JPH09183730A - Medicine containing enantiomer of fosfomycin - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a pharmaceutical composition useful as an immunomodulator, an anti-inflammatory agent, a treating agent for intractable infectious diseases or a formation-inhibiting agent or a disrupting agent of biofilm by compounding the enantiomer of fosfomycin or its salt. SOLUTION: This pharmaceutical composition contains the enantiomer of fosfomycin or its salt. The enantiomer of fosfomycin is preferably used in a form of a salt. For example, the sodium salt or the calcium salt is cited as the salt of fosfomycin. When used as a treating agent for an intractable infectious disease, the enantiomer of fosfomycin or its salt is preferably compounded with an antibacterial agent in an amount of 0.5-2mol based on 1mol of the enantiomer or its salt. As the antibacterial agent, e.g. gentamycin or a β-lactam agent is cited. A daily dose of the enantiomer of fosfomycin is preferably 0.2-1g/ adult in the case where it is used as an immunomodulator, an anti-inflammatory agent or a biofilm-formation inhibiting agent, and the medicine is preferably administered in one or several divided doses.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION Field of the Invention The present invention relates to a pharmaceutical application of the enantiomers of fosfomycin, more immunomodulators enantiomers of fosfomycin in particular, anti-inflammatory agents, intractable infectious therapeutic agents, biofilm formation inhibitor It relates to a pharmaceutical use as an agent.

[0002] Fosfomycin (Fosfomycin) is widely known as antibiotics produced by actinomycetes (JP-B 45-9828 Patent Publication). A method for synthesizing this fosfomycin has been established, and a method for optically resolving the racemate obtained by the synthesis has been clarified (Japanese Patent Publication No. 46-43206 and Japanese Patent Publication No. 53-24407).

In addition to antibacterial action, fosfomycin also has an immunomodulatory action (JP-A-59-98094),
It has been found to have a special action such as an anti-allergic action and a symptom-relieving action of intractable diseases which are considered to be involved in biofilm.

However, despite the fact that fosfomycin has these interesting special actions, it has not been possible to clearly grasp the effect thereof due to the presence of antibacterial action which is the main action of fosfomycin. In addition, when fosfomycin is administered in the expectation of exhibiting the above-mentioned special action, if the amount thereof is large, there is a fear of bacterial replacement phenomenon. Therefore, it would be interesting if the antibacterial action and the above special action could be separated.

[0005] In particular, regarding the function of immunoregulation, its mechanism in the body is very complicated, but knowledge in this field is rapidly expanded, and immunomodulators and anti-inflammatory agents which act on these biological defense systems. Have been found to be very useful (see Immunology Illustrated, Nankodo, published March 1, 1986).

Therefore, it can be said that a superior drug having an immunomodulatory action and an anti-inflammatory action is always desired.

Further, in recent years, a so-called intractable infectious disease, which has no therapeutic effect even though the most suitable antibacterial agent is used from the viewpoint of antibacterial activity, has become a problem. The existence of a biofilm (microbial film) has been pointed out as one of the intractable factors. Since it was observed that bacteria adhered to the inside of pipes or rock surfaces in nutrient-poor running water to form biofilms (Annual Review of Microbiology, 35 , 29
9,1981), and biofilms formed by bacteria have been observed on the surface of living tissues (Infection and Immunity, 43 , 359, 1984). On the other hand, biofilms have also been observed in endovascular catheters and the like in the field of medical devices (Kidney International, 35 , 614, 198).
9). It is believed that these bacteria produce alginate, which is mainly composed of polysaccharides, and form biofilms that allow the drug to survive without passing through the membrane (annual. Review of Microbiology, 41 , 435, 1987).

Biofilm formation by bacteria in the living body has come to be regarded as one of the factors for making infectious diseases intractable, and as a result, the interaction between biofilm and various antibacterial agents has been the subject of research. However, as of today, no antibacterial agent has been found that has high permeability to biofilms and can completely kill germs inside. Also, the adjunctive therapy of destroying biofilms is not perfect.

On the other hand, the effects of long-term administration of erythromycin and clarithromycin, which are macrolide antibiotics, were reported on Pseudomonas aeruginosa diffuse panbronchiolitis (Journal of Infectious Diseases, 60 , 45, 1986). ), And clarithromycin has been observed to suppress the production of glycocalyx at low concentrations and also suppress adhesion to tissues (infectious diseases, 21 , 161, 1991). However, the action mechanism is unknown, and it is difficult to say that these methods are always effective.

