JP2002302470A - New quaternary ammonium salt and method for producing the same - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain an antibacterial agent exhibiting high antibacterial activity and a wide antibacterial spectrum and having a high safety to the human body. SOLUTION: The antibacterial agent is a bis-quaternary ammonium salt represented by formula (1) (wherein, R1 and R2 are each a 4-12C alkyl; R3 is a 1-2C alkyl; (k) is an integer of 0-4; X is an inorganic or an organic anion; (n) is a valence of the anionic group X and is 1 or 2; and (m) is 2 when (n) is 1 and is 1 when the (n) is 2). The method for producing the salt is also provided. The compound is especially useful as an antiseptic agent for eye drops.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to a novel quaternary ammonium salt and a method for producing the same, and the compound of the present invention is useful as an antibacterial agent such as a preservative for eye drops.

[0002]

2. Description of the Related Art Quaternary ammonium salts having antibacterial activity have been known for a long time and are still widely used at present. However, in the conventional quaternary ammonium salts, the bactericidal activity and the antibacterial activity are usually reduced by the action of carbohydrates, proteins and lipids, and in the acidic region having a low pH, the effects are reduced on the cell spores. There are drawbacks such as no.

As a method for solving the above-mentioned problem, a structure called a quaternary ammonium in which four alkyl groups or the like are bonded to one nitrogen atom in Japanese Patent Application Laid-Open Nos. Hei 6-321902 and Hei 10-114604 is disclosed. An antibacterial agent comprising two compounds in one molecule, that is, a quaternary ammonium salt has been proposed. The compounds described in these publications are compounds in which two or more methyl groups or ethyl groups are bonded to one nitrogen atom constituting the quaternary ammonium, have high antibacterial activity, and have good antibacterial performance. It is actually used as an antibacterial agent. However, although these compounds have high antibacterial activity, improvement has been desired in that their safety to the human body is not sufficient.

[0004]

An object of the present invention is to provide a novel compound which exhibits high antibacterial activity and a broad antibacterial spectrum and is useful as an antibacterial agent which is highly safe for the human body.

[0005]

Means for Solving the Problems The present invention provides the following formula (1)
Is a quaternary ammonium salt represented by

[0006]

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In the formula (1), R ₁ and R ₂ are each an alkyl group having 4 to 12 carbon atoms, which may be the same or different, and R ₃ is an alkyl group having 1 to 2 carbon atoms. Is a group, k is 0 to 4, X ^n- is an inorganic or organic anionic group, n is a valence of the anionic group X ^n- and is 1 or 2, and m is n It is 2 when 1 and 1 when n is 2. ]

The compounds of the present invention can be prepared by reacting the following [I] (hereinafter abbreviated as the first type), [II] (hereinafter abbreviated as the second type) or [III] (hereinafter abbreviated as the third type). Is obtained by Reaction [I]:

[0009]

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[In the formula (6), k is 0 to 4;
Represents a chlorine atom, a bromine atom or an iodine atom. ] [In the formula (7), R ₁ and R ₂ each represent an alkyl group having 4 to 12 carbon atoms, and R ₃ represents an alkyl group having 1 to 2 carbon atoms. ]

Reaction [II]:

[0012]

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[In the formula (8), R ₁ and R ₂ are each an alkyl group having 4 to 12 carbon atoms, R ₁ and R ₂ may be the same or different, and k is 0 to 0. 4 is represented. ]

In the formula (9), R ₃ represents an alkyl group having 1 to 2 carbon atoms, and Z represents a chlorine atom, a bromine atom, an iodine atom or any of the following formulas (10) and (11). Represent. ]

[0015]

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Wherein R ₆ represents an alkyl group having 1 to 2 carbon atoms. ]

[0017]

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Wherein R ₇ and R ₈ each represent a hydrogen atom, an alkyl group having 1 to 12 carbon atoms, or a carboxyl group. ]

Reaction [III]:

[0020]

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[In the formula (12), R ₁ has 4 to 12 carbon atoms.
R ₃ is an alkyl group having 1 to 2 carbon atoms, and k represents 0 to 4. ]

[In the formula (13), R ₂ has 4 to 12 carbon atoms.
J represents a chlorine atom, a bromine atom, an iodine atom or any of the groups of the above formula (11). ]

[0023]

DETAILED DESCRIPTION OF THE INVENTION A. Novel Compound The present invention is a novel quaternary ammonium salt represented by the following formula (1).

[0024]

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[In the formula (1), R ₁ and R ₂ are each an alkyl group having 4 to 12 carbon atoms, R ₁ and R ₂ may be the same or different, and R ₃ is a carbon atom. Number 1-2
Wherein k is 0 to 4, X ^n- is an inorganic or organic anion group, and n is an anion group X ^n-
Is 1 or 2, and m is 2 when n is 1, and 1 when n is 2. ]

In the above formula (1), as the alkyl group represented by R ₁ and R ₂ , those having 4 to 12 carbon atoms can be used. Those having several 5 to 10 are preferable, and those having 6 to 8 carbon atoms are more preferable. R ₃ may be either a methyl group or an ethyl group, but a methyl group is more preferable for exhibiting high antibacterial performance.

X ^{n- in the} above formula (1) is an inorganic or organic anion group, preferably iodine ion, bromide ion, chloride ion, fluorine ion, iodate ion,
Bromate ion, chlorate ion, periodate ion, perchlorate ion, chlorite ion, hypochlorite ion, nitrate ion, nitrite ion, sulfate ion, hydroxide ion or the following formula (2) And an anionic group represented by (5). The number (m) at which the anion group X ^n- is bonded to the compound of the formula (1) is a number such that the product of n and m is 2, assuming that the valence of the anion group X ^n- is n. , An anionic group X ^n-
Is one in the case of divalent, and two in the case of monovalent.

[0028]

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[In the formula, R ₄ represents an alkyl group or an alkenyl group having 1 to 7 carbon atoms which may have a hydroxyl group or a carbonyl group. ]

[0030]

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Wherein R ₅ is absent (COO bonds directly) or is an alkylene or alkenylene group having 1 to 8 carbon atoms, and these organic groups may have a hydroxyl group. . ]

[0032]

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[In the formula, R ₆ represents an alkyl group having 1 to 2 carbon atoms. ]

[0034]

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Wherein R ₇ and R ₈ each represent a hydrogen atom, an alkyl group having 1 to 12 carbon atoms, or a carboxyl group. ]

B. Production Method of New Compound The quaternary ammonium salt represented by the above formula (1) can be produced by the following three methods.

B-1. First Method of Production The first method of producing a quaternary ammonium salt represented by the above formula (1) comprises a halide of the following formula (6) and a halide of the following formula (7): This is a method of producing by reacting with a tertiary amine.

[0038]

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Wherein k is 0 to 4, Y is a chlorine atom,
Represents a bromine atom or an iodine atom. ]

[0040]

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[Wherein R ₁ and R ₂ each have 4 to 12 carbon atoms.
Wherein R ₁ and R ₂ may be the same or different, and R ₃ represents an alkyl group having 1 to 2 carbon atoms. ]

Compounds of the above general formula (6) include α,
α'-dichloro-o-xylene, α, α'-dichloro-
m-xylene, α, α′-dichloro-p-xylene,
α, α′-dibromo-o-xylene, α, α′-diiodo-m-xylene, 3,6-bis (chloromethyl) durene, 2,4-bis (chloromethyl) -1,3,5-trimethyl Benzene, 1,4-bis (chloromethyl)-
Halides such as 2,5-dimethylbenzene and 2,5-bis (chloromethyl) toluene are exemplified.

The compounds of the formula (7) include N, N-
Dipentyl-N-methylamine, N, N-dihexyl-N
-Methylamine, N, N-diheptyl-N-methylamine, N-butyl-N-hexyl-N-methylamine, N-
Pentyl-N-hexyl-N-methylamine, N-hexyl-N-octyl-N-methylamine, N, N-dipentyl-N-ethylamine, N, N-dihexyl-N-ethylamine, N-pentyl-N- Tertiary amines such as hexyl-N-ethylamine.

