KR930004599B1 - Antiviral or antibacterial compound - Google Patents

Abstract

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Description

[Name of invention]

Antiviral or Antimicrobial Compositions

Detailed description of the invention

Technical Field of the Invention

The present invention relates to novel antiviral and antimicrobial compositions comprising polymerized enzymes with pharmaceutically acceptable carriers, and methods of use thereof.

Background of the Invention

The increase in the number of strains and viral diseases resistant to antibiotics has necessitated a new class of drugs for treating humans and animals. It is well known that many current therapeutic agents and agents used to treat patients suffering from various diseases use enzymes in monomeric form. Enzymes are catalytically active proteins that carry out almost all major life processes. Thus, many enzymes have been isolated individually or in specific combinations for their physicochemical, physiological or biological efficacy.

Among the various enzymes that have been shown to have particular therapeutic efficacy are Lysozyme and Ribonuclease. Lysozyme was discovered and known by Fleming in 1922. Only after 1950 did the enzyme function of lysozyme become known. Subsequently, the compound has been a strong physicochemical, physiological and clinical research topic, but the limits of its biological significance have not yet been determined. So far it has been observed that lysozyme has various therapeutic properties such as antiviral, antimicrobial, anti-inflammatory and antihistamine. Antimicrobial efficacy appears to be based on hydrolysis of beta-1-4-glucoside bonds between N-acetylomuraminic acid and N-acetyloglucosamine contained in bacterial walls .

The presence of lysozyme in phagocytes is also well known. Research in this field has shown that the intracellular lysozyme contained in lysozyme degrades predatory bacteria. In humans, lysozyme was observed to stimulate phagocytosis at physiological concentrations of 10-400 mg / ml.

Other properties of lysozyme are also known. Lysozyme, for example, lowers body temperature, which is thought to be a response to endogenic pyrogens liberated by toxins during the infection process. Lysozyme also appears to be involved in the immune process by stimulating the synthesis of gamma globulins, opsonins and other antibodies. It has also been suggested that lysozyme has a strong anti-inflammatory effect. Despite the known beneficial properties of this lysozyme, numerous studies and the production of pharmaceutical agents based on lysozyme, the use of this enzyme for therapeutic purposes is very limited.

Another group of enzymes that have been studied for various biological efficacy is ribonuclease (RIBONUCLEASE). It is a group of enzymes commonly found in many plants and animals as well as bacterial cells. Investigation into the properties and isolation of the enzyme was first attempted in 1955 by Schmidt & McDonald. In the discovery of this enzyme, it was found that the activity of ribonucleases in cancer tissues was significantly reduced. For example, leukemia-inducing viruses have been found to dramatically reduce the activity of acid ribonucleases in mice. It has also been found that in mice suffering from viral leukemia, the activity of acid ribonucleases in mitochondrial and microsome fractions obtained from spleen tissue is significantly reduced.

The studies cited above suggest that the decreased activity of ribonucleases is somewhat closely related to infection caused by viruses. Therefore, it suggests that ribonucleases may have some degree of antiviral activity. However, there are currently no known reports on the preparation of compositions using this enzyme as an antiviral agent.

One of the main reasons why this potentially beneficial enzyme has not yet been widely used is the cytotoxic effects observed in its monomeric form. Tests with cultured fibroblasts showed both lysozyme and ribonuclease monomers at very low concentrations. The potential beneficial effects of these enzymes could be exploited if effective methods could be developed to suppress this cytotoxicity. Thus, these enzymes are based on lysozyme, ribonucleases, or other similar enzymes that can be effectively used to treat viral or bacterial diseases or other detrimental conditions without the commonly observed cytotoxic effects when used in monomeric form. Development of the composition is required.

Overview of the Invention

According to the present invention, it has been found that antiviral or antimicrobial compositions which exhibit no cytotoxic effect and contain lysozyme, ribonucleases, or other enzymes as active ingredients can be prepared using dimeric forms of the enzyme. . By preparing a composition using lysozyme or ribonuclease as an active ingredient and a pharmaceutically acceptable carrier, numerous infectious diseases can be successfully treated with little cytotoxicity.

