JPH05246858A - Preventive/medicine for mrsa infectious disease - Google Patents

Abstract

PURPOSE:To provide the subject preventive/medicine containing, as active ingredient, nucleic acid constituent(s). CONSTITUTION:The objective preventive/medicine containing, as active ingredient, at least one kind of nucleic acid constituent selected from inosine, guanosine-n'-monophosphate (n' is 2, 3 or 5), uridine, and thymidine. The active ingredient is e.g. of such molar ratio as follows: (1) inosine/cytidine/guanosine-n'- monophosphate(GMP)/uridine/thymidine = 4:4:4:3:1; or 4:0.04:4:3: l; (2) adenosine-n'-monophosphate (AMP)/cytidine-n'-monophosphate(CMP)/GMP/uridine-n'- monophosphate (UMP)/thymidine = 4:4:4:3:1; (3) CMP/GMP/UMP/inosine-n'- monophosphate/thymidine = 4:4:3:4:1; or 2:2:1:2:1. Besides, combined use of arginine and/or glutamine can further enhance the preventive/medicinal effect.

Description

Detailed Description of the Invention

[0001]

The present invention relates to a novel preventive and / or therapeutic agent for MRSA infection, and more specifically, it contains at least one selected from the specific nucleic acid constituents (nucleic acid bases, nucleosides and nucleotides) as an active ingredient. MRS
A infection preventive and therapeutic agent.

[0002]

2. Description of the Related Art MRSA stands for methicillin-resistant Staphylococc.
is a general term for nosocomial infections that produce various toxins and enzymes such as enterotoxins and coagulase, and cause infections in a wide range of organs such as the skin, digestive tract, respiratory system and urinary tract. Known as a pathogen. Also, the M
Unlike exogenous bacterial species such as Pseudomonas bacteria that have been a problem in hospital infections, RSA is a resident human bacterium, so patients and medical staff who are MRSA carriers can enter the hospital. It often exists, which makes it difficult to prevent the MRSA infection from occurring in the clinic. Moreover, since the MRSA has strong resistance to many antibacterial agents, once it develops an infectious disease, it often becomes intractable, which is a major clinical problem.
Currently, there are almost no drugs available for the treatment of A infection, and vancomycin, minomycin, phosphomycin, cephametase, cefzonan, etc. are present, but there is a concern that they will become resistant. There is a strong demand in the art for the development of a prophylactic and therapeutic agent effective for MRSA, which is an alternative to the above.

Therefore, an object of the present invention is to provide a new drug effective for the prevention and treatment of the above MRSA infection.

As a result of intensive studies conducted by the inventors of the present invention as a result of the above objectives, the present inventors promoted protein synthesis by first administering intravenously in the form of a nutritional supplement, that is, especially in the form of an infusion,
Among the series of nucleic acid component compositions found to be useful for nutritional management and maintenance of nitrogen balance [see Japanese Patent Application Laid-Open No. 60-126220], surprisingly, a preventive and therapeutic effect on MRSA infection is exhibited. I found that things exist. Furthermore, the present inventors have found that the preventive and therapeutic effects of MRSA infection of the above composition are further enhanced by the combination of the composition with a specific amino acid, that is, arginine and / or glutamine. The present invention has been completed based on this new finding.

[0005]

That is, according to the present invention,
Inosine, guanosine-n'-monophosphate (GMP)
(N ′ = 2 ′, 3 ′ or 5 ′), and an agent for preventing and treating MRSA infection, which comprises at least one nucleic acid constituent selected from uridine and thymidine as an active ingredient.

The preventive and therapeutic agent for MRSA infection of the present invention comprises:
The above inosine, guanosine-n'-monophosphate (GM
P), at least one selected from uridine and thymidine
It is also possible to use the nucleic acid constituent component of the seed as an essential constituent and further combine it with other nucleic acid constituent constituents (nucleic acid bases, nucleosides and nucleotides) arbitrarily. Each component used in this combination is not particularly different from those constituting the nucleic acid component composition according to the applicant's application, and as a nucleic acid base, adenine, hypoxanthine, guanine, cytosine, uracil, thymine, Orotic acid, etc. and their non-toxic salts (eg sodium salts, etc.)
Can be illustrated. As nucleosides, adenosine,
Examples thereof include nucleosides such as inosine, guanosine, cytidine, uridine and orotidine, deoxyadenosine, deoxyguanosine, deoxycytidine, deoxyribonucleosides such as deoxyuridine and thymidine, and non-toxic salts thereof. Examples of the nucleotides include those in which 1 to 3 phosphates are bound to the above nucleosides and their non-toxic salts. Specific examples of such nucleotides are shown below.

