Pharmaceutical composition containing lysine and enzymes: lysozyme, deoxyribonuclease and/or peroxidase for external treatment and prevention of infections caused by type 1, 2 herpes viruses, and bacterial complications caused by herpetic infection

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to the pharmaceutical industry and represents a composition for the external treatment and prevention of infections caused by the type 1, 2 herpes virus and bacterial complications caused by the herpetic infection, containing lysozyme, peroxidase, povyargol as active ingredients, escin and glycyrrhizinic acid or its salts as anti-inflammatory ingredients, liposomes on the basis of high-active hydrated lecithin in a combination with cholesterol as carriers and pharmaceutically acceptable carriers and excipients, with the ingredients of the composition being taken in certain proportions, wt %.

EFFECT: invention provides extending the range of products for treating and preventing the infections caused by type 1, 2 herpes virus and bacterial complications caused by the herpetic infection.

4 ex, 1 tbl

 

The present invention relates to medicine, namely to pharmaceutical compositions containing lysine and enzymes: lysozyme, desoksiribonukleaza and/or peroxidase, which can be used in medicine for external treatment and prevention of infections caused by herpes virus type 1, 2, and bacterial complications caused by herpes infection.

Known medicines for local use, used for the treatment and prophylaxis of herpes infections, containing antiviral components that are not enzymes, such as cream Zovirax™ (acyclovir); spray Epigenome sex™ (glycyrrhizin acid); ointment: alpizarin, Halpin, flakozid containing plant extracts.

From RU 2379312 renowned pharmaceutical composition comprising a compound based lysine, are used in the treatment and prevention of HIV infection.

Known food additive "Lisit" on the basis of lysine, providing immunostimulating effect and used in prevention and complex treatment of chronic viral infections, especially when urogenital viral infections and cold sores (Herpes simplex and Herpes zoster), (http://www.apollux.com/product-01.htm).

Known lysine (2,6-diaminohexanoic acid) - aliphatic amino acid with strong Foundation properties. Lysine is an essential amino acid, which is part of PR is chicosci any protein, necessary for growth, tissue repair, production of antibodies, hormones, enzymes, albumins. This amino acid has antiviral activity, especially against viruses that cause herpes and acute respiratory infections (retrieved from the Internet: http://ru.wikipedia.org/wiki/%CB%E8%E7%E8%ED).

Known L-lysine ascent, decreasing the activity of lysosomal hydrolases, which prevents the degradation of mucopolysaccharides in the walls of capillaries and connective tissue that surrounds them, and thus normalizes increased vascular tissue permeability and has antiexudative (anti-edema and analgesic action, (http://www.vidal.ru/poisk_preparatov/l-lysine-aescinat.htm).

Known drug Intesol 40 in the form of solution for infusion, containing lysine and used for stimulation of anabolic processes parenteral nutrition. (http://www.vidal.ru/poisk_preparatov/infesol-40.htm).

From RU 2010151556 known pharmaceutical composition containing lysozyme, peroxidase, poviargol and liposomes for the treatment and prophylaxis of viral infections caused by RNA-and DNA-containing viruses, such as influenza, herpes, herpes zoster, papilloma of the person, adenovirus and others, as well as bacterial infections caused by gram-positive and gram-negative microorganisms.

From RU 2006114277 known cosmetic composition comprising a Deux is xeribolus and/or ribonuclease for the prevention of herpes infection.

Known cosmetic products for local use, used for the prevention of herpes containing antiviral components that are not enzymes, such as extracts of medicinal plants and germaniyorganicescoe connection (Lipstick preventive anti-herpes "Antiherpes" (P16 9158 915854 71.100.70) 292472. THE 9124-001-00358210-96), glycyrrhizin acid ointment "Germinal" ointment; ban. 20 ml; No. 77.01.12.915.P. 02970.02.00 from the Institute for atherosclerosis research the Academy of natural Sciences (Russia), INAT-PHARMA (Russia) (http://www.biomedservice.ru/publish/pub16.htm).

Well-known cosmetic enzyme-containing composition, in which the enzyme is selected from the group of proteases, including trypsin, chymotrypsin, bromelain, papain and fitsin, and lysozyme, elastase, α-lipase and α-amylase, which is used for the treatment of acne, skin care and cleansing of the skin (DE 4305460). Described composition does not contain enzymes, nucleases, with direct antiviral activity, and, therefore, cannot be used to prevent viral infections.