Detailed Description of the Preferred Embodiments

Antiviral and antimicrobial compositions of the present invention can be prepared by first obtaining lysozyme and ribonucleases in monomeric form. Lysozyme monomers (Catalog No. 28260) and ribonuclease monomers (Catalog No. 34388) used to prepare compounds prepared according to the present invention are from Serva Feine Biochemica, Gmbh und Cooperation, D69-000, Heidelberg Obtained. These enzyme monomers can be polymerized into dimers by convenient methods commonly used in the art. However, the enzyme polymerization reaction carried out and described by Carlsson et al. (See, eg, Biochemistry Journal 173: 723-737 (1978)) is particularly preferred. Also, the method described by Sorrentino et al. [Reference Example: Eur. J. Biochem. 124: 183-9 (1982). It has been observed that particularly useful ribonuclease dimers that can be used in the compositions of the invention are dimers prepared from pancreatic ribonuclease A isolated from animal pancreatic tissue.

Compositions containing lysozyme or ribonuclease dimers and pharmaceutically acceptable carriers as active ingredients are effective in treating many bacterial and viral diseases without undesirable cytotoxicity. A potentially detrimental action of these enzymes was injecting monomers and dimers of lysozyme and ribonucleases into Green Monkey Kidney (GMK) fibroblast media. Lysozyme monomers were found to be cytotoxic to fibroblasts after 24 hours at concentrations of 0.1 mg / ml and 1.0 mg / ml. After 3 days of culture, the cytotoxic effect of fibroblasts was also observed at 0.01 mg / ml concentration, affecting 50% of the cultured cells. After 5 days, 75% of cultured cells are affected by the cytotoxic activity of lysozyme monomers at 1.0 mg / ml and 0.1 mg / ml concentrations. In comparison, lysozyme dimer showed no cytotoxic effect at any concentration used in this test even after 7 days. These studies indicate that the dimeric form of lysozyme is approximately 100 times less toxic to GMK fibroblasts than the monomeric form.

Studies with ribonucleases have shown similar reductions in toxic effects in the dimeric form. In this study, it was found that ribonucleases at concentrations of 0.0001 mg / ml or less were cytotoxic to GMK fibroblasts in 5-day old media. After 7 days of culture, the ribonuclease monomer at a concentration of 0.01 mg / ml or more showed a cytotoxic effect of 100% of the cultured cells. In contrast, the ribonuclease dimer did not show any cytotoxic effect on GMK fibroblasts at all concentrations even after 7 days. That is, it was observed that the dimeric form of ribonucleases is about 1,000-10,000 times less toxic to fibroblasts than the monomeric form. These tests clearly show that the use of monomeric forms of enzymes such as lysozyme and ribonucleases can substantially eliminate the normally accompanying cytotoxic effects by using the enzyme in dimeric form.

With the disappearance of some of the cytotoxic effects, further studies have shown that the dimeric forms of lysozyme and ribonucleases are very useful for treating viral and bacterial infections. In tests conducted with fertilized eggs and Sendai virus strain, lysozyme dimers of various concentrations were injected into the amniotic membrane. In addition, 2 units of Sendai virus were injected into each fertilized egg. After incubation, amniotic fluid and urinary fluid were collected and compared from infected eggs and control eggs. The lysozyme dimer was found to be able to inhibit replication of Sendai virus cultured in 10-day-old fertilized eggs even at concentrations of 0.01 mg / ml or less. Similar tests using lysozyme and ribonuclease dimers showed that these dimeric enzymes had bacteriostatic effects against the Streptococcus bacteria.

