JPH0940631A - Amidinophenol derivative, method for producing the same, and therapeutic agent for mite allergy - Google Patents

Abstract

(57) [Summary] [Structure] Formula [Chemical Formula 1] (In the formula, R is allyl group, 3-methoxypropyl group, 4-
It represents a piperidyl group, an oxolan-2-ylmethyl group, a carbamoylmethyl group or an ethyl group. ) And the like, a therapeutic agent for mite allergy containing an amidinophenol derivative as an active ingredient, a novel amidinophenol derivative useful as the therapeutic agent for mite allergy, and a method for producing the same. [Effect] Since the amidinophenol derivative used in the present invention has an inhibitory activity against mite protease, it is useful for the treatment and / or prevention of asthma, allergic rhinitis, atopic dermatitis and the like caused by mite.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a mite allergy therapeutic agent containing a certain amidinophenol derivative as an active ingredient. The present invention also relates to an amidinophenol derivative useful as a therapeutic agent for mite allergy and a method for producing the same.

[0002]

BACKGROUND OF THE INVENTION House dust mites inhaled by humans by breathing are known to cause exogenous asthma, allergic rhinitis and atopic dermatitis [J. Allergy, 3
9 , 325 (1967) and ibid, 42 , 14 (1968)].
Recently, Dermatophagoides farinae , a type of house dust mite, and Dermatophagoides pteronais
nyssinus ) and major allergen proteins with molecular weights of 25,000 and 15,000 were isolated [J. Immunol., 125 , 587].
(1980) and Int. Archs Allergy Appl. Immunol., 8
1 , 214 (1986)]. In another report, Dermatophagoides pteronysinus ( Dermatophagoides pterony
Two types of proteases (proteolytic enzymes) have been isolated from ssinus ), and it is considered that the proteolytic activity of these enzymes is closely related to mite allergic disease [Lancet, Jul., 15 , 154 (1989)]. In addition, Dermatophagoides farina
It has been experimentally confirmed that Df-protease, which is a protease isolated from e ), has the ability to produce bradykinin that exacerbates the inflammatory response and enhances vascular permeability [Biochem. Biophys. Acta, 1074 . , 62 (199
1)].

From a comprehensive judgment of these facts, it is speculated that the house dust mite protease promotes the production of bradykinin, resulting in various mite allergic diseases. Therefore, suppressing the protease activity of house dust mite is a treatment for mite allergic diseases (asthma caused by mite, allergic rhinitis, atopic dermatitis, etc.) and / or
Or, it is considered to be useful for prevention.

[0004]

2. Description of the Related Art As substances that inhibit the house dust mite protease, Japanese Patent Laid-Open No. 6-192085 discloses aprotinin,
Potato protease inhibitor, soybean trypsin inhibitor, antipain, leupeptin, guanidino fatty acid derivative, guanidinobenzoic acid derivative and amidinophenol derivative have been reported. In the above-mentioned publication, aprotinin is intravenously administered to guinea pigs to confirm the inhibitory effect on the enhancement of vascular permeability by the house dust mite protease (Df-protease). Also,
The in vitro inhibitory effect of various compounds on Df-protease is measured.

[0005]

[Problems to be Solved by the Invention] Since aprotinin, potato protease inhibitor, soybean trypsin inhibitor, antipain and leupeptin are high molecular weight polypeptides and are difficult to chemically synthesize, the present inventor can easily perform chemical synthesis. Research was conducted with the aim of developing a substance that is a low-molecular compound and that inhibits the house dust mite protease. However, when the compounds whose effects were specifically confirmed in the above-mentioned published specifications were examined for absorbability into blood, it was found that none of them had satisfactory absorbability. It is considered that when the compound has poor absorbability, the therapeutic effect upon oral administration is not sufficiently exerted. Therefore, this time, the present inventor has conducted extensive studies in order to find a compound that strongly inhibits the house dust mite protease and has excellent absorbability. As a result, they have found that certain amidinophenol derivatives, more specifically p- (p-amidinophenoxycarbonyl) -α-methyl-cinnamic acid amides, achieve the object, and completed the present invention.

[0006]

That is, the present invention relates to (1) p- (p-amidinophenoxycarbonyl) -α-methyl-cinnamic acid.
N-allyl-N-ethoxycarbonylmethylamide, p
-(P-amidinoanilinocarbonyl) -α-methyl-
Cinnamic acid / N-allyl-N-ethoxycarbonylmethylamide, p- (p-amidinophenoxycarbonyl) -α
-Methyl-cinnamic acid / N-allyl-N-carboxymethylamide, p- (p-amidinophenoxycarbonyl)-
α-methyl-cinnamic acid ・ N-3-methoxypropyl-N-
Carboxymethylamide, p- (p-amidinophenoxycarbonyl) -α-methyl-cinnamic acid / N-4-piperidinyl-N-carboxymethylamide, p- (p-amidinophenoxycarbonyl) -α-methyl-cinnamic acid / N
-(Oxolan-2-ylmethyl) -N-carboxymethylamide, p- (p-amidinophenoxycarbonyl) -α-methyl-cinnamic acid / N-carbamoylmethyl-
N-carboxymethylamide, and p- (p-amidinophenoxycarbonyl) -α-methyl-cinnamic acid / N-
A remedy for mite allergy containing as an active ingredient one or more compounds selected from ethyl-N-carboxymethylamide, non-toxic salts thereof, and acid addition salts thereof.

