Antimicrobial agent

The present invention relates to the use of substances to achieve antibacterial effects.

Microorganisms such as bacteria and fungi live widely in our living space and inhabit the most diverse types of surfaces. Many microorganisms are pathogens, so their spread and / or control has a special role in public health and public health. When such microorganisms invade our bodies, they can cause life-threatening infections. When such an infection occurs in a hospital, it is called a nosocomial infection.

The starting point is an estimate that it costs tens of billions of euros each year worldwide to eliminate the damage caused by nosocomial infections. Therefore, control of pathogenic microorganisms plays a special role in public health and public health.

Apart from the prevention and / or killing of unwanted microorganisms by antibiotics, preventive measures, such as the creation of spaces unsuitable for the survival of microorganisms, are becoming even more important. Among such preventive measures, the use of silver as an additive to organic and inorganic substances has rapidly increased in importance in the last few years. Here, silver ions interfere with important functions of microorganisms. It is now presumed that silver ions interfere with the enzyme and interfere with its important transport function in the cell. Further effects include diminished structural strength of cells and / or damage to membrane structure. Such effects can result in cell damage and / or cell death. Silver has a very wide range of activity against multi-resistant bacteria. Small doses are sufficient for long-term effects. This is called an oligodynamic effect. Nevertheless, in some cases, organic compounds are added to enhance the effectiveness of silver. It is always important that there are enough silver ions. Therefore, nanoscale silver powder called nanosilver is used to obtain a large particle surface.

Silver has no harmful side effects in a wide range of doses. Only when the accumulation of silver in the body is significantly increased can it cause argyria, an irreversible slate discoloration of the skin and mucous membranes. In addition, elevated silver levels can cause dysgeusia, impaired susceptibility to odors, and brain spasms.

Furthermore, it should be mentioned that, in general, the interaction between nanoscale particles and the human body has not yet been fully studied. An extensive research program has just begun. The antibacterial effect of silver is inadequate in many applications. The effect is only obtained with saline solution up to 0.25 molar concentration. Beyond that, the formation of silver chloride occurs. An important drawback to the use of nanosilver is its inadequate cost. On the one hand, this is due to the high cost of silver, and on the other hand, the processing of silver into nanoparticles is time consuming and costly. Further problems arise in the treatment of nanosilver due to the formation of agglomerated particles, agglomerates, and clusters. This reduces the active surface and, as a result, reduces the antibacterial effect. To prevent this, use nanosilver, TiO₂However, as a result, the production cost increases.

Therefore, many attempts have been made to detect antibacterial, microactive, and bactericidal effects with other metals. For example, copper also has a strong antibacterial effect, but is too cytotoxic. So far, the results of a search on the Internet encyclopedia Wikipedia with the search term oligodynamics show that this effect has been seen in the following metals, in descending order of effectiveness: mercury. , Silver, copper, tin, iron, lead, and bismuth. Two expensive precious metals, gold and osmium, also have this effect.

However, in many applications, in addition to a sufficient antibacterial effect, the active substance is generally required to be biocompatible with no cytotoxicity and thrombus-forming properties. Active substances such as mercury, bismuth, and copper do not have these properties due to their high cytotoxicity and lack of biocompatibility.

Several patented and non-patented documents deal with the fabrication and use of nanosilver. Further metal and inorganic compounds have only been reported sporadically. U.S. Pat. No. 5,520,664 discloses a plastic catheter. The antibacterial effect is achieved by introducing atoms by ion implantation. Metals with antibacterial activity include silver, chromium, aluminum, nickel, tungsten, molybdenum, platinum, iridium, gold, silver, mercury, copper, zinc, and cadmium. However, only silver and copper are mentioned in the examples and specific embodiments.

Japanese Unexamined Patent Publication No. 2001-54320 describes a plastic material containing 0.005 to 1% by weight of a mixture of molybdenum trioxide and silver oxide. The present invention relates to a film composed of an antibacterial resin component and a component that can be used for a partition material for a clean room, an uppermost layer of a flooring material, a lining material, a briefcase, a desk pad, a tablecloth, a packaging bag, a fabric, and the like. Here, the problem is that when an inorganic antibacterial active substance is incorporated, the transparency of a plastic material such as a vinyl chloride resin is lost. Loss of transparency is avoided by adding an oxide of hexavalent molybdenum. From this application, it is easily clear that if the weight ratio of molybdenum trioxide to silver oxide exceeds 95: 5, the antibacterial effect cannot be obtained. Therefore, the antibacterial effect is not due to the molybdenum oxide itself. When the ratio of molybdenum oxide to silver oxide is less than 30:70, that is, when the ratio of molybdenum oxide is small, discoloration of the vinyl chloride resin occurs.

The antibacterial plastic material described in JP-A-2001-04022 contains both an organic component and a metallic component having an antibacterial effect. Silver, platinum, copper, zinc, nickel, cobalt, molybdenum, and chromium are mentioned as metal components having an antibacterial effect. However, in examples and preferred embodiments, again, silver and copper are only stated to be active.

A glaze containing silver molybdate for ceramic parts is disclosed in JP-A-2000-143369. Here, 0.01 to 1% silver molybdate is added and converted to metallic silver. The effect is enhanced by adding 10 to 50% titanium oxide.

The antibacterial effect can also be achieved by a component having a photooxidative effect. Based on this, reactive free radicals are formed, which damage the microorganisms. Japanese Unexamined Patent Publication No. 11-012479 describes an antibacterial plastic material containing organic and inorganic components. Metallic particles such as silver, zinc, and copper, as well as additional compounds such as calcium zinc phosphate, ceramics, glass powder, aluminum silicates, titanium zeolite, ashstone, and calcium carbonate are mentioned as examples of inorganic components. Here, a metal oxide such as zinc oxide, titanium oxide, or molybdenum oxide acts as a catalyst for the photooxidation effect. Therefore, Japanese Patent Application Laid-Open No. 11-012479 clarifies that the antibacterial effect can be obtained only when the photooxidation mechanism acts, that is, the essential condition for obtaining the effect is the action of electromagnetic radiation.

