JP2015081233A - Antibacterial treatment method, and antibacterial treatment agent - Google Patents

Abstract

PROBLEM TO BE SOLVED: To deactivate a microorganism such as a bacterium, a fungus, and algae on a surface of a hard material of housing, furniture, equipment or the like, or to modify a receptor, a carrier and an enzyme that protrude from a cell membrane and a cell wall of the microorganism to inhibit each function.SOLUTION: An antibacterial treatment agent includes as an antibacterial base agent a 2-mercaptopyridine-N-oxide salt, a homologous derivative formed by substituting/modifying another group for hydrogen of a 3-6 position of the pyridine ring of the 2-mercaptopyridine-N-oxide salt, or a homologous derivative formed by substituting a pyrimidine ring for the pyridine ring. The antibacterial treatment agent is added to, impregnated in, or coated on an antibacterial treatment target by uniformly distributing so as to be 1 mg or more per 1 mof a surface area of the antibacterial treatment target.

Description

  The present invention is intended to inactivate microorganisms on the surface of hard materials such as houses, furniture, and fixtures, and to accept bacteria, fungi and algae that protrude on the surface of hard materials that protrude outside the cell membrane / cell wall. The present invention relates to an antibacterial treatment method and an antibacterial treatment drug that inhibit each function by binding to a body / transporter / enzyme and denatured.

Bacteria are evolutionarily classified as old prokaryotes, and are classified into positive and negative by the Gram staining method due to the difference in lipopolysaccharide that constitutes the cell wall. One mold is a eukaryote composed of a cell wall, a cell membrane, a nuclear membrane and a nucleus from the outside of the cell, and is classified as a “fungus” together with mushrooms in microbiology. Therefore, filamentous fungi that do not produce mushrooms among fungi are generally referred to as “mold”.
In the present invention, a drug that exerts algae prevention and antibacterial action by pushing it side by side, regardless of whether it is algae, fungi, or bacteria, and a drug that exerts a synergistic effect although its action site is limited by the base. This is referred to as “secondary agent”. In addition, a series of “sulfur-containing compounds” are collectively referred to as “sulfa drugs” like medicines.

Molds often grow on the surfaces of familiar clothing and hard materials in the living environment, and bacteria multiply. Also, glass windows in pools and bathrooms are condensed with high humidity, and the condensed water that has accumulated in the rail grooves flows down from the structure through the outer wall, so the algae grows along the sluice path along with sunlight and grows green. Often seen to become dirty.
Usually, the source of contamination of these fungi, bacteria and algae is environmental air and facility water.

  In order to eliminate such microbial growth, conventionally, as a symptomatic treatment, decomposition and bleaching have been performed using a so-called “chlorine-based disinfectant” obtained by diluting sodium hypochlorite with water. When the treated fungi are observed with a microscope, for example, most of the koji molds are in a lysed state by leaking cell fluid, and the shape at the time of survival is not stopped. In addition, a method of cleaning or wiping with a drug called “antifungal / antibacterial agent” or applying a surface treatment such as applying, spraying, or impregnating the drug itself to an object subjected to microbial contamination has been adopted.

  However, the conventional method, for example, the above-mentioned “chlorine-based disinfectant” sometimes causes a problem that the surface of the hard material to be cleaned is also bleached or altered. Another type of “antifungal / antibacterial agent” has been aimed at combining a plurality of drugs with significantly different chemical structures to exert a medicinal effect against various bacteria. However, it is generally questionable whether the target synergistic effect is actually exhibited because the microbial inactivation mechanism of the composite agent has not been elucidated in detail.

On the other hand, a compound having a basic skeleton such as sulfanilamide (the formula H ₂ NC ₆ H ₅ SO ₂ NH ₂ ) as an anti-infective internal medicine similar to the antibacterial agent is commonly called “sulfa drug” for human animals. It is publicly known.
This is literally a generic name for preparations containing sulfa (sulfur: S). Thousands of sulfa drugs have been synthesized so far, and they have been very effective in treating infections. Doctors are struggling to select sulfa drugs.
Furthermore, the pharmacological mechanism of diuresis and metabolic acidosis of the sulfanilamide has been studied. If the sulfamide group (—SO ₂ NH ₂ ) is contained in the chemical structure, the action of carbonic anhydrase is inhibited, and the hydrogen of the terminal amino group is reduced. It is now accepted that the inhibitory action disappears when a methyl group (—CH ₃ ) or the like is substituted to become “—SO ₂ NH (CH ₃ )” or the like. The reason is considered that when the structure of the terminal group (substrate) becomes a methyl group, it does not fit with the structure of the active site of the enzyme.

The medicinal components of disinfectant, antibacterial and antifungal agents distributed for each use are significantly different from those of structures that demonstrate the entry / exit of low molecular weight substances such as water and glycerin on the cell surface, protein species detection and enzymatic action. This is because the details could not be clarified.
That is, in the above-mentioned chlorinated disinfectant, it has been found that there is a possibility that through-holes are likely to be expanded by changing hypochlorous acid from non-dissociated to ionic form when passing through a transporter such as aquaporin. It was. Traditionally, this assumption has not been made.
If such a phenomenon occurs, the substance concentration equilibrium inside and outside the cell is broken, and water enters too much from the transporter broken by the osmotic pressure difference, and the cell membrane is ruptured. Or, conversely, dehydration occurs and contracts. The phenomenon of “lysis” is thought to occur in this way.
Alcohol-based disinfectants typified by ethyl alcohol can be regarded as aiming at dehydration characteristics and effects due to blockage of transporter passageways.
On the other hand, sulfa drugs and antibiotics may be regarded as acting on or engaging with receptors (receptors) or enzymes that detect proteins, thereby inhibiting the uptake of metabolites or losing the activity of enzymes. Therefore, the emergence of resistant bacteria is unavoidable.
Furthermore, quaternary ammonium salts, that is, cationic surfactants, can be classified as a type that covers the entire microorganism, inhibits mass transfer, and waits for self-destruction due to metabolic disorders.

