RU2519133C1 - Ointment containing encapsulated triterpenic acid or derivatives thereof - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: invention refers to a composition containing encapsulated triterpenic acid: betulinic acid, ursolic acid or derivatives thereof in the form of salts and esters, or triterpene alcohol - betulin, which may be used in medicine for treating and preventing viral infections caused by DNA and RNA-containing viruses, such as influenza viruses, oncogenic viruses, herpes virus, herpes zoster virus, as well as infections caused by gram-positive and gram-negative bacteria: Staphylococcus spp., Streptococcus spp., Enterococcus spp., Shigella spp., Escherichia spp., Salmonella spp., Proteus spp., Acinetobacter spp., Citrobacter spp., Pseudomonas spp., Serratia spp., Klebsiella spp., Antracoides spp., Cryptococcus spp., pathogenic fungi of the genus Microsporum, Trichophyton, Nocardia, Aspergillus, yeast-like fungi of the genus Candida, including multiresistant strains, as well as Actinomycetes and some pathogenic protozoa: Entamoeba histolytica, Trichomonas vaginalis. The invention presents the composition containing an active ingredient presented by 0.5 wt % of betulin or 0.5 wt % of encapsulated triterpenic acid: betulinic acid, ursolic acid or derivatives thereof in the form of salts and esters and others, and carriers presented by: β-cyclodextrins, fullerene, lecithins and polymers binding to the ingredients to form ingredient-carrier complexes, and excipients.

EFFECT: higher efficacy of using the composition.

3 cl

Description

The present invention relates to medicine, namely to a composition containing encapsulated triterpene acid: betulinic acid, ursolic acid or their derivatives in the form of salts and esters or triterpene alcohol - betulin, which can be used in medicine for the treatment and prevention of viral infections caused by DNA - and RNA-containing viruses such as influenza viruses, oncoviruses, herpes zoster, herpes zoster, and infections caused by gram-positive and gram-negative bacteria: Staphylococcus spp., Streptococcus spp., Enterococc us spp., Shigella spp., Escherichia spp., Salmonella spp., Proteus spp., Acinetobacter spp., Citrobacter spp., Pseudomonas spp., Serratia spp., Klebsiella spp., Antracoides spp., Cryptococcus spp., pathogens genus Microsporum, Trichophyton, Nocardia, Aspergillus, yeast-like fungi of the genus Candida, incl. multiresistant strains, as well as Actinomycetes and some pathogenic protozoa: Entamoeba histolytica, Trichomonas vaginalis.

There are known topical medicines used for the treatment and prevention of herpes virus infections, containing antiviral components that are not enzymes, for example Zovirax ™ cream (acyclovir) (http://www.vidal.ru/poisk_preparatov/zovirax~27174.htm); capsules and lyophilisate for the preparation of a solution for intravenous administration of Phosphogliv ™ (phospholipids and trisodium salt of glycyrrhizic acid) (http://www.vidal.ru/poisk_preparatov/phosphogliv.htm); Epigen-intim ™ spray (glycyrrhizic acid) (http: //www.vidal. ru / poisk_preparatov / epigen-intim.htm); ointments: alpizarin (http://alpizarin.ru/), helepin (http://www.vidal.ru/poisk preparatov / helepin-d_20830.htm). flacoside (http://ru.wikipedia.org/wiki/%D4%EB%E0%EA%EE%E7%E8%E4) containing plant extracts.

The known drug Interferon alpha-2a, identical to human leukocyte interferon alpha-2 and having immunomodulating, antiviral and antitumor effects (Reference "Medicines" edited by MA Klyuev, 2001).

