Mycobacterium bactericide

Abstract

Provided is a mycobacterium bactericide. The mycobacterium bactericide includes chlorous acid water. The free chlorine concentration of the chlorous acid water (as Cl) may be 50 ppm or more. A biofilm may be formed with the mycobacterium. The mycobacterium may be Mycobacterium intracellulare, Mycobacterium avium, Mycolicibacterium fortuitum subsp. fortuitum, Mycobacterium runyonii, Mycobacterium abscessus, Mycobacterium kansasii, M. avium subsp. paratuberculosis, or Mycobacterium ulcerans. A medicine containing chlorous acid water is provided for preventing and / or treating MAC lung disease.

Description

Technical Field

[0001] The present disclosure relates to an agent for killing Mycobacterium bacteria, a drug for preventing and / or treating pulmonary MAC disease. The present disclosure also relates to a method for killing Mycobacterium bacteria, a method for preventing and / or treating pulmonary MAC disease. The present disclosure also relates to electrolyzed water containing chlorous acid for killing Mycobacterium bacteria, electrolyzed water containing chlorous acid for preventing and / or treating pulmonary MAC disease. The present disclosure also relates to an agent for killing bacteria forming biofilms, a method for killing bacteria forming biofilms, and electrolyzed water containing chlorous acid for killing bacteria forming biofilms.

Background Art

[0002] In recent years, the incidence of pulmonary MAC disease has been increasing in developed countries and has become a problem. Since Mycobacterium, the causative bacterium of pulmonary MAC disease, forms biofilms, it is known that the bacterium is difficult for drugs to exert an effect and is likely to survive.

[0003] Electrolyzed water containing chlorous acid has chlorous acid (HClO2) as a main active ingredient. Electrolyzed water containing chlorous acid has attracted attention as an antibacterial agent, a bactericidal agent, a disinfectant, a sterilizing agent, and further an antiviral agent and a food additive: a sterilizing agent. The present inventors have found electrolyzed water containing chlorous acid and a method for producing the same, and have filed an application after confirming the bactericidal effect against Escherichia coli (Patent Document 1).

Prior Art Documents

Patent Documents

[0004]

Patent Document 1

Summary of the Invention

Means for Solving the Problems

[0005] As a result of intensive studies, the present inventors have discovered that electrolyzed water containing chlorous acid is effective in killing Mycobacterium bacteria, and have discovered that electrolyzed water containing chlorous acid is effective in killing Mycobacterium bacteria that have formed biofilms, thereby completing the present disclosure. The present inventors have also discovered that electrolyzed water containing chlorous acid is effective in preventing / treating pulmonary MAC disease, thereby completing the present disclosure.

[0006] For example, the present invention provides the following items. (Item 1) A Mycobacterium bactericidal agent containing electrolyzed water containing chlorous acid. (Item 2) The bactericidal agent according to Item 1, wherein the free chlorine concentration (as Cl) of the electrolyzed water containing chlorous acid is 50 ppm or more and 60,000 ppm or less. (Item 3) The bactericidal agent according to Item 1, wherein the free chlorine concentration (as Cl) of the electrolyzed water containing chlorous acid is 100 ppm or more and 60,000 ppm or less. (Item 4) The bactericidal agent according to any one of Items 1 to 3, wherein the Mycobacterium bacteria form biofilms. (Item 5) The bactericidal agent according to any one of Items 1 to 4, wherein the Mycobacterium bacteria are non-tuberculous mycobacteria. (Item 6) The bactericidal agent according to any one of Items 1 to 5, wherein the Mycobacterium bacteria are Mycobacterium intracellulare, Mycobacterium avium, Mycolicibacterium fortuitum subsp. fortuitum, Mycobacterium runyonii, Mycobacterium abscessus, Mycobacterium kansasii, M. avium subsp. paratuberculosis or Mycobacterium ulcerans. (Item 7) An agent for preventing and / or treating pulmonary MAC disease containing electrolyzed water containing chlorous acid. (Item 8) The agent according to item 7, wherein the free chlorine concentration (as Cl) of the chlorous acid water is 50 ppm or more and 60,000 ppm or less. (Item 9) The agent according to item 7, wherein the free chlorine concentration (as Cl) of the chlorous acid water is 100 ppm or more and 60,000 ppm or less. (Item 9A) The method according to any one of items 7 to 9, comprising the features described in any one or more of the above items. (Item 10) A method for killing Mycobacterium bacteria using chlorous acid water. (Item 11) The method according to item 10, wherein the Mycobacterium bacteria are contacted with the chlorous acid water in the absence of organic matter. (Item 12) The method according to item 11, wherein the free chlorine concentration (as Cl) of the chlorous acid water is 50 ppm or more and 60,000 ppm or less. (Item 12A) The method according to any one of items 10 to 12, comprising the features described in any one or more of the above items. (Item 13) The method according to item 10, wherein the Mycobacterium bacteria are contacted with the chlorous acid water in the presence of organic matter. (Item 13A) The method according to item 13, comprising the features described in any one or more of the above items. (Item 14) The method according to item 13, wherein the free chlorine concentration (as Cl) of the chlorous acid water is 100 ppm or more and 60,000 ppm or less. (Item 14A) The method according to item 14, comprising the features described in any one or more of the above items. (Item 15) A method for preventing and / or treating pulmonary MAC disease in a subject, comprising administering an effective amount of chlorous acid water to the subject or contacting Mycobacterium that causes pulmonary MAC disease. (Item 15A) The method according to item 15, comprising the features described in any one or more of the above items. (Item 16) Hypochlorous acid water for killing Mycobacterium bacteria. (Item 16A) Hypochlorous acid water according to item 16, comprising the features described in any one or more of the above items. (Item 17) Hypochlorous acid water for preventing and / or treating pulmonary MAC disease. (Item 17A) Hypochlorous acid water according to item 17, comprising the features described in any one or more of the above items. (Item 18) A bactericide for killing biofilm-forming bacteria, comprising hypochlorous acid water. (Item 18A) The bactericide according to item 18, comprising the features described in any one or more of the above items. (Item 19) A method for killing biofilm-forming bacteria, comprising the step of contacting an effective amount of hypochlorous acid water with biofilm-forming bacteria. (Item 19A) The killing method according to item 19, comprising the features described in any one or more of the above items. (Item 20) Hypochlorous acid water for killing biofilm-forming bacteria. (Item 20A) Hypochlorous acid water according to item 20, comprising the features described in any one or more of the above items. (Item 21) Use of hypochlorous acid water in the manufacture of an agent for killing Mycobacterium bacteria. (Item 21A) The use according to item 21, comprising the features described in any one or more of the above items. (Item 22) Use of hypochlorous acid water in the manufacture of a medicament for preventing and / or treating pulmonary MAC disease. (Item 22A) The use according to item 221, comprising the feature(s) described in any one or more of the above items. (Item 23) The use of electrolyzed water in a medicament for killing bacteria that form biofilms. (Item 23A) The use according to item 23, comprising the feature(s) described in any one or more of the above items.