Adenosine-n'-monophosphate (AMP),
Adenosine-n'-diphosphate (ADP), adenosine-
n'-triphosphate (ATP), inosine-n'-monophosphate (IMP), inosine-n'-diphosphate (IDP), inosine-n'-triphosphate (ITP), guanosine-n '
-Monophosphate (GMP), guanosine-n'-diphosphate (GDP), guanosine-n'-triphosphate (GTP),
Cytidine-n'-monophosphate (CMP), Cytidine-n '
-Diphosphate (CDP), cytidine-n'-triphosphate (C
TP), uridine-n'-monophosphate (UMP), uridine-n'-diphosphate (UDP), uridine-n'-triphosphate (UTP), etc. (where n'is 2 '. 3'or 5 ').

Deoxyadenosine-n'-monophosphate (d
AMP), deoxyadenosine-n'-diphosphate (dA
DP), deoxyadenosine-n'-triphosphate (dAT
P), deoxyguanosine-n'-monophosphate (dGM
P), deoxyguanosine-n'-diphosphate (dGD
P), deoxyguanosine-n'-triphosphate (dGT
P), deoxycytidine-n'-monophosphate (dCM
P), deoxycytidine-n'-diphosphate (dCD
P), deoxycytidine-n'-triphosphate (dCT
P), thymidine-n'-monophosphate (TMP), thymidine-n'-diphosphate (TDP), thymidine-n'-triphosphate (TTP) and the like (where n'is 3 '. Or 5 ').

In the following description of the above nucleotides, the abbreviations in parentheses (IUPAC-
In accordance with IUB regulations or symbols conventionally used in the field), the indication shall be adopted.

The preventive and therapeutic agent for MRSA infection of the present invention comprises:
Preferably, at least one nucleic acid constituent selected from inosine, GMP, uridine and thymidine is further combined with the above-exemplified nucleic acid constituents (nucleic acid bases, nucleosides and nucleotides) as an active ingredient. Is good. The following are examples of particularly preferable combinations of active ingredients.

Cytosine / Thymidine / GMP / UMP / I
MP, thymine / inosine / AMP / CMP / GMP, A
MP / CMP / GMP / UMP / TMP, AMP / CM
P / dGMP / UTP / IMP, cytosine / uridine /
AMP / CMP / UMP, adenine / cytosine / inosin / UMP / TDP, thymine / CMP / dATP / dG
MP / UTP, thymidine / AMP / CMP / GMP / U
MP, thymidine / CMP / GMP / UMP / IMP, cytosine / thymidine / GMP / UMP / IMP and inosine / cytidine / GMP / uridine / thymidine.

Particularly preferred components and their compounding ratios in the case of the combination of the above-mentioned 5 kinds of active ingredients are as follows.

Inosine: Cytidine: GMP: Uridine:
Thymidine = 4: 4: 4: 3: 1 (molar ratio, the same below) and 4: 0.04: 4: 3: 1, AMP: CMP: GM.
P: UMP: thymidine = 4: 4: 4: 3: 1 and CM
P: GMP: UMP: IMP: thymidine = 4: 4: 3:
4: 1 and 2: 2: 1: 2: 1.

It is known that the above-mentioned AMP changes into IMP in the body, and therefore such AMP and IMP
Can replace each other in whole or in part.

Furthermore, the combination of the active ingredients of the pharmaceutical preparation of the present invention is not limited to the above five cases, and on the basis of this, other nucleic acid constituents are further added to make six or more kinds. Of course, it is possible. Among the combinations of 6 or more, preferable examples include cytosine / inosine / AMP / UMP / GMP / IMP, adenine / inosine / thymidine / CMP / UMP / IMP, and dAMP /
ATP / GMP / UDP / IMP / dCMP, guanosine / inosine / uridine / UMP / IMP / dGMP,
AMP / CMP / dGMP / UTP / TMP / IMP,
Thymidine / CMP / GMP / UMP / IMP / TMP,
Cytosine / Thymidine / UMP / CMP / dAMP / dG
MP, ATP / dCMP / GMP / UDP / TTP / I
MP etc. (6 types) can be illustrated.