(2) p- (p-amidinophenoxycarbonyl) -α-methyl-cinnamic acid · N-, which is a novel compound among the compounds used for the above-mentioned therapeutic agent for mite allergy
4-piperidinyl-N-carboxymethylamide, p-
(P-amidinophenoxycarbonyl) -α-methyl-
Cinnamic acid N- (oxolan-2-ylmethyl) -N-carboxymethylamide, p- (p-amidinophenoxycarbonyl) -α-methyl-cinnamic acid N-carbamoylmethyl-N-carbamoylmethyl-N-carboxymethyl The present invention relates to amides, non-toxic salts thereof, and acid addition salts thereof, and (3) a method for producing the novel compound.

Some of the compounds used for the therapeutic agent for mite allergy of the present invention are disclosed in Japanese Patent Application Nos. 5-252178 and 6-319471, and their corresponding European Patent Publication Nos. 588655 and 656349. It is described in the specification. In these specifications, it is described that the above compound has a phospholipase A ₂ inhibitory activity and is useful for the treatment of allergic diseases based on the activity. Further, some of the compounds used in the present invention are described in Japanese Patent Application No. 6-250158, and it is described that they have an antagonistic action against leukotriene B ₄ and are useful for the treatment of allergic diseases. . However, this information is
It does not suggest that the phospholipase A ₂ inhibitor compound and the leukotriene B ₄ antagonist have inhibitory activity against mite protease.

The compounds used as the active ingredient of the therapeutic agent for mite allergy in the present invention are listed below. Compound 1:

Embedded image p- (p-amidinophenoxycarbonyl) -α-methyl-cinnamic acid / N-allyl-N-ethoxycarbonylmethylamide (Japanese Patent Application No. 5-252178 and European Application Publication)
Compounds described in Example 1i of 588655),

Compound 2:

Embedded image p- (p-amidinoanilinocarbonyl) -α-methyl-cinnamic acid / N-allyl-N-ethoxycarbonylmethylamide (compound described in Example 2ee of Japanese Patent Application No. 6-250158),

Compound 3:

Embedded image p- (p-amidinophenoxycarbonyl) -α-methyl-cinnamic acid / N-allyl-N-carboxymethylamide (compound described in Example 4 of Japanese Patent Application No. 5-252178 and European Patent Application Publication 588655),

Compound 4:

[Chemical 6] p- (p-amidinophenoxycarbonyl) -α-methyl-cinnamic acid.N-3-methoxypropyl-N-carboxymethylamide (described in Example 3 of Japanese Patent Application No. 6-319471 and European Patent Publication 656349). Compound),

Compound 5:

[Chemical 7] p- (p-amidinophenoxycarbonyl) -α-methyl-cinnamic acid · N-4-piperidinyl-N-carboxymethylamide,

Compound 6:

Embedded image p- (p-Amidinophenoxycarbonyl) -α-methyl-cinnamic acid / N- (oxolan-2-ylmethyl) -N
-Carboxymethylamide,

Compound 7:

Embedded image p- (p-amidinophenoxycarbonyl) -α-methyl-cinnamic acid / N-carbamoylmethyl-N-carboxymethylamide,

Compound 8:

Embedded image p- (p-amidinophenoxycarbonyl) -α-methyl-cinnamic acid / N-ethyl-N-carboxymethylamide (compound described in Japanese Patent Application No. 5-252178 and European Patent Application Publication 588655, Example 5u).

Although the above compounds 5, 6 and 7 are novel compounds, they can be synthesized by the method described in Japanese Patent Application No. 5-252178 or European Patent Publication No. 588655 or other known methods. it can. An example is shown below.

Compounds 5, 6, and 7, that is, the following structural formula (I)

[0019]

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The compound represented by the formula (wherein R represents a 4-piperidyl group, an oxolan-2-ylmethyl group or a carbamoylmethyl group) has the general formula (II)

[0021]

[Chemical 12]

(In the formula, t-Bu represents a tert-butyl group and R has the same meaning as described above.) The butyl ester group of the compound represented by the formula (1) is hydrolyzed. Hydrolysis of the t-butyl ester group can be performed, for example, by using an inert organic solvent (methylene chloride, chloroform, methanol,
Dioxane, ethyl acetate, anisole, etc.) in an organic acid (trifluoroacetic acid, etc.) or an inorganic acid (hydrochloric acid, etc.) or a mixture thereof at a temperature of 0 to 90 ° C.

The compound represented by the formula (II) has the general formula (II
I)

[0024]

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(Wherein all symbols have the same meanings as described above), and a compound of the general formula (IV)

[0026]

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It is produced by subjecting a compound represented by: to an esterification reaction. The esterification reaction is known,
Examples include (1) a method using an acid halide, (2) a method using a mixed acid anhydride, and (3) a method using a condensing agent.

Among these methods, the method (3) is a method in which, for example, a carboxylic acid and an alcohol are added to a tertiary amine in an inert organic solvent (chloroform, methylene chloride, dimethylformamide, diethyl ether, etc.) or without a solvent. With or without (pyridine, triethylamine, dimethylaniline, dimethylaminopyridine, etc.),
As a condensing agent, 1,3-dicyclohexylcarbodiimide (DCC), 1-ethyl-3- [3- (dimethylamino) propyl] carbodiimide (EDC), 2-chloro-1-methylpyridinium iodine, etc. are used, and 0-40
The reaction is carried out at ℃. These (1),
Both of the reactions (2) and (3) are preferably carried out under an inert gas (argon, nitrogen, etc.) atmosphere under anhydrous conditions.

