JPH10251239A - New 4h-3,1-benzoxazin-4-one derivative - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide the subject new compound applicable by various administration methods such as oral administration and intravenous injection, exhibiting an action stronger than the action of conventional agent for the treatment of pancreatitis and having a trypsin-inhibiting action as well as elastase- inhibiting action and chymotrypsin-inhibiting action. SOLUTION: The objective compound is expressed by the formula I [R<1> and R<2> are each independently a halogen, COR<3> (R<3> is H or a 1-4C alkyl), COOR<4> (R<4> is H or a 1-4C alkyl), etc.; (n) is 0-2] or its salt, e.g. 2-acetyl-6-[2-(4- guanidinobenzoyloxy)benzamido]benzoic acid 4-methoxybenzylmethanesulfonic acid salt. The compound of the formula I can be produced e.g. by bonding 4-guanidinobenzoic acid to an anthranylic acid derivative to obtain the compound of the formula II and finally cyclizing the product. As an alternative, it can be produced by condensing a cyclized 4H-3,1-benzoxazin-4-one derivative (the formula III) to a 4-guanidinobenzoic acid derivative.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to a novel 4H-3,1
A benzoxazin-4-one derivative or a salt thereof, a method for producing the derivative, a pharmaceutical composition containing at least one of the derivative and a salt thereof as an active ingredient,
And a compound useful as a synthetic intermediate of the derivative and a method for producing the compound.

[0002]

2. Description of the Related Art Proteolytic enzymes, particularly serine proteases, which are endopeptidases, are involved in maintaining the homeostasis of living organisms, such as degradation of food, metabolism of unnecessary proteins, blood coagulation / fibrinolysis, and complement reaction. Some examples of these are trypsin, chymotrypsin, elastase, enterokinase, etc. as serine proteases involved in food digestion, and in the blood coagulation system, various blood coagulation factors, thrombin, activation Protein C, plasma kallikrein, etc., plasmin in fibrinolytic systems, tissue plasminogen activator, urokinase, etc.
C1r, C1s, C2, Factor B, Factor D,
For phagocytosis such as C3 convertase, neutrophil elastase, cathepsin G, chymase, tryptase and the like are known.

[0003] These activities are regulated and regulated in the living body by a large amount of serine protease inhibitor present in tissue or plasma, and the qualitative or quantitative abnormality of this inhibitor causes inflammation or thrombosis of each tissue. It causes various diseases such as bleeding. Specific examples in which elastase is particularly involved include rheumatoid arthritis, arteriosclerosis, shock, allergy, pancreatitis, nephritis, certain skin diseases, thrombosis, disseminated intravascular coagulation (DIC), and adult respiratory distress syndrome. (ARDS), pulmonary fibrosis, emphysema, sepsis, reflux esophagitis and the like are known.

Among these diseases, for example, pancreatitis,
The pathogenesis of acute pancreatitis is thought to be autolysis of the pancreas caused by pancreatic enzymes that have escaped from the parenchymal cells of the pancreas into the stroma. Trypsin is an important initiator of pancreatitis, and this enzyme is Activation of other enzymes in turn activates other enzymes. However, subsequent pancreatic hemorrhage and necrosis have been attributed to the action of elastase and phospholipase rather than trypsin ("Therapeutics", 21).
Volume, p. 648 (1988)).

[0005] Further, when the condition becomes severe, the coagulation function is enhanced, and the DIC is prepared. At this time, if the induction of cytokines continues for some reason or DIC develops, neutrophil activation occurs ("Bile and Pancreas", Vol. 12,
747 (1991)).

[0006] In fact, in patients with severe pancreatic disease accompanied by multiple organ failure (MOF), the amount of neutrophil elastase is more than 10 times higher than normal. This indicates that neutrophil activation and lysosomal enzyme release are indeed taking place. Neutrophils attack and destroy the accumulated important organs ("Bile and Pancreas", Vol. 9, p. 1685 (1988)).
Thus, in recent years, acute pancreatitis and its more severe
Involvement of elastase in addition to trypsin has been suggested.

Hitherto, many studies have been made on various 4-guanidinobenzoic acid derivatives as typical low-molecular-weight protease inhibitors, especially trypsin inhibitors.
To date, nafamostat mesilate (FUT) and camostat mesylate (Foipan®) have been used as therapeutic agents. However, even with these therapeutic agents, it is hard to say that the therapeutic effects on pancreatitis are sufficiently satisfied. Further, as the only orally administrable therapeutic agent for pancreatitis, only foilpan is recognized.

Research on 4-guanidinobenzoic acid derivatives mainly focused on the treatment of acute pancreatitis based on trypsin inhibitory activity and the treatment of DIC as an antiplasmin agent. Have also been reported to have elastase inhibitory activity. For example, derivatives described in JP-A-63-165357 and JP-A-4-46148 are exemplified. However, the effects of the compounds disclosed therein are not yet satisfactory and have not yet been approved as pharmaceuticals.

[0009] On the other hand, as an example of research on elastase inhibitors, there is a 4H-3,1-benzoxazin-4-one derivative. For example, 2-alkyl-4H-3,1 such as Teshima
-Report of benzoxazin-4-one derivatives "Journal of Biological Chemistry" (J. B.
iol. Chem. ) 257, 5085-5091
Page (1982), Hedstrom et al., "Biochemistry," Volume 23, 1
753-1759 (1984), 2-ethoxy-
The 4H-3,1-benzoxazin-4-one derivative is
Radha Krishnan et al. "Journal of Molecular Biology" (J. Mol. Biol.) 19
8, 417-424 (1987), 4H-3,
The mechanism of elastase inhibition of 1-benzoxazin-4-one derivatives has been reported. Further, JP-A-60-1694
No. 69, 4H-3,1-benzoxazine-4-
On-amino derivatives, and JP-A-62-30770, also disclose 2-oxy derivatives.

Recently, 4H-3,1-benzoxazine-4-H has been used as a selective neutrophil elastase inhibitor.
2,7-diamino derivatives of on derivatives (WO88 / 09
790), 2-aminomethyl derivatives (Japanese Unexamined Patent Publication No.
148249) and a derivative having an amino acid amide bond at the 6-position (US Pat. No. 5,428,021).

Further, WO96 / 07648 discloses that
It is disclosed that a 4H-3,1-benzoxazin-4-one derivative having a substituted phenyl group, guanidinoalkyl, amidinoalkyl group or the like introduced at the 2-position has a C1r protease inhibitory action. However, this technique does not mention the activity against elastase, and is a technique mainly based on anti-complementary action.

However, any of the above 4H-3,
Even with the 1-benzoxazin-4-one derivative, the therapeutic effect on pancreatitis is not satisfactory and none has been approved as a pharmaceutical. Further, in the above-mentioned prior arts and others, there is no disclosure of a compound in which a 4-guanidinobenzoic acid phenyl ester skeleton and a 4H-1,3-benzoxazin-4-one skeleton are hybridized in the same molecule.

[0013]

SUMMARY OF THE INVENTION An object of the present invention is to cope with various administration forms such as oral or intravenous administration, and has a stronger effect than conventional therapeutic agents for pancreatitis.
An object of the present invention is to provide a 1-benzoxazin-4-one derivative and a salt thereof. Also, other diseases based on trypsin, chymotrypsin and elastase, such as rheumatoid arthritis, arteriosclerosis, shock, acute circulatory failure,
Certain skin diseases such as sepsis, allergy, nephritis, atopic dermatitis, thrombosis, disseminated intravascular coagulation (DI
C), a benzoxazinone derivative and a salt thereof which are effective for the prevention and treatment of adult respiratory distress syndrome (ARDS), pulmonary fibrosis, emphysema, reflux esophagitis and the like.
Another object of the present invention is to provide a method for producing these compounds and a pharmaceutical composition containing them, and further provide an intermediate useful for producing the compounds and a method for producing the same.

[0014]

Means for Solving the Problems The present inventors have made intensive studies in view of the above circumstances, and found that 4-guanidinobenzoic acid phenyl ester skeleton and 4H-3,1-benzoxazin-4-one. By creating a new skeleton in which the skeleton and the skeleton are bound in the same molecule, a group of compounds having a better profile as a protease inhibitor, that is, a trypsin inhibitory activity, an elastase inhibitory activity and a chymotrypsin inhibitory activity, are obtained. The inventors have found that these compounds exhibit excellent effects in treating various diseases such as pancreatitis and have high safety with few side effects, and thus completed the present invention.