The drug of the present invention includes, in addition to the above-mentioned nucleic acid constituents and combinations thereof as the active ingredient, further combinations of at least one selected from arginine and glutamine. The combined use of the specific amino acids further enhances the desired MRSA preventive and therapeutic effects exhibited by the above-mentioned nucleic acid constituents or a combination thereof. Such a specific amino acid can be administered as a single agent by blending a predetermined amount in a formulation consisting of the above-mentioned respective nucleic acid constituents and a combination of the above, or prepared in a formulation form separate from the above formulation. Thus, it can be co-administered with the preparation. In any case, the amount of the specific amino acid used in combination is usually about 1 to 100 times by weight, more preferably 2 to 100 times the weight of the active ingredient consisting of each nucleic acid constituent or a combination thereof.
It is preferably selected from the range of about 30 times by weight.

The agent of the present invention can be prepared by singly mixing the predetermined compounds as the above-mentioned active ingredients, alone or in an appropriate combination or, if necessary, further combining with the above-mentioned specific amino acid, and simply mixing them. Is its route of administration,
It is shaped or prepared into an appropriate dosage form (dosage unit form) depending on the administration method and the like. Examples of the dosage unit form include liquid forms such as injections suitable for intravenous administration, oral forms, powders suitable for tube administration and topical administration, tablets, pills, granules, powders,
Solutions, suspensions, emulsions, capsules, suppositories, syrups,
Ointment etc. can be illustrated. The preparation of such dosage unit forms is
It can be carried out using a suitable pharmaceutical carrier according to a conventional method. Examples of the carrier for the preparation include diluents and excipients such as a filler, a filler, a binder, a moisturizer, a disintegrant, a surface active agent and a lubricant, which are usually used, depending on the use form of the preparation. it can.

The specific amino acid preparation as the above-mentioned action enhancer can also be prepared in the same suitable dosage form as above.

More specifically, in the case of molding in the form of tablets, examples of the pharmaceutical carrier include lactose, sucrose, sodium chloride, glucose, urea, starch, calcium carbonate, kaolin, crystalline cellulose, silicic acid and potassium phosphate. Excipients, water, ethanol, propanol, simple syrup, glucose solution, starch solution, gelatin solution, carboxymethylcellulose, hydroxypropylcellulose, methylcellulose, polyvinylpyrrolidone and other binders, sodium carboxymethylcellulose, carboxymethylcellulose calcium, low-substituted hydroxy Propyl cellulose, dry starch, sodium alginate, agar powder, laminaran powder, sodium hydrogen carbonate,
Disintegrators such as calcium carbonate, polyoxyethylene sorbitan fatty acid esters, sodium lauryl sulfate, surfactants such as stearic acid monoglyceride, disintegration inhibitors such as sucrose, stearin, cocoa butter, hydrogenated oil, etc.
Absorption promoters such as primary ammonium base and sodium lauryl sulfate, humectants such as glycerin and starch, starch,
Lactose, kaolin, bentonite, adsorbents such as colloidal silicic acid, purified talc, stearate, boric acid powder, lubricants such as polyethylene glycol, and the like can be used. Further, the tablet may be a tablet coated with an ordinary coating, for example, a sugar-coated tablet, a gelatin-coated tablet, an enteric-coated tablet, a film-coated tablet, a double tablet, or a multilayer tablet.

In the case of molding in the form of pills, as a pharmaceutical carrier, for example, glucose, lactose, starch, cocoa butter,
Excipients such as hardened vegetable oil, kaolin and talc, gum arabic powder, tragacanth powder, binders such as gelatin and ethanol, disintegrating agents such as laminaran and agar can be used.

When molded into the form of suppositories, polyethylene glycol, cocoa butter, higher alcohols, esters of higher alcohols, gelatin, semi-synthetic glycerides, etc. can be used as pharmaceutical carriers.