La-Cree ™ Lip Balm is a cosmetic product containing licorice extract and bisabolol, used to soften the skin of the lips. (http://vertex.spb.ru/catalog/68/34/la-kri_balyzam_dlya_gub.html)

Known lip balm Uriage Barierderm, containing shea butter, borage oil, unsaponifiable fractions of avocado oil, stearyl glycyrrhetinate, ascorbyl palmitate, tocopherol, used to repair damaged lip skin. (http://www.krason.ru/catalog-bariederm/1740-3620-balzam-dlya-gub.aspx?perf=1)

Known drug Glycyram (ammonium glycyrrrhizinate), used in the form of tablets for the treatment of bronchial asthma and adrenal hypofunction. (http://www.vidal.ru/poisk_preparatov/glycyram-tablets-0.05_27575. htm)

Known combination drug Hexalysis, containing as antimicrobial components - biclotimol and lysozyme, enoxolone isolated from glycyrrhizic acid as an anti-inflammatory component. (http://www.vidal.ru/poisk_preparatov/hexalyse~22640.htm)

There are known topical cosmetics used for the prevention of herpes containing antiviral components that are not enzymes, for example, extracts of medicinal plants and germanium-organic compounds (Antiherpetic preventative lipstick (P16 9158 915854 71.100.70) 292472. TU 9124-001- 00358210-96), glypirrhizic acid "Herpinat" ointment; ban. 20 ml; No. 77.01.12.915.P.02970.02.00 from the Research Institute of Atherosclerosis of the Russian Academy of Natural Sciences (Russia), INAT-PHARMA (Russia) (http://www.biomedservice.ru/publish/pub16.htm)

A cosmetic enzyme-containing composition is known where the enzyme is selected from the group of proteinases, including trypsin, chymotrypsin, bromelain, papain and ficin, as well as lysozyme, elastase, α-lipase and α-amylase, which is used to treat acne, care for skin and skin cleansing (DE 4305460). The described composition does not contain nuclease enzymes with direct antiviral activity, and therefore cannot be used to prevent viral infections.

Known medicines and cosmetics for topical use, used for the treatment and prevention of scars, ulcers and wounds, containing enzymes, for example, Iruxol ™ ointment, containing the enzyme: clostridiopeptidase A and antibiotic: chloramphenicol, (http://www.rlsnet.ru/tn_index_id_1605 .htm); Karipazim ™ powder containing enzymes: papain and lysozyme, for the prevention of scars, (http://www.caripazymum.ru/instruction.html); Fermencol gel containing a complex of collagenolytic proteases (http://fermencol.ru/); Moricrol balm containing the enzyme: moricrazu (http://ekogoods.ru/catalog/11-balzam_%ABmorikrol%BB), Sodermix cream containing the enzyme: superroxiddismutase (http://sodermix.ru/).

Known pharmaceutical preparation "Deoxyribonuclease", produced in the form of lyophilized powder in hermetically sealed vials and ampoules of 0.005; 0.01; 0.025 and 0.05 g. This drug is prescribed for herpetic keratitis, adenoviral conjunctivitis, to reduce viscosity and improve the evacuation of sputum and pus in bronchiectasis and is used topically in the form of a 0.2% solution on isotonic sodium chloride solution (Reference "Medicines" edited by M.A. Klyuyev, 2001).

Known pharmaceutical preparation "Ribonuclease", produced in the form of lyophilized powder in hermetically sealed vials of 0.01 g. This drug is prescribed for diseases of the respiratory tract with difficult to separate sputum, periodontal disease, gingivitis, purulent wounds, trophic ulcers, tick-borne encephalitis, thrombophlebitis. topical application is sprinkled on the wound surface in the amount of 0.025-0.05 g (Reference "Medicines" edited by MA Klyuev 2001).

Known betulinic acid (3β-hydroxy-20 (29) -lupaen-28-oic acid) is a natural pentacyclic triterpenoid. It is contained in the bark of some plant species, mainly fluffy birch (Betula pubescens), from which it got its name. Betulinic acid and its derivatives have anti-inflammatory, antitumor and anti-HIV activity. (Pisha E. Method for selectivity inhibiting melanoma, using betulinic acid. Nat. Med. 1995. Vol.1. P.1046-1051 Fujioka T. Compounds and methods of use to treat HIV infections. J. Nat. Prod. 1994. Vol. 57. P.243-247.)