Known toothpaste with the enzyme-containing composition, in which the enzyme is selected from the group of proteases, including papain (Whitening toothpaste "Rembrandt" (USA), as well as pasta, containing lysozyme, glucose oxidase, lactoperoxidase and lactoferrin (toothpaste Biotene (Finland), which is used for the treatment of xerostomia.

The world is Yong drug "Lysozyme", produced in the form of a hermetically sealed vials of 0.05, 0.1 and 0.15 g of providing bacteriolytic action and has the ability to stimulate nonspecific reactivity of the organism.

A known film-forming composition biodegrading antiseptic "ARGALL" containing poviargol and used to treat wounds and burns.

Well-known pharmaceutical drug "Desoksiribonukleaza", produced in the form of lyophilized powder in a hermetically sealed vials and ampoules 0,005; 0,01; 0.025 and 0.05, This drug is prescribed for herpetic keratitis, adenoviral conjunctivitis, to reduce viscosity and improve evacuation of sputum and pus when multiple bronchiectasis and is used topically in the form of a 0.2% solution of isotonic sodium chloride solution.

Well-known pharmaceutical drug "Ribonuclease", produced in the form of lyophilized powder in a hermetically sealed vials of 0.01, This drug is prescribed for diseases of the respiratory tract with detachable hard sputum, periodontitis, gingivitis, festering wounds, trophic ulcers, tick-borne encephalitis, thrombophlebitis, the local application sprinkled the wound surface in the amount of 0.025 to 0.05,

Pharmaceutical compositions containing lysine and enzymes: lysozyme, desoksiribonukleaza and/or peroxidase for the outer the treatment and prevention of infections caused by the herpes virus type 1, 2, and bacterial complications caused by herpes infection is not described.

Thus, the present invention is to provide pharmaceutical composition comprising lysine and enzymes: lysozyme, desoksiribonukleaza and/or peroxidase, which can be used for external treatment and prevention of infections caused by herpes virus type 1, 2, and bacterial complications caused by herpes infection.

According to the present invention proposed a pharmaceutical composition for topical use containing as an active ingredient is from 0.01 to 5.00 wt.% lysine, 0.01 to 5.00 wt.% lysozyme, 0.01 to 5.00 wt.% deoxyribonuclease and/or 0.01 to 5.00 wt.% peroxidase, as the carrier is 0.01 to 20.0 wt.% liposomes or β-cyclodextrins, or fullerenes, and pharmaceutically acceptable polymeric carriers or excipients.

As a preferred embodiment of the claimed invention proposed pharmaceutical composition specified above, optionally containing one or more than one anti-inflammatory ingredient.

Most preferred is a pharmaceutical composition for topical treatment and prevention of infections caused by herpes virus type 1, 2, and bacterial complications caused by herpes infection, with the holding lysine, lysozyme, desoksiribonukleaza, peroxidase, poviargol as active ingredients, as anti-inflammatory ingredients extract of thuja, escin and glycyrrhizinic acid or its salts, as carriers - liposomes on the basis of the high level of hydrogenated lecithins and pharmaceutically acceptable carriers and excipients, in the following ratio, wt.%:

lysinefrom 0.01 to 5.00
desoksiribonukleazafrom 0.01 to 5.00
lysozymefrom 0.01 to 5.00
peroxidasefrom 0.01 to 5.00
poviargolfrom 0.01 to 5.00

liposome-based highly active

hydrogenated products of lecithinsfrom 0.01 to 5.00
glycyrrhizin acid or its saltsfrom 0.01 to 5.00
extract of thujafrom 0.01 to 5.00
escinfrom 0.01 to 5.00
carbomerfrom 0.01 to 5.00
emulsifierfrom 0.01 to 5.00
preservative0,01-1,00
the pH regulator0,01-1,00
water demineralizedrest