Capsules are usually prepared by mixing the active ingredient of the present invention with the various pharmaceutical carriers exemplified above and filling hard gelatin capsules, soft capsules and the like.

When the drug of the present invention is prepared as an injection such as a solution, emulsion or suspension, it is preferable that they are sterilized and isotonic with blood. As the diluent, for example, water, ethyl alcohol, macrogol, propylene glycol, ethoxylated isostearyl alcohol, polyoxylated isostearyl alcohol, polyoxyethylene sorbitan fatty acid esters and the like can be used. In this case, a sufficient amount of sodium chloride, glucose, glycerin, etc. for adjusting the isotonic solution may be contained in the drug of the present invention, and a usual solubilizer, buffer, soothing agent, etc. may be added. You may add.

When the paste, cream, gel or the like is formed into an ointment, white diluents such as petrolatum, paraffin, glycerin, cellulose derivatives, polyethylene glycol, silicone and bentonite can be used as diluents. Furthermore, a colorant, a preservative, a flavoring agent, a flavoring agent, a sweetening agent and the like and other pharmaceuticals can be contained in the drug of the present invention, if necessary.

The preparation of the present invention can also be prepared in the form of an infusion solution, and the preparation of the present invention in the form of such an infusion preparation contains sugars such as glucose, fructose, xylitol, sorbitol and maltose, lipids, vitamins, electrolytes, Trace elements and the like can be added and blended, and if necessary, stabilizers, pH adjusters and the like can be added and blended.

The amount of the active ingredient to be contained in the drug of the present invention is not particularly limited and may be appropriately selected from a wide range, but it is usually about 0.5 to 10% by weight in the pharmaceutical preparation. Is good. In particular, the infusion formulation of the present invention has a pH of 3.0 to 8.0, preferably 5.0 to 7.5, and a total active ingredient concentration of about 0.5 to 10 w / v%, preferably Is suitably prepared in the range of 2 to 8 w / v%.

The administration method of the above-mentioned pharmaceutical preparation is not particularly limited, and it is determined according to various preparation forms, patient's age, sex and other conditions, degree of disease and the like. For example, tablets, pills, solutions, suspensions, emulsions, granules, capsules, etc. are orally administered, and injections are intravenously administered alone or in a mixture with a normal replenishing solution such as glucose, amino acid, etc. Independently, it is intramuscularly, intradermally, subcutaneously or intraperitoneally administered, and the suppository is intrarectally administered.

The dose of the preparation of the present invention is appropriately selected according to the usage, age of the patient, sex and other conditions, degree of disease and the like, but usually the amount of the active ingredient is about 5 to 12 mg per 1 kg of body weight per day. The dosage should be 1 per day
It can be administered in 4 divided doses. In addition, the formulation of the present invention in the form of an infusion is generally about 10 to 2 per adult per day.
The dose may be about 50 ml, preferably about 20 to 100 ml.

The thus-prepared preventive and therapeutic agent for MRSA infectious disease of the present invention can exert a very excellent effect in the preventive and therapeutic treatment of MRSA infectious disease based on the use of an active ingredient which has not been heretofore known.

[0030]

EXAMPLES In order to explain the present invention in more detail, preparation examples of the preventive and therapeutic agents of the present invention will be given below as Examples, and then pharmacological test examples carried out on the active ingredient of the present invention will be given.

[0031]

Example 1

[0032]

[Table 1]

[0033] Pure crystals of each nucleic acid component having the above composition were added to distilled water for injection and dissolved by stirring. Then, 0.3 g of sodium bisulfite was added as a stabilizer, and hydrochloric acid was added as a pH adjuster. The pH was adjusted to about 7.4 using. Then, the obtained nucleic acid component aqueous solution is aseptically filtered, filled in an infusion container, replaced with nitrogen, the container is closed, and the MRS of the present invention is sterilized in an autoclave at 105 ° C. for 40 minutes.
A preventive and therapeutic agents (total nucleic acid component concentration in free form 8 w / v
%) Was prepared.

[0034]

Example 2

[0035]

[Table 2]

The preventive and therapeutic agent for MRSA of the present invention having the above composition is prepared in the same manner as in Example 1 except that pH is adjusted to about 7.3 using acetic acid and sterilization is performed in an autoclave at 110 ° C. for 40 minutes. (Free nucleic acid component concentration 8 w / v%) was prepared.