<p> The availability of inexpensive substances with antibacterial effects is becoming even more important. These characteristics are especially important when many people are gathered and crowded, or when there is a high demand for hygiene, as is the case in hospitals, medical practice, sanatoriums, and public facilities. Here, reduction of nosocomial infections is particularly important. It is estimated that infection can occur in 0.5% of all hip transplants and 2-4% of all knee osteoarthritis. Especially with catheters, the risk of infection is high. In addition to this, there are also demands for the control and / or prevention of microbial habitat and growth in many more areas of application.</p><p> In addition to the availability of antibacterial substances, when the antibacterial active substance is subsequently incorporated into a composite material made into a corresponding article such as a catheter implant, filter, tube, container, cable, etc. It is also of further interest that the effect does not diminish.</p><p> In general, plastic materials are inexpensive to manufacture and easy to handle. Therefore, it is particularly preferable in many applications. However, here, since characteristics such as flexibility and / or rigidity and working stress differ depending on the type of plastic material, there is a problem that different types of plastic materials need to be used for different applications. Not all plastic materials are suitable, for example, for all applications, for example, catheters or infusion bags also need to have some flexibility, as opposed to, for example, implants or waste containers. .. Therefore, for all types of plastic materials used, whether the antibacterial material maintains its effect in relation to the corresponding plastic material, and / or in order to obtain its effect, the situation. Depending on the situation, it is necessary to test how it should be used with plastic materials. However, this results in a time-consuming, expensive, and extensive series of tests, that is, new tests for each desired application, resulting in increased manufacturing costs.</p><p> Therefore, an object of the present invention is to provide an active substance having a high antibacterial effect as compared with nanosilver. This active substance should have very little cytotoxicity and thrombus formation and should also have high general biocompatibility for medical applications. In addition, this active substance should have high cost-benefit and favorable processing properties. Furthermore, this material not only has an antibacterial effect in the form of nanoscale particles (particle size less than 100 nm), but also antibacterial in the form of non-respirable particles (particle size over 500 nm) and / or in compressed form. If it has an effect, it is advantageous. In addition, it is desirable to make a composite material that contains an antibacterial active substance, has many uses, and which this substance fully maintains its effect within the composite material.</p>

<p> This complex object is an inorganic substance that causes the formation of hydrogen cations upon contact with an aqueous medium in order to obtain an antibacterial effect, which is characterized by containing molybdenum and / or tungsten. Is achieved by using.</p><p> Whereas currently available inorganic active substances use a trace action effect, that is, a damaging effect of metal cations on living cells, the present invention reduces the pH value of the medium in contact with the substance. It utilizes the formation of hydrogen cations that cause it. Here, the free protons have a very small radius, so they immediately bind to water molecules and oxonium ions (H).₃O⁺) Is formed. If the concentration ratio allows, oxonium ions may bind to some water molecules. Therefore, H⁺In addition to H⁺The cation formed by the reaction between water and water and its hydrate are also called hydrogen cations. These are oxonium ions (H)₃O⁺) In addition to Zundel cation (H)₅O₂⁺) And Eigen cation (H)₉O₄⁺).</p><p> Molybdenum oxide reacts with water, for example, and molybdic acid (H).₂MoO₄), Which is H again₂React with O and H₃O⁺, And MoO₄^-Or MoO₄^2-To form. Tungsten trioxide is also H₂Tungstic acid with O (H₂WO₄), Which is H₂React with O and H₃O⁺, And WO₄^-Or WO₄^2-To form. According to Arrhenius, hydrogen cations mediate acidic properties. The pH value is the negative common logarithm of the value of hydrogen ion concentration in moles / liters. In a completely neutral aqueous solution, hydrogen ions and OH^-(Hydroxyd) ion has the same value (10)^-7It is mol / liter) and has a pH value of 7. Here, when a substance forms a hydrogen cation when it comes into contact with an aqueous medium, the value of the hydrogen cation increases, and thus the aqueous medium becomes acidic.</p>

<p> Here, surprisingly, it was found that a substance that forms a hydrogen cation when in contact with an aqueous medium has a very good antibacterial effect. It is also an important advantage that this substance is substantially not consumed. This is especially true when the material has low solubility in aqueous media. The solubility is preferably less than 0.1 mol / liter. The solubility of molybdenum oxide and tungsten oxide is less than 0.02 mol / liter. Therefore, the antibacterial effect exists almost infinitely from the viewpoint of time.</p><p> The aqueous medium may be, for example, water, a solution, or a suspension. Examples of solutions are body fluids and suspension liquid tissues. Here, it is sufficient that the aqueous medium exists in the form of a thin film on the surface of the substance. The effects of the present invention are already achieved in film thicknesses in the nanometer range, as are adsorbents. Therefore, the effects of the present invention have already occurred when the substance is exposed to air. Due to the formation of hydrogen cations, the pH value is usually lowered to &lt;6, preferably &lt;5. The acidic environment created by this causes the death of microorganisms.</p>

The effects of substances according to the present invention have been studied by a series of extensively designed tests. Here, the antibacterial effect and, in part, cytotoxicity and thrombus formation were also investigated. As shown in the examples, molybdenum and tungsten-containing materials whose surfaces are oxidized or which are present in an oxidized state are particularly effective. Molybdenum can exist in different oxidative states (VI, V, IV) and participates in the redox process to form relatively weak complexes with physiologically important compounds. Although molybdenum has a vital biochemical role, it does not bind to physiologically important compounds to a degree strong enough to have a significant inhibitory effect on metabolic processes. Therefore, it is not toxic to the human body. Molybdenum is a simple molybdenum date ion [MoO₄]^2-It is necessary to proceed from the assumption that it is ingested and transported by animals and plants in the form of. This [MoO₄]^2-Anions can penetrate the cell membrane without damaging the cell. Therefore, it is necessary to proceed from the assumption that molybdenum is not cytotoxic. Furthermore, the thrombus-forming effect is not known. Therefore, molybdenum is also suitable for medical applications. Tungsten-containing materials also exhibit a high antibacterial effect. At this time, no explicit statement can be made regarding thrombus formation, as the first study suggested some thrombus-forming effect. It is necessary to further clarify whether this is a property peculiar to tungsten or depends on the state of treatment.

Apart from molybdenum and tungsten-containing materials, antibacterial effects associated with lower pH values were also found in niobium oxide, magnesium oxide, and silicon carbide.

The method used to characterize the antibacterial effect is described in detail in the professional essay shown below: Fremdkorper-assoziierte Infektionen in der Intensivmedizin-Therapie und Praevention, JPGuggenbichler, Antibiotika Monitor 20 (3), 2004, pages 52-64 -Inzidenz und Pravention Fremdkorper-assoziierter Infektionen, JPGuggenbichler, Biomaterialien 5 (4) 2004, pages 228-236.