Thus, compositions prepared from lysozyme or ribonuclease dimers according to the present invention can be used to treat a variety of viral and bacterial infections. The compositions can be prepared in a variety of forms, and administration of these enzyme-carrier compositions can be performed internally or externally to a person or animal suffering from a disease in need of treatment. In the case of internal diseases such as otitis, mastitis, gastrointestinal or vaginal tract infections, the dimer compositions of the present invention are suitably prepared to provide oral, intravenous, parenteral, suppository, or other ways in which dimer solutions can reach the site of infection. Can be administered. In the case of an external disease such as a viral or bacterial skin disease, an infected wound, herpes, or a sexually transmitted disease with trauma, the composition of the present invention may be topically administered to a patient in a variety of suitable forms.

Thus, the preferred form of the composition of the present invention is determined by the nature of the disease or infectious disease to be treated. For external treatment, the compositions can be administered in various forms such as ointments, lotions, solutions, oils and the like. If internal application is required, it can be used in a suitable form such as drops, tablets, solutions, capsules, dentrifices and the like. Thus, the specific form of the composition applied to the patient determines the nature of the pharmaceutically acceptable carrier used in the dimer enzyme composition. Among the many suitable carriers that can be used are hydrophilic bases, physiologically acceptable salt solutions, water, ointments, powders and the like.

Enzymatic therapy for viral or bacterial infections without a cytotoxic action is provided by administering to a human or animal patient an effective amount of the aforementioned dimeric composition. An effective amount means an amount necessary to provide antiviral or antimicrobial efficacy. The amount required for effective treatment may vary in each case depending on the nature of the disease to be treated and the type of dimer composition provided. In general, the compositions of the present invention are administered at a dosage of 0.01 to 50.0 mg / kg body weight, particularly in a range of 1.0 to 2.0 mg / kg body weight. For topical treatment, ointments prepared using about 4.0 mg dimer in about 200 ml of a solution of water, paraffin and propylene glycol were effective when applied 4-5 times a day. Doses in this range should be sufficient to treat many viral or bacterial diseases without the cytotoxic effects that may accompany treatment with enzyme monomers.

The following examples are presented merely to illustrate the invention and in any case are intended to limit the scope of the invention.

Example 1

A comparative study was conducted regarding the cytotoxic effects of monomers and dimers of lysozyme and pancreatic ribonuclease A on the medium of green monkey nephropathy (GMK) fibroblasts. Monomers and dimers at a concentration of 0.0001 to 1.0 mg / ml are applied to the fibroblast medium. The medium is then incubated for 7 days and then cytotoxic. The results of this test are shown in Tables 1 and 2.

As can be seen from Table 1, lysozyme monomers at concentrations of 0.1 mg / ml and 1.0 mg / ml were found to be cytotoxic to fibroblasts after 24 hours. After 3 days of culture, even at a concentration of 0.01 mg / ml, monomers have a cytotoxic effect on tricot cells, affecting 50% of cultured cells. After 5 days, 75% of cultured cells are affected by the cytotoxic activity of lysozyme monomer at 1.0 mg / ml and 0.1 mg / ml concentrations.

In contrast, the dimeric form of lysozyme shows no cytotoxic effect at any concentration used in the test even after 7 days of incubation. Thus, this study demonstrates that lysozyme dimer is about 100 times less toxic to GMK than lysozyme monomer.

As can be seen from Table 2, the monomers of pancreatic ribonuclease A were found to be cytotoxic to GMK fibroblasts in 5-day old media even at low concentrations of 0.0001 mg / ml. After 7 days of culture, at a concentration of 0.01 mg / ml or higher, the cytotoxic effect of pancreatic ribonuclease A is strong enough to remove 100% of cultured cells.

Similar to the case of lysozyme dimers, the dimeric form of pancreatic ribonuclease A shows no cytotoxic effect on GMK fibroblasts and at any concentration used in the test for 7 days. Thus it indicates that pancreatic ribonuclease A dimer is about 1,000 to 10,000 times less toxic to GMK fibroblasts than pancreatic ribonuclease A monomer.