In order to eliminate the above-mentioned drawbacks of the conventional method, the present invention has been devised by clarifying the structural commonality of microorganisms and elucidating the action mechanism of the mercaptopyridine oxide system. That is, chemical substances that can easily bind to proteins that are commonly present on the cell wall and membrane surface, without affecting the nuclear membrane and chromosomes existing inside the cell wall and cell membrane, can be selected from a myriad of chemical species. Selected based on the mechanism of action.
In other words, it is not aimed at lysis where the shape of the carcass disappears, and if it is a base that acts at the entrance level of the transporter, the grounds that the possibility of appearance of the above-mentioned resistant bacteria should be minimized based on.

In the same 6-membered ring material as benzene, one carbon (C) is substituted with nitrogen (N) is pyridine, and the two are substituted with pyrimidine. The derivatives have been synthesized for a long time, and their pharmacological action and toxicity have been investigated. Also, benzoyl peroxide [molecular formula (C ₆ H ₅ CO) ₂ O ₂ ] and sodium methoxide [same CH ₃ ONa] are reacted, and neutralized and extracted with an organic solvent, and then pyridine is added with perbenzoic acid. It is well known that oxidation results in pyridine-N-oxide pyridine-N-oxide.

Further, sodium which is an alkali metal is 2-mercaptopyridine [molecular formula (C ₅ H ₄ N) SH] in which hydrogen (H) at the ortho (o-) position of the pyridine ring, ie, 2-position hydrogen (H) is substituted with a mercapto group (—SH). When the salt is oxidized in the same manner as described above, 2-mercaptopyridine-N-oxide sodium monohydrate [molecular formula C ₅ H ₄ NaNOS · H ₂ O] crystals are obtained, which is also well known.

On the other hand, enzymes are not limited to vesicles in cells. As described above, protein chains acting as enzymes together with receptors and transporters also protrude outside the cell membrane and cell wall of microorganisms.
For example, chymotrypsin, which is a kind of digestive enzyme, is a peptide in which leucine (abbreviation Leu), valine (valid) and cysteine (cys) are peptide-bound, and naturally, a mercapto group (—SH) as an active group on the cysteine branch. ).
The through-hole structure of ion channels, aquaporins and aquaglyceroporins, which have been studied rapidly in recent years, is a cylinder composed of cysteine, arginine (abbreviated Arg), phenylalanine (same Phe) and histidine (same His). It has been found to be a chain protein. That is, this also contains cysteine and has a mercapto group (-SH) in the branch of the bond chain as in the above-described enzyme.

Furthermore, in the human body, when mercapto groups of cysteine branch groups are close to each other, they are relatively easily oxidized to form a disulfide bond (-SS-). Actually, mucins and antibodies in which polypeptides are aligned or bent and α-β chain constant parts of T cells are also cross-linked by the bonds.
Incidentally, “cystine”, which is often found in hair, is a dimerized cysteine dimer with a disulfide bond. Insulin, a kind of hormone, also has a disulfide bond portion and a portion that remains a mercapto group.

However, Non-Patent Document 1, which can be said to be a textbook of modern pharmacology, has no description referring to a chemical reaction between a mercapto group of a cysteine branch group and an antibacterial agent.
The reaction in the body is a reaction in a body fluid containing carbohydrates, lipids, proteins and electrolytes, that is, an aqueous solution. Therefore, it was difficult to isolate and analyze the product and reproduce the reaction, which could not be explained. For these reasons, there are only a few parts in other microbiology-related non-patent literatures that mention the SH group oxidation of enzyme proteins or nucleoproteins.

Therefore, the antibacterial property of o-phenylphenol [OPP; formula C ₆ H ₄ (OH) C ₆ H ₅ ], which was developed as a fungicide and preservative for citrus fruits and bananas, is different from that of conventional phenols and cresols. The explanation of the extended interpretation based on structural similarity has been completed. Therefore, it has never been explained by its action on receptors, transporters and enzymes.
Thiabendazole [TBZ; molecular formula C ₁₀ H ₇ N ₃ S], which has the same application, is also a known substance, but its structure is more complicated than that of the above-mentioned OPP, and the mechanism of action is still not clear. In spite of this, many compounds in which the compound is further made into a complex-type drug have been commercialized, and there are many patent applications.

Many azole antifungal agents inhibit the function of enzymes (for example, cytochrome P450). The azole series to which TBZ also belongs is further classified into imidazole series and triazole series, and each antifungal drug is listed in Non-Patent Document 1, but most are external drugs. The only oral or intravenous administration example in the imidazole system is miconazole.
Of course, TBZ has no pharmaceutical approval. Therefore, since the antibacterial composition based on the action mechanism of the compound cannot be narrowed down, this type of drug discovery places emphasis on antibacterial testing, picks up substances that show medicinal effects with priority on results, and establishes a compound drug formulation. There is a tendency to concentrate on.

On the other hand, even if the above-mentioned proteins forming the receptor / transporter / enzyme are constructed with peptide bonds of amino acids whose order is different from the type, there is no significant difference in the amount or type of each amino acid residue. The situation and microorganisms that have no cysteine cannot exist in reality.
Therefore, any compound that acts on the mercapto group of cysteine should be effective against most microorganisms. This is because, when bound to a transporter, the through hole is capped, and when bound to a receptor or an enzyme, the shape and size of the active site are modified.
Paradoxically, when the antibacterial agent has no medicinal effect, it is interpreted that the microorganism is limited to a factor that inhibits binding to cysteine.