The active ingredient of the claimed pharmaceutical compositions are lysine, enzymes: desoksiribonukleaza, lysozyme, peroxidase, poviargol, extract arborvitae, glycyrrhizin acid and escin. Lysozyme catalyzes the hydrolysis-1,4 - linkages between N-acetylmuramic acid and 2-acetamido-2-deoxy-D-glucose (N-acetylglucosamine) contained in mucopolysaccharides, mucopeptide and chitin in the cell walls of bacteria, fungi and protozoa, has a pronounced antimicrobial (destroys the cell wall of most gram-positive and some gram-negative bacteria), mycocide (delays the development of microscopic fungi, and wound-healing effect. Lysine, as antagonista arginine, inhibits the synthesis of viral proteins. Desoksiribonukleaza (DNA-Aza) has the ability to hydrolyze the DNA of the herpes viruses, adenovirus and other DNA-containing viruses without damaging the DNA of cells of the microorganism. Peroxidase has the ability to oxidize glycoproteins, supercaps the s viruses and thus, to prevent adsorption of viruses on the surface of the membranes of infected cells, and promotes recycling of infected cells by macrophages. Poviargol thanks immobilized silver ions prevents the development of viral and secondary bacterial infection. Extract of thuja has a pronounced anti-inflammatory and antimicrobial action. For the preparation of compositions used substance extract of thuja, poviargol and lysine content of the basic substance is not less than 97% and the enzyme substance: "Desoksiribonukleaza", "Lysozyme", "Peroxidase".

The desoksiribonukleaza, lysozyme and peroxidase in the form of these drugs is not used for the treatment and prevention of viral and bacterial infections due to their difficulty suction and low level of consumer properties, as well as lack of stability at a moisture content of over 10% and in solution, with the loss of enzyme activity during the day.

Unexpectedly found that lysozyme, desoksiribonukleaza, peroxidase and liposomes can be incorporated into a stable pharmaceutical composition for topical application to preserve the activity of these enzymes in a long time, despite the fact that this composition contains a significant amount of water (up to 90%).

The pharmaceutical composition the present invention are generally made by using common methods, using solid or liquid cosmetically acceptable carriers or excipients selected from emulsifiers, dispersing agents, preservatives, flavoring agents, pH regulators, polymeric carriers and other excipients, which are suitable for preparation of compositions for local application. The pharmaceutical composition of the present invention can be prepared in the form of a gel, cream, cream-gel, cream, foam, jelly, paste, ointment and the like.

Additional advantages of the specified composition is the ability to penetrate biological barriers and, consequently, rapid absorption, ease of use, and ease of dispensing.

The rapid penetration due to the binding of enzymes with liposomes based on the high level of hydrogenated lecithins, which are used in cosmetology in pharmaceuticals, for example, AssawinTMgel, anti-aging drugs of the skin, for skin care preparations before and after sun preparations for cosmetics ampoule.

In addition, the active ingredients can be used in combination with one or more than one anti-inflammatory ingredient, such as an extract of thuja, escin and glycyrrhizin acid, providing an additional advantage of the claimed composition.

Declared formats viteska the composition has a stability in relation to the activity of enzymes included in its composition during long-term storage, which is achieved by immobilization of enzymes on polymeric carriers contained in the specified composition. As mentioned polymeric carriers can be used any polymer capable of contacting the enzyme with the formation of complexes of the enzyme-carrier. Examples of such polymeric carriers are polyacrylates, polyglutamate, polyvinylpyrrolidone, polietilenglikolya ether chitosan, gum and sodium carboxymethylcellulose.

The activity of the claimed compositions is due to the activity of its constituent enzymes. Discovered that the pharmaceutical composition for topical application containing lysozyme from 0.01 to 5.00 wt.% and peroxidase from 0.01 to 5.00 wt.%, is stable in relation to the activity of enzymes in its long-term storage, while the activity of these enzymes, prepared in the form of a solution, decreases rapidly. Therefore, the claimed composition can be used for effective prevention of viral infections caused by herpes viruses types 1, 2, and bacterial complications, such as coccal infections and other

The present invention is illustrated by the following examples.