These reactions are carried out in a suitable organic solvent in 50
It can be carried out at a temperature of from 0C to 120C. With respect to the halide of the above formula (6), the third compound of the above formula (7)
The amine is used in an amount of 2 moles or more, for example, 2.0 moles of the compound of the formula (7) per mole of the compound of the formula (6).
It may be used in a ratio of mol to 2.3 mol.

As a reaction solvent, methanol, ethanol,
Alcohols such as n-propanol, a mixed solution of water and alcohol, and aromatic organic solvents such as benzene, toluene and xylene are preferably used. The reaction temperature is generally from 1 hour to 4 hours at 80 ° C. or higher.
The reaction is completed at 0 hours.

B-2. Second Method for Production The second method for producing the quaternary ammonium salt represented by the above formula (1) comprises a tertiary amine represented by the following formula (8) and a tertiary amine represented by the following formula (9) )) By reaction with a quaternizing agent.

[0047]

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[Wherein R ₁ and R ₂ each have 4 to 12 carbon atoms.
Wherein R ₁ and R ₂ may be the same or different, and k represents 0 to 4. ]

[0049]

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[In the formula, R ₃ represents an alkyl group having 1 to 2 carbon atoms, and Z represents a chlorine atom, a bromine atom or an iodine atom or any of the following formulas (10) and (11). ]

[0051]

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[Wherein, R ₆ represents an alkyl group having 1 to 2 carbon atoms. ]

[0053]

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[Wherein, R ₇ and R ₈ each represent a hydrogen atom, an alkyl group having 1 to 12 carbon atoms, or a carboxyl group. ]

The compounds of the above formula (8) include N, N, N ',
N'-tetrapentyl-o-xylylenediamine, N, N'-
Dibutyl-N, N'-dihexyl-o-xylylenediamine, N, N, N ', N'-tetrahexyl-m-xylylenediamine, N, N, N', N'-tetraheptyl-p-xylylene Diamine, N, N, N ', N'-tetrahexyl-2,3,5,6
-Tetramethyl-p-xylylenediamine, N, N'-dibutyl-N, N'-dihexyl-2,3,5,6-tetramethyl-p-xylylenediamine, N, N'-dihexyl-N, N '
-Dioctyl-2,3,5,6-tetramethyl-p-xylylenediamine, N, N, N ', N'-tetrapentyl-1,
3,5 trimethyl-m-xylylenediamine, N, N, N ',
N'-tetrahexyl-2,4dimethyl-p-xylylenediamine, N, N, N ', N'-tetrahexyl-2-methyl-p
-Xylylenediamine and the like.

Examples of the compound of the above formula (9) include alkyl halides such as methane chloride, ethane chloride, methane bromide, ethane bromide and methane iodide, dialkyl sulfates such as dimethyl sulfate, and methyl p-toluenesulfonate. And the like.

These reactions are carried out in a suitable solvent at 50 ° C to 1 ° C.
It can be carried out at a temperature of 20 ° C. Since many quaternizing agents are deactivated by heat or the like, the above formula (8)
It is desirable to use the quaternizing agent compound of the above formula (9) in excess of the tertiary amine. It is preferably at least 4 moles, more preferably at least 6 moles.

As a reaction solvent, alcohols such as methanol, ethanol, n-propanol and 2-methanoxyethanol, or a mixed solution of water and alcohol,
Further, aprotic solvents such as N, N-dimethylformamide, N-methylformamide, nitromethane, nitroethane and acetonitrile are preferably used.

As for the reaction atmosphere, synthesis can be performed in the air, but the reaction is more preferably performed in a nitrogen atmosphere. If the reaction temperature is generally 80 ° C. or higher, the reaction is completed in 1 hour to 40 hours.

The above reaction is carried out in an autoclave under pressure in the presence of a suitable solvent, preferably 10 to 10 times.
It can also be carried out at a temperature of 50 to 100 ° C. at 0 MPa (megapascal). The reaction time is usually from 5 hours to 120 hours.
Can be time.

The tertiary amine of the above formula (8) can be obtained, for example, by reacting the halide of the above formula (6) with the following formula (15).

[0062]

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(Wherein R ₁ and R ₂ each have 4 to 12 carbon atoms)
Wherein R ₁ and R ₂ may be the same or different. )

The compound of the above formula (15) includes N, N-
Secondary amines such as dipentylamine, N, N-dihexylamine, N-butyl-N-hexylamine and N-butyl-N-octylamine are exemplified.

The tertiary reaction of the above formulas (6) and (15) is carried out in a suitable organic solvent such as an alcohol, a mixed solution of an alcohol and water or an aromatic organic solvent at a reaction temperature of 50 ° C. The reaction can be carried out under the conditions of -120 ° C and a reaction time of 1 hour to 48 hours. The preferred use ratio of the secondary amine of the above formula (15) to the halide of the above formula (6) is such that the compound of the above formula (15) is 4 to 6 mol per 1 mol of the compound of the above formula (6). Yes, especially 4.
A ratio of 1 mol to 4.4 mol is preferred. As a result of the above tertiary reaction, a tertiary amine which is a target intermediate product and a raw material amine hydrochloride as a by-product are produced.

The tertiary amine, which is the target intermediate product, and the raw material amine hydrochloride can be separated by any method, but can be separated by extraction or the like. Furthermore, it can be easily performed by utilizing the difference due to the solubility of the organic solvent.

The raw material amine hydrochloride as a by-product can be regenerated into the raw material amine. N in raw material amine hydrochloride of by-product
When a base such as an aOH aqueous solution is acted on, a water-insoluble organic layer is formed. A secondary amine as a raw material is obtained from this organic layer by a general separation / purification method. This regenerated second
The amine can be used again as a raw material.

B-3. Third Production Method A third production method capable of producing the quaternary ammonium salt represented by the above formula (1) is to prepare a tertiary amine represented by the following formula (12): This is a method of treating with a quaternizing agent of the following formula (13).

[0069]

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Wherein R ₁ is an alkyl group having 4 to 12 carbon atoms, R ₃ is an alkyl group having 1 to 2 carbon atoms, and k is 0
~ 4. ]

[0071]

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[Wherein, R ₂ represents an alkyl group having 4 to 12 carbon atoms, and J represents any one of a chlorine atom, a bromine atom, an iodine atom and a group represented by the above formula (11). ]

The compound of the above formula (12) includes N, N'-
Dipentyl-N, N'-dimethyl-o-xylylenediamine, N, N'-dihexyl-N, N'-dimethyl-o-xylylenediamine, N, N'-dipentyl-N, N'-diethyl-o −
Xylylenediamine, N, N'-dipentyl-N, N'-dimethyl-m-xylylenediamine, N, N'-dipentyl-N, N '
-Dimethyl-p-xylylenediamine, N, N'-dihexyl-N, N'-dimethyl-2,3,5,6-tetramethyl-
p-xylylenediamine, N, N'-dipentyl-N, N'-dimethyl-1,3,5 trimethyl-m-xylylenediamine, N, N'-dihexyl-N, N'-dimethyl-2,4 Dimethyl-p-xylylenediamine and the like.

The compounds of the above formula (13) include butane chloride, pentane chloride, hexane chloride, decane chloride,
Alkyl halides such as nonane bromide, decane bromide, pentane iodide, heptane iodide, butyl p-toluenesulfonate, pentyl p-toluenesulfonate, hexane p-toluenesulfonate, and dodecane p-toluenesulfonate; And alkyl sulfonates.

The reaction between the compound of the formula (12) and the compound of the formula (13) can be carried out almost according to the second method. That is, as the solvent, alcohols, a mixed solution of water and alcohol, and an aprotic solvent such as N, N-dimethylformamide are suitably used, and the reaction can be performed without a solvent. The reaction atmosphere, reaction temperature and reaction time can be carried out in the same manner as in the second method. The above equation (1
The use ratio of the quaternizing agent of the above formula (13) to the tertiary amine of 2) is such that the compound of the above formula (13) is used in 2 moles or more per 1 mole of the compound of the above formula (12). well,
The ratio may be 2.0 mol to 2.3 mol. The above reaction can be carried out in an autoclave in the presence of a suitable solvent as in the case of the second reaction.

