weighing 665 mg.

Formulation Example 3 An aerosol solution is prepared containing the following ingredients: Weight Active ingredient 0.25 Ethanol 25.75 Propellant 22 (chlorodifluoromethane) 74.00 Total 100.00 The active ingredient and ethanol are mixed, this mixture is added to a portion of Propellant 22, cooled to -30°C, and transferred to a filling machine. The required amount is then dispensed into a stainless steel container and diluted with the remaining propellant. A bubble unit is attached to the container.

Formulation Example 4 Tablets containing 60 mg of active ingredient are prepared as follows: Active ingredient 60 mg Starch 45 mg Microcrystalline cellulose 35 mg Polyvinylpyrrolidone (10% solution in water) 4 mg Sodium carboxymethyl starch 4.5 mg Magnesium stearate 0.5 mg Talc 1 mg Total 150 mg The active ingredient, starch, and cellulose are passed through a No. 45 mesh U.S. sieve and thoroughly mixed. An aqueous solution of polyvinylpyrrolidone is mixed with the resulting powders, and the mixture is then passed through a No. 14 mesh U.S. sieve. The granules so obtained are dried at 50° C. and passed through a No. 18 mesh U.S. sieve. The granules, previously passed through a No. 60 mesh U.S. sieve, are then sieved. The sodium carboxymethyl starch, magnesium stearate, and talc, previously passed through a sieve, are then added to the granules which, after mixing, are compressed in a tablet machine to yield tablets each weighing 150 mg.

Formulation Example 5 Capsules, each containing 80 mg of active ingredient, are prepared as follows: Active ingredient 80 mg Starch 59 mg Microcrystalline cellulose 59 mg Magnesium stearate 2 mg Total 200 mg The active ingredient, starch, cellulose, and magnesium stearate are blended, passed through a No. 45 mesh U.S. sieve, and packed into 200 hard gelatin capsules.
Fill in mg increments.

Formulation Example 6 Suppositories containing 225 mg of active ingredient are prepared as follows: Active ingredient 225 mg Saturated fatty acid glyceride 2000 mg Total 2225 mg The active ingredient is passed through a No. 60 mesh U.S. sieve and suspended in the saturated fatty acid glyceride which has been melted previously with the minimum necessary heating. The mixture is then poured into 2 g nominal molds and allowed to cool.

Formulation Example 7 A suspension containing 50 mg of active ingredient is made as follows: Active ingredient 50 mg Sodium carboxymethylcellulose 50 mg Syrup 1.25 ml Benzoic acid solution 0.10 ml Flavor q.v. Color q.v. Purified water q.v. total 5 ml. The active ingredient is passed through a No. 45 mesh U.S. sieve and mixed with the sodium carboxymethylcellulose and syrup to form a smooth paste. The benzoic acid solution and flavor, diluted with some of the water, are added and stirred. Sufficient water is then added to produce the required volume.

Formulation Example 8 An intravenous formulation is prepared as follows: Active ingredient 100 mg Saturated fatty acid glyceride 1000 ml A solution of the above ingredients is typically administered intravenously to a patient at a rate of 1 ml per minute.

Formulation Example 9 A lyophilized formulation (1 vial) is prepared as follows: Active ingredient 127 mg Sodium citrate dihydrate 36 mg Mannitol 180 mg The above ingredients are dissolved in water to give an injection solution with an active ingredient concentration of 10 mg/g. The first freezing step is performed at -40°C for 3 hours, and the heat treatment step is performed at -10°C for 1 hour.
The freeze-dried preparation is then dried at 0°C for 60 hours, and a refreezing step is carried out at -40°C for 3 hours. Then, the first drying step is carried out at 0°C and 10 Pa for 60 hours, and the second drying step is carried out at 60°C and 4 Pa for 5 hours. In this way, a freeze-dried preparation can be obtained.

INDUSTRIAL APPLICABILITY The compound of the present invention has an _sPLA2 inhibitory effect.
It inhibits _LA2- mediated fatty acid (e.g., arachidonic acid) release and is effective in treating septic shock and the like.