TABLE 1

Cytotoxicity of Lysozyme Monomer and Lysozyme Dimer

This test is performed three times on medium of GMK (green monkey kidney) fibroblasts.

0-cytotoxicity = 0%

1-cytotoxicity = 25%

2-cytotoxicity = 50%

3-cytotoxicity = 75%

4-cytotoxicity = 100%

TABLE 2

Cytotoxicity of Pancreatic Ribonuclease A Monomers and Dimers

This test is performed three times on medium of GMK (green monkey kidney) fibroblasts.

0-cytotoxicity = 0%

1-cytotoxicity = 25%

2-cytotoxicity = 50%

3-cytotoxicity = 75%

4-cytotoxicity = 100%

Example 2

In terms of antiviral efficacy, lysozyme dimers were studied. In this experiment, lysozyme dimers are injected into 10 day old chicken embryos at concentrations of 10.0 mg / ml, 1.0 mg / ml, 0.1 mg / ml, 0.01 mg / ml and 0.001 mg / ml. Sendai virus strain (titre erythrocyte titer (titre) = 1: 128HA) is added to each concentration of dimer in the amount of 2 hemagglutination units. After administration of the lysozyme dimer and the virus, the eggs are incubated at 37 ° C. for 72 hours. After incubation, amniotic fluid and urinary fluid are collected from infected eggs and subjected to erythrocyte agglutination test by microscopic method using Takatsy set and hen blood cells. Repeat this experiment. The results obtained with this experimental trial are shown in Table 3. As can be seen in Table 3, lysozyme dimers inhibit the replication of Sendai virus cultured in 10 day old embryos even at a concentration of 0.01 mg / ml.

TABLE 3

Effect of Lysozyme Dimer on Sendai Virus Strains

+++-complete suppression, 1: 256

++-limit suppression, 1: 32

1. The test is performed on Sendai virus strains. Viral hemagglutination titer is 1: 128HA.

2. Provide a 10 day old fertilized egg as an experimental model.

3. Add the same amount of Sendai virus-2 hemagglutinin unit to each concentration of lysozyme dimer. At the same time, a control test for lysozyme dimers is started to confirm that the control is erythrocyte cohesive—the result is negative. Four hen eggs are infected with amnion using a virus of 22HA units and a series of lysozyme dimers. The eggs are incubated at 37 ° C. for 72 hours.

4. After incubation, the amniotic fluid and the ureter are collected from the infected eggs. The erythrocyte aggregation test is performed by erythrocyte agglutination microscopy using the Takachi test set and hen erythrocytes.

Example 3

The effects of lysozyme and dimeric forms of pancreatic ribonuclease A on bacteria are tested against several pathogenic strains collected from cows with mastitis. Three strains [Streptococcus agalactiae, S. sp. S. dysgalactiae, S. The effect of different concentrations of lysozyme dimers on S. liberis is shown in Table 4. The test results show that all three Streptococcus strains are sensitive to the activity of lysozyme dimers. It is affected by dimer activity at low concentrations of 1.25 mg / ml. This is most clearly seen in the case of Riveris. The bacteriostatic effect against other strains of Streptococcus was observed to begin at a concentration of about 10 mg / ml.

Table 5 shows the effects of ribonuclease A dimers and lysozyme dimers on pathogenic bacterial strains obtained from human patients. As can be seen from the table, pancreatic ribonuclease A dimers are particularly suitable for bacterial strains Pseudomonas aeruginosa, Escherichia coli, and Proteus vulgaris at concentrations ranging from about 5 to 10 mg / ml. vulgarus) most effective. Bacterial strains of Staphylococcus and Streptococcus have also been found to be sensitive to lysozyme dimers at concentrations of about 5 to 10 mg / ml. Susceptibility testing is generally carried out in accordance with accepted international principles recommended by the WHO.

TABLE 4

The minimum inhibitory concentration (MIC) (mg / ml) -bacterial strain of lysozyme dimer is cultured in a sample collected from cows suffering from mastitis.