The tertiary amine of the above formula (12) can be obtained, for example, from the reaction of the halide of the above formula (6) with the following formula (16).

[0077]

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[Wherein, R ₁ represents an alkyl group having 4 to 12 carbon atoms, and R ₃ represents an alkyl group having 1 to 2 carbon atoms. ]

Examples of the compound of the above formula (16) include secondary amines such as N-pentyl-N-methylamine, N-hexyl-N-methylamine and N-hexyl-N-ethylamine.

The tertiary reaction of the above formula (6) and the above formula (16) is substantially similar to the reaction of the above formula (6) and the above formula (15). Or in an appropriate organic solvent such as an aromatic organic solvent at a reaction temperature of 50.
C. to 120.degree. C. and a reaction time of 1 hour to 48 hours. The preferred usage ratio of the secondary amine of the above formula (16) to the halide of the above formula (6) is as follows:
The compound of 6) is 4 mol to 6 mol, particularly preferably 4.1 mol to 4.4 mol. Separation of the intended intermediate tertiary amine from the starting amine hydrochloride and regeneration of the starting amine hydrochloride as a by-product can be carried out in substantially the same manner as in the case of the reaction of the compound of the formula (6) with the formula (15). .

B-4. Method for Purifying Compounds The compounds produced by the above three methods can be easily purified, if necessary, by usual separation and purification means, for example, column chromatography separation and recrystallization operation. I can do it.

B-5. Exchange of Anion Group The quaternary ammonium salt produced by the above-mentioned three methods can change an anion group contained therein into another specific anion group by ion exchange. . Ion exchange can be easily performed, for example, by treating in a column filled with an anion exchange resin.

That is, the quaternary ammonium salt in the present invention has a quaternary ammonium salt represented by the following formula (14) as a cationic group. The anion group when synthesized by ion exchange, iodine ion, bromide ion, chloride ion nitrate ion, nitrate ion, nitrite ion, chlorate ion, chlorite ion, hypochlorite ion or the above formula It can be produced by substituting any of the anionic groups (2) to (5).

[0084]

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[Wherein R ₁ and R ₂ each have 4 to 12 carbon atoms.
R ₃ is an alkyl group having 1 to 2 carbon atoms, R ₁ , R ₂ and R ₃ may be the same or different, and k represents 0 to 4. ]

C. Antibacterial Activity The compound of the formula (1) of the present invention obtained as described above can be used for various bacteria, as shown in Test Examples 1 and 2 below.
It has a broad antibacterial spectrum against fungi.

The compound of the formula (1) of the present invention exhibits strong bactericidal activity when the alkyl chain (R ₁ , R ₂ ) has a carbon number of 5 to 10, especially 6 to 8.

The compound of the formula (1) of the present invention is 1/10 to 1/100 of that of a conventional commercially available quaternary ammonium salt.
Germicidal activity with a minimum bactericidal concentration of Therefore,
The compound of the formula (1) of the present invention exerts a bactericidal effect equivalent to that of a conventional fungicide at a much lower concentration than a conventional commercially available fungicide of the same type.

D. Safety of Compound The quaternary ammonium salt of the present invention is an extremely safe compound having an LD ₅₀ (rat) of 2000 mg / kg or more in an oral toxicity test in rats. Furthermore, when the hemolytic activity of the compound of the formula (1) of the present invention was measured, conventional quaternary ammonium salts such as benzalkonium chloride, JP-A-6-321924, and
The hemolytic activity was at least 10 times lower than that of the quaternary ammonium salt introduced in, for example, -114604. Thus, the compound of the present invention of the formula (1) has extremely low toxicity to the human body.

E. Uses of Compounds The compounds of the present invention can be used in a wide variety of fields as antibacterial agents, for example, antibacterial and deodorant textile products, leather products, building materials,
Wood, paint, adhesive, plastic, film, paper,
It is useful as an antibacterial agent in pulp, metal processing oil, food, pharmaceuticals, medical / environmental disinfectants, eye drops, detergents, cosmetics, stationery, agricultural chemicals, livestock and the like. Although the compound of the present invention exhibits excellent antibacterial properties by itself, it can be used by appropriately supporting it on a solid or liquid carrier. Further, if necessary, other components such as a surfactant may be blended and used as emulsions, wettable powders, granules, powders, sprays, aerosols and the like. When the compound of the present invention is used as an antibacterial agent, a preferred compounding ratio is 0.0001 to 100% by weight, more preferably 0.001 to 10% by weight, based on the total weight of the antibacterial agent.

F. Use as Preservatives for Eye Drops Conventional preservatives for eye drops include compounds having a cationic group such as quaternary ammonium salts and guanidine, alcohols, aminobenzoate esters and sorbic acid. Quaternary ammonium salts, especially benzalkonium chloride, are generally used because of their high antiseptic properties.

However, when these preservatives are added to the eye drops, the liquid of the eye drops may become cloudy depending on the type of other compound to be added together as a drug component of the eye drops. Furthermore, for benzalkonium chloride, 0.01
% Has been reported to cause damage to the cornea, so the amount is limited to a range where safety is not an issue.

As described above, if the conventional preservative is blended at a low concentration, the problem of white turbidity does not occur, but the original preservative effect becomes insufficient.
Various improvements have been proposed for improving the preservative effect and preventing cloudiness (JP-A-2-311417, JP-A-6-4091).
0).

As a result of intensive studies, the present inventors have found that, as shown in Test Example 1 below, the compound of the present invention represented by the above formula (1) (hereinafter abbreviated as compound (1) of the present invention) It has been found that various compounds used as active ingredients of eye drops and insoluble substances are not generated, and they are extremely effective as a preservative for obtaining transparent eye drops. That is, the preservative for eye drops of the present invention contains the compound (1) of the present invention as an active ingredient.

Examples of the compound for a drug component of eye drops which do not cause cloudiness even when formulated with the compound (1) of the present invention include the following. That is, compounds having a carboxyl group such as sodium hyaluronate, dipotassium glycyrrhizinate, vilenoxine, lysodium chloride, sodium cromoglycate, carboxyvinyl polymer, chondroitin sulfate, sodium dimethylisopropylazulene sulfonate, sodium colistin methanesulfonate, sodium metal Compounds having a sulfonic acid group such as dexamethasone sulfobenzoate; compounds having a phosphonyl group such as flavin adenidine dinucleotide; and pilocarpine hydrochloride. All of these compounds have heretofore been unable to be compounded as a drug component because of the formation of white turbidity when mixed with benzalkonium chloride.

The preservatives for eye drops of the present invention include, in addition to the above-mentioned drug components, various components used in eye drops as required, for example, active ingredients such as anti-inflammatory agents, vitamins and antihistamines, pH adjusters Even if additives such as a buffer, a tonicity agent, a solvent and the like are incorporated, the preservative effect is not affected. Compound (1) of the present invention was prepared in Test Example 3 described below,
As shown in 4 and 5, it has a broad bactericidal spectrum against various bacteria and fungi, and also exhibits higher bactericidal activity and antiseptic power than the conventional general-purpose quaternary ammonium salt benzalkonium chloride. I do.

The compounding ratio of the compound (1) of the present invention in the eye drops is usually from 0.0005 to 0.0005 of the total amount of the eye drops.
0.1% by weight, preferably 0.002 to 0.02
%. If it is less than 0.0005%, the preservative effect of the preservative may be insufficient, and if it is more than 0.1%, it is economically disadvantageous.