In a first aspect of the present invention, the following formula (I)

Embedded image (Wherein R ¹ and R ² are each independently a halogen atom, an alkyl group having 1 to 4 carbon atoms, an alkoxy group having 1 to 4 carbon atoms, a formula COR ³ (wherein R ³ is a hydrogen atom or Represents an alkyl group having 1 to 4 carbon atoms.), A formula COOR
⁴ (wherein, R ⁴ represents a hydrogen atom or an alkyl group having 1 to 4 carbon atoms), an alkylthio group having 1 to 4 carbon atoms, an alkylsulfonyl group having 1 to 4 carbon atoms, and a par having 1 to 4 carbon atoms. Represents a fluoroalkyl group, an alkyl group having 1 to 4 carbon atoms substituted by an alkoxy group having 1 to 4 carbon atoms, and n represents 0 to
Represents an integer of 2, and when n is 2, each R ² may be the same or different. ) Or a salt thereof.

Preferred combinations of substituents in the compound represented by the above formula (I) are shown below, but the present invention is not limited thereto. In the combination of n, R ¹ and R ² , when n is 0, R ¹ is a chlorine atom, a bromine atom, a methyl group, an ethyl group, a propyl group, a methoxy group, an ethoxy group, an acetyl group, an ethoxycarbonyl group (R ⁴ is an ethyl group), a methylthio group, a methanesulfonyl group, a trifluoromethyl group or a methoxymethyl group, and the bond of the 4-guanidinobenzoyloxy group is preferably an ortho position of a phenyl group at the 2-position of the benzoxazine ring.

When n is 1, the type of R ¹ and the bonding position of 4-guanidinobenzoate are the same as when n is 0, R ² is a halogen atom, and an alkoxy group having 1 to 4 carbon atoms ( preferably (wherein, R ³ is (preferably a hydrogen atom or an alkyl group having 1 to 4 carbon atoms R ³ is a methyl group) methoxy) or formula COR ³ preferably has a), even of R ² The substitution position is preferably at the 6-position or the 8-position of the benzoxazine ring.

When n is 2, the types of R ¹ and R ² and 4
- coupling position guanidino benzoic acid ester is the same as when n is 1, the substitution position of R ^2, 6 and 8 positions of the benzoxazine ring. The compound of the present invention forms an acid addition salt with a suitable inorganic or organic acid, and among them, a hydrochloride or a methanesulfonate is preferable.

The compounds of the present invention may exist as optical isomers when the substituents of R ¹ and R ² have an asymmetric carbon atom. Separation and acquisition of these isomers can be performed by those skilled in the art by ordinary techniques, and the compounds of the present invention include both isomers and mixtures. The compounds of the present invention and salts thereof may also form various solvates with pharmaceutically acceptable solvents such as water, ethanol, glycerin or acetic acid. These solvent substances are also included in the compound of the present invention.

In a second embodiment of the present invention, the following formula (VI)
II)

Embedded image (Wherein, the definitions of the substituents represented by R ¹ and R ² and n are the same as those of the formula (I), and A represents a carboxyl protecting group) or a deprotected product thereof (Free carboxylic acid) or salts thereof. Here, the deprotected form is also specifically represented by the following formula (IX).

[0021]

Embedded image(Where R¹, R^TwoDefinition of a substituent represented by
Is the same as in formula (I). ) These are the first of formula (I)
Useful as an intermediate for obtaining the final compound. In addition,
In the compound of formula (VIII) and its deprotected form,
R as in the compound of (I) ¹, R^TwoStructural Variations Based on Substituents of
However, all isomers and mixtures are
Included in the invention. Compound of formula (VIII) and its deprotection
The body and its salts are also pharmaceutically acceptable solvents, such as
Water, ethanol, glycerin or acetic acid
It may form a solvate. These solvates are also compounds of the present invention.
include.

The compound of the formula (VIII) and its deprotected product also have a potent serine protease inhibitory activity similarly to the compound of the formula (I), so that the compound caused by a serine protease such as trypsin, chymotrypsin or elastase can be treated. The therapeutic application of is expected.

A third embodiment of the present invention relates to a compound of the following formula (VIII):

Embedded image (Wherein the definitions of the substituents represented by R ¹ and R ² and n are the same as those in formula (I), and A represents a carboxyl protecting group) or a salt thereof, if desired. A method for producing a compound of the formula (I) or a salt thereof, wherein A is deprotected and an intramolecular cyclization reaction is carried out.

A fourth aspect of the present invention is a compound represented by the following formula (VI):

Embedded image (Wherein the substituents represented by R ¹ and R ² and the definitions of n and A are the same as those of the formula (VIII)) and a salt thereof, and a compound represented by the following formula (VII)

Embedded image (Wherein Y represents a hydroxyl group or a halogen atom). A process for producing a compound of the formula (VIII) or a salt thereof, characterized by reacting with a 4-guanidinobenzoic acid derivative represented by the following formula:

A fifth aspect of the present invention is a pharmaceutical composition containing, as an active ingredient, at least one of the compound represented by the formula (I) or a salt thereof. According to a sixth aspect of the present invention,
It is a prophylactic / therapeutic agent for pancreatitis or reflux esophagitis containing at least one of the compound represented by the formula (I) or a salt thereof as an active ingredient.

In the embodiment of the present invention, the formula (I)
In the definition of R ¹ , R ² , R ³ or R ⁴ in each of the formulas (X) to (X), the alkyl group, alkoxy group, alkylthio group, alkylsulfonyl group, perfluoroalkyl group, and alkoxy-substituted alkyl group have 1 to 1 carbon atoms. There is a notation of 4, which is a carbon number of 1
It is intended to mean both straight or branched chains of the four.
As for the alkoxy-substituted alkyl group, the number of carbon atoms in the alkoxy group and the partial alkyl group portion can independently be 1 to 4.

[0027]

BEST MODE FOR CARRYING OUT THE INVENTION Hereinafter, the present invention will be described in detail. The compound of the present invention has the following formula (I)

Embedded image (Wherein R ¹ and R ² are each independently a halogen atom, an alkyl group having 1 to 4 carbon atoms, an alkoxy group having 1 to 4 carbon atoms, a formula COR ³ (wherein R ³ is a hydrogen atom or Represents an alkyl group having 1 to 4 carbon atoms.), A formula COOR
⁴ (wherein, R ⁴ has the same meaning as described above), and has 1 carbon atom
An alkylthio group having 1 to 4 carbon atoms, an alkylsulfonyl group having 1 to 4 carbon atoms, a perfluoroalkyl group having 1 to 4 carbon atoms, and an alkyl group having 1 to 4 carbon atoms substituted with an alkoxy group having 1 to 4 carbon atoms; n represents an integer of 0 to 2;
Wherein each R ² may be the same or different) or a salt thereof.

Of these, R¹About the halogen atom
May be a chlorine atom or a bromine atom,
As the alkyl group, methyl, ethyl or propyl
Group, a methoxy group as an alkoxy group having 1 to 4 carbon atoms
Or ethoxy group, formula COR ^ThreeAn acetyl group, formula C
OOR^FourAs an ethoxycarbonyl group, having 1 to 4 carbon atoms
As alkylthio, a methylthio group, an alkyl having 1 to 4 carbon atoms
A methanesulfonyl group as a killsulfonyl, having 1 to 1 carbon atoms
Trifluoromethyl as the perfluoroalkyl group of 4
Groups or alkoxyalkyl groups having 1 to 4 carbon atoms
A toxicoxy group is preferred.

[0029] For R ² is a halogen atom (preferably chlorine atom), an alkoxy group having 1 to 4 carbon atoms (preferably methoxy), or the formula COR ³ (preferably acetyl) is preferred, the substitution of R ² The position is preferably the 6-position or the 8-position of the benzoxazine ring, and more preferably the 8-position. n is preferably any of 0 to 2;
^1, the preferred value by the substitution position of the type and R ² substituents of R ² are changed.

The bonding position of the 4-guanidinobenzoyloxy group is preferably the ortho position of the 2-position phenyl group on the benzoxazine ring.

As the acid addition salt, various inorganic acid salts or organic acid salts can be mentioned. For example, salts with inorganic acids such as hydrochloride, hydrobromide, phosphate, sulfate, acetate, oxalate, citrate, tartrate, maleate, alginate, p-toluene sulfone Examples thereof include salts with organic acids such as acid salts, methanesulfonates and salicylates, salts with acidic amino acids such as glutamates and aspartates, and hydrochlorides and methanesulfonates are preferred.

The compound of the formula (I) of the present invention can be prepared by the following process A
Alternatively, it can be synthesized by Production Method B. Production method A is a production method in which 4-guanidinobenzoic acid is bound to an anthranilic acid derivative to form compound (IX), and finally cyclized. On the other hand, the production method B is a production method in which the previously cyclized 4H-3,1-benzoxazin-4-one derivative (X) is condensed with the 4-guanidinobenzoic acid derivative. The present invention is not limited to the following production method.