[0037]

Example 3

[0038]

[Table 3]

The MRS of the present invention having the above composition was prepared in the same manner as in Example 1 except that the pH was adjusted to about 6.4 using hydrochloric acid.
A preventive and therapeutic agent (total nucleic acid component concentration in free form 4 w / v
%) Was prepared.

[0040]

Example 4

[0041]

[Table 4]

An MRSA preventive and therapeutic agent of the present invention having the above composition (total nucleic acid component concentration in free form: 3.4 w / v%) having the above composition was prepared in the same manner as in Example 1 except that a pH adjuster was not used.

[0043]

[Example 5] As a nucleic acid constituent pure crystal, inosine 2.
7 g and 4.1 g of 5'-GMP-2Na were sifted through a 60-mesh sieve, then mixed evenly and filled in a glass container to prepare the MRSA preventive and therapeutic agent of the present invention in powder form. Was prepared. Inosine and 5'-G of this product
The molar ratio of MP-2Na was about 1: 1.

This product can be appropriately prepared into a form for enteral administration at a desired concentration by dissolving it in an appropriate amount of purified water at the time of use.

[0045]

[Test Example 1] Preventive effect test against MRSA-infected mice 49 4-week-old Balb / c female mice were bred for 30 days on a 20% casein diet, and 25 of them were used for prevention and treatment of MRSA infection of the present invention. The agent (1) administration group (Example 1 administration group) was used, and the remaining 24 animals were used as a control group (physiological saline administration group 1). Once a day, from the start of breeding, 0.35 ml of the reagent and physiological saline were intraperitoneally administered to the mice of both groups, and the body weight was measured.

Further, in place of the above-mentioned MRSA infection preventive and therapeutic agent (1) of the present invention, the following MRSA infection preventive and therapeutic agent of the present invention (2) 0.35 ml (this is referred to as Example 4 administration group) and The same test was repeated by intraperitoneally administering 0.35 ml (the same as the injection solution administration group of Table 5) of (3) to each of the 25 mice. For the equality test, 25 animals were separately set as a control group (physiological saline administration group 2).

Agent for Preventing and / or Treating MRSA Infection of the Present Invention (2) ... Drug of Example 4 Agent for Preventing and / or Treating MRSA Infection of the Present Invention (3) ... Example 4
Injection solution having the formulation shown in Table 5 below, which was prepared in the same manner as

[0048]

[Table 5]

Staphylococcus aureus (Staphylococcus aureu)
As s), the 8985N strain of bacteria isolated from the patient (confirmed methicillin resistance) was maintained on an agar medium, and then BH
Substituting in I broth and maintaining at 37 ° C. for 18 to 24 hours, centrifuging (10000 rpm, 15 minutes) from the culture solution to collect the bacteria, suspending them in physiological saline, and then measuring 1.0 at an absorbance of 660 nm. The one prepared in the above was used. The number of bacteria in this solution is Eklund a
nd Lankford) measurement result of 2.1 × 10
It was ⁸ CFU / ml.

On the 10th day from the start of the breeding of the mice, 0.3 ml of the above bacterial solution was administered (inoculated) into the tail vein of each group of mice. Thereafter, the survival rate of mice and the morbidity rate due to changes in body weight and the like were determined. In addition, all mice that survive on the 20th day after bacterial administration were sacrificed, spleen, kidney, and heart were aseptically collected, homogenized with physiological saline, and the number of bacteria in each organ sample was measured by the plate dilution method. At the same time, the weight of each organ was measured. In addition, the number and weight of the bacteria in the organs were measured in the same manner for the individuals who had already died by the 20th day of bacterial infection.

The results of the survival rate after MRSA infection of the mice of each group obtained by using the reagent preparations of (1), (2) and (3) are shown in FIG. 1 [vertical axis: survival rate (% ), Abscissa: days after inoculation]. In the figure, the graph (1) is the MRSA of the present invention.
Infectious disease preventive and therapeutic agent (1) administration group (Example 1 administration group)
The graph (3) shows a group for administering the preventive and therapeutic agent for MRSA infection of the present invention (2) (administration group of Example 4), and the graph (4) shows a preventive and therapeutic agent for MRSA infection according to the present invention (administered by injection solution of Table 5). Groups) are shown respectively.