[0098]

The present invention will be described more specifically below with reference to examples and comparative examples. G-1. Synthesis Examples Hereinafter, synthesis examples of the compound of the present invention will be described. In the following description, abbreviated a synthesized compound, such as "x ₁ -BAMx ₂ -x ₃ x _4". x ₁ : a number indicating the bonding position of the quaternary ammonium group on the benzene ring. (2 at the ortho position, 3 at the meta position, 4 at the para position) x ₂ : A symbol corresponding to the value of k in the general formula (1).
(P for 0, T for 3 and D for 4) x ₃ : A number from 4 to 12 indicating the carbon number of the long alkyl group bonded to the quaternary ammonium. x _4: symbol that corresponds to the type of anion. (C for chlorine,
B for bromine, I for iodine, S for sulfate

<Example 1> As a halogen compound, α,
α′-dichloro-p-xylene 20 mmol as a tertiary amine N, N-dihexyl-N-methylamine 42 mmol,
100 ml of ethanol as a solvent was charged into 300 ml of each reaction vessel and reacted under heating and reflux for 5 hours, and then ethanol as a solvent was removed under reduced pressure to obtain crude crystals.
The crude crystals were recrystallized in a diethyl ether solvent, and dried under reduced pressure to give a white compound, 1,4-bis (N, N-dihexyl-N-methylammoniomethyl) phenylenedichloride (abbreviated as 4BAMP-6C). 0.02 g was obtained. The yield of the target compound was 70%.

[0100] Of the resulting 4BAMP-6C¹H-NMR (solvent CD_ThreeO
D) The analysis results are as follows (unit is δppm; the same applies hereinafter): 0.94 (12H, t, J = 6.8Hz), 1.40 (24H, s), 1.85 (8H, br s), 3.02
(6H, s), 3.31 (8H, m), 4.62 (4H, s), 7.71 (4H, s)₃₄H₆₆N_TwoCl_TwoResults of elemental analysis
Was shown below.

[0101]

[Table 1]

The above analysis confirmed that the obtained substance was the target compound. For a sample 4BAMP-6C obtained above, oral toxicity: Measurement of the (LD ₅₀ rat), was larger than 2000 mg / kg.

<Example 2> In Example 1, N, N-dihexyl-N-methylamine was used instead of N, N-dihexyl-N-methylamine as the tertiary amine.
The same operation as in Example 1 was carried out except that N, N-dioctyl-N-methylamine was used, and a white compound 1,4-bis (N, N-dioctyl-N-methylammonio) as a target compound was obtained. Methyl) phenylenedichloride (4BAMP-8C
7.10 g). The yield of this compound is 52%
Met. ¹ H-NMR analysis of 4BAMP-8C obtained above (solvent CD ₃
OD) results are as follows. 0.91 (12H, t, J = 7.2Hz), 1.38 (40H, s), 1.85 (8H, br s), 3.05
(6H, s), 3.31 (8H, m), 4.68 (4H, s), 7.74 (4H, s) The results of elemental analysis with the molecular formula C ₄₂ H ₈₂ N ₂ Cl ₂ are shown below.

[0104]

[Table 2]

The above analysis confirmed that the substance obtained was the target compound.

Example 3 Example 1 was the same as Example 1 except that α, α′-dichloro-m-xylene was used instead of α, α′-dichloro-p-xylene as the halogen compound. The reaction was carried out under the conditions to obtain white crude crystals. The crude crystals had high hygroscopicity and immediately became liquid. After washing with hexane and drying under reduced pressure, 12.21 g of 1,3-bis (N, N-dihexyl-N-methylammoniomethyl) phenylenedichloride (abbreviated as 3BAMP-6C) as a target compound was obtained. Was. The yield of the target compound was 89%. Of 3BAMP-6C obtained above
^{The result of 1} H-NMR analysis (solvent CD ₃ OD) is as follows. 0.91 (12H, t, J = 6.4Hz), 1.38 (40H, s), 1.85 (8H, br s), 3.05
(6H, s), 3.31 (8H, m), 4.68 (4H, s), 7.50 (1H, m), 7.71 (2H,
m), 7.68 (1H, m) This result indicates that 3BAMP-6C is the target compound.

<Example 4> A pale yellow liquid was obtained in the same manner as in Example 1 except that the halogen compound was changed to α, α'-dichloro-o-xylene. After washing with hexane and drying under reduced pressure, a white liquid compound 1,2-bis (N, N-dihexyl-N-methylammoniomethyl) phenylenedichloride (2BAMP) as a target compound is obtained.
-18C). The yield of the target compound was 89%. The results of ¹ H-NMR analysis (solvent CD ₃ OD) of the above 2BAMP-6C are as follows. 0.91 (12H, t, J = 6.8Hz), 1.38 (40H, s), 1.85 (8H, br s), 3.05
(6H, s), 3.31 (8H, m), 4.68 (4H, s), 7.52 (2H, m), 7.72 (2H, m) This result indicates that 2BAMP-6C is the target compound .

Example 5 N, N, N ', N'- as a tertiary amine
20 mmol of tetrapentyl-p-xylylenediamine,
120 mm methyl p-toluenesulfonate as quaternizing agent
ol, 100 g of ethanol as a solvent, 30 g each
In a 0 ml reaction vessel, heat and reflux under a nitrogen atmosphere.
Let react for hours. The solvent ethanol is removed, and the mixture is washed with water and ether. The solvent was removed under reduced pressure, and the obtained solid was recrystallized with an acetonitrile solvent and dried under reduced pressure.
White solid compound 1,4-bis (N, N-
Dipentyl-N-methylammoniomethyl) phenylene p-toluenesulfonate (abbreviated as 4BAMP-5S)
0.79 g was obtained. The yield of the target compound was 68%. ¹ H-NMR analysis of the 4BAMP-5S obtained above (solvent CD ₃ O
The result of D) is as follows. 0.94 (12H, t, J = 7.2Hz), 1.36 (16H, s), 1.81 (8H, br s), 2.37
(6H, s), 2.98 (6H, s), 3.13 (8H, m), 4.44 (4H, s), 7.23 (4H, d,
J = 7.6 Hz), 7.70 (8H, s + d, J = 5.6 Hz) The results of elemental analysis using the molecular formula C ₄₄ H ₇₂ N ₂ S ₂ O ₆ are shown below.

[0109]

[Table 3]

The results of these analyzes indicate that 4BAMP-5S is the target compound.

<Embodiment 6> In Embodiment 5, the third
N, N, N ', N'-tetrapentyl-p-xylylene
N, N, N ', N'-tetrahexyl-p- instead of diamine
Except that xylylenediamine was used,
Perform the same operation to obtain a white solid compound as the target compound.
1,4-bis (N, N-dihexyl-N-methylammonium
Omethyl) phenylene p-toluenesulfonate (4B
12.63 g of AMP-6S) was obtained. Yield of target compound
Was 75%. Of the 4BAMP-6S obtained above¹H-NMR
Analysis (solvent CD_ThreeOD) results are as follows.

0.91 (12H, t, J = 6.8 Hz), 1.38 (24H, s), 1.79 (8
H, br s), 2.34 (6H, s), 2.98 (6H, s), 3.31 (8H, m), 4.57 (4H,
s), 7.23 (4H, d, J = 8.0 Hz), 7.68 (8H, s + d, J = 5.6 Hz) The results of elemental analysis using the molecular formula C ₄₈ H ₈₀ N ₂ S ₂ O ₆ are shown below. .

[0113]

[Table 4]

These results indicate that 4BAMP-6S is the target compound. Further, the melting point of 4BAMP-6S
It was 160.6 ° C. as measured by SC. The oral toxicity (LD ₅₀ : rat) of the sample 4BAMP-6S obtained in Example 6 was measured to be 2000 mg / k.
g, which proved to be a substance with low toxicity.

<Embodiment 7> In Embodiment 6, the third embodiment
Example 6 except that N, N, N ', N'-tetraoctyl-p-xylylenediamine was used in place of N, N, N', N'-tetrahexyl-p-xylylenediamine as the amine. The same operation as described above was carried out to obtain the desired white solid compound 1,4.
-Bis (N, N-dioctyl-N-methylammoniomethyl) phenylene p-toluenesulfonate (4BAMP-8
12.58 g was obtained. The yield of 4BAMP-8S is 66
%Met. The results of ¹ H-NMR analysis (solvent CD ₃ OD) of 4BAMP-8S obtained above are as follows.