In the following reaction route, R ¹ ,
R ² and n represent the same definition as in the above formula (I), and Acyl
Is an acyl group or an alkoxycarbonyl group represented by an acetyl group, a benzoyl group and a methoxycarbonyl group. A represents a protecting group for a carboxyl group, for example, methyl, ethyl, t-butyl, benzyl or 4-methoxybenzyl group, and P represents a protecting group for a phenolic hydroxyl group, for example, acetyl, pivaloyl, methoxycarbonyl, Y represents a phenacyl group, a methoxymethyl group or a benzyl group, and Y represents a hydroxyl group or a carboxyl group-activating group such as a halogen atom or a mixed acid anhydride residue. Each manufacturing method will be described below in order.

(Production method A)

Embedded image

[0035]

Embedded image

In reaction 1, compound (II) is reacted with preferably at least 2 equivalents of compound (III) in the presence of a base in an inert solvent or without solvent at -20 to 100 ° C. to give compound (IV) Get. As the base, an organic base such as pyridine, triethylamine or N, N-dimethylaniline, or an inorganic base such as sodium hydrogencarbonate or sodium carbonate can be used. As the inert solvent, halogenated hydrocarbons such as dichloromethane, chloroform or 1,2-dichloroethane, ethers such as diethyl ether, tetrahydrofuran (THF) or dioxane, or N, N-dimethylformamide (DMF), N, N N-dimethylacetamide (DM
A) or amides such as 1,3-dimethyl-2-imidazolidinone are used.

In the reaction 2, the compound (IV) and preferably at least two equivalents of an alcohol (AOH (where A represents an alcohol residue such as a methyl, ethyl, t-butyl, benzyl or 4-methoxybenzyl group) Is reacted in an inert solvent at 10 to 100 ° C. in the presence of a base to obtain compound (VI), which is 1,8-diazabicyclo [5.4.0] -7-undecene (DBU). And organic bases such as triethylamine or diisopropylethylamine, metal alkoxides such as potassium t-butoxide, sodium methoxide or sodium ethoxide, etc. Examples of the inert solvent include dichloromethane, chloroform or 1,2-dichloroethane. Such as halogenated hydrocarbons, diethyl ether, THF or dioxane Such ethers or DMF, DMA
Alternatively, amides such as 1,3-dimethyl-2-imidazolidinone can be used.

In reaction 3, compound (VI) is obtained using compound (II ') and compound (V) by a conventional peptide synthesis method. For example, 4th edition experimental chemistry course 2
Vol. 2 (edited by The Chemical Society of Japan, 1992, published by Maruzen Co., Ltd.), Chapter 2, "Amino Acids and Peptides". In reaction 4,
Compound (VIII) is obtained from compound (VI) and compound (VII) according to a conventional method for synthesizing an active ester. For example,
"Protective Groups in Organic
Synthesis ”(Protective Groups in Organic Synthes
is), TW Green (TWGreene), etc., 1991,
John Willy and Sons Company (Joh
n, Willy & Sons Company) in accordance with the method described in Chapter 5.

In reaction 5, compound (IX) is obtained using compound (VIII) according to the usual ester deprotection conditions. For example, "Protective Groups in
Organic Synthesis "(Protective Groups in O
rganic Synthesis (supra) can be performed according to the method described in Chapter 5. In reaction 6, compound (IX) is used to obtain compound (I) according to ordinary dehydration reaction conditions. For example, carbodiimides such as N, N'-dicyclohexylcarbodiimide, acid anhydrides such as acetic anhydride, inorganic acids such as sulfuric acid, alkyl halocarbonates and bases, or acid chlorides and bases are used in various methods. It can be carried out.

(Production method B)

Embedded image

In reaction 7, compound (II) and compound (III ') are reacted according to the method of reaction 1 (production method A) to give compound (IV')
Get. In reaction 8, compound (X) is obtained using compound (IV ') according to the conditions for a general phenol formation reaction by deprotection. For example, it can be carried out according to the method described in Chapter 3 of "Protective Groups in Organic Synthesis" (supra). In reaction 9, compound (VI ') is obtained using compound (IV') according to general ester hydrolysis conditions. For example, the method is performed according to the method described in “Experimental Chemistry Lecture, Vol. 22, Vol. 22, Chapter 1,“ Carboxylic Acids and Derivatives ”, or in a range of room temperature to reflux temperature in water or water containing an organic solvent. be able to. As the organic solvent, a water-soluble solvent such as acetone, methanol or ethanol can be used.

Reaction 10 was carried out using compound (VI ') at -20
Reaction with concentrated sulfuric acid at １００100 ° C. gives compound (X).
Reaction 11 is performed according to reaction 10 using compound (VI) obtained by Production Method A to obtain compound (X). Reaction 1
2. Compound (I) is obtained by subjecting compound (X) to compound (VII) according to reaction 4 (Production method A).

The starting material, anthranilic acid derivative (II) or (II '), was synthesized by a method known in the literature. For example, Baker et al., "Journal of
Organic Chemistry "(J. Org. Che
m. ), Vol. 17, p. 141 (1952), that is, a synthesis method from an isatin derivative. Or, like a packet, "Journal of the American Chemical
Society ”(J. Am. Chem. Soc.) 9
9, p. 3734 (1981),
A method for reducing an aromatic nitro derivative to anthranilic acid,
Or, Stephen et al., "Journal of Organic Chemistry" (J. Org. Chem.) 5
0, p. 718 (1985) and Stephen et al.
"Journal of Organic Chemistry"
(J. Org. Chem.) 56, 6666 (19
1991), that is, a method of carboxylating an aniline derivative. After completion of the reaction, the compound of formula (I) or the acid addition salt thereof obtained as described above is subjected to column chromatography, recrystallization,
It may be subjected to an isolation operation such as extraction.

Next, the pharmacological action of the representative compounds of the present invention will be described, but the present invention is not limited thereto. In addition, the test compound number used for the following test referred to the Example number.

EXPERIMENTAL EXAMPLE 1 Inhibitory Activity of the Compound of the Present Invention on Various Serine Proteases The compound of the present invention (test drug) has human purulent sputum elastase (HSE), porcine pancreas elastase (PPE), and bovine pancreas. Trypsin (Bovine Pancreas Trypsine: TRY) and Bovine Pancreas Chymotrypsin (CH)
The inhibitory activity against Y) was measured by the method described below.

1) Measurement of HSE inhibitory activity Reaction solution: 12% dimethyl sulfoxide, 0.5 M sodium chloride, 0.1% sodium azide, 0.1 M Tris-HCl buffer (pH 7.5) Enzyme: from Elastin Products Purchase Synthetic substrate: MeOSuc-Ala-Ala-Pro-Val-pNA. 0.7 H ₂
O (purchased from Cosmo Bio) Autoanalyzer: COBAS MIRA from Roche Measuring method: 2 μg / ml HS using autoanalyzer
E and a reaction solution containing various measured concentrations of the test drug at 37 ° C.
, And the mixture was reacted for 2 minutes after the addition of the synthetic substrate to a final concentration of 0.25 mM. The initial reaction rate was calculated from the change in absorbance at 405 nm during the reaction, and the 50% inhibitory concentration (IC ₅₀ ) value was determined.

2) Measurement of PPE inhibitory activity Reaction solution: 12% dimethyl sulfoxide, 0.5 M sodium chloride, 0.1% sodium azide, 0.1 M Tris-HCl buffer (pH 7.5) Enzyme: purchased from Sigma Synthetic substrate: Suc-Ala-Ala-Ala-pNA (purchased from Protein Research Promotion Association) Autoanalyzer: COBAS MIRA from Roche Measurement method: 2 μg / ml PP using autoanalyzer
The reaction solution containing E and various measured concentrations of the test drug was
The mixture was preliminarily heated at 0 ° C. and reacted for 10 minutes after the addition of the synthetic substrate to a final concentration of 0.025 mM. 405n during reaction
The initial reaction rate was calculated from the change in absorbance at 50 m.
The inhibitory concentration (IC ₅₀ ) value was determined.

3) Measurement of TRY inhibitory activity Reaction solution: 10 mM calcium chloride, 130 mM sodium chloride, 0.1% bovine serum albumin, 20 mM Tris-HCl buffer (pH 7.5) Enzyme: purchased from Sigma Synthetic substrate: Bz- L-Arg-pNA ・ HCl (purchased from Protein Research Promotion Association) Autoanalyzer: Roche COBAS MIRA Measurement method: 2.5 μg / ml using an autoanalyzer
Reaction solution containing TRY and test drugs at various concentrations
Pre-warmed at 7 ° C. to bring the synthetic substrate to a final concentration of 0.025 mM
After the addition, the reaction was carried out for 8 minutes. 405n during reaction
The initial reaction rate was calculated from the change in absorbance at 50 m.
The inhibitory concentration (IC ₅₀ ) value was determined.