EXAMPLE 1

Pharmaceutical composition in the form of a gel-based lysozyme, peroxidase, powergel and liposomes for topical treatment is the prevention of infections caused by the herpes virus type 1, 2, and bacterial complications caused by herpes infection, has the following composition:

lysozyme0,01
peroxidase0,01
poviargol0,01
liposomes0,01
glycyrrhizin acid or its salts0,01
escin0,01
carbomer0,01
emulsifier0,01
preservative0,01
the pH regulator0,01
water demineralized99,90

The above composition is prepared by the following method. In demineralized water dissolve lysozyme, peroxidase, poviargol, glycyrrhizinate acid or its salts, escin, carbomer, a pH regulator. In the resulting gel consistently contribute emulsifier and preservative. The resulting gel is homogenized and vacuum.

<> EXAMPLE 2

Pharmaceutical composition in the form of a gel-based lysozyme, peroxidase, powergel and liposomes for topical treatment and prevention of infections caused by herpes virus type 1, 2, and bacterial complications caused by herpes infection, has the following composition:

lysozyme5,00
peroxidase5,00
poviargol5,00
liposomes5,00
glycyrrhizin acid or its salts5,00
escin5,00
carbomer5,00
emulsifier5,00
preservative1,00
the pH regulator1,00
water demineralized58,00

The above composition is prepared as in EXAMPLE 1.

EXAMPLE 3

Evaluation of the stability of enzymes in the compositions for local application

Since the main is the lack of enzymes is their instability during long-term storage of ready medicinal forms the basis for the expiration date was the comparative assessment of the preservation of enzyme activity during storage in solution and in the form of a composition for topical use (for example, gel)

Activity of lysozyme and peroxidase was determined by the following methods.

1) determining the activity of lysozyme was carried out according to the method of O. Bukharin Century, based based on measurement of optical density by adding enzyme to the bacterial suspension. In a test tube filled with 0.4 ml of phosphate buffer (pH 6.2) and 0.1 ml of the investigated solution and 2 ml of a standardized suspension of micrococci. The mixture is incubated for 30 minutes at 37°, then measure its optical density on FEC-M at λ=540 nm.

Per unit activity of lysozyme take the least amount of enzyme which under the influence of it on the substrate in terms of experience analyzes this amount of culture suspension Mycrococcus lysodeikticus, which leads to an increase in optical density at a wavelength of 540 nm by one unit.

2) Determination of the activity of peroxidase was carried out according to the method ABTS. The method is based on determination of oxidized ABTS formed as a result of oxidation by hydrogen peroxide in the presence of peroxidase. Per unit activity of peroxidase take the least amount of enzyme that triggers when exposed to its substrate ABTS for 10 minutes at 37°C conque is such a large number of oxidized ABTS, which leads to an increase in optical density at a wavelength of 414 nm in a single unit.

3) the Activity of dnaase was determined by the following method.

The determination of the activity of dnaase was carried out according to the method of Konitsa. Per unit activity of dnaase take the least amount of enzyme which under the influence of it on the substrate in terms of experience frees this number colorstorm products, which leads to an increase in optical density at a wavelength of 260 nm by one unit. In the control and two experimental tubes contribute 0.5 ml of substrate (DNA) and 0.3 ml of 0.2 M Tris-buffer (pH 7). In the experimental pre-warmed tubes contribute 0.2 ml of an enzyme solution. Sample thermostatic for 30 minutes at a temperature of 37.5°C. Then the samples are added 1 ml) cooled in an ice bath 1 n solution of perchloric acid (HClO4), in the control and 0.2 ml of an enzyme solution. The tubes are shaken and incubated in an ice bath 16-25 minutes. The formed precipitation was separated by centrifugation at 4000 rpm for 10 minutes. From the supernatant layer selected 0.5 ml of solution, add 2.5 ml of water and measure the optical density at λ=260 nm.

The results are shown in the following table.

Table
Researched about azzy Main ingredientsThe storage stability
123
Gel No. 1According to Example 1The activity of 95% for lysozyme and 80% for peroxidase and deoxyribonuclease within 1 year of observation
Gel No. 2According to Example 2The activity of 96% for lysozyme, 78% for peroxidase, 82% for desak (Serioulsy within 1 year of observation
The solution of the native enzymeActivity 40% for lysozyme, 30% for peroxidase and 10% deoxyribonuclease for 1 day

As can be seen from the table, the introduction of the enzymes in the composition for local application allows to obtain a stable pharmaceutical composition for topical application in the prevention of viral infections caused by herpes viruses types 1, 2. Despite the fact that with the introduction of the enzymes in the composition of these songs is a minor loss of enzyme activity, with further storage for 1 year, enzymes do not alter its activity due to the fixation of labile the structure of the enzyme liposomes and polymer carrier.