[0007] In the present disclosure, it is intended that the above one or more features may be provided in combination in addition to the explicitly stated combinations. Further embodiments and advantages of the present disclosure will be recognized by those skilled in the art upon reading the following detailed description as necessary.

Advantages of the Invention

[0008] According to the present disclosure, mycobacteria can be easily killed. Even mycobacteria that form biofilms can be killed.

Brief Description of the Drawings

[0009]

Figure 1

Figure 2

Figure 3

Figure 4

Figure 5

Figure 6

Figure 7

Figure 8

Modes for Carrying Out the Invention

[0010] The following provides a more detailed description of the present disclosure. Throughout this specification, it should be understood that singular expressions include the concepts of their plural forms unless otherwise specified. Therefore, singular articles (e.g., "a", "an", "the" in English, etc.) should be understood to include the concepts of their plural forms unless otherwise specified. Also, terms used in this specification should be understood to be used in the meanings commonly used in the relevant field unless otherwise specified. Thus, unless otherwise defined, all technical and scientific terms used in this specification have the same meaning as commonly understood by those skilled in the art to which this disclosure pertains. In case of conflict, this specification (including definitions) shall prevail.

[0011] Abbreviations used in this specification have their conventional meanings within the scope of the relevant field unless otherwise specified.

[0012] References to "about" a value or parameter in this specification include variations that target the value or parameter itself. Unless otherwise specifically stated, for example, "about X" includes "X" itself and values that allow an error of ±10% thereof.

[0013] In this specification, "hypochlorous acid water" is an aqueous solution containing hypochlorous acid (HClO2) used as a bactericide, which can stably maintain hypochlorous acid (HClO2) over a long period of time. When a sample of hypochlorous acid water is measured with a spectrophotometer, an absorption part containing acidic hypochlorite ions (H + +ClO2 - ) showing a peak around 260 nm and an absorption part containing chlorine dioxide (ClO2) showing a peak around 350 nm can be simultaneously confirmed in the UV spectrum, that is, when showing a double peak, the presence of hypochlorous acid water can be recognized.

[0014] Chlorous acid water can be produced by the methods disclosed in International Publications WO2008 / 026607, WO2014 / 188310, WO2014 / 188311, WO2014 / 188312, WO2015 / 093062, and WO2017 / 170904.

[0015] "Chlorous acid water" was designated as a food additive on February 1, 2013, and is a disinfectant with chlorous acid (HClO2) as the main active ingredient. Chlorous acid (HClO2), the main active ingredient of this "chlorous acid water", is a metastable chemical substance and is recognized as a food additive: processing aid as a particularly safe substance by the US Department of Agriculture (USDA) and the US Food and Drug Administration (FDA). The chlorous acid water used in this disclosure can be either a substance corresponding to a food additive or a substance not corresponding thereto, and may be provided as a pharmaceutical, quasi-drug, or miscellaneous product.

[0016] Moreover, "chlorous acid water" can exhibit a strong bactericidal effect even in the presence of organic substances. In the National Institute of Health Sciences (commonly known as the NIHS), in the "Investigation on the Inactivation Conditions of Norovirus in 2015", it received a high evaluation that "only chlorous acid water could be inactivated below the detection limit under all load conditions." Along with the amendment of the Enforcement Regulations of the Food Sanitation Act, the listing of chlorous acid water has been progressing in order of occurrence of large-scale food poisoning incidents, such as in the "Cooking Manual for Large-Scale Cooking Facilities" and the "Hygiene Regulations for Pickled Foods".

[0017] Chlorous acid water can also be applied as a pharmaceutical as a Class 2 disinfectant. It was approved as a pharmaceutical of Class 2 disinfectant in 2019 with a view to its release in 2020. For example, various guidelines such as the "Q&A Regarding Norovirus" and the "Guidelines for Infectious Disease Control in Nurseries" under the jurisdiction of the Ministry of Health, Labour and Welfare, and related manuals such as the "Manual for Infection Control in Facilities for Supporting the Care of the Elderly", and the addition and revision work to each hygiene regulation such as the "Hygiene Regulations for Boxed Lunches and Prepared Dishes" are also being carried out at any time. It is a substance supplied to a wide range of markets in the food hygiene and environmental hygiene markets in Japan.