0.88 (12H, t, J = 6.8 Hz), 1.36 (24H, s), 1.80 (8
H, br s), 2.35 (6H, s), 3.02 (6H, s), 3.31 (8H, m), 4.65 (4H,
s), 7.23 (4H, d, J = 8.0 Hz), 7.72 (8H, s + d, J = 6.0 Hz) Also, the results of elemental analysis with the molecular formula C ₅₆ H ₉₆ N ₂ S ₂ O ₆ Indicated.

[0117]

[Table 5]

These results indicate that 4BAMP-8S is the target compound.

<Example 8> N, N'-dibutyl-N, N'-dimethyl-p-xylylenediamine as a tertiary amine 20 mm
ol, as a quaternizing agent, 120 mmol of hexyl iodide
And 100 g of ethanol as a solvent is charged into a 300 ml reaction vessel, and reacted under heating and refluxing under a nitrogen atmosphere for 10 hours. After removing the solvent ethanol, recrystallizing with acetonitrile and drying under reduced pressure, the target white compound 1,
6.86 g of 4-bis (N-butyl-N-hexyl-N-methylammoniomethyl) phenylenediodide (4BAMP-4, abbreviated as 6I) was obtained. The yield of the target compound is 49
%Met. The results of ¹ H-NMR analysis (solvent CDCl ₃ ) of 4BAMP-4 and 6I obtained above are as follows. 0.91 (6H, t, J = 7.2Hz), 1.10 (6H, t, J = 7.6Hz), 1.36 (16H, s),
1.86 (4H, br s), 2.45 (4H, br s), 3.13 (6H, s), 3.32 (4H, m),
3.64 (4H, m), 5.34 (8H, s), 7.75 (4H, s) The result of elemental analysis represented by the molecular formula C ₃₀ H ₅₈ N ₂ I ₂ is shown below.

[0120]

[Table 6]

These results indicate that 4BAMP-4, 6I is the target compound.

<Example 9> The same procedure as in Example 8 was carried out except that octyl iodide chloride was used instead of hexyl iodide as the quaternizing agent. , 4-bis (N-butyl-N-octyl-N-methylammoniomethyl) phenylenediodide (abbreviated as 4BAMP-4,8I) was 6.56.
g was obtained. The yield of the target compound was 43%. The results of ¹ H-NMR analysis (solvent CDCl ₃ ) of 4BAMP-4 and 8I obtained above are as follows. 0.88 (6H, t, J = 7.2Hz), 1.09 (6H, t, J = 7.6Hz), 1.34 (24H, s),
1.84 (4H, br s), 2.44 (4H, br s), 3.13 (6H, s), 3.28 (4H, m),
3.64 (4H, m), 5.30 (8H, s), 7.73 (4H, s) The results of elemental analysis for the molecular formula C ₃₄ H ₆₆ N ₂ I ₂ are shown below.

[0123]

[Table 7]

These results indicate that 4BAMP-4 and 8I are the target compounds.

<Example 10> As the halogen compound,
20 mmol of 6-bis (chloromethyl) durene was used as a tertiary amine in an amount of 42 mmol of N, N-dibutyl-N-methylamine.
As a solvent, 300 ml each of 100 ml of ethanol.
After charging in a reaction vessel and reacting for 5 hours under heating and reflux,
The crude ethanol was obtained by removing the solvent ethanol under reduced pressure. The crude crystals were washed with ethyl acetate, recrystallized from a mixed solution of acetonitrile and ethyl acetate, and dried under reduced pressure to give a white compound 1,4-bis (N, N
-Butyl-N-methylammoniomethyl) -2,3,
5,6-tetramethylphenylenedichloride (BAMD-4
3.3 g). The yield of the target compound is 32
%Met. The results of ¹ H-NMR (solvent CD ₃ OD) analysis of the obtained BAMD-4 are as follows. 0.92 (12H, t), 1.38 (8H, br s), 1.78 (8H, br s), 2.35 (12H,
s), 3.03 (6H, s), 3.48 (8H, m), 5.15 (4H, s) The results of elemental analysis with the molecular formula C ₃₀ H ₅₈ N ₂ Cl ₂ are shown below.

[0126]

[Table 8]

From the above analysis, it was confirmed that the obtained substance was the target compound.

<Embodiment 11> In Embodiment 10, the third
The amine is N, instead of N, N-dibutyl-N-methylamine.
The same operation as in Example 1 was performed except that N-dipentyl-N-methylamine was used, and a white compound 1,4-bis (N, N-dipentyl-N-methylammoniomethyl) as a target compound was obtained. 2.4 g of -2,3,5,6-tetramethylphenylenedichloride (abbreviated as BAMD-5) was obtained. The yield of this compound was 21%. Of the BAMD-5 obtained above
^{The result of 1} H-NMR analysis (solvent CD ₃ OD) is as follows. 0.91 (12H, t), 1.38 (16H, br s), 1.80 (8H, br s), 2.36 (12H,
s), 3.03 (6H, s), 3.51 (8H, m), 5.14 (4H, s) and the results of elemental analysis with the molecular formula C ₃₄ H ₆₆ N ₂ Cl ₂ are shown below.

[0129]

[Table 9]

The above analysis confirmed that the substance obtained was the target compound.

<Embodiment 12> In Embodiment 10, the third embodiment
The amine is N, instead of N, N-dibutyl-N-methylamine.
The same operation as in Example 1 was carried out except that N-dihexyl-N-methylamine was used, and a white compound 1,4-bis (N, N-dihexyl-N-methylammoniomethyl) as a target compound was obtained. 4.2 g of -2,3,5,6-tetramethylphenylenedichloride (abbreviated as BAMD-6) was obtained. The yield of this compound was 33%. BMD- obtained above
The result of ¹ H-NMR analysis (solvent CD ₃ OD) of ^No. 6 is as follows.

0.91 (12H, t), 1.38 (24H, brs), 1.79 (8H, br
s), 2.35 (12H, s), 3.03 (6H, s), 3.49 (8H, m), 5.16 (4H, s) and the results of elemental analysis of the molecular formula C ₃₈ H ₇₄ N ₂ Cl ₂ are shown below. .

[0133]

[Table 10]

The above analysis confirmed that the substance obtained was the target compound. Sample BA obtained above
For MD-6, oral toxicity: Measurement of the (LD ₅₀ rat), has become greater than 2000mg / kg.

<Thirteenth Embodiment> In the tenth embodiment, the third
The amine is replaced with N, N-dibutyl-N-methylamine
The same operation as in Example 1 was performed except that -octyl-N-hexyl-N-methylamine was used, and a white compound 1,4-bis (N-octyl-N-hexyl-N- Methylammoniomethyl) -2,3,5
2.3 g of 6-tetramethylphenylenedichloride (abbreviated as BAMD-6, 8) was obtained. The yield of this compound was 17%. The results of ¹ H-NMR analysis (solvent CD ₃ OD) of BAMD-6 and 8 obtained above are as follows.

0.93 (12H, t), 1.39 (32H, brs), 1.80 (8H, br
s), 2.37 (12H, s), 3.06 (6H, s), 3.52 (8H, m), 5.19 (4H, s) and the results of elemental analysis as molecular formula C ₄₂ H ₈₂ N ₂ Cl ₂ are shown below. .

[0137]

[Table 11]

The above analysis confirmed that the substance obtained was the target compound.

<Embodiment 14> In Embodiment 10, the third
The amine is N, instead of N, N-dibutyl-N-methylamine.
The same operation as in Example 1 was performed except that N-dioctyl-N-methylamine was used, and a white compound 1,4-bis (N, N-dioctyl-N-methylammoniomethyl) as a target compound was obtained. 2.0 g of -2,3,5,6-tetramethylphenylenedichloride (abbreviated as BAMD-8) was obtained. The yield of this compound was 14%. Of the BAMD-8 obtained above
^{The result of 1} H-NMR analysis (solvent CD ₃ OD) is as follows.

0.92 (12H, t), 1.38 (40H, brs), 1.81 (8H, br
s), 2.38 (12H, s), 3.05 (6H, s), 3.54 (8H, m), 5.18 (4H, s) The results of elemental analysis for the molecular formula C ₄₆ H ₉₀ N ₂ Cl ₂ are shown below. .