In particular, the roll-out culture method described therein has been shown to be useful for testing antibacterial effects. Here, a sample of the active substance is added to the bacterial suspension for a specific period of time, eg, 3 hours. Bacteria grow on the surface. After this time, the sample is rolled across a so-called agar plate and placed in sterile saline. This procedure is repeated several times every 3 hours. Repeating this rollout operation at 3-hour intervals provides information about whether a bacterial reduction or killing effect is occurring and the extent of that effect. This method can be used to test various microorganisms, bacteria, and viruses. Tests for demonstrating the effects of substances according to the invention were carried out individually on the reference strains Pseudomonas aeruginosa, Escherichia coli, and Staphylococcus aureus. Silver and copper were used as reference substances.

As mentioned above, it was possible to obtain the best results with substances containing molybdenum and tungsten. Here, it is extremely important for the present invention that molybdenum oxide or tungsten oxide is formed in the boundary region of the active substance mainly composed of molybdenum or tungsten. If this oxide is not formed to a sufficient degree or in the corresponding form, the antibacterial effect will not appear. This is also the reason why molybdenum and tungsten have not been used as antibacterial active materials until recently.

The antibacterial effect can be adjusted by pre-oxidation with heat, advantageously at temperatures above 300 ° C. Pre-oxidation can also be carried out chemically or electrochemically. This pre-oxidation is required for solid molybdenum and tungsten samples. Here, it was revealed that the substance pre-oxidized by annealing has a better antibacterial effect as compared with the oxide film formed in situ. Pre-oxidation needs to be carried out especially when the conditions at the time of use do not cause sufficient oxidation at all. Here, it is also a decisive condition that the specific surface area of the oxide film is large.

In addition to pure molybdenum and pure tungsten, compounds and alloys of these substances that are sufficiently stable and form an oxide film on their surface are also effective. Examples of the molybdenum compound having an antibacterial effect include molybdenum carbide, molybdenum nitride, molybdenum silicate, and molybdenum sulfide. Molybdenum, molybdenum oxide, and the above-mentioned substances are also commercially available in a very fine form having a particle size of &lt;1 μm by the Fisher method. Among suitable molybdenum alloys, 0.1 to 1% by weight La₂O₃Mo containing, 0.5% by weight Ti, 0.08% by weight Zr, 0.01 to 0.04% by weight of C containing Mo, 5 to 50% by weight of Re containing, and 1.2% by weight of Hf, 0.02 to 0.15% by weight. Must mention Mo containing C in.

Such alloys form an oxide film with antibacterial activity on the surface. In the case of tungsten, a tungsten material that forms an oxide film in situ or by the annealing described above is also effective. In addition to oxidized pure tungsten, tungsten oxides are effective. Here, Tungsten Trioxide Blue (WO_2.84) And WO₃Is specifically mentioned. 0.1 to 1% by weight La₂O₃Tungsten alloys of W containing 1 and W containing 1 to 26% by weight Re also have excellent antibacterial effects. Tungsten carbide, tungsten silicate, and tungsten sulfide are particularly suitable among the possible tungsten compounds that form an oxide film on the surface.

Silver produces a sufficient antibacterial effect only when it is present in a very finely ground state, whereas a substance according to the present invention has an antibacterial effect even when it is present in a compressed and dense form. Studies have shown that increasing the surface area further enhances the effect. Therefore, the presence of this material in a porous form can be advantageous for many applications.

The effect is also obtained when this material is present as a layer or as a component of the layer. A layer of molybdenum oxide and a layer of tungsten oxide, and / or a layer of molybdenum and a layer of tungsten that are oxidized by pre-oxidation if they are oxidized in situ or do not undergo sufficient oxidation in situ, are also particularly advantageous. Was shown to be. This layer can be deposited on top of a plastic material, ceramics, or metal. Particularly suitable deposition processes are thermal vapor deposition, sputtering, chemical vapor deposition, electroprecipitation, and electric arc deposition. The molybdenum oxide layer is, for example, molybdenum hexacarbonyl (Mo (CO)).₆) Can be produced by chemical vapor deposition by decomposition under atmospheric pressure. Organometallic CVD (MOCVD) is also possible. Here, as an organometallic compound, for example, molybdenum acetylacetonate (MoO)₂(CH₃COCH₂COCH₂)₂) Can be used. Molybdenum oxide film can be prepared using such an organometallic compound at a temperature in the range of about 400 to 500 ° C. MoO₃Not only Mo₉O₂₆And Mo₄O₁₁Can also be detected. The antibacterial effect is promoted when the particle size is &lt;1 μm and the layer thickness is in the range of several μm. The films of molybdenum oxide and tungsten oxide can also be deposited by means of reactive electron-beam deposition. Such films also have a very fine particle structure with pores sized in the range of 50-100 nm.

Electrophoresis and sol-gel processes also need to be mentioned as particularly suitable deposition methods.

When depositing layers on metals, common implant materials such as titanium, iron, and cobalt, and alloys thereof are preferred. ZrO with a purity of over 99% by weight, even if it is a ceramic substrate₂And Al₂O₃It is preferable to proceed from established materials such as. Layers can also be deposited on glass or glass ceramics.

As mentioned above, the effect is enhanced if the substance has as large a surface as possible with respect to the aqueous medium. Particularly excellent results can be obtained when the layer has a sponge-like porous structure having a pore size of 50 to 900 μm. Such a porous structure can be made, for example, in the form of a slurry or by depositing an antibacterial active substance from the gas phase and optionally subsequent annealing. Large surface areas can also be obtained when the layers are present in the form of island-like, substantially unconnected aggregated particles. It is particularly advantageous when such island-shaped agglomerated particles cover 40 to 90% of the surface of the substrate. The preferred size of the individual aggregated particles of material is less than 5 μm. The use of a substance according to the invention in powder form may be sufficient in many applications. Therefore, it is advantageous when the powder is very fine particles, that is, the particle size by the Fisher method is &lt;5 μm, preferably less than 1 μm.

It was possible to obtain the best results with metal composites and / or composite powders. Here, the composite material may also exist in the form of a composite powder. Such metal composites include additional metals that are chemically more noble, in addition to the materials according to the invention. Here, the formation of hydrogen cations is promoted by interaction with metals that are chemically more noble. Even a mixture of a substance according to the present invention with a nobler metal has an antibacterial effect if the sample has not undergone pre-oxidation.

Here, the chemically more noble metal is preferably silver, copper, tin, and alloys thereof. As the metal composite material, Mo-Ag, Mo-Cu, Mo-Sn, W-Ag, W-Cu, and W-Sn are particularly advantageous. Here, the content of Mo and / or W was preferably 10 to 90 atomic%, and it was possible to obtain the best results at 30 to 80 atomic%.