───────────────────────────────────────────────────────── Continued from the front page (51) Int.Cl. ⁷ Identification Symbol FI A61P 9/04 A61P 9/04 9/10 9/10 101 101 11/00 11/00 11/06 11/06 17/02 17/02 17/06 17/06 27/16 27/16 29/00 101 29/00 101 37/08 37/08 43/00 111 43/00 111 (81) Designated States EP(AT,BE,CH,CY, DE,DK,ES,FI,FR,GB,GR,IE,I T, LU, MC, NL, PT, SE, TR), OA (BF, BJ, CF, CG, CI, CM, GA, GN, GW, ML, MR, NE, SN, TD, TG), AP (GH, GM, KE, LS, MW, MZ, SD, SL, SZ, TZ , UG, ZW), UA (AM, AZ, BY, KG, KZ, MD, RU, TJ, TM), AE, AG, AL, AM, AT, AU, AZ, BA, BB, BG, BR, BY, B Z, CA, CH, CN, CR, CU, CZ, DE, DK ,DM,DZ,EE,ES,FI,GB,GD,GE, GH, GM, HR, HU, ID, IL, IN, IS, JP, KE, KG, KR, KZ, LC, LK, LR, LS, LT, LU, LV, MA, MD, MG, MK, MN, MW, MX, MZ, NO, NZ, PL, PT, RO, RU, SD, SE, SG, SI, SK, SL, TJ, TM, TR, TT, TZ, UA, UG, US, UZ, VN, YU, ZA, ZW (Note) This publication is based on the publication of the internationally published Japanese patent application (Japanese utility model registration application) in accordance with Article 184-10, Paragraph 1 of the Patent Act (Article 48-13, Paragraph 2 of the Utility Model Act), and is unrelated to this publication.

Claims (16)