Therefore, if the action of fosfomycin on the biofilm can be separated from the antibacterial action, it can be said that it is useful for establishing a method for treating intractable infectious diseases that are believed to be related to the biofilm.

[0011]

SUMMARY OF THE INVENTION The present inventors have now surprisingly found that, while the fosfomycin enantiomer has substantially no antibacterial action, it also provides immunomodulation, anti-inflammatory, intractable infection treatment, and biotechnology. It has been found to be useful in inhibiting or breaking film formation.

Accordingly, the present invention provides a pharmaceutical composition comprising an enantiomer of fosfomycin or a salt thereof.

According to another aspect of the present invention, an immunomodulator, an anti-inflammatory agent, a therapeutic agent for intractable infections, and a biofilm formation inhibitor or destruction comprising an enantiomer of fosfomycin or a salt thereof. An agent is provided.

[0014]

DETAILED DESCRIPTION OF THE INVENTION Enantiomer of fosfomycin Enantiomer of fosfomycin, namely ((+) (cis-1,2-
epoxypropyl) phosphonic acid) is Japanese Patent Publication No. 46-43
It can be easily obtained according to the production method described in JP-B No. 206 and JP-B No. 53-24407.

The enantiomer of fosfomycin is preferably used in the form of a salt, and examples of the salt form include pharmacologically acceptable salts such as sodium salt, calcium salt and trometamol salt. In the present specification, unless otherwise specified, the enantiomer of fosfomycin is used as a concept including its salt.

Efficacy / Effects of Fosfomycin Enantiomer The enantiomer of fosfomycin has an immunoregulatory effect of regulating and normalizing immune function in mammals including humans. The term "modulation" is used herein to mean to restore and maintain the immune system of the mammal, including human, in a normally balanced state.

Therefore, the enantiomer of fosfomycin is
First, it is useful in the treatment of mammals, including humans, who are immunocompromised.

In addition, the enantiomer of fosfomycin is used in the in-vivo system such as infections caused by bacteria, mold, mycoplasma, etc., chronic infections such as sepsis, tuberculosis, leprosy and acute and chronic viral infections. Useful for treating diseases. It is preferably used in combination with antibiotics in the treatment of such diseases.

Further, the enantiomer of fosfomycin has an anti-inflammatory or anti-allergic effect and can be used for the treatment and / or prevention of inflammatory diseases or allergic diseases.

Furthermore, the enantiomer of fosfomycin regulates and normalizes the immune function and is therefore useful for the treatment of autoimmune diseases in which harmful antibodies are present such as systemic osteoarthritis. It is also useful for treating conditions where the immune response is below normal, such as neoplasms and organ transplants.

Although the details of its mechanism of action are not clear, it is expected that the enantiomer of fosfomycin may assist the general immunity in the body by stimulating and assisting cellular immunity. More specifically, it is considered that the immune function is regulated or normalized by selectively controlling the production of certain cytokines. IL-1 is a cytokine predicted to be selectively regulated in its production by the enantiomer of fosfomycin.
α, IL-1β, IL-1ra, INFα, INFγ,
IL-2, IL-6, IL-10, GM-CSF, IL
-8, GROα, MIP-1α, RANTES and the like. In addition, it is expected that the enantiomer of fosfomycin enhances its immune function by enhancing antibody production in mammals including humans. Details of the production control function of these cytokines and the action of enhancing antibody production are described further in the Examples below.

Further, the enantiomer of fosfomycin has an action of inhibiting the formation of biofilm or destroying the biofilm once formed. Therefore, it is useful for the treatment of intractable infectious disease, which has conventionally been thought to be difficult to treat by the formation of the biofilm. Although the mechanism of action of inhibiting biofilm formation or destroying biofilm is not clear, fosfomycin is sialyl-Lewi ^sX (sL ^eX ) which is a ligand of selectin known to be a cell adhesion factor against bacteria. It is considered that by inhibiting the expression of the bacterium as a surface antigen, the bacteria are prevented from adhering to the tissue and the bacteria from adhering to each other.