If this material should not be cytotoxic or thrombogenic, it is advantageous to use silver. However, these properties are completely irrelevant in many applications. Here, as an example, the fitting of a sanitary room is mentioned. Here, copper can be used instead of the expensive silver, which outperforms silver in its antibacterial effect. Even if Mo-Cu or W-Cu powder is added to other materials, a very high effect can be obtained. In addition, the metal composite can also exist in a compressed form as a layer or, for example, a porous shaped body. The production of compressed composites can advantageously be carried out by the infiltration technique.

In addition, substances according to the invention can be used in the production of antibacterial plastic materials.

Here, a composite material containing the substance of the present invention is particularly important when the composite material contains one or more materials, the at least one of which is formed from a crosslinkable polymer mixture. Includes a polymer matrix. The polymer mixture preferably comprises an unsaturated polyolefin (A) having a total carbon-carbon double bond to 1000 carbon atoms greater than 0.37.

It has been found that this composite material can be used for multiple purposes.

The use of unsaturated polyolefins in the polymer mixture makes the polymer mixture crosslinkable. This is preferably caused by the double bonds present in the polymer mixture. In that case, the degree of cross-linking can be adjusted through these double bonds based on the number of carbon-carbon double bonds in the polyolefin and further in the polymer mixture. However, the degree of cross-linking determines the flexibility and / or stiffness of the polymer. When the degree of polymer cross-linking is high, the rigidity is also high as compared with a polymer having a low degree of cross-linking. Therefore, composites according to the present invention can be used in the most different applications.

In addition, the crosslinkable polymer mixture preferably comprises an additional polymer (B).

Here, the term "total amount of carbon-carbon double bonds" associated with the term "unsaturated polyolefin (A)" relates to double bonds derived from vinyl, vinylidene, and / or trans-vinylidene groups. The amount of any type of double bond is measured according to the process described in the experimental section of European Patent No. 1731566.

The cross-linking properties of the polymer mixture can be controlled by introducing double bonds, thereby adjusting to the desired degree of cross-linking.

In various applications, the total carbon-carbon double bond content is preferably at least 0.40 with respect to 1000 carbon atoms. The content of 0.45 to 0.80 with respect to 1000 carbon atoms is of particular importance.

Further, the total content of vinyl groups in the unsaturated polyolefin is preferably higher than 0.11 with respect to 1000 carbon atoms. Here, a particularly preferable range is 0.15 to 0.80 with respect to 1000 carbon atoms, but it may be higher than this.

It is known that two types of vinyl groups are found in the polymer. One is the result of a so-called chain transfer agent, which is produced by the β-cleavage reaction of secondary radicals during the polymerization process. The second is preferably produced in the present invention by polymerization between at least one olefin monomer and at least one polyunsaturated monomer.

Both types of vinyl groups can be included in the polymer mixture of the present invention. However, the content of vinyl groups formed by polymerization between at least one olefin monomer and at least one polyunsaturated monomer is preferably at least 0.03/1000 carbon atoms. The content of 0.06 to 0.40 / 1000 carbon atoms is preferable.

In the present invention, the unsaturated polyolefin may be both unimodal and multimodal, for example bimodal, with a density of 0.860 to 0.960 g / cm.³, Preferably 0.880 to 0.955 g / cm³Especially preferably 0.900 to 0.950 g / cm.³Is.

Furthermore, unsaturated polyolefins are preferably made by polymerization from olefin monomers, preferably ethylene and propylene, and at least one polyunsaturated monomer.

Here, the polymerization can be carried out according to any known method, but radical polymerization under high pressure, which is described in detail in International Publication No. 93/08222, is preferably used.

Further, the unsaturated polyolefin preferably contains at least 60% by weight of ethylene monomer. A content of at least 70% by weight is more preferred, a content of at least 80% by weight is particularly preferred, and at least 90% by weight is most preferred.

The polyunsaturated comonomer is preferably a diene. Diene: -Free of heteroatoms, containing at least 8 carbon atoms, at least 4 carbon atoms present between unconjugated double bonds, and at least one of these double bonds present at the end Monomer having a carbon chain, The following-Equation I:<chemistry num="1"><img file="JP5437809B2_D0001.tif" /></chemistry>[In the formula, R1 and R2 are alkyl groups consisting of 1 to 4 carbon atoms and alkoxy groups also having 1 to 4 carbon atoms, which may be different or similar, and n = 1- 200] Siloxane represented by, and The following-Formula II:<chemistry num="2"><img file="JP5437809B2_D0002.tif" /></chemistry>[In the formula, R is-(CH₂)_m-O- or-(CH₂CH₂O)_n-Or-CH₂-C₆H₁₀-CH₂-O-, m is 2 to 10, t and n are 1 to 5] Α, ω-divinyl ether, represented by Those selected from the group consisting of are particularly preferred.

Dienes can be used in any possible combination.

Dienes used in the present invention and their preparation are described in more detail in WO 93/08222, WO 96/35732, and WO 97/45465, with reference to them. ..

Dienes are 1,7-octadiens; 1,9-decadiens; 1,11-dodecadiens; 1,13-tetradecadiens; tetra-methyldivinyldisiloxane; divinylpoly (dimethylsiloxane); and 1,4-butadiene diene. It is particularly preferred to be selected from the group consisting of vinyl ethers or combinations thereof.

In addition to polyunsaturated monomers, C such as propylene, 1-butene, 1-hexene, and 1-nonene, for example.₃-C₂₀Alpha-olefins, or additional comonomer such as polar comomeros such as alkyl acrylates, alkyl methacrylates, and vinyl acetate can be used for the polymerization.

However, the polar monomer content in the unsaturated polyolefin (A) is less than 150 micromoles, preferably less than 125 micromoles, and particularly preferably less than 100 micromoles.

In addition, the polymer mixture preferably comprises an additional copolymer (B). The copolymer (B) is preferably polar.

In addition, the polar copolymer (B) can contain the above compounds, such as unsaturated polyolefins, and thus the corresponding number of carbon-carbon double bonds. This further enhances the crosslinkability of the polymer mixture.

Here, the content of the carbon-carbon double bond in the polar copolymer is at least 0.15 with respect to 1000 carbon atoms. The content is preferably 0.20 to 0.35 with respect to 1000 carbon atoms.

However, polar copolymers are characterized by containing polar monomer units, i.e., at least 500 micromoles, preferably 700 micromoles, particularly preferably 900 micromoles, most preferably polar copolymers per gram of polar copolymers. It contains an amount of 1100 micromoles per gram.