In summary, Patent Document 1 and Patent Document 2 each include 6 systems including 5 systems of nitrile, pyridine, haloalkylthio, organic iodo, and thiazole, in addition to 2-mercaptopyridine-N-oxide salt. A prescription for a composite antibacterial agent is described that requires selection of one or more compounds.
However, mercaptopyridine oxides should be an independent system, and it is not preferable to include them in one kind of pyridine or sulfa drugs for the reasons described above. This is because mercaptopyridine and mercaptopyridine oxide have completely different mechanisms of action. Therefore, it is not possible to perform an equivalent drug efficacy evaluation for each rough strain.
Furthermore, in the case of a composite agent, since the action and effect are generally completely different if the base and the secondary agent react, consideration for complex formation becomes indispensable.
As specific examples, Non-Patent Document 2 exemplifies several types of metal complexes with respect to the substrate of the present invention, and raises a question about the effect of Patent Document 1 aimed at the effect as a composite agent. Even if the wetted material is made of metal, there is a high possibility that the medicinal effect will be lost, which may cause a synergistic effect as well as an offset effect.

The reaction between chlorinated disinfectants and proteins has been elucidated to some extent for hypochlorous acid and proteins in saliva (eg mucins). For example, in the present inventors' research, the reaction is as fast as the disulfide bond reaction described above. On the other hand, when chlorinated to form organic chloramine, the bond between nitrogen and chlorine is strong, and even when iodide or potassium bromide (KI, KBr) is added, the bond is reversely dissociated and iodine (I ₂ ). It has also been found that it takes a long time of about 30 minutes to liberate bromine (Br ₂ ).
Furthermore, in the study of sterilization by ultrasonic waves, it has been found that even when the killing rate is 99%, most of the koji molds remain in the shape, and only the ciliated portions are disturbed by the ultrasonic waves.
That is, in order to inactivate microorganisms, it is not an essential condition to destroy the main body (such as bacterial cells) and to leak cell fluid.

JP 2006-52205 A Japanese Patent No. 3526919 (registered on February 27, 2004)

NEW Pharmacology (3rd edition), Nanedo Co., Ltd. (1996) C.A.Doose etal., J.Chhromatogr.1052 (2004) 103-110

Any compound that denatures a protein without blocking the structure of the receptor / transporter / enzyme that the microorganism has, or exerts an enzyme activity, and that acts to block the groove or through-hole of the protein structure. Even if the number is large, it should be effective with a relatively low concentration of the drug.
The present invention has been vaguely recognized in the drug discovery process of sulfa drugs and the like so far, "the active ingredient of the enzyme and the receptor is adsorbed and blocked, and the function of both is inhibited to inactivate the microorganism. The evaluation of the concept of “plan” was downgraded to “efficacy as an adjunct to the base”.
That is, the originality of the present invention is that “inactivation of a wide range of microbial species is based on a medicinal component capable of forming a disulfide (covalent) bond stronger than adsorption, not only for enzymes and receptors but also for transporters. The introduction of a completely new concept of “should be”.
Therefore, searching for compounds that have a minimum growth inhibitory concentration (MIC value) of 25 mg / L or less for most microorganisms, that is, antibacterial bases, and composite prescription design have become problems.

Although some methionine in sulfur-containing amino acids other than cysteine, since the end of the branch is a methylthio group rather than a mercapto group (-SCH _3), the reactivity of this group is extremely reduced. Therefore, the antibacterial agent composition aiming at coupling | bonding with a methylthio group was excluded from the choice from the beginning.
Moreover, it is an organic compound having the same mercapto group that easily binds to the mercapto group of cysteine, and it is an oxidizing active group present in the vicinity that promotes disulfide bonds by dehydrogenating from both mercapto groups. Accordingly, the oxygen possessed by N-pyridine oxide inevitably becomes the top candidate for the oxidation active group.
That is, in the presence of a strong oxidizing agent such as hypochlorous acid (HOCl) or hydrogen peroxide (H ₂ O ₂ ), the active group other than the mercapto group is oxidized to increase the consumption amount. Cannot achieve its intended purpose of not aiming.

Even when antibacterial treatment is performed on the surface of the hard material in clothing and living environment, the presence of water, that is, water, is indispensable for the combination of falling bacteria and fungal spores and the antibacterial agent. Therefore, it is desirable that the antibacterial agent is water-soluble in order to cause disulfide bonds with most of the bacterial species, and the bacterial species exhibiting a medicinal effect is extremely narrow when it is fat-soluble.
When the chemical is added to a building material or the like, the amount that is not exposed increases, so that it is necessary to increase the adsorption specific surface area and to reduce the addition amount. That is, it is intentionally easy to get wet. Such condition setting is one of the problems.

Furthermore, when an antibacterial treatment such as impregnation or coating is performed, an unreacted antibacterial base may flow out to the natural world and cause secondary phytotoxicity if dissolved in water. Therefore, one of the challenges is to bind firmly to cysteine in microbial proteins that are close to each other and prevent them from flowing out.
Excessive addition, impregnation and application to an antibacterial treatment object lead to an increase in the residual concentration of unreacted substances, so even if the upper limit of the addition amount / rate is not set in the claims of the present invention, the outflow Based on the results of volume and rate experiments, there is a natural restriction when prescribing. Further, since it is necessary to avoid damaging the original characteristics of the antibacterial object, the amount and rate of addition are naturally limited in this respect as well.