Bacteria capable of inhibiting or destroying biofilm formation by enantiomers of fosfomycin include Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus spp., Pseudomonas aeruginosa, Klebsiella spp., Escherichia coli, Enterococcus spp. Bacteria such as Bacterium, Synthobacter, and Serratia can be mentioned.

By administering the enantiomer of fosfomycin, it is possible to inhibit the formation of the biofilm or to destroy the biofilm that has been formed.
Thus, by administration of the appropriate antibiotic, concomitantly with or subsequent to the administration of this enantiomer of fosfomycin,
Treating intractable infections that may involve biofilms, such as chronic respiratory infections such as diffuse panbronchiolitis due to Pseudomonas aeruginosa, chronic urinary tract infections such as intractable prostatitis, or It can be prevented.

In addition to fosfomycin, ofloxacin, gentamicin, imipenem / cilastatin are used as antibiotics in combination with enantiomers of fosfomycin as an immunomodulator or anti-inflammatory agent or biofilm formation inhibitor or destroyer in the present invention. , New quinolone drugs such as ceftazicin, clarithromycin,
Examples include aminoglycoside drugs, β-lactam drugs, macrolide drugs and the like.

According to a preferred embodiment of the present invention, it is preferable to administer an antibacterial agent in an amount of 0.1 to 5 mol, and more preferably 0.5 to 1 mol, per 1 mol of fosfomycin enantiomer.
2 moles.

Pharmaceutical Composition The enantiomer of fosfomycin may be directly administered to mammals including humans, but usually it is preferably administered as a pharmaceutical composition containing it as an active ingredient.

Pharmaceutical compositions containing the enantiomer of fosfomycin as an active ingredient are administered orally and parenterally (eg, intravenous injection,
It can be administered to humans and non-human animals by any route of administration (intramuscular injection, subcutaneous administration, rectal administration, transdermal administration). Therefore, the pharmaceutical composition according to the present invention is made into a suitable dosage form depending on the administration route, specifically, mainly by intravenous injection,
Injections such as intramuscular injection, capsules, tablets, granules, powders,
Oral agents such as pills, fine granules, troches, rectal agents,
Various preparations such as oily suppositories and aqueous suppositories can be prepared. These various preparations include commonly used excipients, fillers, binders, wetting agents, disintegrating agents, surfactants, lubricants, dispersants, buffers, preservatives, solubilizing agents, preservatives. , A flavoring agent, a soothing agent, a stabilizer and the like can be used for the production. Examples of the non-toxic additives that can be used include lactose, fructose, glucose, starch, gelatin, magnesium carbonate, synthetic magnesium silicate, talc, magnesium stearate, methyl cellulose,
Examples include carboxymethyl cellulose or a salt thereof, gum arabic, polyethylene glycol, syrup, petrolatum, glycerin, ethanol, propylene glycol, citric acid, sodium chloride, sodium sulfite, sodium phosphate and the like.

The dose of the enantiomer of fosfomycin is appropriately determined depending on the individual case in consideration of symptoms, age, sex, etc., but as an immunomodulator or an anti-inflammatory agent,
Usually about 0.1 to 5 g / day, preferably 0.2 / day
~ 1 g, which is administered once or several times a day. In order to inhibit or destroy biofilm formation, it is usually about 0.1 to 5 g, preferably 0.2 to 1 g per day for an adult, and this is administered once or several times a day. Further, the dose of fosfomycin when the enantiomer of fosfomycin and the antibiotic are administered simultaneously or sequentially may be the same as above.

[0030]

Example 1 In sepsis, many cytokines (especially TNF, IL-
1 and IL-6) play an important role, and it is considered that inflammation is greatly involved in the pathological condition.
The anti-inflammatory effect of fosfomycin enantiomer sodium salt was examined as follows in the pathological condition of sepsis using a mouse Pseudomonas aeruginosa endogenous infection model.

SPF ddY mouse (4-6 weeks old, male)
200 mg / kg of ampicillin was preliminarily administered to the above to disturb the intestinal resident bacteria, and then Pseudomonas aeruginosa strain D4 was suspended in 0.45% saline and given as drinking water to settle in the intestines. Then, cyclophosphamide 150-200 mg / k
g was administrated twice (the second administration was the third day after the first administration) to make it into a granulocytopenic state, causing endogenous sepsis due to Pseudomonas aeruginosa colonized in the intestinal tract. Twelve hours after the second administration of cyclophosphamide, the amount of enantiomer sodium salt of fosfomycin shown in Table 1 below was intraperitoneally administered, and further three times at 24-hour intervals (that is, The enantiomer sodium salt of fosfomycin was administered 4 times in total), and the life and death of the mice were observed over time. The test was performed with 10 animals per group.