The compound represented by the general formula (IV) is a known compound. Further, the compound represented by the general formula (III) can be easily synthesized from a known compound by a known method (see Examples described later). In each reaction in the present specification, the reaction product is a conventional purification means, for example, distillation under normal pressure or reduced pressure, high performance liquid chromatography using silica gel or magnesium silicate, thin film chromatography, or column chromatography, or It can be purified by a method such as washing and recrystallization. Purification may be performed for each reaction or may be performed after completion of several reactions.

[0030]

[Salt and Acid Addition Salt] Suitable salts of the compound used in the present invention include salts of alkali metals (sodium, potassium, etc.), alkaline earth metals (calcium, magnesium, etc.).
Salt, ammonium salt, pharmaceutically acceptable organic amine (tetramethylammonium, triethylamine, methylamine, dimethylamine, cyclopentylamine, benzylamine, phenethylamine, piperidine, monomethanolamine, diethanolamine, tris (hydroxymethyl) aminomethane , Lysine, arginine, N-
Methyl-D-glucan etc.) and the like.

Suitable acid addition salts include inorganic acid salts such as hydrochlorides, hydrobromides, sulfates, phosphates, nitrates,
Or acetate, trifluoroacetate, lactate, tartrate, oxalate, fumarate, maleate, citrate, benzoate, methanesulfonate, ethanesulfonate, benzenesulfonate, Toluene sulfonate,
Organic acid salts such as isethionate, glucuronate and gluconate may be mentioned. Preferred are hydrochloride, methanesulfonate, acetate and trifluoroacetate.

[0032]

BEST MODE FOR CARRYING OUT THE INVENTION Each substance used in the present invention has a strong inhibitory activity against mite protease and an inhibitory effect on inflammation caused by mite protease, and further has good absorbability and extremely low toxicity. Because there are few
It is useful for treating and / or preventing a mite allergic disease to mammals including humans, especially humans, such as asthma, allergic rhinitis, atopic dermatitis and the like caused by mites.

It has been confirmed that the toxicity of the compounds used in the present invention is extremely low. For example, the LD ₅₀ value for intravenous (iv) administration is
117 mg / kg (mouse; male), 134 mg / kg
(Mouse; female), 2 for compound 3 methanesulfonate
50 mg / kg or more (mouse; male), 231 mg / kg
(Mouse; female). Also, with a single oral administration,
There were no deaths at 1000 mg / kg (rat) of the compound 1 acetate, the compound 3 methanesulfonate and the compound 8 acetate. Therefore, it can be judged that all the substances used in the present invention are safe and suitable for use as a medicine.

The substance used in the present invention is usually administered systemically or locally, orally or parenterally. The dose varies depending on the age, body weight, symptoms, therapeutic effect, administration method, treatment time, etc., but is usually 1 mg to 1000 mg per adult, preferably 50 mg to 500 mg, once to several times per day. Orally administered orally, per adult, in the range of 0.1 mg to 100 mg once a day, intravenously once to several times a day, or 5
A 20% ointment is applied to the affected area once to several times a day.
Of course, as described above, since the dose varies depending on various conditions, a dose smaller than the above dose range may be sufficient in some cases, or a dose exceeding the range may be necessary in some cases.

Solid compositions for oral administration according to the present invention include tablets, powders, granules and the like. In such solid compositions, the one or more active substances comprise at least one inert diluent, such as lactose, mannitol, glucose, hydroxypropylcellulose, microcrystalline cellulose, starch, polyvinylpyrrolidone, aluminum metasilicate. It is mixed with magnesium acid. The composition may contain, in a conventional manner, additives other than an inert diluent, for example, a lubricant such as magnesium stearate, a disintegrant such as calcium cellulose glycolate, a stabilizer such as lactose, glutamic acid or asparagine. A solubilizing agent such as an acid may be contained. If necessary, the tablets or pills may be coated with a film of a gastric or enteric substance such as sucrose, gelatin, hydroxypropylcellulose, hydroxypropylmethylcellulose phthalate, or may be coated with two or more layers. Further, the capsule may be made of an absorbable substance such as gelatin.

Liquid compositions for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, elixirs and the like, commonly used inert diluents, For example, it contains purified water and ethanol. The composition may contain, in addition to the inert diluent, adjuvants such as wetting agents and suspending agents, sweetening agents, flavoring agents, fragrances, and preservatives.
Other compositions for oral administration include sprays which contain one or more active substances and are formulated in a manner known per se.

Injections for parenteral administration according to the present invention include sterile aqueous or non-aqueous solutions, suspensions and emulsions. Aqueous solutions and suspensions include, for example, distilled water for injection and physiological saline. Examples of the water-insoluble solutions and suspensions include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, alcohols such as ethanol, and polysorbate 80. Such compositions may also contain adjuvants such as preserving, wetting, emulsifying, dispersing, stabilizing (eg, lactose) and solubilizing agents (eg, glutamic acid, aspartic acid). These are sterilized by, for example, filtration through a bacteria retaining filter, blending of a bactericide or irradiation. They can also be used to produce sterile solid compositions which are dissolved in sterile water or sterile injectable solvents before use. The ointment for parenteral administration according to the present invention can be manufactured by a conventional method.