Means for solving the above-described problems are as follows.
(1) 2-mercaptopyridine-N-oxide salt, or a homologous derivative in which hydrogen at the 3-6 position of the pyridine ring of the 2-mercaptopyridine-N-oxide salt is substituted or modified with another group, or An antibacterial treatment agent containing a homologous derivative in which the pyridine ring is substituted with a pyrimidine ring as an antibacterial base is uniformly distributed so as to be 1 mg or more per 1 m ² of the surface area of the antibacterial treatment target, and added to the antibacterial treatment target. An antibacterial treatment method characterized by performing impregnation or coating.
(2) The antibacterial agent is a sulfa drug compound that inhibits the functions of enzymes and receptors on cell walls and cell membranes of bacteria, fungi, and algae to produce a synergistic antibacterial effect. The antibacterial treatment method according to (1), wherein the upper limit of an equivalent amount is included.
(3) The antibacterial treatment agent is characterized by further containing an equivalent amount of the antibacterial base as an upper limit with a complex of iodine and an organic polymer having a weaker oxidizing power than hypochlorous acid as a second adjunct. The antibacterial treatment method according to (2).
(4) 2-mercaptopyridine-N-oxide salt, or a homologous derivative in which hydrogen at the 3-6 position of the pyridine ring of the 2-mercaptopyridine-N-oxide salt is substituted or modified with another group, or An antibacterial treatment agent comprising, as an antibacterial base, a homologous derivative in which the pyridine ring is substituted with a pyrimidine ring.
(5) The antibacterial agent is a sulfa compound that inhibits the functions of enzymes and receptors on the cell walls and cell membranes of bacteria, fungi, and algae to produce a synergistic antibacterial effect. The antibacterial treatment agent according to (4), characterized in that it contains an upper limit of an equivalent amount.
(6) The antibacterial treatment agent is characterized by further containing an equivalent amount of the antibacterial base as an upper limit with a complex of iodine and an organic polymer having a weaker oxidizing power than hypochlorous acid as a second adjunct. The antibacterial treatment agent according to (5).

According to the above means (1), the 2-mercaptopyridine-N-oxide salt has a mercapto group at the ortho position (2 position) with respect to the nitrogen N (1 position) in the chemical structure. Inhibits all functions of enzymes, receptors and transporters that protrude outside the cell wall.
Among the amino acids constituting the protein structure, a disulfide bond is formed in the vicinity of a mercapto group which is a branch group of cysteine. The oxygen atom coordinated to the nitrogen atom of the pyridine ring promotes oxidation (dehydrogenation) at this time.

Therefore, a quick disulfide bond cannot be expected with 2-mercaptopyridine which is not oxidized or is not salted with sodium or potassium. That is, two chemical structural conditions are indispensable: being an oxide and being a salt with an alkali metal in order to impart water solubility. Therefore, it is not preferable to simply enumerate homologous derivatives as “pyridine-based”.
The above-mentioned minimum growth inhibition concentration (MIC value) is indicated by the drug concentration unit “mg / L” in the medium. On the other hand, the antibacterial treatment of the present application is addition, impregnation or application to an object. In this case, since the drug that comes into contact with microorganisms such as bacteria and fungi is the surface of the hard material, unlike the case of culturing, the drug concentration in the deep layer need not be considered as long as the drug is contained in a thin layer on the surface. .
Assumptions for ensuring consistency between the MIC value in the medium test and the drug concentration in the surface treatment are shown below.
The density of the antibacterial treatment base of 25 mg / m ² corresponds to approximately 25 mg / kg and an aqueous solution concentration of 25 mg / L on the assumption that an aqueous solution having a specific gravity of 1 is stretched on the surface of the treatment object with a film thickness of 1 mm. Assuming that the microbe to be inactivated has a slightly larger size of 10 μm, the thin film volume at a film thickness of 0.04 mm = 40 μm, which is four times this, is 40 cm ³ / m ² . If 1 L of a liquid with a drug concentration of 25 mg / L is spread with a film thickness of 40 μm, the liquid 1 L = 1000 cm ³ , and thus the surface area of the spread liquid is 1000/40 m ² = 25 m ² , and thus the drug density Becomes 1 mg / m ² .
The solvent after adding, impregnating or applying to the object is substantially evaporated. The lower limit of the thickness at which the hard material surface is uniformly wetted is naturally affected by the surface of the hard material and the surface tension / absorption rate of the solvent. Therefore, in the present application, the thickness of the impregnated or coated film before drying is considered to be about 40 μm, and if the drug density is 1 mg / m ² or more, “the blank where the drug does not adhere to the hard material surface” cannot be formed. The lower limit of the drug density was “1 mg per 1 m ² of surface area of the treatment target”. That is, the numerical value is an index for “uniformly distributing” when the drug concentration cannot be made uniform as in a solution, and the actual concentration in a chemical solution for antibacterial / antifungal treatment is more than 25 mg / L Slightly increase the margin of accuracy.
Accordingly, the surface area that can be treated is naturally different between application and immersion. The on-site effect is determined by a culture method such as a stamp method or a wiping method, or ATP (adenosine triphosphate) measurement. However, this judgment method has not yet been standardized internationally.
In addition, in a cytotoxicity test conducted using the available V79 cells and the base alone, the IC ₅₀ value (colony formation 50% inhibitory concentration) was about 0.05 mg / mL = 50 mg / L. This result means that the base reacts with the enzyme other than the enzyme of the microorganism and inhibits the growth regardless of the origin of the cell. However, the IC ₅₀ value is 20 times the concentration (2.5 mg / L) of the sulfa drug (di-n-butylthiocarbamate zinc complex) used as a test control, and its inhibitory power is weaker than that of the control.
In addition, an antibacterial base is a salt of N-oxide (-N-oxide) in which mercaptopyridine (2-Mercaptopyridine) or mercaptopyrimidine (2-Mercaptopyrimidine) is oxidized in terms of chemical structure, and a mercapto group (-SH). Corresponds to the one in the 2-position relative to the nitrogen (N) of the pyridine ring or pyrimidine ring to which the oxygen atom is coordinated.
N-oxide may be a tautomer having the structure of “1-Hydroxy-2-pyridinethione” or “1-Hydroxy-2-Pyrimidinethione”. Characteristic absorption is seen around 330 nm.
The powder or granular substance is a salt with an alkali metal typified by sodium or potassium, and dissociates into ions when prepared as a liquid.