The results were as shown in Table 1 below.

Table 1 Survival rate 8 days after the final administration of cyclophosphamide Fosfomycin enantiomer Na salt 10 mg / kg administration group 60% 2 mg / kg administration group 80% 0.4 mg / kg administration group 80 % Untreated control group 30% Further , a test was conducted by using fosfomycin sodium salt in the same manner as above, and the life and death of mice were observed over time. In this experiment, 16 animals per group were used.

The results were as shown in Table 2 below.

Table 2 Survival rate 5 days after the final administration of cyclophosphamide Fosfomycin Na salt 5 mg / kg administration group 55% untreated control group 15%

Example 2 The influence of the previously administered enantiomer sodium salt of fosfomycin on the pathological condition of sepsis was examined as follows. First, in the same manner as in Example 1, Pseudomonas aeruginosa was established in the intestinal tract, and the fosfomycin enantiomer was adjusted to 5, 1, 0.2 mg / mouse with physiological saline every 12 hours for 4 consecutive days. Each sodium salt was administered intraperitoneally. After that, cyclophosphamide 150-200mg / kg
Dose was given twice (the second dose was 3 days after the first dose). The subsequent life and death of the mouse was observed over time. The experiment was conducted with 10 animals per group.

The results were as shown in Table 3 below.

Table 3 Survival rate 3 days after the final administration of cyclophosphamide Fosfomycin enantiomer Na salt 5 mg / kg administration group 30% 1 mg / kg administration group 10% 0.2 mg / kg administration group 0 % Untreated control group 0%

Example 3 The effect of fosfomycin enantiomers on various functions of human monocytes was examined in vitro as follows. After separating mononuclear cells from human peripheral blood by the Ficoll / Hypaque method, the monocytes were collected by the plastic dish attachment method. Cytokine production was performed after culturing for 24 hours in the coexistence of LPS and fosfomycin enantiomer sodium salt,
The culture supernatant was collected and the production of TNF-α and IL-6 was measured by an ELISA kit. In addition to enantiomers of fosfomycin, Clarithromycin (CAM), Dexamethason
e (DX) was used as a control drug.

The results were as shown in Tables 4 to 6 below.

Table 4 Concentration of Na salt of enantiomer of fosfomycin TNF-α production amount IL-6 production amount (μg / ml) (ng / ml) (ng / ml) 0 2.9 39 1.6 2 0.044 8 1.8 50 40 1.3 55 Table 5 Concentration of CAM Production of TNF-α Production of IL-6 (μg / ml) (ng / ml) (ng / ml) 0 2.9 39 1.6 1.7 33 8 1.5 30 30 40 1.3 27 Table 6 Concentration of DX Production amount of TNF-α Production amount of IL-6 (μg / ml) (ng / ml) (ng / ml) ) 0 2.9 39 1.6 1.5 20 8 1.1 11 40 1.0 8 Regarding the cytokine production ability, the enantiomer of fosfomycin strongly suppressed the production ability of TNF-α, but IL-
No inhibitory effect was observed on the 6 productivity. This is due to the enantiomer of fosfomycin proinflammator
It suggests that the production inhibitory effect of y cytokine is selective.

Example 4 The antibody production-enhancing effect of fosfomycin enantiomers was divided into 5 groups of 6-week-old male C3H / HeN mice as follows.
Each group consisted of 8 animals, and each was administered with the enantiomer of fosfomycin at the dose and administration route shown in the table below. Five days after the administration, the mice were dislocated and killed, and their spleens were extracted. The extracted spleen was used as a spleen cell suspension and protein I plaque method (Eur.J.Immunol. 6: 588,1976) was used to express I in the spleen.
The number of gM antibody-producing cells was measured. That is, 25 μl of spleen suspension (total volume 5 ml), 25 μl of protein A
Coated sheep red blood cells (6 fold dilution), 25 μl guinea pig complement (6 fold dilution), 25 μl anti-mouse IgM antiserum (10 fold)
(Double dilution) is mixed with 0.5% agar (0.2 ml) at the same time, spread in a petri dish, put on a cover glass, and 37 ° C.
It was cultured for 4 hours. The number of hemolytic plaques (PFC) that appeared thereafter was measured, and the number of IgM antibody-producing cells (hemolytic plaque forming cells) per spleen was calculated.