[0018] "Hypochlorous acid water" with hypochlorous acid as the main active ingredient has a sterilizing power equivalent to or stronger than that of other chlorine-based agents such as "hypochlorous acid water" and "sodium hypochlorite". Although its reactivity is mild and it does not have an instantaneous sterilizing effect (instantaneous efficacy), it has an accurate sterilizing power while maintaining a mild reactivity. Moreover, "hypochlorous acid water" has the characteristic of maintaining a stable sterilizing power, and in an environment contaminated with a large amount of organic matter, which has been said to be the most difficult for conventional chlorine oxide-based agents, it can exert a sterilizing effect slowly but surely and accurately. (Sterilizing power against microorganisms lurking in dirt).

[0019] Therefore, it can exert an inactivating effect on resistant bacteria (such as heat-resistant bacteria whose resistance is enhanced by forming spores and drug-resistant bacteria that are no longer effective against antibiotics), which have been difficult to sterilize and have been a challenge until now, as well as on fungi such as mold and yeast, and furthermore on viruses (including non-enveloped viruses). "Hypochlorous acid water" does not need to be adjusted during use, so no dedicated generating device is required, and anyone can use it whenever they want, anywhere, and it is also safe.

[0020] Moreover, regarding the effect of "hypochlorous acid water" in the presence of organic matter, it is also published on the website of the Ministry of Health, Labour and Welfare and in the "Investigation Report on the Inactivation Conditions of Norovirus in 2015 (Food Hygiene Management Department, National Institute of Health)".

[0021] "Hypochlorous acid water" does not need to be prepared during use, so no dedicated generating device is required, and anyone can use it whenever they want, anywhere, and it is also safe.

[0022] In this specification, "hypochlorous acid water preparation" refers to a preparation prepared using hypochlorous acid water as the active ingredient. Depending on the use, additional components, pH, hypochlorous acid content, free chlorine concentration, etc. can be adjusted.

[0023] In this specification, "hypochlorous acid water as the active ingredient" refers to hypochlorous acid water used as the active ingredient. Hypochlorous acid water can be prepared by the method as described in this specification.

[0024] The "hypochlorous acid water preparation" used in this disclosure may not cause any harm because it has low irritation even when directly contacting animals. When the hypochlorous acid water preparation used in this disclosure is applied to the skin, there is no corrosive reaction of the skin, no erythema or edema. When applied to the eyes, the cornea, luster, and conjunctiva are normal, and it may not show skin sensitization.

[0025] In this specification, "in the absence of organic substances" means that in addition to the complete absence of organic substances, it also indicates that they are substantially non-existent (including below the threshold value). It refers to all states that are not "in the presence of organic substances".

[0026] In this specification, "free chlorine", "free chlorine concentration", or "free residual chlorine concentration" is the value measured by Appendix 3 of the "Inspection Method for Free Residual Chlorine and Combined Chlorine Determined by the Minister of Health, Labour and Welfare Based on Article 17, Paragraph 2 of the Enforcement Regulations of the Waterworks Law" (hereinafter referred to as the colorimetric method (DPD indicator)), and is the value obtained by the oxidation of the DPD indicator.

[0027] In this specification, "nontuberculous mycobacteriosis" refers to an infectious disease caused by nontuberculous mycobacteria excluding Mycobacterium tuberculosis and Mycobacterium leprae. It is also called atypical mycobacteriosis. The main causative bacteria include Mycobacterium avium, Mycobacterium intracellulare, Mycobacterium kansasii, Mycobacteruim shinjukuense, etc.

[0028] In this specification, "pulmonary MAC disease" refers to pulmonary Mycobacterium avium complex disease, which means an infection in the lungs among nontuberculous mycobacterioses.

[0029] (Preferred Embodiment) Preferred embodiments of the present disclosure will be described below. It is understood that the embodiments provided below are for better understanding of the present disclosure, and the scope of the present disclosure should not be limited to the following description. Therefore, it is obvious that those skilled in the art can make appropriate modifications within the scope of the present disclosure with reference to the description in this specification. It is also understood that the following embodiments can be used alone or in combination with each other.

[0030] (Mycobacterium bactericide) In one aspect of the present disclosure, a Mycobacterium bactericide containing electrolyzed water is provided. In the bactericide of the present disclosure, the free chlorine concentration (as Cl) of the electrolyzed water can be at least 50 ppm. In the bactericide of the present disclosure, the free chlorine concentration (as Cl) of the electrolyzed water can be at least 100 ppm. In the bactericide of the present disclosure, the free chlorine concentration (as Cl) of the electrolyzed water can be 50 ppm, 100 ppm, 150 ppm, 200 ppm, 250 ppm, 300 ppm, 350 ppm, 400 ppm, 450 ppm, 500 ppm, 600 ppm, 700 ppm, 800 ppm, 900 ppm, 1000 ppm or more, and can be 60000 ppm, 50000 ppm, 40000 ppm, 30000 ppm, 20000 ppm, 10000 ppm, 5000 ppm or less. The free chlorine concentration (as Cl) can be any value between these values.

[0031] In the bactericide of the present disclosure, it can be used when the Mycobacterium forms a biofilm. Examples of Mycobacterium that forms a biofilm include Mycobacterium intracellulare.

[0032] In the bactericide of the present disclosure, the Mycobacterium can be a non-tuberculous mycobacterium.

[0033] In the bactericide of the present disclosure, the Mycobacterium bacteria can be Mycobacterium intracellulare, Mycobacterium avium, Mycolicibacterium fortuitum subsp. fortuitum, Mycobacterium runyonii, Mycobacterium abscessus, Mycobacterium kansasii, Johne's bacillus (M. avium subsp. paratuberculosis) or Mycobacterium ulcerans.