According to the above-mentioned means (2), the single component base material synergistically inhibits the functions of enzymes and receptors on the cell wall and cell membrane with respect to bacteria, fungi, and algae having a relatively high minimum growth inhibitory concentration. Provides antibacterial effect. The sulfa drugs used as an auxiliary agent, unlike the base, formulate a compound that inhibits the functions of the above-described enzymes and receptors. The transporter is characterized by having a through-hole, and the action mechanism is different from that of the base in that the sulfa drug cannot be expected to block the through-hole.
A compound having a dimethylaminosulfonyl group [—SO ₂ N (CH ₃ ) ₂ ], a diiodomethylsulfone group [—SO ₂ (CHI ₂ )] or a sulfamide group (—SO ₂ NH ₂ ) in the chemical structure is Since it contains sulfur (S) in common, it enters the above-mentioned chemical species generically called “sulfa drugs”. All can be summarized as those aimed at inhibiting enzyme activity.
More limitedly, sulfanilamide or a homologous derivative is effective in mating with an active site such as carbonic anhydrase possessed by a microorganism and delaying the rate of dehydration or the like that results in an equilibrium reaction. Also from this fact, the mechanism of action is completely different from the base of the above-mentioned means (1).
Therefore, the composite agent of a base and an auxiliary agent exhibits a synergistic effect by making the action mechanism different regarding antibacterial properties. Since the combination of the base and the secondary agent reacting during storage must be avoided, the choice of secondary agent is naturally narrowed.
The silver / copper ion-supported inorganic agent described in the above-mentioned Patent Document 1 is likely to form a complex with the base, as can be guessed in Non-Patent Document 2, and to have a countervailing effect.
The original aim of the supplement is synergistic effect. A sulfa drug having a structure that fits into the active site of a microbial enzyme is an option of the secondary agent in the above-mentioned means (2).

There are other reasons for using a sulfa drug other than the base as a secondary agent. Receptors that detect the concentration of lipids (such as cholesterol) in human blood cannot detect lipids themselves, and therefore, in the form of lipid proteins, fit with amino acid moieties and give a signal for taking lipids into vascular constituent cells. In addition, the α-β chain of sensitized T cells that are cellular immunity and the tip variable part of an antibody (B cell) that is humoral immunity are also detected by the amino acid structure of the antigen protein. Therefore, a general sulfa drug does not fit entirely in the groove of the enzyme active site, but partially fits it, so it is considered that the probability of fitting with a receptor other than the enzyme is low. It is.
From the above viewpoint, it is considered that the salazosulfapyridine (also known as sulfasalazine) selected in the examples of the present invention acts on the enzyme fitting part and amino acid transporter (transporter) protein to inhibit the metabolism of microorganisms. It is done.
The compound with mercaptopyridine also has a pyridine ring in the material structure, so that there is no need to worry about the occurrence of an offset effect when they react with each other.

According to the above-mentioned means (3), for example, an iodine complex of polyvinylpyrrolidone known as an antibacterial agent (abbreviation: popidone-iodo) is considered that iodine is coordinated and added to a lone pair of pyrrolidone ring nitrogen. It exhibits antibacterial and antifungal synergistic effects through a mechanism of action different from mercaptopyridine and sulfa drugs.
In addition, the pyrrolidone ring described above is different from a five-membered ring and a pyridine ring is a six-membered ring. In addition, the pyrrolidone ring is carbonyl in which one of carbons covalently bonded to nitrogen is oxidized, and there is a difference that is not a resonance structure like a pyridine ring, but there is no significant difference in chemical action.

On the other hand, the mechanism of action of free iodine is not significantly different from that of free chlorine such as hypochlorous acid. Hypochlorous acid exerts a powerful action up to lysis, but iodine that has been slowly released does not have much action.
Therefore, popidone-iodine has long been used for disinfection of fingers and surgical sites and surface disinfection of mucosal wound sites and fever sites.
From the above viewpoint, even if the popidone-iodide selected in the examples of the present invention is a triple complex with the drugs of the above-mentioned means (1) and (2), an offset effect due to the mutual reaction between the drugs is recognized. There wasn't.

  Microorganisms on the surface of hard materials in clothing and living environment are food chain, unlike the pioneer in the colony of the bacteria group that makes up the so-called “slime” (slime city) (the basis for the living environment) Few. Therefore, fungi, bacteria or algae compete with each other, and an inactivation method aimed at breaking the food chain in a slimy symbiotic society in an aqueous system cannot be taken.

Further, the 2-mercaptopyridine-N-oxide salt is an existing chemical substance commercially available under the name of a manufacturer's component “pyrithione sodium”, for example, for use in preventing scale buildup. Moreover, the acute oral toxicity The _{LD 50} was 2000 mg / kg or more (rat), it does not correspond to the hazardous component.
The above-mentioned thiabendazole is also a sulfa drug in a broad category, but since suspicion has not been solved in terms of teratogenicity, in the present invention, the TBZ-like structure is excluded from the choice of the secondary agent from the beginning. Originally, the history of sulfa drugs that have been studied as pharmaceuticals is long, so even if you do not stick to thiabendazole, refer to the accumulated chemical technology information and refer to “the acute oral toxicity LD ₅₀ is large or non-toxic”. Select and suffice.