[0141]

[Table 12]

The above analysis confirmed that the substance obtained was the target compound.

<Example 15> In Example 10, 2,4-bis (chloromethyl) -1,3,3 was used as the halogen compound instead of 1,4-bis (chloromethyl) durene.
The reaction was carried out under the same conditions as in Example 1 except that 5-trimethylbenzene was used, and the target compound, white compound 2,4-bis (N, N-dibutyl-N-methylammoniomethyl) -1,3, was obtained. 0.7 g of 2,5-trimethylbenzene dichloride (abbreviated as BAMT-4) was obtained. The yield of the target compound was 7%. The results of ¹ H-NMR analysis (solvent CD ₃ OD) of BAMT-4 obtained above are as follows.

0.91 (12H, t), 1.38 (8H, brs), 1.79 (8H, br)
s), 2.35 (9H, s), 3.03 (6H, s), 3.49 (8H, m), 5.16 (4H, s), 6.8
2 (1H, s) The results of elemental analysis for the molecular formula C ₂₉ H ₅₆ N ₂ Cl ₂ are shown below.

[0145]

[Table 13]

This result shows that BAMT-4 is the target compound.

<Embodiment 16> In Embodiment 15, the third embodiment
The amine is N, instead of N, N-dibutyl-N-methylamine.
The same operation as in Example 15 was carried out except that N-dipentyl-N-methylamine was used, and a white compound 2,4-bis (N, N-dipentyl-N-methylammoniomethyl) as a target compound was obtained. 0.8 g of -1,3,5-trimethylbenzene dichloride (abbreviated as BAMT-5) was obtained. The yield of this compound was 7%. ¹ H-NMR of BAMT-5 obtained above
The results of the analysis (solvent CD ₃ OD) are as follows.

0.90 (12H, t), 1.36 (16H, brs), 1.78 (8H, br
s), 2.35 (9H, s), 3.05 (6H, s), 3.47 (8H, m), 5.14 (4H, s), 6.8
2 (1H, s) The results of elemental analysis for the molecular formula C ₃₃ H ₆₄ N ₂ Cl ₂ are shown below.

[0149]

[Table 14]

The above analysis confirmed that the obtained substance was the target compound.

<Embodiment 17> In Embodiment 15, the third
The amine is N, instead of N, N-dibutyl-N-methylamine.
The same operation as in Example 15 was carried out except that N-dihexyl-N-methylamine was used, and a white compound 2,4-bis (N, N-dihexyl-N-methylammoniomethyl) as a target compound was obtained. 1.4 g of -1,3,5-trimethylbenzene dichloride (abbreviated as BAMT-6) was obtained. The yield of this compound was 11%. ¹ H-NM of BAMT-6 obtained above
The results of R analysis (solvent CD ₃ OD) are as follows.

0.91 (12H, t), 1.38 (24H, brs), 1.79 (8H, br
s), 2.35 (9H, s), 3.03 (6H, s), 3.49 (8H, m), 5.16 (4H, s), 6.8
1 (1H, s) The results of elemental analysis for the molecular formula C ₃₇ H ₇₂ N ₂ Cl ₂ are shown below.

[0153]

[Table 15]

The above analysis confirmed that the substance obtained was the target compound.

<Embodiment 18> In Embodiment 15, the third embodiment
The amine is replaced with N, N-dibutyl-N-methylamine
The same operation as in Example 15 was carried out except that -octyl-N-hexyl-N-methylamine was used, and a white compound 2,4-bis (N-octyl-N-
Hexyl-N-methylammoniomethyl) -1,3,5
1.1 g of trimethylbenzene dichloride (abbreviated as BAMT-6,8) was obtained. The yield of this compound was 8%. The results of ¹ H-NMR analysis (solvent CD ₃ OD) of BAMT-6,8 obtained above are as follows.

0.92 (12H, t), 1.39 (32H, brs), 1.79 (8H, br
s), 2.37 (9H, s), 3.06 (6H, s), 3.51 (8H, m), 5.18 (4H, s), 6.8
4 (1H, s) The results of elemental analysis for the molecular formula C ₄₁ H ₈₀ N ₂ Cl ₂ are shown below.

[0157]

[Table 16]

The above analysis confirmed that the substance obtained was the target compound.

<Embodiment 19> In Embodiment 15, the third embodiment
The amine is N, instead of N, N-dibutyl-N-methylamine.
The same operation as in Example 15 was carried out except that N-dioctyl-N-methylamine was used, and a white compound 2,4-bis (N, N-dioctyl-N-methylammoniomethyl) as a target compound was obtained. 1.2 g of -1,3,5-trimethylbenzene dichloride (abbreviated as BAMT-8) was obtained. The yield of this compound was 8%. ¹ H-NMR of BAMT-8 obtained above
The results of the analysis (solvent CD ₃ OD) are as follows.

0.93 (12H, t), 1.40 (40H, brs), 1.80 (8H, br)
s), 2.36 (9H, s), 3.04 (6H, s), 3.51 (8H, m), 5.19 (4H, s), 6.8
5 (1H, s) The result of elemental analysis for the molecular formula C ₄₅ H ₈₈ N ₂ Cl ₂ is shown below.

[0161]

[Table 17]

The above analysis confirmed that the substance obtained was the target compound.

G-2. Test Examples Hereinafter, test examples relating to the antibacterial property and antiseptic activity of the compound of the present invention will be described. For comparison, first, four kinds of conventional antibacterial agents were prepared.

<Comparative Example 1> As Comparative Example 1, benzalkonium chloride, a commercially available quaternary ammonium salt-based antibacterial agent, was prepared.

<Comparative Example 2> As Comparative Example 2, an antibacterial agent proposed in JP-A-10-114604 was prepared as follows. Α, α'-dichloro- as a halogen compound
A reaction vessel was charged with 20 mmol of p-xylene, 42 mmol of N, -dodecyl-N, N-dimethylamine as a tertiary amine, and 100 ml of ethanol as a solvent, and these were reacted under heating to reflux for 5 hours. The crude ethanol was obtained by removing the solvent ethanol under reduced pressure. The crude crystals were recrystallized from a diethyl ether solvent and dried under reduced pressure to give a white compound 1,4-bis (N-dodecyl-).
8.02 g of (N, N-dimethylammoniomethyl) phenylenedichloride (abbreviated as 4BADMP-12C) was obtained. The yield of the target compound was 70%. Of 4BADMP-12C obtained in the above ¹ H-
The results of NMR analysis (solvent CD ₃ OD) are as follows.

0.93 (6H, t, J = 6.8 Hz), 1.30 (36H, br), 1.85 (4
H, br s), 3.25 (12H, s), 3.60 (4H, m), 5.30 (4H, s), 7.85 (4H,
s) The results of elemental analysis performed with the molecular formula C ₃₆ H ₇₀ N ₂ Cl _{2 are} as follows.

[0167]

[Table 18]

The results of these analyzes indicate that 4BADMP-12C is the target compound.

Comparative Example 3 As Comparative Example 3, an antibacterial agent proposed in JP-A-6-321902 was prepared as follows. 20 mmol of 4-mercaptopyridine was dissolved in 50 ml of ethanol, 10 mmol of 1,6-dibromohexane was added dropwise with stirring, and the reaction was continued for 12 hours under reflux with heating. After cooling the solution, the resulting white precipitate was filtered. The obtained precipitate was dissolved in 50 ml of water, and 1N-
After the solution was adjusted to pH 11 by dropwise addition of an aqueous NaOH solution, extraction was repeated three times using diethyl ether. Molecular sieve 3A 1/16 for ether layer
(Wako Pure Chemical Industries, Ltd.) and dried overnight, after which the ether was removed to give a pale yellow solution compound. After 50 ml of DMF was added to this compound and dissolved, 40 mmol of octyl iodide was added, and the mixture was reacted under heating and reflux conditions for 24 hours. After completion of the reaction, the light yellow solid obtained by removing DMF as a solvent was recrystallized with an acetonitrile solvent and dried under reduced pressure to give a white solid compound 4,4 ′-(1,6-dithiohexamethylene) -bis-. 4.40 g of (1-octylpyridinium iodide) (abbreviated as 4MHO) was obtained. The yield of the target compound was 56%. ¹ H of 4MHO obtained in the above
The results of -NMR (solvent CD ₃ OD) analysis are shown below.