[0038]

EXAMPLES AND EXPERIMENTAL EXAMPLES Hereinafter, the present invention will be described in more detail with reference to Reference Examples, Examples and Experimental Examples, but the present invention is not limited to these Examples and Experimental Examples. Reference Example 1: 1- (tert-butoxycarbonyl) -4-hydroxypiperidine

Embedded image

4-hydroxypiperidine hydrochloride (8.6
A suspension of g) in methylene chloride (150 ml) was cooled to 0 ° C., triethylamine (14 ml) and di-tert-butyl-dicarbonate (20.7 ml) were added, and the solution was stirred at room temperature for 2 hours. The reaction solution was poured into chilled 1N hydrochloric acid, the two layers were separated, the aqueous layer was extracted with chloroform, the organic layer was washed successively with 1N hydrochloric acid and brine, dried over sodium sulfate, and concentrated under reduced pressure. 20.7 g of the title compound having the following physical data was obtained as a crude product. TLC (ethyl acetate: n-hexane = 1: 1): Rf =
0.29.

Reference Example 2: 1- (tert-butoxycarbonyl) -4-oxopiperidine

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A methylene chloride solution (80 ml) of oxalyl chloride (7.9 ml) was cooled to −78 ° C., and anhydrous dimethyl sulfoxide (hereinafter DMSO) was added under an argon atmosphere.
Abbreviated. ) (6.4 ml) in methylene chloride (20 m
1) was added dropwise and stirred for 5 minutes. The hydroxy compound (produced in Reference Example 1, 20.7 g) dissolved in methylene chloride (40 ml) was added dropwise to this solution over 10 minutes, and the mixture was further stirred for 25 minutes. Triethylamine (63 ml) was added to the reaction mixture, the mixture was returned to room temperature, stirred for 20 minutes, water was added, and the obtained yellow solution was separated into two layers.
The aqueous layer was extracted with chloroform.

The organic layer was washed 3 times with 1N hydrochloric acid, a saturated sodium hydrogen carbonate solution and brine successively, dried over sodium sulfate, the solvent was distilled off under reduced pressure, and the obtained brown solid was washed with n-hexane. And the title compound 1 having the following physical properties
4.0 g was obtained as a white powder. TLC (ethyl acetate: n-hexane = 1: 1): Rf =
0.44.

Reference Example 3: 1- (tert-butoxycarbonyl) -4- (tert-butoxycarbonylmethylamino)
Piperidine

Embedded image

Oxo compound (synthesized in Reference Example 2, 3.0 g)
And glycine t-butyl ester hydrochloride (2.5 g) in methanol (50 ml) under argon atmosphere, triethylamine (2.3 ml) and 10% palladium-carbon (3 ml).
(00 mg) was added, and the mixture was stirred under a hydrogen atmosphere at room temperature overnight. Palladium-carbon was filtered over Celite, and the filtrate was concentrated and dried to give the title compound as a crude product having the following physical properties. TLC (ethyl acetate: n-hexane = 2: 1): Rf =
0.39.

Reference Example 4: p- (methoxycarbonyl)-
α-Methyl-cinnamic acid / N- [1- (tert-butoxycarbonyl) piperidin-4-yl] -N- (tert-butoxycarbonylmethyl) amide

Embedded image

A solution of p- (methoxycarbonyl) -α-methylcinnamic acid (2.64 g) in thionyl chloride (20 ml) was added to 1 part.
After refluxing at 00 to 110 ° C. for 45 minutes, excess thionyl chloride was distilled off under reduced pressure, and the residue was azeotroped twice with 20 ml of toluene to completely remove thionyl chloride to prepare a white solid of acid chloride. The white solid was dissolved in 10 ml of methylene chloride under an argon atmosphere. This solution was added dropwise to a mixed solution of an ice-cooled amine compound (produced in Reference Example 3) in methylene chloride (15 ml) and pyridine (15 ml). The resulting yellow solution was stirred at room temperature for 30 minutes,
It was poured into chilled 1N hydrochloric acid. The aqueous layer was extracted with chloroform, and the organic layers were combined and washed with a saturated sodium hydrogen carbonate solution to remove p- (methoxycarbonyl) -α-methyl-cinnamic acid, followed by washing with water and drying with sodium sulfate. 4.1 g of the title compound having the following physical data after concentration under reduced pressure
Was obtained as a crude product (yellow oil). TLC (ethyl acetate: n-hexane = 1: 1): Rf =
0.42.

Reference Example 5: p-carboxy-α-methylcinnamic acid.N- [1- (tert-butoxycarbonyl) piperidin-4-yl] -N- (tert-butoxycarbonylmethyl) amide

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Methyl ester compound (produced in Reference Example 4,
1N to 4.1 g of 1,4-dioxane solution (20 ml)
Sodium hydroxide solution (7 ml) was added, and the mixture was stirred at room temperature overnight. Pour the reaction solution into ice water and adjust the pH to 1N with hydrochloric acid.
It was adjusted to 1 and extracted with ethyl acetate. After washing with water, drying over sodium sulfate and concentration under reduced pressure, the title compound
1.46 g was obtained. The obtained compound was used for the next reaction without purification.

Reference Example 6: p- (p-amidinophenoxycarbonyl) -α-methyl-cinnamic acid.N- [1- (tert
-Butoxycarbonyl) piperidin-4-yl] -N-
(Tert-Butoxycarbonylmethyl) amide

Embedded image

Carboxylic acid derivative (manufactured in Reference Example 5, 1.46)
Dicyclohexylcarbodiimide (0.91 g) was added to a pyridine solution (20 ml) of g) and 4-amidinophenol hydrochloride (0.55 g), and the mixture was stirred at room temperature overnight. The resulting white precipitate was filtered, and the filtrate was concentrated to give an oily substance which was purified by column chromatography to give 1.43 g of the title compound having the following physical data. TLC (chloroform: methanol: acetic acid = 10: 2:
1): Rf = 0.43.