In addition, the antimicrobial base alone of the above-mentioned means (1) increases the MIC value, but is found to be effective against all 57 types of microbial strains detected from general buildings.
This is nothing other than evidence that a mercapto group is present in the cysteine branch group that is always held in common by microorganisms, and that the surface of the microorganism cell is denatured by disulfide bonding with the mercapto group of the base of the present invention. It is a feature that completely separates the mechanism from the function-inhibiting action due to the blockage of the enzyme active site assumed in the secondary agent.

1 is a basic molecular structure diagram of an antibacterial base according to an embodiment of the present invention. It is a molecular structure figure of the pyrimidine ring substance concerning an embodiment of the invention. It is a molecular structure figure of an example of the 1st auxiliary agent concerning embodiment of this invention, sulfasalazine. It is a molecular structure diagram of popidone iodine, an example of the second auxiliary agent according to the embodiment of the present invention. It is an ultraviolet-spectroscopy spectrum figure in the monomer-dimer coexistence of the base concerning embodiment of this invention. It is explanatory drawing of the coating process method concerning embodiment of this invention. It is explanatory drawing of the immersion treatment method concerning embodiment of this invention. It is a comparison table | surface which shows the classification | category and scientific name of the microorganism species concerning embodiment of this invention. It is a list of microbial species detected from the general building concerning an embodiment of the invention. However, only No. and scientific name are listed.

  Hereinafter, with reference to the microbial Japanese / scientific name comparison table of FIGS. 1 to 7 and FIG. 8, the microbial inactivation base and the treatment method on the hard surface of the clothes and living environment of the present invention will be described.

The basic molecular structure of the antibacterial base of the present invention shown in FIG. 1 is a pyridine oxide skeleton (circle in the six-membered ring represents π orbital electron resonance as in benzene), as described above. It has a mercapto group (-SH) in the ortho position (2-) with respect to nitrogen N (position 1). In addition, tautomeric structures of the symbols “A” and “B” in FIG. 1 are conceivable. In particular, in a polar solvent typified by water, a dimer having the same “C” structure coexists with the monomer.
Accordingly, in the ultraviolet spectrum of FIG. 5, the absorbance at about 330 nm possessed by only the monomer is measured at a plurality of concentrations, a calibration curve is prepared, and based on this, the concentration of the base can be measured.
Before dissolution, a stable base is formed in the form of a salt with an alkali metal such as sodium or potassium. Even if the ring structure of the base is replaced with the structure shown in FIG. 2, there is no difference in action.
In addition, sulfasalazine was selected as the first adjunct (sulfa agent) and used as an example. In the case of a liquid preparation, sulfasalazine has a low solubility in a polar solvent, so the mixing amount was close to the saturation concentration, but the concentration ratio was substantially about 2% of the base.
Furthermore, popidone iodine was selected as the second adjunct (iodine agent) and used as an example. In this case, since the solubility is the same as that of the base, the equivalent amount to the base was set as the upper limit.

Therefore, as Example 1, a preliminary experiment was conducted for the purpose of proving the reason for the base of the base.
First, E. coli was cultured and prepared to 6.0 × 10 ⁹ CFU / mL. 1 mL of the strain water was added to 200 mL of water having a base concentration of 50.14 mg / L. As a result, the concentration before reaction / adsorption was slightly diluted and calculated as “50.14 × 200/201 = 49.89 mg / L (standard deviation: 0.01)”.
After contacting the base and Escherichia coli for about 10 minutes and then filtering with a 0.45 μm filter made of a material that does not react and adsorb to the base, the concentration of the base was measured to be 49.1 mg / L. Therefore, the concentration decreased to “49.89−49.16 = 0.73 mg / L” (pH = 7.04), and compared to the original 49.89 mg / L, “0.73 / 49.89≈0.0. “015 = 1.5%”.
In the same manner, E. coli was cultured and adjusted to 1.0 × 10 ⁵ CFU / mL. The base was contacted with E. coli as described above. The concentration before the reaction / adsorption was slightly diluted and calculated as “49.14 × 200/201 = 48.90 mg / L (standard deviation: 0.01)”.
Further, after the contact, the concentration decreased by “48.90−48.36 = 0.54 mg / L”. It was a “significant decrease” of “0.54 / 48.90≈0.011 = 1.1%” with respect to the original 48.90 mg / L.

This result shows that the mercapto group of cysteine, which is possessed by microorganisms and not disulfide-bonded, binds to 2-mercaptopyridine and does not fit with the outer shell enzyme of the microorganism. It is a proof that it has not been liberated.
In addition, since the molecular diameter of the base component is much larger than that of water or hypochlorous acid, it cannot pass through the transporter and enter the cell. Therefore, even if the microorganism dies, it does not enter a lysed state, and the shape before life is substantially left.
That is, it has been proved that the base of the present invention reacts or strongly adsorbs on the cell surface regardless of whether microorganisms are alive or dead, and has an antimicrobial / antifungal premise.