Further, in FIG. 1, as a graph (2), the results of the physiological saline administration groups 1 (24 animals) and 2 (25 animals) are shown together as a control group.

From FIG. 1 above, the survival rate of mice in the control group was 25% in the saline-administered group 1 (6 out of 24 mice).
And 16% in physiological saline-administered group 2 (4 out of 25), which was 20.4% in total (10 out of 49), whereas the present group (Example 1 administration group) 72% (18 out of 25), 80% in the present invention group (Example 4 administration group)
(20 out of 25) and the group of the present invention (Table 5 injection solution administration group)
Was 64% (16 out of 25), which were all significantly high. This is also clear from the fact that the survival rate 20 days after inoculation of each of the groups of the present invention with respect to the control group was statistically treated according to the χ 2 -test, and all were p <0.01. From this, the preventive effect of the drug of the present invention on MRSA infection was clarified.

Further, the weight measurement results of each of the surviving and deceased mice obtained using the reagent composition of the above (1) [vertical axis: body weight (g), horizontal axis: days after inoculation] are shown. ,
It is shown in FIG. 2 in comparison with those of the control group.

In the figure, graph (1) represents the present invention group (surviving individuals), graph (2) represents the present invention group (dead individuals), graph (3) represents the control group (surviving individuals), and graph ( 4) shows a control group (dead individual), respectively.

The recovery of the surviving mice in both groups from MRSA infection can be seen from the recovery process of weight loss. From the figure, in both groups, surviving animals showed an increase in body weight from about 10 days after infection. It can be seen that the recovery from MRSA infection has occurred.
On the other hand, in both groups, it is found that the weight loss after infection continues to die in the case of death.

Further, in Table 6 below, in the case of using the reagent composition of the above (1), at the time of death of a mouse (surviving individual) and a dead mouse (dying individual) that survived for 20 days after MRSA infection, The results of measuring the number of bacteria in each organ (spleen, kidney, heart) and the weight (g) of each organ are shown.

[0058]

[Table 6]

From the table, the number of kidney bacteria of the dead individuals was 2.0 × 10 ⁸ CFU or more in both groups, but it was 2.0 × 10 ⁶ CFU or less in the surviving individuals, and that of the heart and spleen. It can be seen that the numbers of bacteria are extremely low in both the dead and surviving individuals in both groups, and that the organ weights of the spleens in both groups are more than twice as large as in the dead individuals. Based on these findings, there is an increase in immune response in surviving individuals.

From the above results, it is clear that the preventive and therapeutic agent for MRSA infection of the present invention can significantly increase the resistance of an individual to MRSA infection.

[0061]

[Test Example 2] Therapeutic effect test on MRSA-infected mice 69 4-week-old Balb / c female mice were bred for 30 days on a 20% casein diet, and on the 10th day from the start of breeding,
MRSA was inoculated in the same manner as in Test Example 1, and from the day after the bacterial inoculation, 35 test mice (invention group) were prepared to prevent and treat MRSA infectious disease of the present invention prepared in Example 1 (the above-mentioned reagent agent). (1)) and the other 34 animals (control group) were each intraperitoneally administered with 0.35 ml of physiological saline once a day.

Table 7 shows the results of comparative examination of the survival rates of the mice in each group on the 20th day after the inoculation of the above bacteria.

[0063]

[Table 7]

From Table 7, the survival rate of the mice of the present invention was significantly higher than that of the mice in the control group, and the therapeutic effect of the agents of the present invention on MRSA infection is clear.

Further, in the above, the same test was carried out using the following reagent agents (2) to (9) instead of the reagent agent (1) as the preventive and therapeutic agent for MRSA infection of the present invention. ..