[0034] (Killing of biofilm-forming bacteria) In one aspect, the present disclosure provides an agent and method for killing bacteria that form biofilms.

[0035] In the bactericide for killing biofilm-forming bacteria of the present disclosure, the free chlorine concentration (as Cl) of the electrolyzed water can be at least 50 ppm. In the bactericide of the present disclosure, the free chlorine concentration (as Cl) of the electrolyzed water can be at least 100 ppm. In the bactericide of the present disclosure, the free chlorine concentration (as Cl) of the electrolyzed water can be 50 ppm, 100 ppm, 150 ppm, 200 ppm, 250 ppm, 300 ppm, 350 ppm, 400 ppm, 450 ppm, 500 ppm, 600 ppm, 700 ppm, 800 ppm, 900 ppm, 1000 ppm or more, and can be 60000 ppm, 50000 ppm, 40000 ppm, 30000 ppm, 20000 ppm, 10000 ppm, 5000 ppm or less, and the free chlorine concentration (as Cl) can be any value between these values.

[0036] In the bactericide of the present disclosure, examples of bacteria that form biofilms include bacteria of the genus Mycobacterium (e.g., Mycobacterium intracellulare), bacteria of the genus Pseudomonas (e.g., P. aeruginosa), bacteria of the genus Staphylococcus, bacteria of the genus Streptococcus (e.g., S. mutans), bacteria of the genus Legionella, bacteria of the genus Helicobacter, and bacteria of the genus Mycobacterium.

[0037] In the bactericide of the present disclosure, the bacteria that form biofilms can be non-tuberculous mycobacteria.

[0038] In the bactericide of the present disclosure, the bacteria that form biofilms can be of the genus Mycobacterium (e.g., Mycobacterium intracellulare).

[0039] (Bacterial killing method) In one aspect of the present disclosure, a method for killing Mycobacterium bacteria or bacteria that form biofilms using chlorous acid water is provided.

[0040] In the method of the present disclosure, it can be carried out in the absence of organic matter. In this method, the free chlorine concentration (as Cl) of the chlorous acid water can be at least 50 ppm. The free chlorine concentration (as Cl) of the chlorous acid water can be 50 ppm, 100 ppm, 150 ppm, 200 ppm, 250 ppm, 300 ppm, 350 ppm, 400 ppm, 450 ppm, 500 ppm, 600 ppm, 700 ppm, 800 ppm, 900 ppm, 1000 ppm or more, and can be 60,000 ppm, 50,000 ppm, 40,000 ppm, 30,000 ppm, 20,000 ppm, 10,000 ppm, 5000 ppm or less, and the free chlorine concentration (as Cl) can be any value between these values.

[0041] In the method of the present disclosure, the method may be in the presence of an organic substance. In this method, the free chlorine concentration (as Cl) of the chlorous acid water may be at least 100 ppm. The free chlorine concentration (as Cl) of the chlorous acid water may be 100 ppm, 150 ppm, 200 ppm, 250 ppm, 300 ppm, 350 ppm, 400 ppm, 450 ppm, 500 ppm, 600 ppm, 700 ppm, 800 ppm, 900 ppm, 1000 ppm or more, and may be 60,000 ppm, 50,000 ppm, 40,000 ppm, 30,000 ppm, 20,000 ppm, 10,000 ppm, 5000 ppm or less, and the free chlorine concentration (as Cl) may be any value between these values.

[0042] (Treatment and prevention of pulmonary MAC disease) In one aspect of the present disclosure, there is provided an agent for preventing and / or treating pulmonary MAC disease containing chlorous acid water.

[0043] In the agent of the present disclosure, the free chlorine concentration (as Cl) of the chlorous acid water may be at least 50 ppm. In the agent of the present disclosure, the free chlorine concentration (as Cl) of the chlorous acid water may be at least 100 ppm. In the agent of the present disclosure, the free chlorine concentration (as Cl) of the chlorous acid water may be 50 ppm, 100 ppm, 150 ppm, 200 ppm, 250 ppm, 300 ppm, 350 ppm, 400 ppm, 450 ppm, 500 ppm, 600 ppm, 700 ppm, 800 ppm, 900 ppm, 1000 ppm or more, and may be 60,000 ppm, 50,000 ppm, 40,000 ppm, 30,000 ppm, 20,000 ppm, 10,000 ppm, 5000 ppm or less, and the free chlorine concentration (as Cl) may be any value between these values.

[0044] In one aspect of the present disclosure, there is provided a method for preventing and / or treating pulmonary MAC disease in a subject, which includes the step of administering an effective amount of chlorous acid water to the subject and / or contacting the subject with Mycobacterium bacteria.

[0045] (Chlorous acid water and its production examples) The electrolyzed water used in the present disclosure has the characteristics found by the inventors. Electrolyzed water produced by any method such as known production methods described in the above-mentioned documents can be used. As a typical composition, for example, a composition containing 61.40% electrolyzed water, 1.00% potassium dihydrogen phosphate, 0.10% potassium hydroxide, and 37.50% purified water can be formulated and used (sold by the applicant. 72% electrolyzed water corresponds to 30,000 ppm chlorous acid.), but is not limited thereto. This agent reduces the attenuation of chlorous acid due to contact with organic substances under acidic conditions, while maintaining the bactericidal effect. Also, it has the characteristic of slightly generating chlorine gas and suppressing the amplification of the odor of the mixture of chlorine and organic substances.

[0046] In one embodiment, the electrolyzed water of the present disclosure can be produced by adding sulfuric acid or an aqueous solution thereof in an amount and concentration capable of maintaining the pH value of an aqueous sodium chlorate solution at 3.4 or less and reacting to generate chloric acid, and then adding hydrogen peroxide in an amount equal to or more than the amount required for the reduction reaction of the chloric acid.