Example 1 p- (p-amidinophenoxycarbonyl) -α-methyl-cinnamic acid.N- (4-piperidinyl) -N-carboxymethylamide.2 trifluoroacetic acid salt

[Chemical 21]

Trifluoroacetic acid (20 ml) was added to the anisole solution (20 ml) of the t-butyl ester compound (1.43 g produced in Reference Example 6), the mixture was stirred at room temperature for 5 hours, and then concentrated to give an ether residue. Solidified with to give a white powder. This is washed repeatedly with chloroform and ether,
After drying, 1.07 g of the title compound having the following physical data was obtained. TLC (ethyl acetate: acetic acid: water = 3: 1: 1): Rf =
0.10; IR (cm ^-1 ): ν = 3372, 3097, 1737, 1676, 1609, 14
84, 1439, 1268, 1206, 1181, 1137, 1069, 1016, 838,
723; NMR (CD ₃ OD): δ = 8.22 (2H, d, J = 8Hz), 7.92 (2H, d, J
= 8Hz), 7.7-7.4 (4H.m), 6.70 (1H.s), 4.5-4.0 (3H.br),
3.6-3.4 (2H.m), 3.2-3.0 (2H.m), 2.3-1.9 (7H.br).

Reference Example 7: p- (methoxycarbonyl)-
α-Methyl-cinnamic acid / N- (oxolan-2-ylmethyl) -N- (tert-butoxycarbonylmethyl) amide

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The same procedure as in Reference Example 4 except that N- (oxolan-2-ylmethyl) -N- (tert-butoxycarbonylmethyl) amine was used in place of the amine compound (produced in Reference Example 3) in Reference Example 4. By the method of 4., 4.69 g of the title compound having the following physical data was obtained as a crude product (yellow oil). TLC (ethyl acetate: n-hexane = 1: 1): Rf =
0.45.

Reference Example 8: p- (p-amidinophenoxycarbonyl) -α-methyl-cinnamic acid.N- (oxonlan-2-ylmethyl) -N- (tert-butoxycarbonylmethyl) amide

Embedded image

Instead of the methyl ester compound (produced in Reference Example 4) in Reference Example 5, a methyl ester compound (Reference Example 7) was used.
2.69 g of the title compound having the following physical data was obtained in the same manner as in Reference Example 5 → Reference Example 6 using 4.69 g of the compound prepared in 1.). TLC (chloroform: methanol: acetic acid = 10: 2:
1): Rf = 0.42.

Example 2: p- (p-amidinophenoxycarbonyl) -α-methyl-cinnamic acid.N- (oxolan-2-ylmethyl) -N-carboxymethylamide.methanesulfonic acid hydrochloride

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Trifluoroacetic acid (10 ml) was added to (20 ml) of an anisole solution of t-butyl ester compound (produced in Reference Example 8, 2.79 g), stirred at room temperature for 14 hours, concentrated, and solidified with ether. The white powder thus obtained was washed with chloroform and ether and dried. The obtained compound was dissolved in 10 ml of acetic acid, and methanesulfonic acid (0.29
ml) was added and concentrated, and the obtained oily substance was solidified with ether to obtain 2.49 g of the title compound having the following physical data. TLC (chloroform: methanol: acetic acid = 10: 2:
1): Rf = 0.3; IR (cm ⁻¹ ): ν = 3370, 1737, 1687, 1606, 1480, 14
12, 1267, 1210, 1177, 1060, 1015, 887, 785, 562, 5
37; NMR (CD ₃ OD): δ = 8.21 (2H, d, J = 8Hz), 7.92 (2H, d, J
= 8Hz), 7.6-7.45 (4H, m), 6.73 and 6.65 (1H, s, rotame
r), 4.5-4.3 (1H, m), 4.3-4.0 (2H, br), 4.0-3.7 (3H, m),
3.7-3.5 (1H, br), 2.70 (3H, s), 2.17 and 2.10 (3H, s, ro
tamer), 2.2-1.8 (3H, m), 1.8-1.4 (1H, m).

Reference Example 9: 4- (tetrahydropyranyloxymethyl) benzaldehyde

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A solution of 1- (tetrahydropyranyloxymethyl) -4-hydroxymethyl-benzene (7.38 g) and triethylamine (32 ml) in DMSO (50 m).
1), DMS of sulfur trioxide pyridine complex (17 g)
O solution (50 ml) was added, the mixture was stirred at room temperature for 30 minutes, and ice water was added. The reaction solution was extracted with ethyl acetate, 1N hydrochloric acid,
The extract was washed successively with water, saturated sodium hydrogen carbonate solution and brine, dried over magnesium sulfate and concentrated under reduced pressure to give the title compound (7.51 g) having the following physical data. TLC (ethyl acetate: n-hexane = 1: 1): Rf =
0.816.