EXAMPLE 4

Evaluation of antiviral and antibacterial activity of the composition

To evaluate antiviral and antibacterial activity, a study was conducted of the effectiveness of the composition of EXAMPLE 1 in comparison with the antiviral drug Acyclovir cream. The study involved 39 patients aged 18 to 45 years, with clinical manifestations of herpes infection. On the floor, the group was divided as follows: male - 11 female - 28. Disease duration ranged from 2 years to 12 years. While the vast majority of patients reported mild and moderate for herpes infection - 2-3 times a year 14, 4-6 times per year - at 22 and 6-8 times a year in 3 patients. Among women in the observed group of Orofacial manifestations were observed in 18, genital localization in 10 man. In the group of men in 7 patients registered with genital herpes, and 4 observed was noted defeat in the face zone. With 7 men and 2 women - 5) was observed joining manifestations of herpes conditionally pathogenic bacterial flora with the development of complications such as strepto-stphiladelphia.

Most patients were registered small area of damage, ranging from 0.5 to 1.0 cm in 29 cases, the centers of area of 1.5 cm in 9 observed.

In the comparison group find the of n 11 patients with manifestations of herpetic infection receiving outdoor therapy cream Acyclovir". On the floor of the comparison group were divided as follows - men - 7 women, 4 men. Manifestations of herpes all men were in the genital location, 1 women herpes also had genital localization, and 3 women were observed Orofacial herpes. By severity 8 patients in the control group had a mild case, 3 - moderate severity.

Before treatment and 4 weeks after end of treatment all patients performed the following studies:

Survey and inspection

- Study of the material from the lesion using the polymerase chain reaction for detection of herpes simplex virus human

- Examination of scrapings from the urogenital tract to detect chlamydia, Miko and ureaplasmas (with lesions of the urogenital tract)

Testing serum for syphilis; research on HIV after conducting a pre-test interview

- The clinical analysis of blood

All patients after obtaining informed consent for external treatment was assigned the composition of EXAMPLE 1, 4-6 applications per day, three patients with severe were simultaneously conducted systemic therapy drugs acyclic nucleoside 5 days. In the comparison group patients received external is arapey cream Acyclovir" 6 applications per day.

The treatment ended 31 patients. 8 patients after remission did not report for follow-up. Most of the patients (28 people) reported good tolerability of the drug. 3 people in the process of the drug was noted quite pronounced burning sensation after applying the cream on the lesions. In the process of observation none of the patients was not observed allergic reactions to the composition of EXAMPLE 1.

The positive effect from the use of the composition of EXAMPLE 1 was observed in all patients and was expressed in the acceleration of epithelialization of the foci and the impact on opportunistic bacterial flora with a resolution of infectious complications.

While examining patients of the main group on the background of treatment positive clinical effect was observed for most of the observed in the form of cessation of new lesions on average 3±1,2 day, the beginning of epithelization 6±2,1 day, complete epithelialization was observed to 11±2,3 day. The epithelization time depended on the size of the initial lesion and the degree of severity of herpes infection. Positive therapeutic effect was observed in patients with bacterial complications of herpes infection, probably due to part of the existing components. Noteworthy is the fact that quite good results is s obtained in patients with severe herpes infections, where combination therapy was carried out. At the same time epithelialization of the lesions in these patients were compared with a group of patients with a mild case of herpes.

In the group of patients that used the cream Acyclovir obtained similar results in terms of the cessation of new lesions (3±1.4 days), by the time the epithelization (6±2.3 days), then as full epithelialization of the foci of lesions occurred more slowly and was 14±2.1 days. Physical examination conducted before treatment, abnormalities not identified as persons of the core group and the comparison group.

Thus, a study on a small group of patients, allows to draw the following conclusions:

1. Found that using the compositions of EXAMPLE 1 are possible in the treatment of lesions caused by herpes virus 1, 2 type.