[0047] In another embodiment, the electrolyzed water of the present disclosure can be produced by adding sulfuric acid or an aqueous solution thereof in an amount and concentration capable of maintaining the pH value of an aqueous sodium chlorate solution at 3.4 or less and reacting to generate chloric acid, and then adding hydrogen peroxide in an amount equal to or more than the amount required for the reduction reaction of the chloric acid to generate an aqueous solution containing chlorous acid, and then adding any single substance of inorganic acid or inorganic acid salt, or two or more single substances or a combination thereof, and adjusting the pH value within the range of 2.9 to 8.5.

[0048] Furthermore, in another embodiment, the chlorous acid water of the present disclosure is obtained by adding sulfuric acid or an aqueous solution thereof in an amount and concentration capable of maintaining the pH value of an aqueous sodium chlorate solution at 3.4 or less, reacting them, generating chloric acid, and then adding hydrogen peroxide in an amount equal to or greater than the amount required for the reduction reaction of the chloric acid to generate an aqueous solution of chlorous acid. Then, an inorganic acid or an inorganic acid salt or an organic acid or an organic acid salt, either alone or in combination of two or more thereof, or a combination of these, is added, and the pH value is adjusted within the range of 3.2 to 8.5 to produce the chlorous acid water.

[0049] Furthermore, in another embodiment, the chlorous acid water of the present disclosure is obtained by adding sulfuric acid or an aqueous solution thereof in an amount and concentration capable of maintaining the pH value of an aqueous sodium chlorate solution at 3.4 or less, reacting them, generating chloric acid, and then adding hydrogen peroxide in an amount equal to or greater than the amount required for the reduction reaction of the chloric acid to generate an aqueous solution of chlorous acid. Then, an inorganic acid or an inorganic acid salt, either alone or in combination of two or more thereof, or a combination of these, is added to the aqueous solution, and then an inorganic acid or an inorganic acid salt or an organic acid or an organic acid salt, either alone or in combination of two or more thereof, or a combination of these, is added, and the pH value is adjusted within the range of 3.2 to 8.5 to produce the chlorous acid water.

[0050] Also, in another embodiment, in the above method, the inorganic acid can be carbonic acid, phosphoric acid, boric acid, or sulfuric acid.

[0051] Furthermore, in another embodiment, the inorganic acid salt can be a carbonate, an inorganic hydroxide, a phosphate, or a borate.

[0052] Also, in another embodiment, as the carbonate, sodium carbonate, potassium carbonate, sodium bicarbonate, or potassium bicarbonate can be used.

[0053] Furthermore, in another embodiment, as the inorganic hydroxide, sodium hydroxide, potassium hydroxide, calcium hydroxide, or barium hydroxide can be used.

[0054] Furthermore, in another embodiment, as the phosphate, disodium hydrogen phosphate, sodium dihydrogen phosphate, trisodium phosphate, tripotassium phosphate, dipotassium hydrogen phosphate, or potassium dihydrogen phosphate can be used.

[0055] Also, in another embodiment, as the borate, sodium borate or potassium borate can be used.

[0056] Furthermore, in another embodiment, as the organic acid, succinic acid, citric acid, malic acid, acetic acid, or lactic acid can be used.

[0057] Furthermore, in another embodiment, as the organic acid salt, sodium succinate, potassium succinate, sodium citrate, potassium citrate, sodium malate, potassium malate, sodium acetate, potassium acetate, sodium lactate, potassium lactate, or calcium lactate can be used.

[0058] In a method for producing an aqueous solution containing chlorous acid (HClO2) that can be used as a bactericidal agent, chloric acid (HClO3) obtained by adding sulfuric acid (H2SO4) or an aqueous solution thereof to an aqueous solution of sodium chlorate (NaClO3) to make it acidic is added with an amount of hydrogen peroxide (H2O2) necessary to convert it to chlorous acid by a reduction reaction, thereby producing chlorous acid (HClO2). The basic chemical reactions of this production method are represented by the following Formulas A and B.

Chemical formula

[0059]

Chemical formula

[0060] At this time, chlorine dioxide gas (ClO2) is generated (Formula C), but by coexisting with hydrogen peroxide (H2O2), chlorous acid (HClO2) is produced through the reactions of Formulas D to F.

[0061] By the way, the generated chlorous acid (HClO2) has the property that multiple chlorous acid molecules decompose with each other, or decompose into chlorine dioxide gas or chlorine gas at an early stage due to the presence of chloride ions (Cl - −), hypochlorous acid (HClO), and other reduction products. Therefore, in order to make it useful as a bactericidal agent, it is necessary to prepare it so that the state of chlorous acid (HClO2) can be maintained for a long time.

[0062] Therefore, by adding an inorganic acid, inorganic acid salt, organic acid, or organic acid salt, either alone, or two or more alone, or a combination thereof, to the chlorous acid (HClO2), chlorine dioxide gas (ClO2), or an aqueous solution containing these obtained by the above method, a transition state is created and the decomposition reaction is retarded, so that chlorous acid (HClO2) can be stably maintained for a long time.

[0063] In one embodiment, an inorganic acid or inorganic acid salt, specifically a carbonate or inorganic hydroxide, added alone, or two or more alone, or a combination thereof, to the chlorous acid (HClO2), chlorine dioxide gas (ClO2), or an aqueous solution containing these obtained by the above method can be used.