The results were as shown in Table 7 below.

Table 7 Dose Dose Route IgMPFC ± S. E. FIG. / Spleen 60 mg / kg administration group Intraperitoneal 87,900 ± 2,192 *) 30 mg / kg administration group Intraperitoneal 102,250 ± 3,281 *) 60 mg / kg administration group Oral 116,850 ± 5,708 *) 30 mg / kg administration group Oral 147,900 ± 6,413 *) Control group 48,975 ± 2,924 **) In the table, *) shows a significant difference at a t-test of 5%, and **) is the average. Values ± standard error.

Example 5 Pseudomonas aeruginosa biofilm was formed in vitro by a known method (infection, 21, 161, 1991). That is, a piece of Teflon was put in a physiological saline solution containing rabbit plasma, and this was inoculated with Pseudomonas aeruginosa. When left at 37 ° C for 6 days, a Pseudomonas aeruginosa biofilm was formed on the surface of Teflon. To this, add 50μ of the enantiomer sodium salt of fosfomycin.
g / ml, ofloxacin or gentamicin 10 μg /
The amount of viable bacteria in the biofilm was measured by adding so that the amount became ml.

The results are shown in FIG.

Example 6 The action of fosfomycin enantiomers on the sL ^eX antigen on the surface of Streptococcus pyogenes was examined by the ELISA method as follows.

Streptococcus pyogene
s ) 19615 strain in brain heart infusion liquid medium (BHI, Difco, Detroit, USA) for 18 hours,
After culturing under the conditions of 37 ° C. and 5% CO ₂ , they were centrifugally washed with Dulbecco's phosphate buffered saline (PBS). The turbidity at 660 nm was 0.6 (2.
0 × 10 ⁸ cfu / ml), fosfomycin enantiomer sodium salt was added to 10 μg / ml, and the mixture was incubated for 1 hour at 37 ° C., 5% CO ₂ The test bacterial solution was used. ELISA of 50 μl of this bacterial solution
Plate (Corning Glass Works No.25801, New York,
(USA) and dispensed to dryness. Next, PBS containing 1.0% bovine serum albumin (1% BSA-
After adding 300 μl of PBS) to each well, the plate was left at room temperature for 60 minutes. Micro Plate Washer (Model
1550, Biorad Lab., Richmond, USA) and then washed with anti-sL ^eX monoclonal antibody (mAb SNH-3)
(200 μg / 100 μl, 100-fold diluted solution, Wako Pure Chemical Industries, Ltd.) was added and the mixture was reacted at room temperature for 120 minutes. After washing again, horseradish peroxidase labeled goat anti-mouse I
gM (Kirkegaard & Perry Labs., Inc., Gaithersburg,
USA) (2000 times dilution) 50 μl was added to each well,
The reaction was carried out at room temperature for 60 minutes. After washing again in the same manner, H
₂ O ₂ and 2,2'-azino-di-3-ethylbenzothiazoline- (6)-
sulfonate (ABTS, Kirkegaard & Perry Labs., Inc., Gaith
50 μl of a mixed solution (Ersburg, USA) was added and reacted at room temperature. ELISA intensity (Model 2550, Biorad Lab., Richmon
d, USA). As a control, the antigen, the second antibody, and the substrate mixed solution were reacted.

The results are shown in Table 8 below.

Table 8 sL ^ex expression intensity (OD 414 nm) Fosfomycin enantiomer-added group 0.58 + 0.036 Control group 0.86 + 0.021