2. The drug is well tolerated by patients.

3. Side effects registered in 3 patients who participated in this study and resulted in burning sensation in the area of application of the composition of EXAMPLE 1, an allergic reaction to the drug in the study were not met.

4. Perhaps the use of the composition of EXAMPLE 1 in bacterial complications of herpes infection that is caused by the presence in its composition of ingredients with proteomika the m action.

5. The use of a composition of EXAMPLE 1 promotes more rapid epithelialization of the lesions compared with the use of cream Acyclovir".

6. The composition of EXAMPLE 1 can also be used in combined therapy of patients with severe recurrent herpes infection, contributing to more rapid epithelialization.

Pharmaceutical composition for topical treatment and prevention of infections caused by herpes virus type 1, 2, and bacterial complications caused by herpes infection, containing as active ingredients lysozyme, peroxidase, poviargol, as anti-inflammatory ingredients escin and glitzirrizinovu acid or its salts, as carriers - liposomes on the basis of the high level of hydrogenated lecithins in combination with cholesterol and pharmaceutically acceptable carriers and excipients, in the following ratio, wt.%:

lysozymefrom 0.01 to 5.00
peroxidasefrom 0.01 to 5.00
poviargolfrom 0.01 to 5.00
liposome-based
highly hydrogenated lecithin the from 0.01 to 5.00
glycyrrhizin acid or its saltsfrom 0.01 to 5.00
escinfrom 0.01 to 5.00
carbomerfrom 0.01 to 5.00
emulsifierfrom 0.01 to 5.00
preservative0,01-1,00
the pH regulator0,01-1,00
water demineralizedrest



 

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EFFECT: invention ensures obtaining stable pharmaceutical compositions.

1 cl, 7 ex, 1 tbl

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to a method for preparing a magnet-sensitive lipid composition. The declared method involves preparing a therapeutic agent containing cisplatin, a source of phospholipids and a magnetism carrier representing pirocarbon-coated ferric nanoparticles to be exposed to UV radiation. The above phospholipid source is the pharmacopeial drug 'Phosphogliv' with the composite prepared in the micellar form.

EFFECT: providing the uniformity and thermodynamic stability of the composite prepared over a short optimum period of time with using no organic solvents.

1 dwg, 2 ex

FIELD: biotechnologies.

SUBSTANCE: liposomes are used as a matrix for activated ferment - horseradish peroxidase. To 5 mg of horseradish peroxidase oxygenated with a periodate method there added is 1 ml of liposome suspension in 0.01 M solution of carbonate-bicarbonate buffer at pH 9.5, and subjected to ultrasonic treatment during 1 min. Then it is incubated for 1 hour; immobilised with immunoglobulins in concentration of 5 mg during 2 hours at the temperature of 22±4°C; stabilised with 5 mg of sodium borane with further gel-chromatographic cleaning.

EFFECT: invention allows obtaining liposomal-immunoperoxidase conjugate for indication of infectious agents in an immunoenzymometric analysis and increasing service life of a preparation up to 6 years.

1 tbl, 3 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to pharmaceutical industry and represents a thermally-sensitive liposome for treating cancer in a subject, containing at least one phosphatidylcholine, at least one phosphatidylglycerol, monostearoyl phosphatidylcholine, pegylated phospholipid and an active substance specified in a group consisting of taxotere, docetaxel and carboplatin wherein the liposome has a gel to liquid transition point from approximately 39°C to approximately 45°C.

EFFECT: invention provides storage-stability for a long period of time of one month or more at low temperatures.

9 cl, 8 ex, 8 dwg

FIELD: medicine.

SUBSTANCE: group of inventions relates to medicine, namely to therapeutic treatment of gout and prevention of accompanying side effects and complications in patients. method of prevention and prediction of risk of infusion responses in case of intravenous introduction of pegylated uricase to patients with gout consists in determination of uric acid content in patient's blood serum with the following indication to cessation of therapy at the level of approximately 4 mg/dl. Claimed method also makes it possible to identify risk group of patients, who receive pegylated uricase, with reduction of treatment efficiency due to formation of antibodies and higher risk of infusion response development.

EFFECT: group of inventions provides timely cessation of treatment to prevent infusion responses in patients with gout in treatment with pegylated uricase.

37 cl, 2 tbl, 6 dwg

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