4) Measurement of CHY inhibitory activity Reaction solution: 20 mM calcium chloride, 10% N, N-dimethylformamide, 100 mM Tris-HCl buffer (pH
7.5) Enzyme: Purchased from Sigma Synthetic substrate: Bz-L-Tyr-pNA / HCl (purchased from Protein Research Promotion Association) Autoanalyzer: Roche's COBAS MIRA Measurement method: 3 μg / ml using an autoanalyzer CH
Reaction solution containing Y and various measured concentrations of the test drug was
The mixture was preliminarily heated at ℃, and reacted for 15 minutes after the addition of the synthetic substrate to a final concentration of 0.1 mM. The initial reaction rate was calculated from the change in absorbance at 405 nm during the reaction, and the 50% inhibitory concentration (IC ₅₀ ) value was determined.

Table 1 shows the results. In addition, the camostat mesylate was used for the comparative example. Table 1 ───────────────────── IC ₅₀ (μg / ml) Example ELA PPE TRY CHY ────────────── ─────── 51 0.56 0.09 0.04 0.20 52 0.45 0.45 0.05 1.30 53 0.97 2.36 0.06 2.80 54 0.10 0.15 0.09 0.13 55 0.25 0.44 0.06 1.70 56 0.96 8.30 0.08 7.30 57 1.00 4.11 0.02 2.10 58 3.18 9.00 0.08 2.80 59 0.41 5.40 0.07 0.95 65 0.20 0.27 0.08 0.22 66 0.05 0.58 0.07 1.00 67 0.09 0.31 0.11 0.17 68 0.25 0.80 0.09 3.00 69 1.20 3.10 0.05 1.10 70 0.09 0.72 0.10 0.48 71 0.20 0.44 0.16 0.15 72 4.50 1.60 0.04 1.00 73 3.20 4.80 0.10 1.20 Comparative example> 30 ＞ 30 0.02 ＞ 30 ─────────────────────

Although the compound of the present invention has a trypsin inhibitory action similar to that of camostat mesylate, it has been found that, unlike camostat mesylate, it has a strong inhibitory action on other elastases and chymotrypsin.

Experimental Example 2: Effect on mouse ethionine pancreatitis model MHC (ICR) female mice were bred for 4 days on normal solid feed for 2 days using 4- to 5-week-old female mice, and then fed a choline-deficient diet for 2 days. Day 24 was fasted for 24 hours. 0.5% on day 6
A choline-deficient diet containing DL-ethionine was given for 24 hours to cause pancreatitis. On the seventh day, I switched to a choline-deficient diet again,
The amylase activity (PAMY) of the pancreas extirpated on the eighth day was measured.

The test drug was dispersed in distilled water or a 0.5% carboxymethylcellulose aqueous solution at 100 mg / kg for two days on days 6 and 7, and then orally administered twice a day. Control group (test drug non-administration group)
Administered distilled water instead of the drug, and the normal group was kept on a normal diet. The activity value was calculated according to the following formula. Activity value (%) = (control PAMY-drug PAMY / control PAMY-
Normal PAMY) × 100 The results are shown in Table 2. In addition, camostat mesylate was used as a comparative example.

Table 2 ──────────────── Examples Therapeutic effect (%) ──────────────── 51 86 52 55 53 47 55 80 56 54 57 90 58 55 59 94 61 65 63 41 64 42 67 64 68 62 69 61 70 89 72 48 75 64 Comparative Example 5 ────────────────

All of the compounds of the present invention showed a remarkable therapeutic effect on pancreatitis by oral administration in a mouse ethionine pancreatitis model. In addition, the action strength was significantly stronger than that of camostat mesylate.

EXPERIMENTAL EXAMPLE 3: Toxicity Experiment Each compound of Examples 51, 54, 60, 65, 71 and 74 of the present invention was purified by Wist
As a result of oral administration to ar male rats at 1 g / kg for 14 consecutive days, no apparent toxicity was observed.

As is clear from the above description and the results of the experimental examples, the compounds of the present invention have a broad and strong inhibitory effect on trypsin, chymotrypsin and elastase which are involved in the cause or exacerbation of pancreatitis.
Further, the compound of the present invention has a strong inhibitory effect in an animal ethionine pancreatitis model. Furthermore, the compounds of the present invention show efficacy in a rat caerulein model.
The compounds of the present invention also show excellent efficacy in a rat reflux esophagitis model. Furthermore, the compound of the present invention is effective by oral administration, has extremely low toxicity, and is excellent in safety. The intermediate compounds (VIII) and (I) of the present invention
In X), the test of Experimental Example 1 or Experimental Example 2 described above showed almost the same activity as the compound of the formula (I). No clear findings were found regarding toxicity.

The compound of the present invention can be used for the prevention and treatment of pancreatitis, for example, improvement of acute symptoms of pancreatitis (acute pancreatitis, acute exacerbation of chronic pancreatitis, postoperative acute pancreatitis, acute pancreatitis after pancreatography, traumatic pancreatitis), chronic pancreatitis It is effective in preventing relapse of pancreatitis or treating chronic pancreatitis, or preventing and treating reflux esophagitis. The compounds of the present invention may further comprise other diseases based on serine proteases, especially trypsin, chymotrypsin and elastase, such as shock, acute circulatory failure, sepsis,
Rheumatoid arthritis, arteriosclerosis, allergy, hay fever, nephritis, certain skin diseases (including atopic dermatitis), thrombosis, disseminated intravascular coagulation (DIC), adult respiratory distress syndrome (ARDS), lung It can also be used in the prevention and treatment of fibrosis, emphysema and the like. Also, the intermediate compounds (VIII) and (IX) of the present invention can be used as a medicine for the above-mentioned diseases based on serine proteases, similarly to the compound of the formula (I). In addition, 4-guanidinobenzoic acid ester skeleton and 4H-3,1-benzoxazine-
In the compound of the present invention in which a 4-one skeleton was bound in the same molecule, a stronger effect on the treatment of pancreatitis than that exhibited by the single administration of the compound having each partial structure was confirmed. Also, in the comparison of the water solubility of each drug, the water solubility was significantly improved as compared with the compound having each partial structure.

The compound of the present invention or an acid addition salt thereof is usually orally administered to humans and other animals as a drug, but parenterally (for example, intravenous administration, intramuscular administration, subcutaneous administration, rectal administration, Vaginal administration, transdermal absorption, transmucosal absorption, etc.) can also be used.

The pharmaceutical composition of the present invention is a compound of the formula (I) of the present invention.
It is sufficient that at least one compound represented by the formula (1) is included, and it is prepared in combination with a pharmaceutically acceptable carrier. More specifically, excipients (eg, lactose, sucrose, mannitol, crystalline cellulose, silicic acid), binders (eg, crystalline cellulose, saccharides (mannitol, sucrose), dextrin, hydroxypropylcellulose (HPC), hydroxypropyl Methylcellulose (HPMC), polyvinylpyrrolidone (PVP), macrogol), lubricant (eg, magnesium stearate, calcium stearate, talc), coloring agent, flavoring agent, disintegrant (eg, corn starch, carboxymethylcellulose), preservative Agents, tonicity agents, stabilizers (eg, sugars, sugar alcohols), dispersants, antioxidants (eg, ascorbic acid, butylhydroxyanisole (BHA), propyl gallate, dl-
α-tocopherol), buffer, preservative (eg, paraben, benzyl alcohol, benzalkonium chloride), fragrance (eg, vanillin, l-menthol, rose oil), solubilizer (eg, cholesterol, triethanolamine) ), Suspending or emulsifying agents, and suitable pharmaceutically acceptable carriers or solvents can be combined with the compounds of the present invention to give various dosage forms.

Examples of the dosage form include capsules, pills, tablets, granules, fine granules, powders, suspensions, emulsions,
Internal solutions such as limonade, elixir and syrup, external solutions such as inhalants, sprays, aerosols and coatings, ophthalmic / nasal solutions, patches, ointments, lotions, liniments, cataplasms Preparations, suppositories, aqueous or non-aqueous injections, emulsion or suspension injections, and solid injections which are dissolved, emulsified or suspended before use. In dosage forms for oral administration, enteric preparations can be prepared by using an appropriate coating agent.