[0064] In another embodiment, an inorganic acid or an inorganic acid salt, specifically a carbonate or an inorganic hydroxide, is added alone or in combination of two or more alone or in combination thereof to an aqueous solution, and an inorganic acid, an inorganic acid salt, an organic acid or an organic acid salt is added alone or in combination of two or more alone or in combination thereof, and such can be used.

[0065] In addition, in yet another embodiment, an inorganic acid, an inorganic acid salt, an organic acid or an organic acid salt added alone or in combination of two or more alone or in combination thereof is added to the aqueous solution produced by the above method, and such can be used.

[0066] Examples of the inorganic acid include carbonic acid, phosphoric acid, boric acid or sulfuric acid. Examples of the inorganic acid salt include carbonates, inorganic hydroxides, phosphates or borates. More specifically, examples of the carbonate include sodium carbonate, potassium carbonate, sodium hydrogen carbonate, potassium hydrogen carbonate, examples of the inorganic hydroxide include sodium hydroxide, potassium hydroxide, calcium hydroxide, barium hydroxide, examples of the phosphate include disodium hydrogen phosphate, sodium dihydrogen phosphate, trisodium phosphate, tripotassium phosphate, dipotassium hydrogen phosphate, potassium dihydrogen phosphate, and examples of the borate include sodium borate, potassium borate. Further, examples of the organic acid include succinic acid, citric acid, malic acid, acetic acid or lactic acid. Examples of the organic acid salt include sodium succinate, potassium succinate, sodium citrate, potassium citrate, sodium malate, potassium malate, sodium acetate, potassium acetate, sodium lactate, potassium lactate or calcium lactate.

[0067] When an acid and / or its salt is added, temporarily Na + +ClO2 - ⇔ Na-ClO2 or K + +ClO2 - ⇔ K-ClO2 or H + +ClO2 -⇔ A transition state such as H-ClO2 is created, which can slow down the progress of chlorous acid (HClO2) to chlorine dioxide (ClO2). As a result, it becomes possible to maintain chlorous acid (HClO2) for a long time and produce an aqueous solution containing chlorous acid (HClO2) with little generation of chlorine dioxide (ClO2).

[0068] The decomposition in an acidic solution of chlorite is represented below.

[0069] [Chemical formula]

[0070] As represented by this formula, the decomposition rate of an aqueous chlorite solution at a certain pH increases as the pH decreases, that is, as the acid becomes stronger. That is, the absolute reaction rates of reactions (a), (b), and (c) in the above formula increase. For example, the proportion of reaction (a) becomes smaller as the pH decreases, but the total decomposition rate fluctuates greatly, that is, it becomes large. Therefore, the generation amount of chlorine dioxide (ClO2) also increases as the pH decreases. For this reason, the lower the pH value, the faster the sterilization and bleaching, but the work becomes difficult due to the irritating and harmful chlorine dioxide gas (ClO2), and it also has an adverse effect on human health. In addition, the reaction of chlorous acid to chlorine dioxide proceeds quickly, chlorous acid becomes unstable, and the time for maintaining the bactericidal power is extremely short.

[0071] Therefore, when adding the above inorganic acid, inorganic acid salt, organic acid or organic acid salt to an aqueous solution containing chlorous acid (HClO2), the pH value is adjusted within the range of 2.9 to 8.5 from the viewpoints of suppressing the generation of chlorine dioxide and balancing the bactericidal power.

[0072] (Problems in comparing and evaluating the antimicrobial effects of chlorous acid water and sodium hypochlorite) In comparing and evaluating the antimicrobial effects of chlorous acid water and sodium hypochlorite, there are issues regarding the concentration expression of chlorine oxides. There are available chlorine concentration and free chlorine. The antimicrobial effect depends on free chlorine, which is the source of oxidizing power. For sodium hypochlorite, the relationship between free chlorine and available chlorine concentration is approximately 1:1. However, for chlorous acid water, the available chlorine concentration does not coincide with free chlorine as in the case of sodium hypochlorite. Therefore, when comparing the bactericidal powers of both agents on the same footing, it is necessary to compare using the oxidizing power that represents the antimicrobial effect, that is, free chlorine, rather than the available chlorine concentration.

[0073] Generally, the oxidizing power of chlorine oxide agents is determined by measurement methods that utilize colorimetric methods such as the DPD method and the TMB method. However, there is no standard for measuring free chlorine in chlorous acid water like there is for sodium hypochlorite. Therefore, a calibration curve is created by setting 1 mg / L of free chlorine (as Cl) in sodium hypochlorite as the oxidizing power 1. Note that the oxidizing power can be expressed in terms of free chlorine (as Cl). When attempting to compare with the same free chlorine, although the free chlorine in sodium hypochlorite is generated from Cl radicals, since the free chlorine in chlorous acid water has HClO2 as the source of generation, if evaluated based on a corresponding standard, it becomes difficult to compare. Therefore, by calculating with oxidizing power 1 = free chlorine (as Cl) 1 mg / L as in the case of sodium hypochlorite and using the same standard, comparison and evaluation can be performed on the same footing as sodium hypochlorite.

[0074] As a method for measuring free chlorine (as Cl), for the sample, a buffer solution and DPD indicator are added, and the absorbance is measured at a wavelength of 510 nm using a spectrophotometer. For the measurement of free chlorine (as Cl) in the presence of organic substances, measurement is performed at a wavelength of 655 nm using a TMB reagent, and the concentration is determined from the measured value. Also, as a method for the bactericidal effect confirmation test, the free chlorine (as Cl) of the test agent is prepared by the DPD method. Each agent is brought into contact with a bacterial solution containing organic substances, and after a certain period of time has elapsed, it is neutralized using sodium thiosulfate, and the viable cell count of this neutralized solution is confirmed. [Chemical formula]

[0075] (Method of Using the Disinfectant / Agent of the Present Disclosure) The disinfectant / agent of the present disclosure can be used for environmental preparation (infection prevention) in hospitals and the like. It can be directly applied to facilities such as floors, walls, and doors, as well as medical instruments. It is also possible to directly apply it to animals including humans.