Example 7 The action of fosfomycin enantiomers on the sL ^eX antigen on the surface of S. pyogenes biofilm was examined by the fluorescent antibody method as follows. Streptococcus
pyogenes ) 19615 strain for 18 hours in brain heart infusion liquid medium (BHI, Difco, USA)
After culturing under the conditions of 7 ° C. and 5% CO ₂ , they were centrifugally washed with Dulbecco's phosphate buffered saline (PBS). After adjusting to 2.0 × 10 ⁷ cfu / ml with fresh brain heart infusion liquid medium, cell desk (SUMILON) is immersed in this bacterial solution and incubated at 37 ° C for 4 days, and biofilm is deposited on the cell desk surface. Formed. Fosfomycin enantiomer sodium salt was added to the mixture at 10 μg / ml and allowed to act for 1 hour. After washing this cell desk, it was reacted with an anti-sL ^eX monoclonal antibody (mAb SNH-3) at room temperature for 1 hour. After washing again, goat anti-mouse IgM labeled with fluorescein isothiocyanate was added and reacted at room temperature for 1 hour. After washing again in the same manner, using the ACAS570 laser cytometer,
The expression intensity of sL ^eX was measured at 488 nm. As a control,
The cell desk without addition of the fosfomycin enantiomer was measured.

In the control, the fluorescence on the surface of the cell desk was observed on a relatively entire surface, and the intensity was very strong. On the other hand, the fluorescence intensity was weak and the expression of sL ^eX was weakened on the surface of the cell desk acted by the fosfomycin enantiomer.

Example 8 Minimum inhibitory concentration (MIC) of enantiomers of fosfomycin
Was measured according to a conventional method. As a control, the MIC of fosfomycin was also measured. The results are as shown in Table 9 below.

Table 9 MIC (μg / ml) strain FOM (+) FOM S. aureus 209P JC-1> 200 25 S. epidermidis ATCC14990> 200 3.13 E. coli NIHJ JC-2> 200 25 C. freundii GN346 / 16> 200 6.25 S.marcescens No.1> 200 3.13 P.aeruginosa GN10362> 200 In the table 3.13 , FOM (+): fosfomycin enantiomer, FOM: fosfomycin.

Acute Toxicity Test The enantiomer sodium salt of fosfomycin had an LD ₅₀ of 2000 mg / kg in mouse (male) intraperitoneal administration, and its toxicity was extremely low as was the case with fosfomycin sodium salt.

[Brief description of the drawings]

FIG. 1 shows the enantiomer sodium salt of fosfomycin,
It is a figure which shows the change of the viable cell number in a biofilm when ofloxacin or gentamicin is used together.

─────────────────────────────────────────────────── ─── Continuation of the front page (51) Int.Cl. ⁶ Identification number Reference number within the agency FI Technical display location // C07M 9:00 (72) Inventor Tetsuro Hara 760 Moshioka-cho, Kohoku-ku, Yokohama-shi, Kanagawa Akira Yoji Confectionery Co., Ltd., Pharmaceutical Research Laboratory (72) Inventor Koji Omoto, 760, Shimooka-cho, Kohoku-ku, Yokohama-shi, Kanagawa Meiji Confectionery Co., Ltd. (72) Inventor Hiroyuki Kobayashi Hamura, Nishitama-gun, Tokyo 1221-5 Machikawasaki (72) Yoichiro Miyake 3-4-20 Sumiyoshi, Tokushima City, Tokushima Prefecture (72) Inventor Katsuhiko Hirota 512-54 Enokise, Kawauchi Town, Tokushima City, Tokushima Prefecture (72) Inventor Ishi Shigeaki Saka 3-6 Yatayamacho, Yamatokoriyama City, Nara Prefecture

Claims (9)

[Claims] 1. A pharmaceutical composition comprising an enantiomer of fosfomycin or a salt thereof. 2. The pharmaceutical composition according to claim 1, which is an immunomodulator. 3. The pharmaceutical composition according to claim 1, which is an anti-inflammatory agent. 4. The pharmaceutical composition according to claim 1, which is a therapeutic or preventive agent for allergic diseases. 5. The pharmaceutical composition according to claim 1, which is a therapeutic agent for intractable infections. 6. The pharmaceutical composition according to claim 1, which is a biofilm formation inhibitor or a disrupting agent. 7. The pharmaceutical composition according to claim 1, further comprising an antibacterial agent. 8. An antibacterial agent is contained in an amount of 0.1 to 5 mol with respect to 1 mol of the enantiomer of fosfomycin or a salt thereof.
The pharmaceutical composition according to claim 7. 9. The biofilm is Staphylococcus aureus,
9. The method according to claim 6, which is formed by Staphylococcus epidermidis, streptococcus, Pseudomonas aeruginosa, Klebsiella, Escherichia coli, enterococci, Enterobacter, Citrobacter, or Serratia bacteria. The described pharmaceutical composition.