0.81 (6H, t, J = 7.2 Hz), 1.25 (20H, m), 1.71 (8
H, m), 3.32 (8H, m), 4.78 (4H, m) 7.80 (8H, m) The results of elemental analysis with the molecular formula C ₃₂ H ₅₄ N ₂ S ₂ I _{6 are} as follows.

[0171]

[Table 19]

Comparative Example 4 As Comparative Example 4, an antibacterial agent proposed in JP-A-2000-95763 was synthesized and used as follows. 20 mmol of 4-mercaptopyridine was dissolved in 50 ml of acetone, 10 mmol of xylylene dichloride was added dropwise with stirring, and the reaction was continued under reflux with heating for 5 hours. After cooling the solution, the resulting white precipitate was filtered. The obtained precipitate was washed with acetone, dissolved in 50 ml of pure water, and a 0.5N aqueous solution of NaOH was added dropwise to adjust the solution to pH 10. Then, extraction was repeated three times with toluene. After the organic layer was washed with water and dried, toluene was removed under reduced pressure to obtain crude crystals. This compound has DMF
After adding and dissolving 50 ml, octyl iodide 40 mm
ol was added and reacted at 100 ° C. for 15 hours. After completion of the reaction, the light yellow solid obtained by removing the solvent DMF was poured into 200 ml of ethyl acetate, the precipitated solid was separated by filtration, recrystallized with a mixed solvent of acetonitrile / ethyl acetate and dried under reduced pressure. , A white solid compound 4,
4.0 g of 4 '-(p-xylyldithio) -bis (1-octylpyridinium iodide) (abbreviated as I-8)
(Yield 50%) was obtained.

G-2-1. Minimum Bactericidal Concentration (MBC) for Bacteria <Test Example 1> According to a general sterile water dilution method, cells in an initial logarithmic growth phase cultured using a nutrient broth were subjected to a bacterial suspension concentration in sterile water. It was adjusted to about 10 ⁶ cell / ml. After dispensing 0.5 ml of the serially diluted drug solution, inoculate 0.5 ml each of the prepared cell suspension, contact at 30 ° C. for 30 minutes, and transfer 0.1 ml of the test solution to 2 ml of nutrient broth. After static culture at 37 ° C. for 24 hours, the MBC value was determined based on the presence or absence of proliferation. Five kinds of Gram-negative bacteria and four kinds of Gram-positive bacteria were used as test bacteria. 4BAMP-6C (Example 1), 4B
AMP-6S (Example 6) was used, and benzalkonium chloride (Comparative Example 1) was used. Table 20 shows the results.

[0174]

[Table 20]

<Test Example 2> BAMD-6 (Example 12), BAMD
-6,8 (Example 13), BAMT-6 (Example 17) and benzalkonium chloride (Comparative Example 1), Gram-negative bacteria
The MBC values for 5 species and 4 Gram-positive bacteria were determined in the same manner as in Test Example 1 above. Table 21 shows the results.

[0176]

[Table 21]

G-2-2. Minimum Growth Inhibitory Concentration (MIC) for Bacteria <Test Example 3> Bacterial solution in stationary phase adjusted to a bacterial suspension concentration of 10 ⁶ cell / ml using nutrient broth according to a general broth dilution method Was added to the serially diluted drug solution, and the mixture was allowed to stand at 37 ° C. for 24 hours, and then the MIC value was determined based on the presence or absence of proliferation. Escheric as test bacteria
hia coli K12 W 3110 was used. As test samples, the compounds obtained in Examples 1 to 9 and the benzalkonium chloride prepared in Comparative Example 1 were used. The results are shown in Table 22.

[0178]

[Table 22]

<Test Example 4> The compounds obtained in Examples 10 to 19 and benzalkonium chloride were tested in Test Example 3.
M for Escherichia coli K12 W 3110
IC values were determined. The results are shown in Table 23.

[0180]

[Table 23]

<Test Example 5> The MIC value was determined by an agar plate dilution method according to the method of the Japanese Society of Chemotherapy. That is, the bacterial suspension concentration was 10 ⁶ cell / ml using a nutrient broth.
Was added to the agar medium containing the drug diluted in two steps and incubated at 37 ° C. for 24 hours, and the MIC value was determined based on the presence or absence of bacterial growth.
The following three types were used as test bacteria. Escherichia Coli K12 W 3110 (E. coli) Psedomonas aeruginosa ATCC 27583 (P. aruginosa) Staphylococcus aureus IFO 12732 (S. aureus) As a test sample, 4BAMP-5C (1,4-bis (N, N-
Dipentyl-N-methylammoniomethyl) phenylenedichloride), 4BAMP-6C (Example 1), 4BAMP-6,8B
(1,4-bis (N-octyl-N-hexyl-N-methylammoniomethyl) phenylenedibromide) and benzalkonium chloride (Comparative Example 1) were used. Table 24 shows the results.
Shown in

[0182]

[Table 24]

From the results of Test Examples 1 to 5, the MBC value or MIC value when the compound of the present invention was used was equal to or smaller than that when benzalkonium chloride was used.
It is apparent that the compounds of the present invention have the same or higher antibacterial activity against bacteria than benzalkonium chloride.

G-2-3. Minimum Growth Inhibitory Concentration (MIC) for Fungi <Test Example 6> According to a general broth dilution method, a spore solution was prepared from a precultured test bacterium in sterile water containing a wetting agent. 1 ml of the serially diluted drug solution and 1 ml of the spore solution were mixed and cultured in an incubator at 27 ° C. for one week, and then the presence or absence of proliferation was determined by turbidity. As test bacteria, Aspergillus niger IFO 6341 (A.nig
er) and Candida albicans ATCC 10231 (C. albicans). As test samples, 4BAMP-5C (1,4-bis (N, N-dipentyl-N-methylammoniomethyl) phenylenedichloride), 4BAMP-6C (Example 1), 4BAMP-C
6,8B (1,4-bis (N-octyl-N-hexyl-N
-Methylammoniomethyl) phenylenedibromide) and benzalkonium chloride (Comparative Example 1) were used. The results are shown in Table 25.

[0185]

[Table 25]

From the results of Test Example 6, the MIC value of the compound of the present invention was lower than that of benzalkonium chloride, and therefore the compound of the present invention was more fungi than benzalkonium chloride. It is clear that the antibacterial activity is high.

G-2-4. Minimum concentration of hemolytic activity against erythrocytes <Test Example 7> Human blood was collected and an equal volume of Alsever solution (glucose 2.05 g, NaCl 0.42 g, sodium citrate 0.8
g, pH 6.1 containing 0.55 g of citric acid) and stored at 4 ° C. Immediately before use in the experiment, the Alsever stored blood was centrifuged to remove the supernatant and the leukocyte layer on the red blood cells. Further, about 10 times the amount of PBS was added to the erythrocytes, and the centrifugal washing was repeated three times. Finally, the same amount of PBS as the erythrocytes was added to obtain 50% erythrocytes. 10 μl of 50% erythrocytes was added to 990 μl of a serially diluted PBS dilution of the drug, which was allowed to act at 37 ° C. for 1 hour, centrifuged, and the supernatant was collected to measure OD540. Separately,
A similar test was carried out without using any drug, and used as a control for 0% hemolysis, and as a control for 100% hemolysis using MilliQ water instead of PBS. As the minimum hemolytic activity concentration of the drug,
The lowest concentration at which the hemolysis concentration was 50% or less was used. Using 4BAMP-6C (Example 1), 4BAMP-5S (Example 5) and 4BAMP-6S (Example 6) as test samples, benzalkonium chloride (Comparative Example 1), 4BADMP-12C (Comparative Example 2) And 4MHO
(Comparative Example 3) was used. The results are shown in Table 26. In Table 26 below, 4BAMP-5C (1,4-bis (N, N-dipentyl-N-methylammoniomethyl) phenylenedichloride) which is a compound of the present invention obtained by synthesizing in the same manner as in Example 1 is shown. Are also shown.