[0076] In addition, references such as scientific literature, patents, and patent applications cited in this specification are hereby incorporated by reference in their entirety to the same extent as if each were specifically recited herein.

[0077] As described above, the present disclosure has been described with reference to preferred embodiments for ease of understanding. Hereinafter, the present disclosure will be described based on examples. However, the above description and the following examples are provided for illustrative purposes only and not for the purpose of limiting the present invention. Therefore, the scope of the present invention is not limited to the embodiments or examples specifically described herein, but is limited only by the claims.

Example

[0078] (Quantitative Method for Hypochlorous Acid Water) Precisely weigh about 5 g of this product, add water to make exactly 100 ml. Accurately measure 20 ml of this sample solution, put it into an iodine flask, add 10 ml of sulfuric acid (1→10), then add 1 g of potassium iodide, immediately seal it tightly and mix well. Pour potassium iodide test solution into the upper part of the iodine flask and let it stand in the dark for 15 minutes. Next, loosen the stopper, pour in the potassium iodide test solution, immediately seal it tightly and mix well, and then titrate the liberated iodine with 0.1 mol / L sodium thiosulfate (indicator: starch test solution). The indicator is added after the color of the solution changes to light yellow. Conduct a blank test separately for correction. 1 ml of 0.1 mol / L sodium thiosulfate solution = 1.711 mg HClO2).

[0079] (Production Example) The hypochlorous acid water preparation used in the following examples was produced as follows. In this specification, hypochlorous acid water may be abbreviated as "hypo-water", but they are synonymous. Component analysis table of chlorous acid water

[0080]

Table 1

[0081] Using this chlorous acid water, a chlorous acid water preparation was produced based on the following formulation.

[0082]

Table 2

[0083]

Table 3

[0084] The "chlorous acid water preparation produced with chlorous acid water" prepared based on the above preparation method was used to measure the concentration of "chlorous acid water" based on the above "quantification method of chlorous acid water", and a buffer solution (phosphate buffer containing dipotassium hydrogen phosphate and potassium dihydrogen phosphate) was used to prepare the chlorous acid water of each example so that the free chlorine concentration described in each example was obtained.

[0085] (Bactericidal effect confirmation test of chlorous acid water and sodium hypochlorite against M. intracellulare) There are various disinfectants and sterilizing agents used for environmental maintenance in hospitals and the like. Among them, as chlorine oxide-based disinfectants and sterilizing agents, sodium hypochlorite, acidified sodium hypochlorite, etc. are used. Generally, sodium hypochlorite is widely used.

[0086] The bactericidal effects of chlorous acid water and sodium hypochlorite against M. intracellulare were confirmed in the absence of organic matter and in the presence of 0.5% bovine serum albumin (hereinafter referred to as BSA), which is an organic matter (Figure 1). At that time, if a bactericidal effect is observed with a reduction effect of 4 logs or more in the number of bacteria, it is considered to have a bactericidal effect.

[0087] First, it was found that even with a short contact time of 1 minute, chlorous acid water exhibits a bactericidal effect at 50 ppm in the presence of organic substances and 100 ppm in the presence of 0.5% BSA. In contrast, sodium hypochlorite shows no bactericidal effect even at 200 ppm of free chlorine (as Cl) in the absence of organic substances, and no bactericidal effect is observed even at 1000 ppm of free chlorine (as Cl) in the presence of 0.5% BSA.

[0088] From this, it was found that chlorous acid water is effective against M. intracellulare.

[0089] (Test for confirming the bactericidal effect against M. intracellulare forming biofilm) Since M. intracellulare produces biofilm, it is said that drugs are less effective. Therefore, it was investigated whether chlorous acid water has a bactericidal effect against M. intracellulare forming this biofilm (Figure 2). Figure 2 shows the results of treating M. intracellulare forming biofilm with sterile water for 30 minutes, chlorous acid water at 200 ppm or more for 30 minutes.

[0090] As a result, when treated with chlorous acid water, the surviving bacteria almost disappeared. From this, it was found that chlorous acid water also shows a bactericidal effect against M. intracellulare in the biofilm.

[0091] (Observation of the surface of M. intracellurare after bactericidal treatment by scanning electron microscope (SEM)) From the results of the bactericidal effect confirmation test, it was found that chlorous acid water is very effective against M. intracellulare. Therefore, as the mechanism of the bactericidal effect, an attempt was made to confirm by what principle it is being sterilized.

[0092] First, to determine how it acts on the bacterial surface, the bacteria after sterilization treatment were observed using a scanning electron microscope in a comparison between electrolyzed water and sodium hypochlorite. In addition, to confirm the state of normal M. intracellulare, when treated with sterilized water, it was found that an extracellular matrix exists behind the bacteria (Figure 3). Therefore, it is considered that this extracellular matrix forms a biofilm.

[0093] Next, when comparing the case of treatment with sodium hypochlorite with the case of treatment with sterilized water, disappearance of the extracellular matrix and slight changes on the bacterial surface were observed (Figure 4). From this, it was found that although no bactericidal effect on M. intracellulare was observed, there is a sufficient possibility that sodium hypochlorite acts on the extracellular matrix and the bacterial surface.

[0094] When treated with electrolyzed water, the surface remained clean and almost no influence on the bacterial surface was observed (Figure 5). However, detachment and aggregation of the extracellular matrix were confirmed, and it is considered that the detachment and aggregation of this extracellular matrix may have a low correlation with the bactericidal effect.