-= No bacterial growth

+ = Bacterial growth

Susceptibility testing is generally carried out in accordance with accepted international principles recommended by the WHO.

TABLE 5

The minimum inhibitory concentration (MIC) (mg / ml) bacterial strain of pancreatic ribonuclease A dimer is cultured in a sample obtained from the patient.

-= Bacteria do not grow

+ = Bacteria grow

MSRA-Methicilin resistant Staphylococcus aureus. Strains are collected from the patient. Susceptibility testing is generally carried out in accordance with accepted international principles recommended by the WHO.

Example 4

The effect of lysozyme dimers and pancreatic ribonuclease A dimers on the proliferation of the K-562 erythrocyte cell system is determined by applying various concentrations of dimers to these cells. The test results are shown in Tables 6 and 7. In short, all concentrations of lysozyme dimer used in the experiments show potent cytopathogenic efficacy on K-562 cells. In addition, as can be seen in Table 7, pancreatic ribonuclease A dimers have similar efficacy in erythroid cell lines only at a concentration of 1.0 mg / ml.

TABLE 6

Effect of Lysozyme Dimer on the Proliferation of K-562 Red Blood Cell System. K-562 cells are used at a concentration of 10 ⁵ ml. After culturing for 24 hours with 5% CO ₂ flowing at 37 ° C, the effect is evaluated.

TABLE 7

Effect of Pancreatic Ribonuclease A Dimer on Proliferation of K-562 Red Blood Cell System. K-562 cells are used at a concentration of 10 ⁵ ml. After culturing for 24 hours with 5% CO ₂ flowing at 37 ° C, the effect is evaluated.

Example 5

The effect of lysozyme dimer on the purulent otitis media in dogs is tested. The study was conducted using 19 different breeds of dogs with various forms of the disease. The disease is characterized by an inflammatory process that lasts about 7-14 days on average, but one dog lasted nine months. Seven dogs were tested for purulent discharge from inflamed ears to confirm the presence of bacterial strains in them prior to treatment. These culture tests include Staphilococcus germs, blue pusbacillis, Pseudomonas aeruginosa, Coccidia species, and various bacilli species. Shows.

Painful dogs use various expressions of pain, such as shaking their heads or trying to touch an infected ear with their feet. These dogs generally have a decreased appetite and a high body temperature (39.2-41.2 ° C). Eighteen of the dogs in this study had never received any medication before. Only one dog who had been infected for nine months had been given antibiotics, but they were not effective in treating the condition.

The dogs are treated with a composition of the present invention consisting of a solution of 20 mg of lysozyme dimer and 25 ml of physiological saline. The composition is applied as a drop, with 10 drops in the inflamed ear 4 to 5 times a day. One day after treatment, noticeable progress was made. The temperature dropped, and the dog noticeably felt more comfortable and found appetite. Between 3 and 6 days of treatment, the symptoms of purulent inflammation disappeared completely. In the case of a dog that had not been successfully treated with antibiotics for nine months before, the disease was completely eliminated after 10 days. Thus lysozyme dimer has been shown to be effective in treating purulent otitis media in dogs.

Example 6

The effect of lysozyme dimers in cows suffering from mastitis is tested. The study uses six cows with mastitis. These cows show symptoms of diseases such as body temperature and loss of appetite above 40.5. All six cows started treatment on the second day of the disease. Milk samples are collected for bacterial testing prior to injecting the composition. The medium was observed to contain microorganisms such as Staphylococcus and Streptococcus agalactiae.

Each cow is infused twice daily with 40 mg of lysozyme dimer in 50 ml physiological saline solution. Only 24 hours later, the body temperature was found to be normal and the appetite returned. After 3 days all cows treated did not show any symptoms of the disease. So the treatment lasted up to 4 days. Subsequently, milk was examined and the pathogens found before treatment disappeared. In addition, no changes were found in milk indicating subclinical mastitis. No cows were found to lose milk production in any of the treated cows. Twenty four hours after lysozyme dimer treatment, no interferences were found in the cow's milk. Therefore, the rapid disappearance of the disease symptom as well as the complete maintenance of the milking amount shows that the lysozyme dimer composition of the present invention can be successfully used for the treatment of bovine mastitis. This treatment will be particularly economically important in the dairy industry, which costs about $ 5.4 billion per year to prevent mammary infections by Staphylococcus and Streptococcus bacterial strains.