The dosage of the compound of the present invention as a drug varies depending on the condition. Usually, when administered to an adult per day, when administered systemically (orally or parenterally such as intravenously) to an adult, The dosage is in the range of about 1 to 2000 mg, preferably in the range of about 10 to 600 mg, but the dose is appropriately adjusted according to the condition of the patient and the like, and is not limited thereto. Oral or parenteral administration may be carried out in a single dose or divided into 2 to 6 doses, or intravenous drip or the like is also possible. In the case of topical administration, it depends on the symptoms.
Per 1 to 150 mg per preparation. An appropriate amount can be administered once or several times a day to the affected part by a method such as application. The amount is sufficient to treat the disease of interest, but the dosage form of the therapeutic agent, the method of administration,
It is preferable to appropriately increase or decrease according to the number of administrations per day, degree of symptoms, body weight, age and the like. The compounds of the present invention can be used in combination with conventional therapeutic agents.

[0063]

EXAMPLES Next, the present invention will be described in more detail with reference to Examples, but the present invention is not limited thereto. In the following description, Examples of the intermediates for producing the compounds of the present invention or the intermediate compounds of the present invention are described as Reference Examples.

Tables 3 to 8 show the physical property data of each compound, and Tables 9 to 14 show the structures. In Tables 9 to 14, Ac is an acetyl group, Me is a methyl group, tBu is t-
A butyl group and PMB represent a 4-methoxybenzyl group, and MsOH represents methanesulfonic acid.

The nuclear magnetic resonance spectrum (NMR) was measured by JEOL JNM-EX270 (manufactured by JEOL Ltd.).
NM-EX270) or the company's Jeol JNM-LA3
00 (JEOL JNM-LA300) was analyzed by infrared absorption spectrum (I
R) is HORIBA F manufactured by HORIBA, Ltd.
The melting point of T-200 was measured by METTLER (M
(Ettler) FP800.

Reference Example 1 Methyl 6- (methoxymethyl) anthranilate 2-methoxymethyl-6-nitrobenzoate (1
To a solution of 7.2 g) in methanol (400 ml) was added 10% palladium-carbon (0.5 g), and the mixture was stirred overnight at room temperature under a hydrogen atmosphere. After the reaction, the catalyst was removed by filtration,
The filtrate was concentrated under reduced pressure. A 1N aqueous sodium hydroxide solution (165 ml) was added to the residue, and the mixture was refluxed for 1 hour. After cooling, the reaction solution was washed with ether (100 ml). Remove the water layer 10
% Sulfuric acid and adjusted to pH 3-4, then ethyl acetate (200
ml × 3). Combine the organic layers and add water (200m
1 × 2) and saturated saline (200 ml), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crystals of the residue were recrystallized from ethyl acetate-hexane to give the title compound (10.2 g) as colorless crystals.

Reference Example 2 Methyl 2-methylthio-6-nitrobenzoate N, N of methyl 2,6-dinitrobenzoate (8.9 g)
-Sodium thiomethoxide (4.34 g) was added to a dimethylformamide (230 ml) solution under ice-water cooling,
Stirred at room temperature for 2 days. After concentrating the reaction solution under reduced pressure, the residue was dissolved in ethyl acetate (2 L), washed with water (500 ml × 2) and brine (500 ml), and dried over anhydrous sodium sulfate. After evaporating the solvent under reduced pressure, the residue was purified by silica gel column chromatography (eluent: hexane / ethyl acetate = 8/1 to 4/1). The fractions containing the target compound were collected and the solvent was distilled off under reduced pressure to give the title compound (4.51).
g) was obtained as a yellow oil.

Reference Example 3 Methyl 6-methylthioanthranilate The compound (8.03 g) of Reference Example 2 in methanol (200
ml) solution was added with 10% palladium-carbon, and the mixture was stirred under a hydrogen atmosphere at room temperature for 1 day. After completion of the reaction, the catalyst was removed by filtration, the filtrate was concentrated under reduced pressure, and the residue was subjected to silica gel column chromatography (eluent: hexane / ethyl acetate = 2).
0/1). The fractions containing the desired product were collected and the solvent was distilled off under reduced pressure to obtain the title compound (4.11 g) as a pale yellow oil.

Reference Example 4 6-Methylthioanthranilic acid To a compound of Reference Example 3 (4.10 g) was added a 1N aqueous sodium hydroxide solution (50 ml), and the mixture was refluxed for 2.5 hours. The reaction solution was adjusted to pH 4 with 5% sulfuric acid, and ethyl acetate (300
ml × 3). Combine the organic layers and add water (200m
1) and saturated saline (200 ml), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was recrystallized from ethyl acetate-hexane to give the title compound (3.29).
g) was obtained as pale brown crystals.

Reference Example 5 2-amino-6-ethoxycarbonylbenzoic acid 2-ethoxycarbonyl-6-nitrobenzoic acid (50
To a solution of g) in ethanol (200 ml) was added 10% palladium-carbon (0.5 g), and the mixture was stirred overnight at room temperature under a hydrogen atmosphere. After completion of the reaction, the catalyst was removed by filtration, and the filtrate was concentrated under reduced pressure. Ether-hexane was added to the residue to give a fine powder, followed by filtration to obtain the title compound (39.2 g) as a yellow powder.

Reference Example 6 2-methoxy-5- (trifluoromethyl) aniline 1-methoxy-2-nitro-4- (trifluoromethyl) benzene (50.0 g) in methanol (700 m
l) 10% palladium-carbon was added to the solution, and the mixture was stirred under a hydrogen atmosphere at room temperature for 1 day. After completion of the reaction, the catalyst was removed by filtration, the filtrate was concentrated under reduced pressure, and the residue was triturated with hexane.
The title compound (41.8 g) was obtained as a pale gray powder.

Reference Example 7 N- [2-methoxy-5- (trifluoromethyl) phenyl] -2,2-dimethylpropionamide Anhydrochlorination of the compound of Reference Example 6 (41.8 g) and triethylamine (30.8 ml) Methylene (525 ml)
The solution was cooled with ice water and pivaloyl chloride (26.6 g)
Was added dropwise over about 1 hour, and the mixture was stirred overnight at room temperature. The reaction suspension was mixed with water (200 ml × 2) and saturated saline (200 ml).
ml), dried over anhydrous sodium sulfate and concentrated to dryness under reduced pressure to give the title compound (59.9 g).

Reference Example 8 2- (2,2-Dimethylpropionamido) -3-methoxy-6- (trifluoromethyl) benzoic acid A solution of the compound of Reference Example 7 (157 g) in anhydrous tetrahydrofuran (1.5 l) was added with argon. Under gas atmosphere, -40 to 40
N-Butyllithium (1.6M hexane solution, 731 ml) was added dropwise at −50 ° C., and at the same temperature for 15 minutes and
The mixture was stirred at 0 to 0 ° C for 1 hour. -10 carbon dioxide gas to the reaction solution
The mixture was introduced at ０0 ° C. for 1.5 hours and at room temperature for 2.5 hours, and stirred at room temperature overnight. The reaction solution was poured into water (5 L) and washed with ethyl acetate (1 L × 2). Add concentrated hydrochloric acid to the aqueous layer to adjust the pH to 1
２2 and extracted with ethyl acetate (2 L × 2). The organic layers were combined, washed with water (500 ml) and saturated saline (500 ml), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. Ether was added to the residue to give a fine powder, which was filtered to give the title compound (126.3 g) as colorless crystals.

Reference Example 9 Methyl 2- (2,2-dimethylpropionamido) -3-methoxy-6- (trifluoromethyl) benzoate Sodium hydrogencarbonate was added to a solution of the compound of Reference Example 8 (106 g) in acetone (1 L). (36.3 g) and dimethyl sulfate (31.4 ml) were added, and the mixture was refluxed for 3.5 hours. The reaction solution was poured into water (1 L) and extracted with ethyl acetate (2 L × 2). The organic layers were combined, washed with water (1 L) and saturated saline (1 L), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. Hexane was added to the residue to give a fine powder, which was filtered to give the title compound (96.9 g) as colorless crystals.

Reference Example 10 Methyl 2-diacetylamido-3-methoxy-6- (trifluoromethyl) benzoate Acetic anhydride (207 m) of the compound of Reference Example 9 (22.0 g) was prepared.
A solution of 1) and acetic acid (104 ml) was refluxed for 15 hours. After the reaction solution was concentrated under reduced pressure, the residue was treated with ether (500 m
l), water (100 ml x 2), saturated aqueous sodium bicarbonate (1
00 ml × 2), water (100 ml × 2) and saturated saline (100 ml), and dried over anhydrous sodium sulfate. The solvent was evaporated under reduced pressure, hexane was added to the residue to give a fine powder, and the residue was filtered to give the title compound (19.9 g) as colorless crystals.