Next, as Example 2, in order to prove that the antibacterial property is widespread regardless of bacteria, fungi, and algae with a base-only formulation, a variety of microbial strains owned by a test contracting organization are used, and 2-mercaptopyridine is used. -Sodium N-oxide was added to 3 types of media so as to be 25 mg / L each, or the concentration was adjusted stepwise to prepare and dispensed into the required number of petri dishes.
One kind of seed strain suitable for each medium was planted in each petri dish and cultured for 7 days under the conditions of 30 ° C. and relative humidity of 95%. And the case where proliferation was not seen was determined as "growth prevention." Moreover, the growth inhibition minimum concentration (MIC value) was determined for each microbial species in the above-described stepwise dilution test. The results thus obtained are summarized and shown in Table 1. From the experimental method, the MIC value exceeding 25 mg / L is not counted.
Furthermore, two kinds of salazosulfapyridine as the first auxiliary agent and povidone-iodo (polyvinylpyrrolidone iodine complex) which is a free halogen agent are selected as the second auxiliary agent, and each is equal in weight. After mixing with powder, add 25 mg / L each of the base following the base in the same manner as described above, or add the same amount if the base is gradually reduced in concentration, and prepare and dispense the medium as described above. Went.
In this case, the culture / judgment method and the MIC value measurement are exactly the same as those for the base alone. In addition, the number of bacterial species to confirm the effectiveness has been greatly increased.

Table 1 summarizes the number of microbial species whose effectiveness was confirmed.

The mercaptopyridine oxide system alone is effective in a wide range of microbial species, and is suitable as a base because it is effective against a wide range of microbial species.
On the other hand, it was found that the synergistic effect obtained was small even when a sulfa agent was used as a base and a composite agent of the same type, an inorganic antibacterial agent or a non-sulfur-containing halogen-based organic antibacterial agent.
This is because the above-mentioned thiabendazole is not included in the numbers listed in Table 1 unless the concentration is 100 mg / L or more, and especially against algae, it does not show medicinal effects even at the concentration. Therefore, thiabendazole is inferior in efficacy compared with a chlorine-based disinfectant that exhibits a remarkable effect on almost all types of microorganisms at a free residual chlorine concentration of 1 mg / L unless adverse effects are taken into consideration.
However, the mercaptopyridine-N-oxide system of the present invention is only combined with one or two kinds of sulfa drugs and is comparable to halogen-based disinfectants, and the average MIC value is one digit higher than that of the base alone. Go down. In addition, it exhibits a medicinal effect that does not interfere with practical use against two bacterial species slightly inferior to chlorine-based disinfectants.

The results of Example 1 and Example 2 show that the product of the present invention can prevent antibacterial / antifungal / algae generation at 1/5 to 1/20 of the MIC value of the sulfa drug, and the basis for that is 2- Each suggests a mercaptopyridine-N-oxide salt.
Since this compound has a mechanism of action different from that of sulfa drugs aimed at inhibiting the function of enzymes and receptors as described above, in fact, it belongs to fungi, Kawaratake, Pseudomonas aeruginosa, Escherichia coli, Staphylococcus aureus and Bacillus subtilis. It exhibits medicinal effects in a wide range of species, including the genus Trenteporia, Chlorella and Schizosrix, which are familiar with algae. However, for Pseudomonas aeruginosa, Bacillus subtilis, and Escherichia coli that have capsules and secrete polysaccharide mucus, about 10 times the concentration of chlorinated disinfectant is required.

FIG. 6 is an explanatory diagram of a method for carrying out the treatment of the present invention on textiles, cloths, nets, leather, rubber / plastic films, etc. in clothing and living environment.
When the antibacterial treatment object 5 is a cloth or a flat plate, the system is constructed so as to rise and fall between the feed rollers 6. A nozzle 4 for spray coating or spraying processing is provided at the apex of the processing object. The chemical solution containing the microorganism-inactivating base or the composite agent 1 is sprayed or applied from the storage container 2 to the surface of the object 5 to be processed by the drug delivery pump 3.

  When the base or composite 1 is dissolved or suspended, the liquid ejected from the nozzle 4 adheres to the surface of the object 5 to be processed and forms a permeation or film to a certain depth together with the solvent. Accordingly, the amount of the medicine discharged is adjusted to such an extent that the surface is wetted so as not to leave a blank for application. When the solvent spontaneously evaporates and is dried to some extent, the drug is concentrated to a film thickness of 40 μm or less, and the antifungal, antibacterial, and algae prevention treatment is completed.

FIG. 7 is an explanatory view of a method for performing treatment by immersing in a chemical solution in which the base or composite agent 1 is dissolved or suspended. If the object 5 is not a continuous plate made of basic materials such as building materials / furniture furniture, painted products, household and industrial products, school supplies, exercise equipment and toys, the object 5 shown in FIG. The processing method is replaced by a belt conveyor.
In any case, it is still the same that the anti-mold, antibacterial and anti-algae treatment is performed by dipping. In this case, as a matter of course, the mechanism of the feed roller 6 is also constructed and organized in a form commensurate with the solid shape of the processing object 5. The process is completed when the liquid is pulled up and dried.
Although the surface wetness of the processing object 5 depends on the surface tension and the chemical solution permeation rate, the film thickness of the drug at the time of drying is approximately 40 μm or less and can be processed without a shortage of the exposed drug density.

In addition, when the processing target object 5 does not have a mirror finish, if the adsorption surface area is expected to be relatively large, the powdery base or composite agent 1 is directly kneaded or electrostatically / powder coated in the manufacturing process of the processing target object 5. It is also possible. However, since the physical and mechanical strength of the product must not be impaired, the amount and rate of addition are naturally limited.
In this case, the medicinal effect of the base or complex 1 is that moisture is adsorbed or taken into the cells of the microorganism, and the reaction with an enzyme or a receptor in an aqueous solution (exactly in an electrolyte solution containing protein or the like). It is thought that it is demonstrated by fitting.
Therefore, the calculation of the amount added to the base or complex 1 is considered to be rate-limiting to the dissolution rate and needs to be about 100 times the MIC value (2500 mg / kg = 0.25% by weight when the product thickness is 1 mm). Become.