Example 7

Canine parvovirus (CPV) infection is treated by oral administration of lysozyme dimer. Of 27 dogs, three to six years old, of different species and weight, veterinarians found a typical combination of parvovirus infections. In addition to dehydration and desensitization, all animals in this test show symptoms of high body temperature (40-41.6 ° C), frequent seizures of severe vomiting, and a number of characteristic malodorous diarrhea. The animals also felt that they were suffering severely. Treatment begins on average between three and five days of infection, depending on how quickly the owner of the animal brings the animal to the livestock hospital. Infected dogs are given 1-2 mg of lysozyme dimer per kilogram of body weight twice daily. Drinkable animals are given lysozyme dimers in drinking water. Non-drinking animals are given medication via a probe with a physiological salt solution.

Of the 27 dogs under treatment, 25 completely recovered their physical health after 3-5 days of treatment. Typically, we observed a significant decrease in the number of lingual and vomiting seizures on the first day, and after two days of treatment, most of the dogs had completely disappeared. In some dogs, these symptoms disappeared after a single dose. No side effects from administration of the lysozyme dimer composition were found in any animal. This clinical trial shows that dogs suffering from parvovirus infection can successfully use the lysozyme dimer composition of the present invention.

Example 8

The effect of the lysozyme dimer composition of the present invention on human skin diseases is tested. The trial was conducted on dozens of patients aged 15 to 35 who suffered from a variety of skin diseases that had traditionally been treated but could not be successfully treated by conventional methods. Diseases belonging to this group include:

1.Forunculosis chronica-2 patient

2. Patients with Cycosis barbae-1

3. Impetigo contagiosa-11 patients

Acne vulgaris-22 patients

5. Patients with Resacea-6

6. Varicose ulcer-12 Patients

Several patients in this group were tested for bacterial culture prior to treatment. In most cases, Staphylococcus Aureus was cultured from the collected material. Treatment consists of administering an ointment containing 4 mg of lysozyme dimer four times daily. Particularly preferred constituents of the ointment are as follows.

Lysozyme Dimer 4.0mg

Acetyl stearoyloxy alcohol 25.0mg

10.0mg paraffin liquid

Span 60 5.0 mg

Tween 60 8.0 mg

Propylene Glycol 10.0mg

Asseptina M 0.3 mg

Asseptina P 0.16 mg

Distilled water is added to make a total of 200 ml.

The various skin conditions of all patients disappeared within 10-12 days, and some patients were observed to eliminate the disease after 3 days. For patients with chronic Forunculosis, treatment continues for 4 to 5 weeks, and for patients with Varicose ulcer, how severe the skin condition is, how long before treatment It usually takes two to twelve weeks to recover, depending on whether you've got it. The results obtained in this study indicate that lysozyme dimers can be used to successfully treat many skin diseases.

Example 9

The lysozyme dimer composition of the present invention is used to treat various infectious diseases of the genitals. Nine female patients aged 25 to 49 years old, namely seven patients with chronic colitis, one patient with Douglas' abscess and one patient with Bartholinitis Treatment at. Patients with chronic vaginitis were given vaginally administered suppositories containing 10 mg of lysozyme dimer in a hydrophilic base 2 ccm. Suppositories were administered twice daily for 7 days. The disappearance of reproductive inflammation was observed in all patients. In addition, leucorrhea and other symptoms disappeared. Patients with Douglas cyst abscess were given 20 mg of lysozyme dimer in 5 ml of 0.95% sodium chloride (NaCl) solution twice daily for four days. This solution was administered directly to the Douglas River. Prior to administration of lysozyme dimer, purulent contents were taken from the Douglas River. Its culture indicates the presence of Streptococcus haemolyticus and Bacterium coli. Within 24 hours of the first dose of lysozyme dimer, the patient's abnormal high temperature had returned to normal. In addition, there were no pain symptoms during this period. No pus was found in the Douglas River 4 days after the second injection of the lysozyme dimer composition. In this case, the disease recurred 3 weeks later, but the progression of disease could be inhibited by administering twice a unit dose of lysozyme dimer to the Douglas River.