(2) Drug of Example 4 (administration of 0.35 ml) (3) Injection solution of the formulation shown in Table 5 prepared in the same manner as in Example 4 (administration of 0.35 ml) (4) Example 1 Inosine injection solution having a total free nucleic acid content of 0.80 w / v% prepared in the same manner as above (inosine content of 0.80 w / v%, 0.35 ml administration) (5) 0.35 ml of the drug of Example 1 and arginine 0.1
Combined administration with 4 g (oral administration) (6) 0.35 ml of the drug of Example 4 and 0.2 glutamine
Combined administration with 8 g (oral administration) (7) 0.35 ml of the drug of Example 4 and 0.1 of arginine
Combined administration with 4 g and glutamine 0.28 g (oral administration) (8) Combined administration with inosine injection solution 0.35 ml (4) above and arginine 0.14 g and glutamine 0.28 g (oral administration) (9) Physiology Administration of saline solution (0.35 ml) The results obtained are shown in Table 8 below.

[0067]

[Table 8]

From Table 8 above, the therapeutic effect of the drug of the present invention on MRSA infection is clear.

[0069]

[Test Example 3] Effect test of orally ingested nucleic acid for resistance to MRSA infection 84 4-week-old Balb / c female mice were divided into 3 groups, and 34 mice were purified with a nucleic acid-free diet (NF, composition below). Then, the other 40 animals were treated with 0.5% RNA feed prepared by adding 0.5% RNA to NF, and the remaining 10 animals were treated with 2.5.
% RNA feed was used for 30 days.

NF feed composition (% by weight) Corn starch 41.5 Casein 25.0 Alpha starch 10.0 Cellulose powder 8.0 Linole salad oil 6.0 AIN76TM salt mixture (manufactured by Oriental Yeast Co., Ltd.) 3.5 Granulated sugar 5.0 AIN76TM vitamin mixture (manufactured by the same company) + choline bitartrate 1.0 total 100.0 On the 10th day from the start of breeding, MRSA bacteria were inoculated into each group of mice in the same manner as in Test Example 2, and from the day after the bacterial inoculation, To each of the NF feed-fed mice and the 2.5% RNA feed-fed mice, 0.35 ml of physiological saline was intraperitoneally administered once daily. In addition, 40 mice fed 0.5% RNA feed were further divided into 2 groups, and one of them (20 mice) was treated with the preventive and therapeutic agent for MRSA infection of the present invention prepared in Example 1 (this Invention group), the other (20) physiological saline,
0.35 ml of each was intraperitoneally administered once daily.

Table 9 shows the results of comparative examination of the survival rates of the mice in each group on the 20th day after the inoculation of the above bacteria.

[0072]

[Table 9]

From Table 9, it is clear that MRSA infection resistance is not observed with orally ingested RNA, but MRSA infection resistance can be enhanced by parenteral ingestion of the drug of the present invention.

[0074]

[Test Example 4] MRSA infection resistance test 65 4-week-old Balb / c female mice were bred for 30 days in a 20% casein diet, divided into 6 groups of 10 to 12 mice, and once a day from the start of breeding. , 0.80% (w / v%, the same applies hereinafter) inosine injection solution, 1.22% 5'-GMP-2N, prepared in the same manner as in Example 1.
a 0.35% each of injection solution, 0.73% cytidine injection solution, 0.55% uridine injection solution and 0.18% thymidine injection solution
ml or 0.35 ml of physiological saline (control group).

MRSA bacteria were inoculated on the 10th day after the start of breeding in the same manner as in Test Example 1, and the survival rates of the mice in each group on the 20th day after the inoculation were compared and examined. The results are shown in Table 10.

[0076]

[Table 10]

From Table 10, it is seen that the survival rate of mice in the control group is higher than that in the group of the present invention (inosine injection solution, 5'-GMP-).
2Na injection solution, cytidine injection solution and thymidine injection solution administration group) had a high value, and in particular, the survival rate of mice in the inosine injection solution administration group was significantly high, which indicates that the agent of the present invention has a therapeutic effect on MRSA infection. it is obvious.