  Select 2-mercaptopyridine-N-oxide sodium monohydrate as the antibacterial base, sulfasalazine (salazosulfapyridine) from sulfa drugs as an auxiliary agent, and popidone-iodo (iodine of polyvinylpyrrolidone) as a free halogen agent After selecting the two types of complex) and treating the hard material surface of the object to be treated, the bacteria, fungi, and algae were cultured by wiping or pressing methods. Of all 57 microbial strains.

The negative bacterial species and the like were compared with the preliminary experiment results of Example 1 and shown in order from the relatively high MIC value as follows.
Pseudomonas eleginosa, a species of Pseudomonas aeruginosa, Cryptococcus latiaras belonging to fungi, Erminsosporium gramineum, Penicillium islandicam, Penicillium lilacinum, Pitcha membana efaciens, Rhodotorula gallinis, Rhodotorula lactosa and It was the only autotrophic bacterium belonging to the bacterium. The MIC value of other types of fungi / bacteria for which the drug “base and complex of two types of complex” of the present invention was determined to be effective was 15 mg / L or less, and the MIC value of algae was generally 10 mg / L. .

  The types of bacteria, fungi, and algae depend on the difference in the chromosomal genes they possess, but the physical structure and shape are determined by the functional differences between the enzyme and the receptor. In this respect, it can be said that the function of the transporter is little involved.

  Therefore, the remarkable effect of the secondary agent alone was limited, and there was a remarkable synergistic effect to lower the MIC value of the base. The medicinal effect of sulfasalazine as a first adjunct was remarkable for Aspergillus spp., Altarnaria spp., Geotricum spp., Mucor spp. And Trichoderma spp. A remarkable synergistic effect was also observed against S. aureus and E. coli.

  In addition, the efficacy of popidone iodine as a second adjunct overlapped with the fungi, Aspergillus genus and Geotricum genus described above, and was remarkable for Botrytis genus, Cladosporium genus, Fusarium genus and Penicillium genus. The synergistic effect on bacteria was about the same as that of sulfasalazine.

  In summary, in order to prevent the growth of bacteria, fungi and algae present on familiar hard surfaces without omission, antibacterial, antifungal and anti-algae agents with different action mechanisms are combined with mercaptopyridine-N-oxide salts. It is possible to cope with the composite preparation of the present invention based on the above and combined with one kind of sulfa drugs and one kind of halogenated complex. In general uses such as daily necessities, inorganic antibacterial agents and composite organic antibacterial agents of more than four types are not required.

  Moreover, when the hard material surface is antibacterial treated, the elution amount is a concern. As described above, the drug remaining on the surface after the treatment according to the present invention is in a substantially dry state, and microorganisms that have come into contact with the drug in the course of treatment are converted into dormant bodies (spores or cysts) or dead bodies. Therefore, the amount of residual drug after treatment is very small, and the amount eluted in the water adsorbed on the surface is negligible. Therefore, the surroundings are similar to competing surfactants and sulfa drugs alone or in combination with antibacterial agents. It is not necessary to consider the outflow to the environment.

  The present invention relates to objects having solid surfaces, such as clothes, textiles, fabrics, nets, leather, rubber, plastics, films, building materials, furniture, paint products, household and industrial products, school supplies, sports equipment, toys, etc. If so, it can be widely used for antibacterial treatment. Moreover, since it is effective against algae, it can be used by spraying on the outer wall of a window where condensation is likely to occur to prevent indigo and green stains.

Claims (6)

2-mercaptopyridine-N-oxide salt, or a homologous derivative in which hydrogen at the 3-6 position of the pyridine ring of the 2-mercaptopyridine-N-oxide salt is substituted or modified with another group, or the pyridine ring An antibacterial treatment agent containing a homologous derivative substituted with a pyrimidine ring as an antibacterial base is uniformly distributed so as to be 1 mg or more per 1 m ² of the surface area of the antibacterial treatment target, and is added to, impregnated or coated on the antibacterial treatment target. An antibacterial treatment method characterized by performing attachment.   The antibacterial agent is equivalent to the antibacterial base with a sulfa drug compound that inhibits the functions of enzymes and receptors on the cell walls and cell membranes of bacteria, fungi, and algae to produce a synergistic antibacterial effect. The antibacterial treatment method according to claim 1, wherein the upper limit is included.   The antibacterial treatment agent is characterized by further containing an equivalent amount of the antibacterial base as an upper limit with a complex of iodine and organic polymer having weaker oxidizing power than hypochlorous acid as a second auxiliary agent. 2. The antibacterial treatment method according to 2.   2-mercaptopyridine-N-oxide salt, or a homologous derivative in which hydrogen at the 3-6 position of the pyridine ring of the 2-mercaptopyridine-N-oxide salt is substituted or modified with another group, or the pyridine ring An antibacterial agent containing a homologous derivative substituted with a pyrimidine ring as an antibacterial base.   The antibacterial agent is equivalent to the antibacterial base with a sulfa drug compound that inhibits the functions of enzymes and receptors on the cell walls and cell membranes of bacteria, fungi, and algae to produce a synergistic antibacterial effect. The antibacterial treatment agent according to claim 4, wherein   The antibacterial treatment agent is characterized by further containing an equivalent amount of the antibacterial base as an upper limit with a complex of iodine and organic polymer having weaker oxidizing power than hypochlorous acid as a second auxiliary agent. 5. The antibacterial treatment agent according to 5.