In patients with bartholinitis, the purulent contents are removed and treated with immediate administration of 20 mg of lysozyme dimer in 1 ccm of 0.9% sodium chloride in a suppurated gland. After 4 days the patient was completely cured. Observation was continued for four months, but there was no recurrence of the disease. The trial shows that lysozyme dimers have a very beneficial therapeutic effect on women with certain infectious diseases in the reproductive area. In addition, lysozyme dimers can be immediately administered to cavities containing pus to treat localized tumors.

Example 10

The effect of lysozyme dimer solution on infected wounds was studied. The study group consisted of 4 patients with postoperative infection wounds: 2 female patients after open surgery, 1 female patient who underwent toe amputation due to necrosis during diabetic angiopathy, and a burger Patients who have had lower extremity cuts by Burger's disease, in all cases, wet and wash 20 mg lysozyme dimer solution in 5 ml of 0.9% NaCl 4 times daily. After open surgery, patients with purulent wounds were fully treated 4 and 6 days later. For other patients, treatment efficacy was achieved after 21 days and 5 months, respectively. This test demonstrates that lysozyme dimer can be used as a treatment to treat postoperative infection wounds without side effects.

Example 11

Clinical observations were performed on patients suffering from Herpes genitalis treated with ribonuclease A dimer compositions. The study group consists of five female patients aged 23 to 36 years. Four of them had the disease for the first time, and one woman had it for the third time. All patients were in the foam phase, a symptom that usually occurs between 3-5 days of disease, and complained of severe pain in the perineal region, especially during urination. In all patients, swelling and labial infections were found, as well as many blister-filled blisters on the mucous labia and the femoral and anal cortex. Inguinal lymph nodes in all patients were enlarged and painful.

Ointments containing ribonuclease A dimers are applied over the affected area of blisters or mucosa 4-5 times a day. Applied ointments include the following ingredients.

Ribonuclease Dimer 4.0mg

Acetyl stearoyloxy alcohol 25.0mg

10.0mg paraffin liquid

Span 60 5.0 mg

Tween 60 8.0 mg

10.0 mg propylene glycol

Asseptina M 0.3 mg

Asseptina P 0.16 mg

Distilled water is added to make 200 ml.

Patients who received dimer treatment substantially reduced pain after a few minutes and at least an hour after first applying the ointment, and pain completely disappeared after 10 to 20 hours. The results of the physical examination show that in four patients the pathological changes have completely disappeared after three days of treatment. In other patients the pathology disappeared completely after 5 days. None of these patients showed any new blisters after applying an ointment containing pancreatic ribonuclease A dimer. The study shows that the ointment containing the dimer of the present invention can be successfully used for the treatment of Herpes genitalis.

Example 12

More than 100 patients of various ages were treated with herpes zoster treatment using the compositions of the present invention. All of these patients had a number of blisters in the upper lip, filled with redness and turbidity. All patients complained of pain in the affected area, and there was tissue tension. Treatment consisted of topical application of an ointment containing pancreatic ribonuclease A dimer 4-5 times a day.

All patients treated without exception said the pain and tension in the tissues disappeared quickly. As observed in patients with genital herpes, the pain completely disappeared within the next few hours. Physical examination shows that the swelling and blisters disappeared within two to three days. In each case, it takes 5 days to completely treat in a completely clean state. In patients who started treatment on the first day of the disease, it was further observed that skin symptoms such as swelling irritation and papules disappeared completely after 24 hours. Patients suffering from frequent relapses were observed to have prolonged time to relapse and aggravated recurrence symptoms. In no case were there any side effects.