Polar copolymers are made from olefins, preferably ethylene, and polar comonomer by polymerization. Here, at least one of the above-mentioned polyunsaturated monomers or a mixture thereof may be present.

Preferably, the polar comonomer is C₃To C₂₀It is a monomer of, for example, a hydroxyl group, an alkoxy group, a carbonyl, a carboxy group, an ester group, or a mixture thereof.

It is also preferred that the monomer unit be selected from the group consisting of alkyl acrylates, alkyl methacrylates, and vinyl acetates.

The comonomer is C₁To C₆Alkyl acrylate, C₁To C₆It is particularly preferable that it is a comonomer selected from the group consisting of alkyl methacrylate or vinyl acetate.

For example, alkyl esters of methacrylic acid such as methyl, ethyl, or butyl methacrylate, or polar monomers from the group consisting of vinyl acetate are considered particularly preferred. Due to its thermal stability, the acrylate type is preferred.

Polar copolymer (B) is the so-called melt flow rate, MFR_{2.16 / 190 ° C}However, it should be preferably 0.5 to 70 g / 10 minutes, more preferably 1 to 55 g / 10 minutes, and most preferably 1.5 to 40 g / 10 minutes.

The crosslinkable polymer mixture is preferably prepared by mixing two components, an unsaturated polyolefin (A) and a polar copolymer (B). A detailed description of the process of making the individual components (A) and (B) is given in European Patent No. 1,731,566.

The crosslinkable polymer mixture preferably contains 5-60% by weight of the polar copolymer, more preferably 8-50% by weight, particularly preferably 10-40% by weight, based on the weight of the crosslinkable polymer mixture. It is preferably 15 to 35% by weight.

Further, the crosslinkable polymer mixture preferably has a total carbon-carbon double bond content of more than 0.30 with respect to 1000 carbon atoms. Particularly preferably, the total content of carbon-carbon double bonds to 1000 carbon atoms is greater than 0.35; greater than 0.40; greater than 0.45, greater than 0.50; greater than 0.55, and particularly greater than greater than 0.60. Here, this quantification is based on the content of vinyl, vinylidene, and trans-vinylidene groups for 1000 carbon atoms for both unsaturated polyolefins (A) and polar copolymers (B).

Here, the content of the vinyl group is preferably 0.05 to 0.45 for the vinyl group with respect to 1000 carbon atoms, more preferably 0.10 to 0.40, and particularly preferably 0.15 to 0.35 with respect to 1000 carbon atoms. ..

The polymer matrix of the present invention is formed by cross-linking the cross-linking polymer mixture described.

This is preferably done with a cross-linking agent. This cross-linking agent regenerates radicals and thus initiates a cross-linking reaction. Compounds with at least one -O-O- or -N = N- bond are particularly preferred cross-linking agents. The use of peroxide is particularly preferred.

For example, di-tert-amyl peroxide; 2,5-di (tert-butylperoxy) -2,5-dimethyl-3-hexane; 2,5-di (tert-butylperoxy) -2,5-dimethylhexane, tert-butylcumyl peroxide; di (tert-butyl) peroxide; dicumyl peroxide; di (tert-butylperoxide-iso-propyl) benzene, butyl-4,4-bis (tert-butylperoxy) valerate; 1,1 -Bis (tert-butylperoxy) -3,3,5-trimethylcyclohexane; tert-butylperoxybenzoate, dienebenzoyl peroxide are suitable peroxides.

The antibacterial active material is preferably mixed prior to the cross-linking reaction to form a composite with the polymer matrix.

Here, the substance according to the present invention is preferably incorporated in a plastic material, particularly the polymer matrix described above, in an amount of 0.1 to 50% by volume. In a particularly advantageous embodiment, it is 3 to 15% by volume.

The cross-linking reaction is carried out under conventional conditions, eg, at a temperature of at least 160 ° C.

The crosslinked polymer matrix preferably has a breaking elongation of less than 175%, which is the so-called hot-set elongation, measured according to the method of IEC60811-2-1, more preferably 100. It is a value less than%, particularly preferably less than 90%. The value of elongation at break correlates with the degree of cross-linking. The lower the value of elongation at break, the higher the degree of cross-linking of the polymer mixture.

The degree of cross-linking and, therefore, the stiffness of the polymer mixture can be controlled by the content of the double bond and the amount of radical initiator, as described above.

In many applications, polyethylene with a high degree of cross-linking is preferable.

Composites according to the invention can be easily processed by injection molding. Granules mixed in an extruder are preferably used in the preparation of injection molded composites, which are simultaneously subjected to a cross-linking reaction. In contrast to the production of polymer matrix composites made with silver nanopowder, when using materials according to the invention, no carrier for the active material is used to avoid the formation of aggregated particles or clusters. It is possible.

The degree of cross-linking adjusted individually, and thus, for example, as further described below, in addition to the possibilities of various applications, the polymer matrix used in the composite is mechanically and thermally stable. Is also good.

As additives to plastic materials and / or polymer matrices, molybdenum oxide, pre-oxidized molybdenum, tungsten oxide, pre-oxidized tungsten, Mo-Cu, W-Cu, Mo-Ag, and W-Ag are plastics. It imparts a very good antibacterial effect to materials and / or polymer matrix composites. The best results were obtained with Mo-Cu, W-Cu, Mo-Ag, and W-Ag. Again, we must proceed from the assumption that the chemically noble metal promotes the oxidation of the non-noble metal, thus producing hydrogen ions. When a composite powder of Mo-Cu, W-Cu, Mo-Ag, or W-Ag is used as an additive, the molybdenum and / or tungsten phase and the copper and / or silver phase are in very small form. It is still important to exist. For example, a composite powder produced by a coating process can be used. The particle size of the composite powder is preferably &lt;5 μm.

The substance according to the present invention may exist in combination with one or more kinds of ceramic materials. The production is carried out by, for example, a hot press method. Alumina, titanium oxide, silicon oxide, silicon carbide, and zirconium oxide are particularly suitable as ceramic phases. Additives according to the present invention include, for example, MoO so that conventional fabrication methods and conditions for ceramics can be used.₃And WO₃Etc., those existing in the highest oxidized state are suitable. In addition to this, metals Mo and W may also be present.