[0078]

[Test Example 5] MR in mouse fed with amino acid-enriched sample
SA infection resistance test 100 4-week-old Balb / c strain female mice were divided into 5 groups, and the 1st group was fed with 20% casein feed. Groups 2 and 3 were fed with 20% casein diet containing arginine and glutamine as amino acids in amounts of 2% and 4%, respectively, and the third group was prepared according to Example 4 of the present invention. 1 daily preventive and therapeutic agent for MRSA infection
Each time, 0.35 ml was intraperitoneally administered, and each animal was raised. Group 4 was fed with 20% casein feed supplemented with 2% arginine as an amino acid, and the preventive and / or therapeutic agent for MRSA infection of the present invention prepared in Example 4 was used once daily for 0.3 times.
5 ml was intraperitoneally administered and raised. Also, the fifth group is 20%
The MRSA of the present invention prepared by feeding a casein feed containing 4% of glutamine as an amino acid and further adjusting it in Example 4.
The preventive and therapeutic agents for infectious diseases were intraperitoneally administered once a day in an amount of 0.35 ml for breeding.

On the 10th day from the start of breeding, MRSA was inoculated once into each group of mice in the same manner as in Test Example 1,
On the 8th day after inoculation, the spleen of each group of mice was collected, homogenized with sterile physiological saline, and the number of viable cells was examined by the surface smearing method using mannitol agar medium. The resistance to MRSA infection was evaluated by considering the following conditions.

The results obtained are shown in Table 11.

[0081]

[Table 11]

From Table 11, the number of bacteria in the spleen of the first group (control) was distributed to 10 ⁵ to 10 ⁶ , whereas in the second group (amino acid combination group), 10 ⁴ to 10 ⁸ cells were distributed. 1 in the third group (amino acid combination group + prophylactic and therapeutic agent administration group of the present invention)
0 to ² or less to 10 ^6, the fourth group (amino acids formulations + present invention prophylactic and therapeutic agent administered group) In 10 ² or less to 10 ^7,
It is also understood that in the fifth group (amino acid combination group + the preventive and therapeutic agent-administered group of the present invention), 10 ⁴ to 10 ⁷ each are distributed. When the cumulative χ ² test of these results was carried out, the numbers of bacteria in the second, third, fourth, and fifth groups were significantly lower than those in the first group.
Group 3 and 4 had significantly lower numbers of bacteria compared to group 4
It is clear that the number of bacteria in the third group is significantly lower than that of the groups 5 and 5. From these facts, when arginine and glutamine are added to normal feed to enhance amino acids, MRS
Resistance to infection by A is enhanced, and the combined use of this amino acid-enriched feed with the preventive and / or therapeutic agent for MRSA infection of the present invention further enhances resistance to MRSA infection, and this effect is especially due to arginine and glutamine as amino acids. It can be seen that it is more prominent even when compared with the case of using the preventive and therapeutic agents of the present invention respectively.

[Brief description of drawings]

FIG. 1 is a graph showing the survival rate of MRSA infection preventive and therapeutic agent-administered group of the present invention and control group (physiological saline-administered group) mouse in the test according to Test Example 1 after MRSA infection.

FIG. 2 is a graph showing the results of measuring the weight of each surviving individual and dead individual of the MRSA infection preventive and therapeutic agent-administered group of the present invention and the control group (physiological saline-administered group) mouse in the test according to Test Example 1.

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Claims (3)

[Claims] 1. An active ingredient comprising at least one nucleic acid constituent selected from inosine, guanosine-n'-monophosphate (GMP) (n '= 2', 3'or 5 '), uridine and thymidine. An agent for preventing and treating MRSA infection, which comprises: 2. The preventive and therapeutic agent for MRSA infection according to claim 1, wherein the active ingredient is selected from the following ingredient compositions. Inosine: Cytidine: Guanosine-n'-monophosphate (G
MP): uridine: thymidine = 4: 4: 4: 3: 1 (molar ratio, the same applies hereinafter) and 4: 0.04: 4: 3: 1, adenosine-n′-monophosphate (AMP): cytidine- n'-
Monophosphate (CMP): Guanosine-n'-monophosphate (G
MP): uridine-n′-monophosphate (UMP): thymidine = 4: 4: 4: 3: 1 and cytidine-n′-monophosphate (CMP): guanosine-n′-monophosphate (GM)
P): uridine-n'-monophosphate (UMP): inosine-n'-monophosphate (IMP): thymidine = 4: 4: 3:
4: 1 and 2: 2: 1: 2: 1 [where n ′ is 2 ′,
3'or 5 ']. 3. The M according to claim 1, further containing at least one selected from arginine and glutamine.
Preventive and therapeutic agent for RSA infection.