Example 13

A clinical study was conducted in six patients with herpes zoster treated with the dimer composition of the present invention. For five patients, the disease developed in a typical manner, but the sixth patient developed a unique complication, discussed below. These patients are treated with an ointment containing pancreatic ribonuclease A dimer, the treatment beginning on day 3 or 4 of the disease. Ointment is applied 4 to 5 times a day.

All patients completely lost pain in the first 24 to 48 hours. After 3-4 days the swelling was observed to dry out, ending the treatment after this period. Within the next few days, the skin condition was completely cured. None of the patients persisted until after the first 24 to 48 hours, and then the disease did not recur in three patients observed over a year. One patient mentioned above had abnormal clinical development of the disease. A 42-year-old woman has been treated for lung cancer using cobalt therapy that severely damaged her immune system. Contrary to the usual symptoms of shingles, blisters were found throughout the body. Treatment with the ribonuclease dimer of the present invention has been very successful in patients with such compromised immunity, and after several days in this woman the blisters have completely disappeared. Successful cases seen in this patient and other patients in the group demonstrate that pancreatic ribonuclease A dimers are particularly effective against herpes zoster caused by Varicella viruses.

Claims (29)

An antiviral or antimicrobial composition comprising as an active ingredient a dimer selected from a lysozyme dimer and a ribonuclease dimer and a pharmaceutically acceptable carrier. The composition of claim 1 wherein the dimer is a lysozyme dimer. The composition of claim 1 wherein the dimer is a ribonuclease dimer. 4. The composition of claim 3, wherein the dimer is pancreatic ribonuclease A dimer. The composition of claim 1, wherein the pharmaceutically acceptable carrier is a hydrophilic base. The composition of claim 1 wherein the pharmaceutically acceptable carrier is a 0.5-1.5% NaCl solution. The composition of claim 1 wherein the pharmaceutically acceptable carrier is water. 8. The composition of claim 7, wherein the concentration of dimer in water is about 0.02 mg / ml. The composition of claim 1 wherein the pharmaceutically acceptable carrier is an ointment. The composition of claim 9 wherein the ointment comprises water, paraffin and propylene glycol. The composition of claim 9 wherein the concentration of dimers in an ointment is about 0.02 mg / ml. The composition of claim 1, wherein the pharmaceutically acceptable carrier is a physiologically acceptable salt solution. The composition of claim 12, wherein the concentration of dimer in the salt solution is about 0.8 mg / ml. The composition of claim 6, wherein the concentration of dimer in NaCl solution is about 4 mg / ml. The composition of claim 5, wherein the concentration of dimer is about 10 mg per 2 ccm of hydrophilic base. The composition of claim 1 wherein the concentration of the dimer is in the range of 0.001 mg / ml to 20.0 mg / ml. The composition of claim 1, which is administered at a dosage of about 0.01 to about 50.0 mg per kg of body weight. The composition of claim 1 which is administered orally. The composition of claim 1, which is administered intravenously. The composition for treating viral or bacterial skin diseases according to claim 1, wherein the composition is administered locally to a human or animal patient. The composition for treating herpes according to claim 1, which is applied locally to the infected site. The composition of claim 1, wherein the composition is treated locally by infection. The composition for treating vaginal infections according to claim 1, wherein the composition is applied locally to the infected area. The composition of claim 23, wherein the infection is vaginitis. The composition of claim 23, wherein the composition is treated with suppositories to treat vaginal infections. The composition for treating otitis media according to claim 1, which is applied to an infected site. 27. The composition of claim 26, wherein said composition is applied to the ear of an infected patient in the form of a drop to treat otitis. The composition for treating mastitis according to claim 1, which is administered to a human or animal patient. 29. The composition of claim 28 for treating mastitis by application to the site of infection by syringe injection.