Therefore, the material combinations shown below are suitable ceramic composites: Al.₂O₃-MoO₃, Al₂O₃-WO₃, ZrO₂-MoO₃, ZrO₂-WO₃, Al₂O₃-Mo-MoO₃, Al₂O₃-W-WO₃, ZrO₂-Mo-MoO₃, ZrO₂-W-WO₃, TiO₂-MoO₃, TiO₂-WO₃, TiO₂-Mo-MoO₃, TiO₂-W-WO₃, SiO₂-MoO₃, SiO₂-WO₃, SiO₂-Mo-MoO₃, And SiO₂-W-WO₃.. Here, MoO₃Or WO₃The advantageous proportion of is 0.001 to 50 mol percent. MoO₃Or WO₃ZrO against₂, Al₂O₃, TiO₂, Or SiO₂The advantageous molar ratio of is 1 to 100.

Due to its high antibacterial effect, there are several advantageous uses for substances according to the present invention. This includes implants and other medical technology instruments. However, with respect to implants, substances according to the invention can be used particularly favorably in catheters, stents, bone implants, tooth implants, artificial blood vessels, and internal artificial organs.

Advantageous applications in the field of catheters include port catheters and bladder catheters. Port catheters typically include a chamber with a silicone membrane and a connected tube. To date, chambers are usually made of plastic material, plastic exterior titanium, or ceramics. Here, the catheter, or the chamber of the catheter, can be made from a substance according to the present invention, or a material containing the substance. However, it is also possible to provide a catheter having a layer according to the present invention, or a portion of the catheter. Very good results can be obtained when the chamber consists of Mo-Ag, with an Ag content of 1-40% by weight. According to prior art, this chamber is also exteriorized with plastic material. Furthermore, it is also advantageous that the plastic material and / or the silicone film contains this material.

Problems due to bacterial contamination can also occur at Luer lock connections, three-way cocks, and cock benches, thus representing the preferred use of materials according to the present invention. ..

In coronary stents, it is advantageous to apply a substance according to the invention by a coating process onto a shape memory alloy stent, such as Nitinol. Substances according to the present invention can also be advantageously used for ureteral stents. Ureteral stents are usually made from polyurethane or silicone. Here, a substance according to the present invention may be added to this polymeric material, or may again be applied as a layer on its surface.

Bone implants come into contact with tissue fluid. Again, a substance according to the present invention can exert its effect. Here, it is advantageous to apply a substance according to the present invention as a layer. An example of a bone implant is the hip joint. The area of the condyle is advantageous to smooth the layer, while the shaft of the implant can be provided with a porous coating. As described above, the substance according to the present invention is also suitable for obtaining an antibacterial effect in an artificial blood vessel or a hernia omentum because it can be easily incorporated into a polymer material. Medical technical applications also include use as a surgical case.

In addition, the substances according to the invention can be used in any kind of medical container. Due to the high risk of contamination by microorganisms, it is advantageous to use substances according to the present invention in nasal spray bottles.

Apart from its pure medical and veterinary potential, there are multiple possible applications in the field of hygiene. This material is suitable as an additive for absorbent sanitary articles or wound coverings. Sanitary products and scratch coatings have polymer fibers or lattices. Here, a substance according to the present invention can be advantageously attached to the surface of the fiber and / or the lattice, or the fiber and / or the lattice may contain this substance.

In addition, the wound coating spray, also known as the "liquid wound coating plaster" on the market today, often has only a short-term antibacterial effect. It has become clear that a substance according to the present invention can be used as an additive thereof in order to enhance its antibacterial effect or to maintain it for a long period of time. Here, when the substance according to the present invention contains molybdenum or is composed of molybdenum, its use is preferable. Molybdenum and / or its compounds and alloys are more preferably used in wound coating sprays at concentrations of 0.05 to 1.0% by volume, especially 0.1 to 0.5% by volume.

Substances according to the invention are also suitable as additives for varnishes, coatings, and adhesives. Here, the benefit is shown when the varnish, coating agent, or adhesive contains 0.01 to 70% by volume of this substance. A particularly preferable range is 0.1 to 40% by volume. MoO₃And / or WO₃Is particularly suitable as an additive for cost-sensitive products. Here, the preferable particle size by the Fisher method is 0.5 to 10 μm. Here, it is possible not to add a precious metal such as silver. However, if a particularly high effect is required, additives based on W-Ag, W-Cu, Mo-Ag, Mo-Cu, Mo-Sn, and W-Sn are suitable. Particles having a preferred size of 0.5 to 10 μm by the Fisher method can be incorporated into a liquid varnish system such as a two-component polyurethane varnish by a conventional dispersion technique.

Apart from its use in the medical and hygiene fields, substances according to the present invention can also be used as additives in personal hygiene products. Advantageous products here include ointments, soaps, dental cleanser compositions, toothpastes, toothpastes, toothbrushes, intertooth cleansing agents, and tooth cleansing chewing gums. chewing gum) is mentioned.

Furthermore, the substance according to the present invention can also be advantageously used as an additive for the filter. Here, in addition to tungsten or molybdenum, metal composites further comprising a nobler phase such as silver, copper, or tin have demonstrated their benefits to a special degree. Here, the filter can again be composed of polymeric fibers containing or coated with this material.

Currently, antibacterial active substances are already used in products such as clothing and shoe insoles. Even in the field of such applications, the low cost as compared with nanosilver can be advantageously utilized. Here, the polymer fiber may contain this substance, or the substance may be present in the form of being deposited on the polymer fiber.

Since the substances according to the invention can be easily mixed with varnishes, coatings and / or plastic materials, products made from them are suitable for furnishing, especially hygiene room fixtures. ing.

Apart from such areas of application, there are many additional areas of application for substances according to the invention, especially for products that are in frequent contact with organisms. These include, for example, switches, fixtures, credit cards, keyboards, cell phone exteriors, coins, banknotes, door handles, and internal equipment in public transport. An even more advantageous use is a component of an air conditioning system. The substance according to the present invention is suitable for, for example, an air conditioner for transportation means such as an automobile. Conventionally, radiating fins made of aluminum alloy can be advantageously coated with a substance according to the present invention. Shafts of building air conditioning systems can also be designed to antibacterial specifications by adding this active substance to the shaft material or by coating the shaft material with it. Humidifiers can also have the corresponding antibacterial properties.

Further, it is preferable to use a substance according to the present invention for cables, particularly cables containing polyurethane.

This is just a list to illustrate possible advantageous uses. In addition, substances according to the invention can be used in all cases where nanosilver has already been used or people are already beginning to consider it. Here, it is necessary to consider that the requirements to be met regarding antibacterial effect, thrombus forming property, and cytotoxicity differ depending on the field of application.

The present invention is further characterized by the following items:

1. An antibacterial active substance containing molybdenum and / or tungsten, and a composite material made of one or more materials, wherein at least one material is a polymer matrix formed from a crosslinkable polymer mixture. A composite material comprising, wherein the crosslinkable polymer mixture contains an unsaturated polyolefin (A) having a total amount of carbon-carbon double bonds to 1000 carbon atoms greater than 0.37.