Reference Example 10: p- (Tetrahydropyranyloxymethyl) -α-methyl-cinnamic acid ethyl ester

[Chemical formula 26]

A solution of ethyl-2- (diethoxyphosphinyl) propionate (10.1 g) in tetrahydrofuran (30 ml) was added at 0 ° C. to sodium hydride (1.7 g).
Was added and stirred for 30 minutes, and a solution of an aldehyde compound (prepared in Reference Example 9, 7.51 g) in tetrahydrofuran (30 m
l) was added and the mixture was stirred at room temperature for 1 hour. Ice water was added to the reaction solution and the mixture was extracted with ethyl acetate. The organic layer was washed with water and brine successively, dried over magnesium sulfate and concentrated under reduced pressure. The residue was purified by column chromatography (n-hexane: ethyl acetate = 4: 1) to obtain 9.01 g of the title compound having the following physical properties. TLC (n-hexane: ethyl acetate = 2: 1): Rf =
0.783.

Reference Example 11: p- (Tetrahydropyranyloxymethyl) -α-methyl-cinnamic acid

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5 N sodium hydroxide (12 ml) and ethanol (50 ml) were added to the ethyl ester compound (manufactured in Reference Example 10, 9.10 g), and the mixture was stirred at room temperature for 4 hours and then at 50 ° C. for 1 hour. 2N hydrochloric acid (30m
l) was added. The reaction mixture was extracted with ethyl acetate, washed successively with water and brine, dried over magnesium sulfate and concentrated under reduced pressure to give the title compound (11.6 g) having the following physical properties. TLC (n-hexane: ethyl acetate = 2: 1): Rf =
0.575.

Reference Example 12: N-carbamoylmethyl-N
-(Tert-Butoxycarbonylmethyl) amine

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1-carbamoylmethylamine (20.1 g)
And an aqueous solution of 1N sodium hydroxide (180 ml) (10
Bromoacetic acid tert-butyl ester (14.7
(ml) in dioxane (350 ml) was added, and the mixture was stirred at room temperature for 1 hr. The reaction mixture was extracted with chloroform, washed with brine, dried over magnesium sulfate, and concentrated under reduced pressure to give the title compound (14.4 g) having the following physical data. TLC (chloroform: methanol = 9: 1): Rf =
0.281.

Reference Example 13: p- (Tetrahydropyranyloxymethyl) -α-methyl-cinnamic acid N-carbamoylmethyl-N- (tert-butoxycarbonylmethyl) amide

[Chemical 29]

Cinnamic acid compound (prepared in Reference Example 11, 1
3.4 g) and an amine compound (produced in Reference Example 12, 14.4 g)
1-ethyl-3-in a pyridine solution (100 ml) of
[3- (Dimethylamino) propyl] carbodiimide (14 g) was added, the mixture was stirred at room temperature overnight, concentrated, ice water was added, and the mixture was extracted with ethyl acetate. 1N hydrochloric acid, water,
It was washed successively with saturated sodium hydrogen carbonate solution and brine, dried over magnesium sulfate and concentrated under reduced pressure, and the residue was subjected to column chromatography (n-hexane: ethyl acetate = 1: 1).
Purification by → 1: 3 → 0: 1) gave 15.9 g of the title compound having the following physical data. TLC (ethyl acetate): Rf = 0.467.

Reference Example 14: p-Hydroxymethyl-α-
Methyl-cinnamic acid / N-carbamoylmethyl-N- (tert
-Butoxycarbonylmethyl) amide

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A 65% acetic acid solution (1%) was added to the tetrahydropyranyloxymethyl derivative (produced in Reference Example 13, 15.9 g).
50 ml) and tetrahydrofuran (15 ml) were added,
The mixture was stirred at 50 ° C for 3 hours, ice water was added, and the mixture was extracted with ethyl acetate. The organic layer was washed successively with water, saturated sodium hydrogen carbonate solution and brine, dried over magnesium sulfate and concentrated under reduced pressure to give the title compound (10.7 g) having the following physical data. TLC (chloroform: methanol = 9: 1): Rf =
0.185.

Reference Example 15: p-formyl-α-methyl-
Cinnamic acid / N-carbamoylmethyl-N- (tert-butoxycarbonylmethyl) amide

[Chemical 31]

In Reference Example 9, instead of 1- (tetrahydropyranyloxymethyl) -4-hydroxymethyl-benzene, a hydroxymethyl compound (prepared in Reference Example 14,
Using 10.7 g) and in the same manner as in Reference Example 9, 9.87 g of the title compound having the following physical properties was obtained as a crude product. TLC (chloroform: methanol = 9: 1): Rf =
0.357.

Reference Example 16: p-carboxy-α-methyl-cinnamic acid.N-carbamoylmethyl-N- (tert-butoxycarbomylmethyl) amide

Embedded image

Formyl body (manufactured in Reference Example 15, 9.87)
g) in acetonitrile solution (50 ml) at 0 ° C., an aqueous solution of sodium dihydrogen phosphate (2.1 g) (15 ml)
Then, add 31% hydrogen peroxide (3 ml) and an aqueous solution of sodium chlorite (5.0 g) (15 ml), and add 2 at room temperature.
Stirred for hours. A saturated sodium carbonate solution and ether were added to the reaction mixture, and the aqueous layer was washed with ether. 1 in the water layer
After adjusting to pH 1 by adding N hydrochloric acid, the product was extracted with ethyl acetate. The organic layer was washed successively with water and brine, dried over magnesium sulfate and concentrated under reduced pressure to give the title compound (8.07 g) having the following physical properties as a crude product. TLC (chloroform: methanol: acetic acid = 10: 2:
1): Rf = 0.757.