[0188]

[Table 26]

<Test Example 8> BAMD-5 (Example 11), BAMD
-6 (Example 12), BAMT-5 (Example 16), BAMT-6 (Example 17), benzalkonium chloride (Comparative Example 1), 4BAD
The minimum hemolytic activity concentration of MP-12C (Comparative Example 2), 4MHO (Comparative Example 3) and I-8 (Comparative Example 4) was measured in the same manner as in Test Example 5. The results are shown in Table 27.

[0190]

[Table 27]

From the results of Test Examples 7 and 8, the compound of the present invention showed a minimum hemolytic activity concentration about 8 to 60 times larger than that of the conventional antibacterial agent. It is clear that the safety to the human body is much better than conventional antibacterial agents.

G-2-5. Interaction with Eye Drop Drug Component <Test Example 9> 1% of Eye Drop Drug Component and Preservative of the Present Invention
A solution was prepared by dissolving in sodium chloride solution and neutralizing the pH with an appropriate amount of dilute hydrochloric acid or sodium hydroxide, and the transparency was visually observed. Sodium chondroitin sulfate (hereinafter abbreviated as SKA), sodium hyaluronate (abbreviated as SHA), sodium cromoglycate (abbreviated as SCA), sodium dimethylisopropylazulene sulfonate (abbreviated as SDPA), flavin adenidinucleotide as a drug component for eye drops (Abbreviated as FA) and 4BAMP-6C (Example 1) and benzalkonium chloride (Comparative Example 1) were used as preservative test samples. Table 28 shows the mixing ratios and the observation results. When the preservative of the present invention was blended, it was transparent, whereas when benzalkonium chloride was blended, cloudiness was produced.

[0193]

[Table 28]

G-2-6. Antiseptic test <Test Example 10> Using a solution formulated according to the formulations shown in Tables 29 and 30 below, described in the United States Pharmacopeia (Pharmacopeia of
the United States XX, 873 (1980)). Observations were 1, 4, 24 hours, 7, 14,
After 21 and 28 days, the antiseptic power was determined. E.coli, P.auru. And S.aureus which are bacteria as test bacteria and fungi
A.niger and C.albicans were used. As test samples, 4BAMP-6C and benzalkonium chloride were used. Table 31 shows the results of the observation time at which the antiseptic effect was confirmed. From this test result, it is clear that the preservative of the present invention exhibits a preservative power in a short time, that is, has a high preservative power, as compared with benzalkonium chloride.

[0195]

[Table 29]

[0196]

[Table 30]

[0197]

[Table 31]

[0198]

The compound of the present invention is a novel quaternary ammonium salt, in which two quaternary ammonium groups are bonded to a benzene ring, and the quaternary ammonium has 4 to 4 carbon atoms per nitrogen atom. It has a structure in which two 12 alkyl groups and one alkyl group having 1 to 2 carbon atoms are bonded. The compound of the present invention is useful as an antibacterial agent, has higher safety to the human body than known antibacterial agents, and has an excellent bactericidal effect and a broad antibacterial spectrum by itself. The compounds of the present invention are particularly useful as preservatives for eye drops.

 ────────────────────────────────────────────────── ─── Continued on the front page (72) Inventor Katsuka Kume 1-Funami-cho, Minato-ku, Nagoya-shi, Aichi F-term in Nagoya Research Institute (reference) 4C076 AA12 BB24 CC10 DD23D DD26Z DD49R FF39 4H006 AA01 AA02 AB29 AC52

Claims (9)

[Claims] 1. A quaternary ammonium salt represented by the following formula (1). Embedded image Wherein R ₁ and R ₂ are each an alkyl group having 4 to 12 carbon atoms, and may be the same or different;
R ₃ is an alkyl group having 1 to 2 carbon atoms, k is 0 to 4, X ⁿ⁻ is an inorganic or organic anion group, and n
Is a valency of the anionic group X ^n- and is selected from either 1 or 2, and m is 2 when n is 1 and 1 when n is 2.
It is. ] 2. The inorganic or organic anionic group
^n- is iodine ion, bromine ion, chloride ion, iodate ion, bromate ion, chlorate ion, periodate ion, perchlorate ion, chlorite ion, hypochlorite ion, nitrite ion, nitrite ion. The quaternary ammonium salt according to claim 1, which is a nitrate ion, a sulfate ion, a hydroxyl group ion or an anion group represented by any of the following formulas (2) to (5). Embedded image [In the formula, R ₄ represents an alkyl group or alkenyl group having 1 to 7 carbon atoms which may have a hydroxyl group or a carbonyl group. ] [Wherein, R ₅ does not exist (COOs are directly bonded)
Or an alkylene group or alkenylene group having 1 to 8 carbon atoms, and these organic groups may have a hydroxyl group. ] [Wherein, R ₆ represents an alkyl group having 1 to 2 carbon atoms. ] [Wherein, R ₇ and R ₈ are each a hydrogen atom, and have 1 to 12 carbon atoms.
Represents an alkyl group or a carboxyl group. ] 3. A compound synthesized by reacting a compound of the following formula (6) with a compound of the following formula (7):
A method for producing a quaternary ammonium salt as described above. Embedded image [In the formula, k is 0 to 4, and Y represents a chlorine atom, a bromine atom or an iodine atom. ] [Wherein, R ₁ and R ₂ are each an alkyl group having 4 to 12 carbon atoms, R ₁ and R ₂ may be the same or different, and R ₃ is an alkyl group having 1 to 2 carbon atoms. Represents a group. ] 4. A compound synthesized by reacting a compound of the following formula (8) with a compound of the following formula (9):
A method for producing a quaternary ammonium salt as described above. Embedded image [Wherein, R ₁ and R ₂ are each an alkyl group having 4 to 12 carbon atoms, R ₁ and R ₂ may be the same or different, and k represents 0 to 4. ] [Wherein, R ₃ represents an alkyl group having 1 to 2 carbon atoms, and Z represents a chlorine atom, a bromine atom, an iodine atom or the following formula (10):
It represents any of the groups of (11). ] [Wherein, R ₆ represents an alkyl group having 1 to 2 carbon atoms. ] [Wherein, R ₇ and R ₈ are each a hydrogen atom, and have 1 to 12 carbon atoms.
Represents an alkyl group or a carboxyl group. ] 5. A compound of the following formula (12) and a compound of the following formula (13)
The method for producing a quaternary ammonium salt according to claim 1, wherein the compound is synthesized by reacting with a compound of the formula (1). Embedded image [Wherein, R ₁ is an alkyl group having 4 to 12 carbon atoms, R ₃ is an alkyl group having 1 to 2 carbon atoms, and k represents 0 to 4. ] [Wherein, R ₂ represents an alkyl group having 4 to 12 carbon atoms, and J represents a chlorine atom, a bromine atom, an iodine atom or the above formula (11)
Represents any of the groups ] 6. An anion group of a quaternary ammonium salt having a cationic group represented by the following general formula [14] produced by the production method according to claim 3, 4 or 5, , Bromide, chloride, iodate, bromate, chlorate, periodate, perchlorate, chlorite, hypochlorite, nitrate, nitrite, sulfate, A method for producing a quaternary ammonium salt, wherein the quaternary ammonium salt is substituted with one kind of anionic group selected from a hydroxyl group ion or any one of the ionic compounds of the above formulas (2) to (5). Embedded image Wherein R ₁ and R ₂ are each an alkyl group having 4 to 12 carbon atoms, and may be the same or different;
R ₃ is an alkyl group having 1 to 2 carbon atoms, and k represents 0 to 4. ] 7. An antibacterial agent comprising the quaternary ammonium salt according to claim 1 as an active ingredient. 8. An ophthalmic preservative comprising the quaternary ammonium salt according to claim 1 as an active ingredient. 9. An eye drop containing the quaternary ammonium salt according to claim 1 or 2 as a preservative.