Reference Example 11 3-methoxy-6- (trifluoromethyl) anthranilic acid A suspension of the compound of Reference Example 10 (9.8 g) and a 2N aqueous sodium hydroxide solution (120 ml) was refluxed for 2 hours. The pH of the reaction solution was adjusted to 2 to 3 with 4N hydrochloric acid under ice water cooling, and the mixture was extracted with ethyl acetate (500 ml × 2). The organic layers were combined, washed with water (100 ml × 2) and saturated saline (100 ml), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. Hexane was added to the residue to give a fine powder, which was filtered to give the title compound (5.10 g) as colorless crystals.

Reference Example 12 N- (2-acetyl-5-methoxyphenyl) -2-
(Hydroxyimino) acetamide To a suspension of chloral hydrate (33.1 g) and sodium sulfate (207 g) in water (550 ml) was added 2'-amino-4'-methoxyacetophenone (32.1 g) in concentrated hydrochloric acid (37 ml). ) And a suspension of water (120 ml) were added and stirred at room temperature for 1 hour. To this was added a suspension of hydroxylamine hydrochloride (40.4 g) in water (120 ml), and the mixture was refluxed for 1 hour. The precipitated insoluble matter was collected by filtration, dissolved in methanol (300 ml), and then dissolved in ethyl acetate (500 ml).
ml) was added, the precipitated insolubles were removed by filtration, and the filtrate was concentrated under reduced pressure. The residue was subjected to silica gel column chromatography (eluent: hexane / ethyl acetate = 4/1 to 1 /
Purified in 1). The fractions containing the desired product were collected and concentrated to dryness under reduced pressure to give the title compound (6.40 g) as pale orange crystals.

Reference Example 13 7-acetyl-2,3-dihydro-4-methoxy-1H
-Indole-2,3-dione The compound of Reference Example 12 (6.20 g) was gradually added to concentrated sulfuric acid (30 ml) at 70 ° C, followed by stirring at 80 ° C for 20 minutes. The reaction solution was poured into ice water (400 ml), and the precipitate was collected by filtration and washed with a small amount of ethanol and ether to give the title compound (5.75 g) as a red-brown powder.

Reference Example 14 3-Acetyl-6-methoxyanthranilic acid A solution of the compound of Reference Example 13 (5.75 g) and a 1.5N aqueous sodium hydroxide solution (86.6 ml) was added with 60 to 7 to give a solution.
At 0 ° C., 30% hydrogen peroxide (9.80 ml) was added little by little, and after the addition was completed, the mixture was stirred at the same temperature for 1.5 hours. The pH of the reaction solution was adjusted to 2 by adding 1N hydrochloric acid, and ethyl acetate (100
ml × 3). Combine the organic layers and add water (100
ml) and saturated saline (100 ml), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: hexane / ethyl acetate = 1/3). The fractions containing the desired product were collected and concentrated under reduced pressure to dryness to give the title compound (1.0 g) as pale yellow crystals.

Reference Example 15 2,3,5-Trimethoxybenzonitrile 1-bromo-2,3,5-trimethoxybenzene (2
4.7 g) and copper cyanide (18.7 g).
A suspension of dimethylformamide (374 ml) was refluxed for 4 hours. The insoluble matter was removed by filtration, and the filtrate was concentrated under reduced pressure.
The residue was subjected to silica gel column chromatography (eluent:
Hexane / ethyl acetate = 97/3 to 19/1). The fractions containing the desired product were collected and concentrated under reduced pressure to dryness to give the title compound (21.0 g) as colorless crystals.

REFERENCE EXAMPLE 16 2,3,5-Trimethoxybenzoic acid The compound of Reference Example 15 (15.8 g) in ethanol (23
0 ml) and water (150 ml) were added with sodium hydroxide (24.0 g) and refluxed for 6 hours. After concentrating the reaction solution under reduced pressure, the residue was dissolved in water (200 ml) and washed with ether (200 ml × 2). The aqueous layer was removed, acidified with hydrochloric acid, and extracted with ethyl acetate (200 ml × 2). The organic layers were combined, washed with water (100 ml) and saturated saline (100 ml), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was crystallized from methylene chloride-hexane and filtered to give the title compound (16.1 g) as colorless crystals.

Reference Example 17 The compound of Reference Example 16 (15.5 g) was added to 2-nitro-3,5,6-trimethoxybenzoic acid nitric acid (d = 1.38, 41.0 ml) at 10 ° C. or lower. Stirred at warm for 20 minutes. The reaction suspension was poured into ice, and the precipitated crystals were collected by filtration. The crystals were sufficiently washed with water to give the title compound (9.07).
g) was obtained as pale yellow crystals.

Reference Example 18 3,5,6-Trimethoxyanthranilic acid 2-nitro-3,5,6-trimethoxybenzoic acid (8.
To a suspension of 5 g) in methanol (76 ml) was added 10% palladium-carbon (0.76 g), and the mixture was stirred under a hydrogen atmosphere at room temperature for 2 days. After completion of the reaction, the catalyst was removed by filtration, and the filtrate was concentrated under reduced pressure. The residue is subjected to silica gel column chromatography (eluent: hexane / ethyl acetate = 3/2 to 1).
/ 3). The fractions containing the desired product were collected and concentrated under reduced pressure, and the residue was crystallized from ethyl acetate-hexane to give the title compound (5.67 g) as yellow crystals.

Reference Example 19 2- (2-acetoxyphenyl) -5-acetyl-4H
-3,1-benzoxazin-4-one 6-acetylanthranilic acid (1.80 g) under ice-water cooling
2- (chloroformyl) phenyl acetate (6.00 g) was added little by little to a pyridine (10 ml) solution, and the mixture was stirred at the same temperature for 1 hour and at room temperature overnight under an argon atmosphere. Ice water (40 ml) was added to the reaction solution, and ethyl acetate (40 ml) was added.
× 2). Combine the organic layers and add 1N hydrochloric acid (40
ml), water (40 ml) and saturated saline (40 ml)
, Dried over anhydrous sodium sulfate and concentrated under reduced pressure. The precipitated crystals were washed with ether to give the title compound (0.98 g) as colorless crystals.

Reference Examples 20 to 26, Reference Examples 28 to 31 and Reference Examples 35 to 43 were synthesized in the same manner as Reference Example 19 using the corresponding anthranilic acid derivatives.

Reference Example 27 2- (2-acetoxyphenyl) -5-methylsulfonyl-4H-3,1-benzoxazin-4-one Under ice-cooling with water, methylene chloride (4.70 g) of the compound of Reference Example 26 (4.70 g) was added. M-chloroperbenzoic acid (7.12 g) was added little by little to the solution (120 ml), and the mixture was stirred at room temperature for 2 hours. The insoluble matter of the reaction suspension was removed by filtration, and the filtrate was washed with a saturated aqueous solution of sodium sulfite (50 ml × 2) and water (50 ml).
1) and saturated saline (50 ml), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. Hexane was added to the residue to give a fine powder, which was filtered and washed with ether to give the title compound (5.08 g) as colorless crystals.

Reference Example 32 2- (2-acetoxyphenyl) -5- (1,2-dibromoethyl) -8-methoxy-4H-3,1-benzoxazin-4-one The compound of Reference Example 36 (9. 0 g) in anhydrous chloroform (130 ml) was added to N-bromosuccinimide (1).
6.5 g) and 2,2'-azobis-isobutyronitrile (174 mg) were added, and the mixture was refluxed for 6 hours. Ethyl acetate (1 L) was added to the reaction solution, and a saturated aqueous sodium hydrogen carbonate solution (500 m
l), water (500 ml) and saturated saline (500 m
1), dried over anhydrous sodium sulfate and concentrated under reduced pressure. Ether was added to the residue, and the mixture was pulverized, filtered, and washed with ether to give the title compound (11.6 g) as pale yellow crystals.

Reference Example 33 2- (2-acetoxyphenyl) -5- (2-bromo-
1-hydroxyethyl) -8-methoxy-4H-3,1
-Benzoxazin-4-one Acetone (450) of the compound of Reference Example 32 (11.0 g)
silver) (3.75 g) was added to the solution of (ml) -water (11 ml) under ice-cooling under light shielding, and the mixture was stirred at room temperature for 1 hour under light shielding. The insolubles of the reaction suspension were removed by filtration, and the filtrate was concentrated under reduced pressure. The residue was dissolved in ethyl acetate (2 L) and water (1
L) and saturated saline (1 L), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: hexane / ethyl acetate = 4/1 to 3/1). The fractions containing the desired product were collected and concentrated to dryness under reduced pressure to give the title compound (2.23 g) as colorless crystals.