Reference Example 17: p- (p-amidinophenoxycarbonyl) -α-methyl-cinnamic acid.N-carbamoylmethyl-N- (tert-butoxycarbonylmethyl) amide.acetate

[Chemical 33]

Using the carboxylic acid body (7.04 g) produced in Reference Example 16 instead of the carboxylic acid body used in Reference Example 6, the title compound (7.80 g) having the following physical properties was obtained in the same manner as in Reference Example 6. Got TLC (chloroform: methanol: acetic acid = 10: 2:
1): Rf = 0.30.

Example 3: p- (p-amidinophenoxycarbonyl) -α-methyl-cinnamic acid.N-carbamoylmethyl-N-carboxymethylamide.trifluoroacetic acid salt

Embedded image

The t-butyl ester compound (7.80 g) produced in Reference Example 17 was used in place of the t-butyl ester compound used in Example 1, and the following physical properties were determined in the same manner as in Example 1. 6.67 g of the title compound are obtained. TLC (ethyl acetate: acetic acid: water = 3: 1: 1): Rf =
0.23. IR (cm ^-1 ): ν = 3328, 3119, 1736, 1669, 1610, 14
93, 1412, 1301, 1272, 1240, 1212, 1189, 1136, 107
2, 1017. NMR (CD ₃ OD): δ = 8.20 (2H, d, J = 8Hz), 7.90 (2H, d, J
= 9Hz), 7.60-7.50 (4H, m), 6.75 (1H, br.), 4.40-4.15 (4
H, m), 2.15 (3H, br.s).

[0079] Experimental Example 1: inhibitory effect on house dust mite protease (in vitro) Experimental method:.... A house dust mite protease (Df, proteases) from the culture extracts of house dust mite (Dermatophagoidesfarinae), Int Arch Allergy Appl Immunol, 91, 8
0 (1990). The purified enzyme [150 units / mg (37 ° C, pH 8)] was stored at -80 ° C in a phosphate buffer solution (PBS) (1 mg / ml). The stored enzyme solution was diluted with cold 1 mM hydrochloric acid before use. The activity of the enzyme depends on the synthetic substrate (Boc-Gln
-Gly-Arg-MCA) at 37 ° C by the method of Kawabata et al. [Eur. J. Biochem., 172 , 17 (1988)].
Was determined fluorescently according to. Various compounds (0-0.2μ
M) in 50 mM Tris-HCl buffer (pH 8.0, 188)
To 0 μl) was added the substrate (0-50 μM, 100 μl) dissolved in dimethylsulfoxide. After adding the enzyme (1.1 nM, 20 μl) dissolved in 1 mM hydrochloric acid, the initial value of hydrolysis was excited at 380 nm, emitted at 440 nm, and measured by a fluorescence spectrophotometer.

The inhibitory effect of each compound on Df-protease was evaluated by changing the molar ratio of each compound to Df-protease. That is, a mixture of Df-protease and each compound was incubated at 37 ° C. for 1 to 3 minutes (the incubation time was changed depending on each compound), and the residual enzyme activity was measured under the same conditions as above. Ki value is Lineweave
Calculated from r-Burk plot. Table 1 shows the results.

[0081]

[Table 1]

Comparative compounds X, Y and Z in Table 1 are the compounds shown below.

Compound X: Compound 18 described in JP-A-6-192085

Embedded image p- (p-amidinophenoxycarbonyl) benzoic acid
2R-benzyloxycarbonyl-1-pyrrolidinylamide,

Compound Y: Compound 19 described in JP-A-6-192085

Embedded image 4-[(4-amidino-2-methoxycarbonyl) phenoxycarbonyl] benzoic acid N-phenyl-N-ethoxycarbonylmethylamide,

Compound Z: Compound 20 described in JP-A-6-192085

Embedded image p- (p-amidinophenoxycarbonyl) -α-methylcinnamic acid / N- [1R-ethoxycarbonyl-2- (3
-Indolyl) ethyl] amide. ─

Regarding the inhibitory effect of Df-protease in vitro , the compounds used in the present invention are disclosed in
It is understood from Table 1 that it is comparable to the amidinophenol derivative described in the specification of 6-192085.

Experimental Example 2: Absorption of Drugs When the compound was orally administered, it was decided to examine the absorption from the ratio of absorption from the digestive tract and transfer to blood. That is, each compound was orally administered, blood was collected after a certain period of time, and the amount of the compound in plasma was measured using the inhibitory activity against trypsin as an index. When examining the absorption amount of a compound using the inhibitory activity against trypsin as an index, it is an essential condition that each compound has the same inhibitory activity. The compounds used in the present invention and the comparative compound have almost the same inhibitory activity (IC) against trypsin.
It is known to have ₅₀ = 1 x 10 ^-7 M).

Experimental method: Trypsin inhibitory activity in blood after oral administration to rats ( ex vivo ) Wistar male rats (8 weeks old, 210 to 2) fasted overnight.
60 g), the drug was dissolved in distilled water to give 100 mg /
Single dose was given orally at kg / 10 ml. Control group (cont
rol) was administered with distilled water. 0 from each animal after administration
Citrate blood collection from the jugular vein after 0.5, 1, 3, 6, 9 and 24 hours (3.8% sodium citrate: blood = 0.1 m
l: 0.9 ml) and centrifuged (4 ° C, 3000 rpm, 10 mi
Plasma was obtained according to n). 2 μg / ml trypsin 0.1 ml and 0.1 mM Bo in 0.1 ml of plasma or distilled water
0.8 ml of c-Phe-Scr-Arg-MCA was added to make the total volume 1 ml, and the mixture was reacted at 37 ° C. for 15 minutes. 1 ml of 30% acetic acid was added, and after the reaction was stopped, 1 ml of distilled water was added, and the fluorescence of liberated 7-amino-4-methyl-coumarin (AMC) was measured with a fluorometer (Ex = 380 nm, Em = 460n).
m). The results are shown in Table 2.