[0095] (Results of pulsed-field gel electrophoresis of DNA after bactericidal treatment against M. intacellulare) In examining the bactericidal mechanism of electrolyzed water, from the SEM results, it is considered that there is a low possibility of acting on the bacterial surface to sterilize. To examine the effect of electrolyzed water on the chromosomal DNA of bacteria, chromosomal DNA was extracted from the bacteria after sterilization treatment, and the state of the chromosomal DNA was confirmed by pulsed-field gel electrophoresis.

[0096] As a result, when treated with sodium hypochlorite, bands were formed at the same position as in the case of water. However, when treated with chlorous acid water, the bands disappeared, indicating that the DNA was fragmented (Fig. 6). From this, it was found that chlorous acid water penetrates into the cells of M. intracellulare and degrades chromosomal DNA, suggesting that damage to chromosomal DNA is one of the bactericidal actions of chlorous acid water.

[0097] (Results of confirmation of bactericidal effect against other mycobacteria) Furthermore, we investigated whether a bactericidal effect could be observed against other Mycobacterium species. In the absence of organic matter, chlorous acid water showed an effect at a free chlorine concentration of 50 ppm or more against other Mycobacterium species, while no effect was observed with 200 ppm of sodium hypochlorite (Fig. 7). The same results as for M. intracellulare were obtained.

[0098] In addition, in the presence of 0.5% BSA, chlorous acid water showed an effect at a free chlorine concentration of 100 ppm or more, while no effect was observed with 1000 ppm of sodium hypochlorite (Fig. 8). These results are the same as those for the bactericidal effect against M. intracellulare. From this, no differences in the bactericidal effect of chlorous acid water against Mycobacterium species were observed in terms of strain or difference.

[0099] (Summary) · Chlorous acid water showed a stronger bactericidal effect against M. intracellulare than sodium hypochlorite. In addition, chlorous acid water also showed sufficient effect against other Mycobacterium species. · Chlorous acid water showed a bactericidal effect against M. intracellulare in biofilms. · No destruction of the cell surface was observed by treatment with chlorous acid water, but a strong fragmentation phenomenon of chromosomal DNA was observed. · Hypochlorous acid water is less likely to be inhibited by organic substances compared to sodium hypochlorite, and it is thought that it may penetrate into cells and kill bacteria by fragmenting bacterial molecules. · It is considered that the DNA after bacterial treatment is fragmented also for mycobacteria other than M. intracellulare.

[0100] (Note) As described above, the present invention has been exemplified using the preferred embodiments of the present invention, but it is understood that the scope of the present invention should be interpreted only by the claims. It is understood that the patents, patent applications, and other documents cited in this specification should be incorporated by reference into this specification as if the contents themselves were specifically described herein. This application claims priority to Japanese Patent Application No. 2019-173414 (filed on September 24, 2019), and the entire contents thereof are incorporated by reference into this specification. It is understood that the patents, patent applications, and other documents cited in this specification should be incorporated by reference into this specification as if the contents themselves were specifically described herein.

Industrial Applicability

[0101] A method for killing mycobacteria is obtained. A method for preventing / treating pulmonary MAC disease is obtained.

Claims

1. A Mycobacterium bactericide containing chlorous acid water.

2. The bactericide according to claim 1, wherein the free chlorine concentration (as Cl) of the chlorous acid water is 50 ppm or more and 60,000 ppm or less.

3. The bactericide according to claim 1, wherein the free chlorine concentration (as Cl) of the chlorous acid water is 100 ppm or more and 60,000 ppm or less.

4. The bactericide according to any one of claims 1 to 3, wherein the Mycobacterium bacterium forms a biofilm.

5. The bactericide according to any one of claims 1 to 4, wherein the Mycobacterium bacterium is a non-tuberculous mycobacterium.

6. The bactericide according to any one of claims 1 to 5, wherein the Mycobacterium bacterium is Mycobacterium intracellulare, Mycobacterium avium, Mycolicibacterium fortuitum subsp. fortuitum, Mycobacterium runyonii, Mycobacterium abscessus, Mycobacterium kansasii, M. avium subsp. paratuberculosis or Mycobacterium ulcerans.

7. A drug for preventing and / or treating pulmonary MAC disease containing chlorous acid water.

8. The drug according to claim 7, wherein the free chlorine concentration (as Cl) of the chlorous acid water is 50 ppm or more and 60,000 ppm or less.

9. The drug according to claim 7, wherein the free chlorine concentration (as Cl) of the chlorous acid water is 100 ppm or more and 60,000 ppm or less.

10. A method for killing Mycobacterium bacteria using chlorous acid water, excluding medical acts on humans. Claim 11 The method according to claim 10, wherein the killing of the Mycobacterium bacteria is performed on a floor, wall, door, or medical instrument. Claim 12 The method according to claim 10 or 11, wherein the Mycobacterium bacteria are contacted with the chlorous acid water in the absence of organic matter. Claim 13 The method according to claim 12, wherein the free chlorine concentration (as Cl) of the chlorous acid water is 50 ppm or more and 60,000 ppm or less. Claim 14 The method according to claim 10 or 11, wherein the Mycobacterium bacteria are contacted with the chlorous acid water in the presence of organic matter. Claim 15 The method according to claim 14, wherein the free chlorine concentration (as Cl) of the chlorous acid water is 100 ppm or more and 60,000 ppm or less. Claim 16 A method for preventing and / or treating pulmonary MAC disease in an animal other than a human, comprising administering an effective amount of chlorous acid water to the animal other than a human or contacting Mycobacterium causing pulmonary MAC disease.

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