Reference Example 34 7- (2-Acetoxybenzamide) -3-bromomethyl-6-methoxy-1 (3H) -isobenzofuranone Following purification of the compound of Reference Example 33, hexane / ethyl acetate = 1/1. The fraction containing the desired product was collected and concentrated to dryness under reduced pressure to give the title compound (3.07
g) was obtained as pale brown crystals.

Reference Example 44 2- (2-acetoxyphenyl) -6-chloro-5,8
-Dimethoxy-4H-3,1-benzoxazine-4-
On a solution of 3,6-dimethoxyanthranilic acid (9.0 g) in methylene chloride (150 ml) was added ice-water cooling to t-butyl hypochlorite (6.38 g) in methylene chloride (50 m2).
l) The solution was added dropwise over 30 minutes and stirred at the same temperature for 5 minutes. Under ice-water cooling, pyridine (50 ml) was added dropwise to the reaction solution over 10 minutes, and 2- (chloroformyl) phenyl acetate (2
1.75 g) in methylene chloride (150 ml)
After dripping over a period of time, the mixture was cooled for 1 hour under ice-water cooling and
Stirred for hours. The reaction solution was 1N hydrochloric acid (500 ml × 2),
Saturated aqueous sodium bicarbonate (500 ml × 2) and saturated saline (25 ml
0 ml x 2), dried over anhydrous sodium sulfate,
It was concentrated under reduced pressure. The residue was purified by silica gel chromatography (eluent: hexane / ethyl acetate = 2/1 to 2/3). The fractions containing the desired product were collected and concentrated under reduced pressure, and the residue was triturated with ethyl acetate-hexane, and filtered to give the title compound (8.2 g) as colorless crystals.

Reference Example 45 4-Methoxybenzyl 2-acetyl-6- (2-hydroxybenzamide) benzoate The compound of Reference Example 19 (3.50 g) and 4-methoxybenzyl alcohol (3.13 ml) in anhydrous tetrahydrofuran (32 ml) )) Solution, under ice-water cooling, 1,8-diazabicyclo [5.4.0] -7-undecene (3.89).
ml), and the mixture was stirred for 10 minutes and further at room temperature for 3 days. The reaction solution was concentrated under reduced pressure, and the residue was treated with ethyl acetate (150 m
1) hydrochloric acid (60 ml), water (60 ml)
And saturated brine (60 ml), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was crystallized from hexane-ether and filtered to give the title compound (4.28 g).
Was obtained as colorless crystals.

The compounds of Reference Examples 46 to 48, Reference Example 50, Reference Examples 52 to 58, and Reference Examples 60 to 69 start from the compounds of Reference Examples 20 to 23, 25 to 31, and 35 to 44. It was synthesized in the same manner as in Reference Example 45 as a raw material.

REFERENCE EXAMPLE 49 2-Chloro-6- (2-hydroxybenzamido) benzoic acid 4-methoxybenzyl 6-chloroanthranilic acid (30 g) in pyridine (43
ml) and methylene chloride (800 ml) solution under ice-cooling with 2- (chloroformyl) phenyl acetate (86).
A solution of g) in methylene chloride (500 ml) was added dropwise over 2 hours under an argon atmosphere, followed by stirring at room temperature for 1 hour. The reaction solution was treated with 1N hydrochloric acid (500 ml × 2) and water (300
ml × 2) and saturated saline (300 ml),
It was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The precipitated crystals are pulverized with ethyl acetate, filtered, washed with ether, and then containing 2- (2-acetoxyphenyl) -5-chloro-4H-3,1-benzoxazine-4-yl containing O-acetylsalicylic acid. On (38 g) was obtained. To a suspension of this compound and 4-methoxybenzyl alcohol (33 ml) in anhydrous tetrahydrofuran (500 ml) was added 1,8-diazabicyclo [5.4.0] -7-undecene (39 ml) under ice-cooling for 10 minutes. And further stirred overnight at room temperature. The reaction solution was concentrated under reduced pressure, the residue was dissolved in ethyl acetate (1.5 l), washed with 1N hydrochloric acid (500 ml), water (500 ml) and saturated saline (300 ml), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. did. The residue was subjected to silica gel chromatography (eluent: hexane /
Ethyl acetate = 19/1 to 10/1). The fractions containing the desired product were collected and concentrated under reduced pressure, and the residue was triturated with hexane and filtered to give the title compound (41 g) as colorless crystals.

Reference Example 51 t-butyl 2- (2-hydroxybenzamido) -6-methoxybenzoate The compound of Reference Example 24 (3.00 g) and molecular sieves 3A (3.60 g) in anhydrous t-butanol (30
ml), 1,8-diazabicyclo [5.4.
0] -7-undecene (2.88 ml) was added and the mixture was refluxed for 9 hours. The insolubles in the reaction suspension were removed by filtration, the insolubles were washed with a small amount of ethyl acetate, and the filtrate and the washing were combined and concentrated under reduced pressure. The residue was dissolved in ethyl acetate (100 ml), washed with 1N hydrochloric acid (50 ml), water (50 ml) and brine (50 ml), dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue is subjected to silica gel column chromatography (eluent: hexane / ethyl acetate = 20/1).
〜１０10 / 1). The fractions containing the desired product were collected and concentrated to dryness under reduced pressure to give the title compound (2.82 g) as colorless crystals.

Reference Example 59 6-acetyl-2- (2-hydroxybenzamide)-
4-methoxybenzyl 3-methoxybenzoate (Method a) The compound of Reference Example 33 (2.0 g) and 4-methoxybenzyl
To a solution of methoxybenzyl alcohol (2.29 ml) in anhydrous tetrahydrofuran (20 ml) was added 1
8-Diazabicyclo [5.4.0] -7-undecene (2.75 ml) was added, and the mixture was stirred at room temperature overnight. After the reaction solution was concentrated under reduced pressure, the residue was dissolved in ethyl acetate (150 ml), washed with 1N hydrochloric acid (60 ml), water (60 ml) and saturated saline (60 ml), and dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the residue was subjected to silica gel column chromatography (eluent: hexane / ethyl acetate = 4 /
1-2 / 3). The desired fraction was collected, concentrated under reduced pressure to dryness to give the title compound (1.41 g) as a pale-yellow powder.

(Method b) The compound of Reference Example 34 (0.59
g) and 4-methoxybenzyl alcohol (0.52
1,8-diazabicyclo [5.4.0] was added to a suspension of anhydrous tetrahydrofuran (30 ml) under ice-water cooling.
-7-Undecene (0.76 ml) was added, and the mixture was stirred at room temperature overnight. After the reaction solution was concentrated under reduced pressure, the residue was treated with ethyl acetate (5.
0 ml), 1N hydrochloric acid (20 ml), water (20 m
l) and saturated saline (20 ml), and dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the residue was subjected to silica gel column chromatography (eluent: hexane / hexane).
Ethyl acetate = 4/1 to 2/3). The fractions of the desired product were collected, concentrated under reduced pressure to dryness, and the title compound (0.41 g)
Was obtained as a pale yellow powder.

Example 1 2-Acetyl-6- [2- (4-guanidinobenzoyloxy) benzamide] benzoic acid 4-methoxybenzyl methanesulfonate The compound of Reference Example 45 (23.8 g) and 4-guanidinobenzoic acid N, N'-dicyclohexylcarbodiimide (12.9 g) was added to a suspension of methanesulfonate (15.6 g) in pyridine (240 ml) under ice-water cooling.
The mixture was stirred overnight at room temperature under an argon atmosphere. The precipitated insoluble matter was removed by filtration, ether (2 L) was added to the filtrate, and the mixture was sufficiently stirred. After standing, the supernatant was removed by decantation. Further add ether (200 ml) and stir well,
Filtration provided the title compound (38.4 g) as a white powder.

Examples 2 to 25 correspond to Reference Examples 46 to 69.
Was synthesized in the same manner as in Example 1 using the compound of the formula (1) as a starting material.

Example 26 2-Acetyl-6- [2- (4-guanidinobenzoyloxy) benzamide] benzoic acid methanesulfonate The compound of Example 1 (3.50 g) was added to anisole (1. 24 ml) and then trifluoroacetic acid (9.
96 ml), and the mixture was stirred at the same temperature for 3 hours under an argon atmosphere. Ether (100 ml) was added to the reaction solution at the same temperature, stirred for about 10 minutes, allowed to stand, and the supernatant was removed by decantation. Add more ether (50 ml)
This operation was repeated. Ether (50 ml) was added to the residue, and the mixture was pulverized sufficiently and then filtered to give the title compound (2.8).
8 g) as a white powder.

Examples 27 to 31 and 33
Example 50 was synthesized in the same manner as in Example 26, using Examples 2 to 6 and Examples 8 to 25 as starting materials.