[0089]

[Table 2]

Comparative compounds X, Y and Z in Table 2 are the same as those listed in Table 1 of Experimental Example 1. It can be seen from Table 2 that the compound of the present invention has a significantly improved absorbability upon oral administration as compared with the amidinophenol derivative described in JP-A-6-192085. In detail, although both trypsin inhibitory activities in vitro are similar, the blood trypsin inhibitory activity (maximum inhibition%) when actually orally administered was 59 to 98 in the group of the compound of the present invention.
%, Whereas 0 in the compound group of JP-A-6-192085
It was ~ 18.9%. That is, the strength of the blood trypsin inhibitory activity directly reflects the good or bad absorbability. As described above, it was revealed that the compound used in the present invention has remarkably excellent absorbability upon oral administration as compared with the compound disclosed in JP-A-6-192085. As shown in Table 1, since the in vitro inhibitory effect against mite proatease is comparable, when actually orally administered, the compound of the present invention exerts a markedly superior therapeutic effect over the comparative compound. It is easy to guess.

─────────────────────────────────────────────────── ─── Continuation of the front page (51) Int.Cl. ⁶ Identification code Internal reference number FI Technical indication C07D 307/14 C07D 307/14

Claims (6)

[Claims] 1. P- (p-amidinophenoxycarbonyl) -α-methyl-cinnamic acid / N-allyl-N-ethoxycarbonylmethylamide, p- (p-amidinoanilinocarbonyl) -α-methyl-cinnamic acid・ N-allyl-N-
Ethoxycarbonylmethylamide, p- (p-amidinophenoxycarbonyl) -α-methyl-cinnamic acid / N-allyl-N-carboxymethylamide, p- (p-amidinophenoxycarbonyl) -α-methyl-cinnamic acid / N −
3-methoxypropyl-N-carboxymethylamide,
p- (p-amidinophenoxycarbonyl) -α-methyl-cinnamic acid · N-4-piperidinyl-N-carboxymethylamide, p- (p-amidinophenoxycarbonyl) -α-methyl-cinnamic acid · N- (oxolane -2-ylmethyl) -N-carboxymethylamide, p- (p-
Amidinophenoxycarbonyl) -α-methyl-cinnamic acid / N-carbamoylmethyl-N-carboxymethylamide, and p- (p-amidinophenoxycarbonyl)
-A-methyl-cinnamic acid / N-ethyl-N-carboxymethylamide, a non-toxic salt thereof, and a mite allergy therapeutic agent containing one or more compounds selected from non-toxic salts and acid addition salts thereof as an active ingredient . 2. p- (p-Amidinophenoxycarbonyl) -α-methyl-cinnamic acid / N-allyl-N-ethoxycarbonylmethylamide, p- (p-amidinoanilinocarbonyl) -α-methyl-cinnamic acid・ N-allyl-N-
Ethoxycarbonylmethylamide, p- (p-amidinophenoxycarbonyl) -α-methyl-cinnamic acid / N-allyl-N-carboxymethylamide, p- (p-amidinophenoxycarbonyl) -α-methyl-cinnamic acid / N −
3-methoxypropyl-N-carboxymethylamide,
p- (p-amidinophenoxycarbonyl) -α-methyl-cinnamic acid / N-4-piperidinyl-N-carboxymethylamide, p- (p-amidinophenoxycarbonyl) -α-methyl-cinnamic acid / N- (oxolane -2-ylmethyl) -N-carboxymethylamide, p- (p-
Amidinophenoxycarbonyl) -α-methyl-cinnamic acid / N-carbamoylmethyl-N-carboxymethylamide, and p- (p-amidinophenoxycarbonyl)
Mite for oral administration containing one or more compounds selected from -α-methyl-cinnamic acid / N-ethyl-N-carboxymethylamide, non-toxic salts thereof, and acid addition salts thereof as an active ingredient Allergy remedy. 3. The active ingredient is p- (p-amidinophenoxycarbonyl) -α-methyl-cinnamic acid / N-allyl-
The therapeutic agent for mite allergy according to claim 1, which is N-ethoxycarbonylmethylamide. 4. The active ingredient is p- (p-amidinophenoxycarbonyl) -α-methyl-cinnamic acid / N-allyl-
The mite allergy therapeutic agent for oral administration according to claim 2, which is N-ethoxycarbonylmethylamide. 5. p- (p-amidinophenoxycarbonyl) -α-methyl-cinnamic acid.N-4-piperidinyl-N
-Carboxymethylamide, p- (p-amidinophenoxycarbonyl) -α-methyl-cinnamic acid / N- (oxolan-2-ylmethyl) -N-carboxymethylamide, or p- (p-amidinophenoxycarbonyl)
-Α-methyl-cinnamic acid / N-carbamoylmethyl-N-
Carboxymethylamides, their non-toxic salts, or their acid addition salts. 6. Formula (II): (In the formula, R represents a 4-piperidyl group, an oxolan-2-ylmethyl group or a carbamoylmethyl group, and t-B
u represents a tert-butyl group. ) Of the compound represented by
-Characterized by hydrolyzing a butyl ester group,
Formula (I) (The symbols in the formula have the same meanings as above.)