2. The composite material according to item 1, wherein the crosslinkable polymer mixture further contains a copolymer (B).

3. The composite material according to item 1 or 2, wherein the mass content of the substance in the composite material is 0.1 to 50% by volume.

4. The composite material according to any one of items 1 to 3, wherein the surface of the substance is at least partially oxidized.

5. The composite material according to any one of items 1 to 4, wherein the substance is molybdenum oxide or tungsten oxide.

6. The composite material according to any one of items 1 to 4, wherein the substance is molybdenum, a molybdenum alloy, and / or a molybdenum compound, and has a layer of molybdenum oxide on its surface.

7. The composite material according to any one of items 1 to 4, wherein the substance is tungsten, a tungsten alloy, and / or a tungsten compound, and has a layer of tungsten oxide on the surface thereof.

8. The above molybdenum compound is 0.1 to 1% by weight of La₂O₃Mo containing, 0.5% by weight Ti, 0.08% by weight Zr, 0.01 to 0.04% by weight of C containing Mo, 5 to 50% by weight of Re containing, or 1.2% by weight of Hf, 0.02 to 0.15% by weight. The composite material according to item 6, characterized in that it is Mo containing C of.

9. The composite material according to item 6, wherein the molybdenum compound is molybdenum carbide, molybdenum nitride, molybdenum silicate, and / or molybdenum sulfide.

10. La of 0.1 to 1% by weight of the above tungsten alloy₂O₃7. The composite material according to item 7, characterized in that it is W containing 1 to 26% by weight of Re.

11. The composite material according to item 7, wherein the tungsten compound is tungsten carbide, tungsten nitride, tungsten silicate, and / or tungsten sulfide.

12. Any one of items 1 to 11, wherein the unsaturated polyolefin (A) of the crosslinkable polyolefin mixture is prepared by polymerization of an olefin monomer and at least one polyunsaturated monomer. The composite material described in the section.

13. The composite material according to item 12, wherein the olefin monomer is ethylene.

14. The composite material according to item 12, wherein the polyunsaturated monomer is a diene.

15. The polyunsaturated components mentioned above a) Contains no heteroatoms, contains at least 8 carbon atoms, contains at least 4 carbon atoms between unconjugated double bonds, and at least one of these double bonds is present at the end. Carbon chain or b) Equation I:<chemistry num="3"><img file="JP5437809B2_D0003.tif" /></chemistry> [In the formula, R1 and R2 are alkyl groups consisting of 1 to 4 carbon atoms and alkoxy groups also having 1 to 4 carbon atoms, which may be different or similar, and n = 1- 200] Α, ω-divinylsiloxane represented by, or c) Equation II:<chemistry num="4"><img file="JP5437809B2_D0004.tif" /></chemistry> [In the formula, R is-(CH₂)_m-O- or-(CH₂CH₂O)_n-Or-CH₂-C₆H₁₀-CH₂-O-, m is 2 to 10, n is 1 to 5] Α, ω-divinyl ether represented by, or d) A mixture of a), b), and / or c), The composite material according to item 14, characterized in that it comprises any of the above.

16. The composite material according to any one of items 1 to 15, wherein the copolymer (B) is polar.

17. The composite material of item 16, wherein the polar copolymer is made by polymerization of one type of olefin and at least one type of polar copolymer.

18. Use of the composite material according to any one of items 1 to 17 in an article in which the growth of microorganisms should be avoided.

19. The use according to item 18, characterized in that the article is a medical product.

20. The product comprises a chamber having a silicone membrane and a connected tube, wherein the chamber and / or tube comprises the composite material according to any one of items 1-17. Use as described in item 19.

21. The use according to item 20, wherein the product is a luer lock connection, a three-way cock, and / or a cock bench.

The present invention will be described in more detail below by way of example.

Table 1 includes items related to sample preparation.

Table 2 shows the effect on Staphylococcus aureus, Table 3 shows the effect on E. coli, and Table 4 shows the effect on Pseudomonas erginosa.