Example 32 2- [2- (4-guanidinobenzoyloxy) benzamide] -6-methoxybenzoic acid methanesulfonic acid salt The compound of Example 7 (1.40 g) in 1,2-dichloroethane (7.00 ml) ) The suspension was added with anisole (1.36 ml) and trifluoroacetic acid (2.57 ml) under ice-water cooling.
, And stirred at the same temperature for 20 minutes and at room temperature overnight under an argon atmosphere. Under ice-water cooling, ether (30
0 ml) and stirred. After standing, the supernatant was removed by decantation, ether (200 ml) was added, and this operation was repeated. Ether (200m
l) was added and the mixture was pulverized sufficiently, followed by filtration to obtain the title compound (1.12 g) as a white powder.

Example 51 5-acetyl-2- [2- (4-guanidinobenzoyloxy) phenyl] -4H-3,1-benzoxazin-4-one methanesulfonic acid salt The compound of Example 26 (2.70 g) ) Pyridine (80m
l) N, N'-Dicyclohexylcarbodiimide (1.95 g) was added to the solution under ice-water cooling, and the mixture was stirred overnight at room temperature under an argon atmosphere. Precipitated insolubles were removed by filtration, and the filtrate was concentrated under reduced pressure to about 1/4. Ether (200 ml) was added to the residue, and the mixture was sufficiently stirred. After standing, the supernatant was removed by decantation. Further ether (100m
l) was added and the mixture was stirred for about 10 minutes, and then filtered to give the title compound (1.63 g) as a milky white powder.

Embodiments 52 to 75 are similar to Embodiment 27.
Compounds No. to No. 50 were synthesized in the same manner as in Example 51 using the starting materials.

[0104]

[Table 1]

[0105]

[Table 2]

[0106]

[Table 3]

[0107]

[Table 4]

[0108]

[Table 5]

[0109]

[Table 6]

[0110]

[Table 7]

[0111]

[Table 8]

[0112]

[Table 9]

[0113]

[Table 10]

[0114]

[Table 11]

[0115]

[Table 12]

[0116]

[Table 13]

[0117]

[Table 14]

[0118]

[Table 15]

[0119]

[Table 16]

[0120]

[Table 17]

[0121]

[Table 18]

[0122]

[Table 19]

[0123]

[Table 20]

[0124]

[Table 21]

[0125]

[Table 22]

[0126]

[Table 23]

[0127]

[Table 24]

[0128]

[Table 25]

[0129]

[Table 26]

[0130]

[Table 27]

[0131]

[Table 28]

[0132]

[Table 29]

[0133]

[Table 30]

[0134]

[Table 31]

[0135]

[Table 32]

[0136]

[Table 33]

[0137]

[Table 34]

[0138]

[Table 35]

[0139]

[Table 36]

[0140]

[Table 37]

[0141]

[Table 38]

[0142]

[Table 39]

[0143]

[Table 40]

[0144]

[Table 41]

[0145]

[Table 42]

[0146]

[Table 43]

[0147]

[Table 44]

Examples of preparations containing the compound of the present invention are shown below, but the present invention is not limited thereto.

(Formulation Example 1 Capsule) Component Ingredient Amount (g) {Compound of Example 72 250 Lactose 735 Magnesium Stearate 15} ──── The above components were weighed and mixed uniformly. Fill 200 mg of the mixed fraction into a suitable hard capsule,
A capsule was produced.

(Formulation Example 2 Tablet) Component Ingredient Amount (g) {Compound of Example 54 100 Lactose 280 Potato Starch 90 Polyvinyl Alcohol 15 Magnesium Stearate 15} ───────── After the above components were weighed, the title compound, lactose and potato starch were uniformly mixed. An aqueous solution of polyvinyl alcohol was added to the mixture, and granules were prepared by a wet granulation method. The granules were dried, mixed with magnesium stearate, and compressed and compressed to produce tablets weighing 100 mg.

(Formulation Example 3 Granules) Component Ingredient Amount (g) {Compound of Example 60 100 Lactose 500 Maize Starch 350 Hydroxypropylcellulose 50} ─────── After the above components were weighed and uniformly mixed, granules were produced by a conventional method.

(Formulation Example 4 Injection) ───────────────────────── Component Use Amount ───────────化合物 Compound of Example 51 100 mg 5% glucose solution for injection 500 mL ───────────────────────化合物 The compound of Example 51 was dissolved in a 5% glucose solution for injection to prepare an injection for intravenous drip.

[0153]

The 4H-3,1-benzoxazin-4-one derivative of the present invention strongly inhibits various serine proteases, particularly trypsin, elastase and chymotrypsin, which are involved in the cause or exacerbation of pancreatitis. Further, the 4H-3,1-benzoxazin-4-one derivative of the present invention exhibits excellent efficacy in various pancreatitis models and reflux esophagitis models. In addition, the derivative of the present invention is effective in oral administration, has very low toxicity and is excellent in safety. Therefore, the derivative of the present invention exerts a potent serine protease inhibitory action even in clinical (human) or animals to prevent and treat pancreatitis, For example, improvement of acute symptoms of pancreatitis (acute pancreatitis, acute exacerbation of chronic pancreatitis, postoperative acute pancreatitis, acute pancreatitis after pancreatography, traumatic pancreatitis), prevention of relapse of chronic pancreatitis or treatment of chronic pancreatitis, or reflux It is effective in preventing and treating esophagitis.

The compounds of the present invention may furthermore be used for other diseases based on serine proteases, especially trypsin, chymotrypsin and elastase, such as shock,
Acute circulatory failure, sepsis, rheumatoid arthritis, arteriosclerosis,
Prevention of allergies, hay fever, nephritis, certain skin diseases (including atopic dermatitis), thrombosis, disseminated intravascular coagulation (DIC), adult respiratory distress syndrome (ARDS), pulmonary fibrosis, emphysema -It can be used effectively in therapy.

Claims (8)

[Claims] (1) The following formula (I): (Wherein R ¹ and R ² are each independently a halogen atom, an alkyl group having 1 to 4 carbon atoms, an alkoxy group having 1 to 4 carbon atoms, a formula COR ³ (wherein R ³ is a hydrogen atom or Represents an alkyl group having 1 to 4 carbon atoms.), A formula COOR
⁴ (wherein, R ⁴ represents a hydrogen atom or an alkyl group having 1 to 4 carbon atoms), an alkylthio group having 1 to 4 carbon atoms, an alkylsulfonyl group having 1 to 4 carbon atoms, and a par having 1 to 4 carbon atoms. Represents a fluoroalkyl group, an alkyl group having 1 to 4 carbon atoms substituted by an alkoxy group having 1 to 4 carbon atoms, and n represents 0 to
Represents an integer of 2, and when n is 2, each R ² may be the same or different. Or a salt thereof. Wherein R ¹ is a chlorine atom, a bromine atom, a methyl group, an ethyl group, a propyl group, a methoxy group, an ethoxy group, an acetyl group, an ethoxycarbonyl group, a methylthio group, a methanesulfonyl group, a trifluoromethyl group or a methoxy methyl The compound according to claim 1, which is a group or a salt thereof. 3. The compound according to claim 1, wherein the 4-guanidinobenzoyloxy group is bonded to the ortho position of the phenyl group at the 2-position of the benzoxazine ring, or a salt thereof. 4. A compound represented by the following formula (VIII): (Wherein, the definitions of the substituents represented by R ¹ and R ² and n are the same as those of the formula (I), and A represents a carboxyl protecting group) or a deprotected product thereof (Free carboxylic acid) or salts thereof. (5) a compound represented by the following formula (VIII): (Wherein the definitions of the substituents represented by R ¹ and R ² and n are the same as those in formula (I), and A represents a carboxyl protecting group) or a salt thereof, if desired. A method for producing a compound of the formula (I) or a salt thereof, wherein A is deprotected and an intramolecular cyclization reaction is carried out. 6. A compound represented by the following formula (VI): (Wherein the substituents represented by R ¹ and R ² and the definitions of n and A are the same as those of the formula (VIII)) and a salt thereof, and a compound represented by the following formula (VII): ] The method according to claim 4, wherein the compound is reacted with a 4-guanidinobenzoic acid derivative represented by the formula (Y represents a hydroxyl group or a carboxyl group activating group).
A method for producing the compound of II) or a salt thereof. 7. A pharmaceutical composition comprising the compound according to claim 1 or a salt thereof as an active ingredient. 8. A preventive / therapeutic agent for pancreatitis or reflux esophagitis, comprising the compound according to any one of claims 1 to 3 or a salt thereof as an active ingredient.