[Claims] [Claim 1] General formula (I): wherein E is N or C-R ⁴ ; when E is N, G is C-R ²⁵ , and when E is C-R ⁴ , G is N; R ¹ is (a) a C1-C20 alkyl, a C1-C20 alkenyl, a C1-C20 alkynyl, a carbocyclic group, or a heterocyclic group; (b) the group represented by (a) substituted with one or more groups, each independently selected from non-interfering substituents;
or (c) -(L ¹ )-R ⁵ (wherein L ¹ is a hydrogen atom, a nitrogen atom, a carbon atom,
a divalent linking group of 1 to 18 atoms selected from an oxygen atom and a sulfur atom, ^R5 being a group selected from (a) or (b); one of ^R3 and ^R4 being -( ^L2 )- (acidic group) (wherein ^L2 is a linking group to the acidic group, and the length of the linking group to the acidic group is 1 to 5), and the other being a hydrogen atom; ring A being a group represented by the formula: (wherein ^R2 is _CONH2 , _CONHNH2 , or _CSNH2 ; ^R18 , ^R19 , ^R20 , ^R21 , R22, ^R23 , ^R26 , and ^R27 are each independently a hydrogen atom or a lower alkyl; and ^R24 and ^R25 are each independently a hydrogen atom, a C1-C6 alkyl, an aryl, a halogen, or an aralkyl), a prodrug thereof, or ^a pharmaceutically acceptable salt thereof, or a hydrate thereof. [Claim 2] General formula (II): wherein ^R24 and ring A are as defined above; J is N or C- ^R8 , and when J is N, G is C- ^R25 ( ^R25 is as defined above), and when J is C- ^R8 , G is N; ^R6 is -( _CH2 )m- ^R9 (m is an integer of 1 to 6, ^R9 is a group represented by the formula (d): (In the formula, a, c, e, n, q, t, and v each independently represent 0, 1, or 2.
R ¹⁰ and R ¹¹ are each independently halogen, C1-C10 alkyl, C
a is an oxygen atom or a sulfur atom; β is —CH ₂ — or —(CH 2 ) ₂ —; γ is an oxygen atom or a sulfur atom; b is an integer of 0 to 3; d is an integer of 0 to 4; f, p, and w are each independently an integer of 0 to 5; r is an integer of 0 to 7; and u is an integer of 0 to 4), or ( _e ) a group represented by C1-C6 alkyl, C1-C6 alkyloxy, C1-C
a group selected from (d) substituted with one or more substituents selected from the group consisting of C1-C6 haloalkyloxy, C1-C6 haloalkyl, phenyl, and halogen); one of ^R7 and ^R8 is a hydrogen atom, and the other is -( ^L3 ) ^-R12 (wherein ^L3 is a group represented by the formula: (wherein M is —CH ₂ —, —O—, —N(R ¹⁵ )—, or —S—; R ¹³ and R ¹⁴ each independently represent a hydrogen atom, a C1-C10 alkyl, an aryl, an aralkyl, a carboxy, or a halogen; and R ¹⁵ represents a hydrogen atom or a C1-C6 alkyl; and R ¹² represents a group of the formula: (wherein R ¹⁶ is a hydrogen atom, a metal, or a C1-C10 alkyl; R ¹⁷ is each independently a hydrogen atom or a C1-C10 alkyl; and h is an integer of 1 to 8), a prodrug thereof, or a pharmaceutically acceptable salt thereof, or a hydrate thereof. 3. The method of claim 1, wherein R ¹ and R ⁶ are of the formula: (wherein R ¹⁰ , R ¹¹ , b, d, f, p, r, u, w, α, β, and γ are as defined above), a prodrug thereof, or a pharmaceutically acceptable salt thereof, or a hydrate thereof according to claim 1 or 2. 4. The method of claim 1, wherein R ¹ and R ⁶ are of the formula: (wherein R ¹⁰ , R ¹¹ , d, p, u, and w are as defined above), a prodrug thereof, a pharmaceutically acceptable salt thereof, or a hydrate thereof according to any one of claims 1 to 3. 5. The compound according to claim 1, wherein R ³ and R ⁷ are —O—(CH ₂ ) m —COOH (wherein m is as defined above) or —O—CH(—R ³⁰ )—COOH (wherein R ³⁰ is a hydrogen atom or a C1
5. The compound according to claim 1, wherein R is -C3 alkyl), a prodrug thereof, a pharmaceutically acceptable salt thereof, or a hydrate thereof. [Claim 6] General formula (III): (wherein G, J, R ¹⁰ , R ³⁰ , ring A, and m are as defined above; ring B is a benzene ring or a cyclohexane ring; and R ³⁸ is a hydrogen atom or a C1-C3 alkyl), a prodrug thereof, or a pharmaceutically acceptable salt thereof, or a hydrate thereof. 7. The compound according to any one of claims 1 to ₆ , wherein ^R2 is -CONH2, a prodrug thereof, a pharmaceutically acceptable salt thereof, or a hydrate thereof. 8. A compound according to any one of claims 1 to 7, wherein R ¹⁸ , R ¹⁹ , R ²⁰ , R ²¹ , R ²² , R ²³ , R ²⁶ , and R ²⁷ are all hydrogen atoms, a prodrug thereof, a pharmaceutically acceptable salt thereof, or a hydrate thereof. 9. A pharmaceutical composition comprising the compound according to any one of claims 1 to 8 as an active ingredient. 10. The pharmaceutical composition according to claim 9, which is an sPLA ₂ inhibitor. 11. The pharmaceutical composition according to claim 9, which is a type I sPLA ₂ inhibitor. 12. The pharmaceutical composition according to claim 9, which is a type II sPLA ₂ inhibitor. 13. The pharmaceutical composition according to claim 9, which is a V-type _sPLA2 inhibitor. 14. The pharmaceutical composition according to claim 9, which is an X-type _sPLA2 inhibitor. 15. Use of a compound according to any one of claims 1 to 8 for the manufacture of a medicament for treating a disease caused by _sPLA2 . 16. A method for treating a disease caused by _sPLA2 in a mammal, including a human, which comprises administering to said mammal a therapeutically effective amount of a compound according to any one of claims 1 to 8.