<p> Table 1 shows the substances tested. Table 1 also includes an overview of starting material composition and sample preparation. The pressing process was performed by die pressing on the samples of W 02, W 03, W 04, W 05, Mo 02, Mo 03, Mo 04, and Mo 05 according to the present invention at a pressing pressure of about 250 MPa. The sintering step was performed on these samples at a temperature of 850 ° C. for 60 minutes in a pure hydrogen atmosphere in a tungsten tube furnace. Non-alloy tungsten (Sample W 09) and non-alloy molybdenum (Sample Mo 09) are subjected to isobaric pressing at 220 MPa and baked at a temperature of 2250 ° C for 4 hours and / or at a temperature of 2100 ° C for 4 hours. After knotting, a roll bending step was performed, and the degree of deformation was about 70%.</p><p> TransOptic, a Buhler acrylic resin usually used to make polished sections, was used as the polymer matrix for making polymer matrix composites. Molybdenum layers were deposited using atmospheric plasma spraying (Samples SL 50, SL 51, SL 52). Here, the thickness of the layer was about 100 μm, and the density of the layer was 85% of the theoretical density. Since this coating process was performed in the air exposure, the oxygen content in the layer was approximately 1.5% by weight. Here, oxygen is mainly MoO₃It existed in the form of. The molybdenum layer was deposited on titanium alloy (SL 50), niobium (SL 51), and intermetallic compound (SL 52).</p><p> Comparative samples include non-alloyed copper (Cu 01), non-alloyed silver (SL 14), 20% by weight copper powder embedded in a plastic matrix (SL 20), 50% by weight copper embedded in a plastic matrix. Powder (SL 20), 20% by weight silver (SL 21) embedded in the plastic matrix, and 50% by weight silver (SL 27) embedded in the plastic matrix were used. In addition, for comparison, the antibacterial effects of a number of additional materials, mainly niobium, tantalum, and titanium, were measured.</p><p> The antibacterial effect was investigated using the rollout culture method described above. The tests were performed separately for Pseudomonas erginosa, E. coli, and Staphylococcus aureus. For this purpose, a sample of the active substance was added to the bacterial suspension. Bacterial growth occurred on the surface. After 3, 6, 9, and 12 hours, the sample was rolled across a so-called agar plate and added to sterile saline. After this rolling step, agar plates were photographed to evaluate their effect on bacterial reduction and / or killing. Photographs and evaluations of the effects of Staphylococcus aureus are shown in Table 2, Escherichia coli in Table 3, and Pseudomonas erginosa in Table 4.</p><p> Here, it can be seen that all substances mainly composed of tungsten or molybdenum have an antibacterial effect at least equal to or partially superior to that of pure silver in a compressed form. Samples containing silver or copper in addition to molybdenum, or samples containing silver or copper in addition to tungsten, have been shown to be particularly effective.</p><p> Polymer matrix composites containing molybdenum oxide or tungsten oxide must also be evaluated as having a good antibacterial effect. Preferably, it is advantageous to use a fine particle powder having a particle size of less than 5 μm by the Fisher method.</p><p> With the exception of samples mixed with copper, tantalum or niobium-based samples have no effect. Linked to good results in Ta-Cu and Nb-Cu is the high antibacterial effect of copper, albeit associated with cytotoxicity.</p><p> The effectiveness of titanium-based comparative samples must also be evaluated as poor.</p><p> Testing with polymer matrix materials has shown that the effect can be controlled by the amount and particle size of molybdenum and / or tungsten powder added. The finer the molybdenum and / or tungsten powder, the higher the effect (SL 33, SL 34). Here, the molybdenum oxide powder has a higher antibacterial effect than the molybdenum metal powder (SL 22, SL 33).</p><p> In addition to the samples listed here, niobium oxide, silicon carbide, and manganese oxide had antibacterial effects attributed to lower pH values.</p><p> In addition, the first test for cytotoxicity was performed. All copper-containing materials have been shown to be cytotoxic. The first test for thrombus formation was also performed. The silver-containing tungsten alloy has a high thrombus-forming property as compared with the silver-containing molybdenum alloy. However, it should be mentioned in a limited way that surface properties also affect the results.</p><p> Tests with water-soluble molybdenum and tungsten salts were also performed to elucidate the mechanism of action. Sodium molybdate (Na) for this purpose₂MoO₄) And Tungsten Molybdate (Na)₂WO₄) Was embedded in a plastic matrix, and the above-mentioned test was performed to measure the antibacterial effect.</p><p> Here, the pH value of the physiological saline did not decrease. The sample had no antibacterial effect. Therefore, after 24-hour age-hardening, the content of the elements dissolved in the saline solution was measured. As expected, this value was very high for water-soluble compounds. For example, in the case of sodium molybdate, the molybdenum content in saline solution is 50 mg / l · cm.²Was measured. For comparison, for antibacterial active substances, this value is 0.1 (Sample SL 18), 0.4 (Sample SL 22), and 0.4 mg / l · cm.²(Sample SL 24). Therefore, the antibacterial effect does not correlate with the content of molybdenum or tungsten in saline.</p><p> Similar results were obtained for sodium tungstate. Here, the content of tungsten in physiological saline is 324 mg / l · cm.²Met. As an example, the value for sample SL 17 is 0.1, the value for sample SL 19 is 0.3, and the value for sample SL 35 is 0.9 mg / l · cm.²Was measured.</p><p> The silver content in physiological saline after age hardening for 24 hours was measured for silver-containing materials such as W 02 and W 03. Here, the value for W 02 is 28.6, and the value for W 03 is 68.2 mg / l · cm.²Met.</p><p> As is known from the literature, silver is Ag⁺It acts antibacterial through the formation of ions. Ag⁺The effect increases as the concentration increases. However, for molybdenum and tungsten, no dependence of its antibacterial effect on the content of molybdenum and / or tungsten in saline could be confirmed. As a result, we need to proceed from the fact that molybdenum and tungsten themselves are not active. Therefore, the pH value of saline was measured after the test. Materials without antibacterial effect, such as tantalum, tantalum-5Ag, tantalum-20Ag, niobium, niobium-5Ag, and niobium-20Ag, were approximately neutral. Pure silver also does not induce a drop in pH at all.</p><p> However, it was confirmed that all the samples according to the present invention lowered the pH value. The pH value of W 09 was 4.8, W 02 was 3.3, W 03 was 3.1, and the sample of tungsten carbide containing 20% by weight of silver was 5.3, Mo 09 was 4.0, Mo 02 was 3.9, and Mo 03 was 3.8. The decrease in pH value is due to the oxonium ion (H).₃O⁺) Due to the formation. This is H₂MoO₄And / or H₂WO₄Formed from the reaction between water and water, MoO₄^-, MoO₄^2-, Or WO₄^-Or WO₄^2-Occurs.</p><p> H₂MoO₄And / or H₂WO₄Is MoO₃And / or WO₃And H₂Reformed from reaction with O and / or dissolved oxygen.</p><p><tables num="1"><img file="JP5437809B2_D0005.tif" /></tables></p><p><tables num="2"><img file="JP5437809B2_D0006.tif" /></tables></p><p><tables num="3"><img file="JP5437809B2_D0007.tif" /></tables></p><p><tables num="4"><img file="JP5437809B2_D0008.tif" /></tables></p><p><tables num="5"><img file="JP5437809B2_D0009.tif" /></tables></p><p><tables num="6"><img file="JP5437809B2_D0010.tif" /></tables></p><p><tables num="1"><img file="JP5437809B2_D0011.tif" /></tables></p><p><tables num="2"><img file="JP5437809B2_D0012.tif" /></tables></p><p><tables num="3"><img file="JP5437809B2_D0013.tif" /></tables></p><p><tables num="4"><img file="JP5437809B2_D0014.tif" /></tables></p><p><tables num="5"><img file="JP5437809B2_D0015.tif" /></tables></p><p><tables num="6"><img file="JP5437809B2_D0016.tif" /></tables></p><p><tables num="7"><img file="JP5437809B2_D0017.tif" /></tables></p><p><tables num="8"><img file="JP5437809B2_D0018.tif" /></tables></p><p><tables num="9"><img file="JP5437809B2_D0019.tif" /></tables></p><p><tables num="10"><img file="JP5437809B2_D0020.tif" /></tables></p><p><tables num="11"><img file="JP5437809B2_D0021.tif" /></tables></p><p><tables num="12"><img file="JP5437809B2_D0022.tif" /></tables></p><p><tables num="13"><img file="JP5437809B2_D0023.tif" /></tables></p><p><tables num="14"><img file="JP5437809B2_D0024.tif" /></tables></p><p><tables num="15"><img file="JP5437809B2_D0025.tif" /></tables></p><p><tables num="16"><img file="JP5437809B2_D0026.tif" /></tables></p><p><tables num="17"><img file="JP5437809B2_D0027.tif" /></tables></p><p><tables num="18"><img file="JP5437809B2_D0028.tif" /></tables></p>