Ursolic acid is known - a pentacyclic triterpene compound, isolated from plant materials, exhibits physiological activity both with external and internal use. Its anti-inflammatory, antitumor and antimicrobial properties allow its use in cosmetics. Ursolic acid and its isomer, accompanying it in most plants, oleanolic acid have been recommended for the treatment and prevention of skin cancer in a number of countries. Both triterpene compounds promote hair growth by stimulating peripheral blood flow in the scalp and activating maternal hair cells. Treating the skin with drugs that include these compounds prevents hair loss and dandruff. For ursolic acid and its derivatives, a wide spectrum of biological activity has been identified: anti-inflammatory, hypocholesterolemic, anti-atherosclerotic, antimicrobial, hypolipidemic, pacemaking. In addition, ursolic acid can be used as a modifier of protein synthesis, as a cosmetic, as an agent against lymphocytic leukemia and an antitumor drug. http://www.nioch.nsc.ru/russ/research/res_6.htm

Known data on the antiviral activity of grinding acid. (http://www.ijpbs.net/vol-2_issue-3/pharma_science/44.pdf)

Derivatives of betulinic acid used to treat viral infections, in particular HIV infection, are known from WO 2012106188, US 2011313191, US 2012101098.

From KR 20110018671 known pharmaceutical compositions containing triterpene acids for the prevention and treatment of cardiovascular disorders caused by hypercholesterolemia.

FROM KR 2010040107 known compositions containing triterpene acid to eliminate wrinkles.

From JP 2011020989, a method for stabilizing triterpenic acid by N-methypyrrolidone is known.

Derivatives of betulinic acid known as antitumor agents are known from RU 2445317.

From WO 2011051520, compositions containing triterpenic acids as an auxiliary component in protein vaccines are known.

An ointment containing a corticosteroid and beta-cyclodextrin with anti-inflammatory and anti-allergic effects is known from RU 2098135, in which the use of beta-cyclodextrin provides an increase in the bioavailability of the drug, prolongation of the action, halving the amount of the active substance and reducing side effects, as well as cheaper product.

From US 20110098261 A1, a composition is known containing a betulinic acid derivative for the treatment and / or prophylaxis of viral infections, namely against the influenza virus. The composition can be made in a form for local use and encapsulated.

From WO 2012106188 A1 a composition is known having antiviral activity, in particular for the treatment of HIV infections, containing betulinic acid. The composition may be in the form of an ointment.

In A.V. Simon. Molecular modification of betulinic acid anti-melanoma drugs and approaches to its solubilization // Abstract, 2004, conclusion, the effectiveness of the inclusion of betulinic acid and its derivatives in liposomes was studied.

From KR 100757043 B1, an ointment containing a triterpene-based nanocapsule (ursolic, oleanolic acid) is known.

From GAO D. Et all. Oleanolic acid liposomes with polyethelene glycol mmodification: promising antitumor drug delivery // Int J Nanomedicine, 2012 Jul 6, 7: 3517, abstract, known capsule containing oleanolic acid, which is a powerful means of delivery.

From US 20010018459 A1 it is known that triterpeic acids (betulinic acid, ursolic acid, oleanolic acid) and their derivatives (esters) have an antifungal effect.

From Kazakova O.V. et all. Synthesis oftriterpenoid acylates - an effective reproduction inhibitors of influenza A (H1N1) and papilloma veruses // Bioorg Khim., 2010 Nov-Dec, 36 (6) 841-8, PMID: 21317951, abstract, it is known that derivatives of oleanolic glyc cirrhizic, ursolic and betulinic acids have an effect against the human papillomavirus.

The difference between the claimed composition from US 20110098261 A1; WO 2012106188 A1; A.B. Simon. Molecular modification of betulinic acid anti-melanoma drugs and approaches to its solubilization // Abstract, 2004, conclusion; KR 100757043 B1; GAO D. Et all. Oleanolic acid liposomes with polyethelene glycol mmodification: promising antitumor drug delivery // Int J Nanomedicine, 2012 Jul 6.7: 3517, abstract; US20010018459 A1; Kazakova O.B. et all. Synthesis of triterpenoid acylates - an effective reproduction inhibitors of influenza A (H1N1) and papilloma veruses // Bioorg Khim., 2010 Nov-Dec, 36 (6) 841-8, PMID: 21317951, abstract, is that it contains additional as an active ingredient, 0.001 wt.% interferon, or 5.0 wt.% lysozyme, or 5.0 wt.% sanguirytrin. The presence of interferon in the composition enhances the antiviral effect of triterpene acid; the presence of lysozyme or sanguirytrin is the antimicrobial effect of triterpene acid, which ultimately provides greater effectiveness of the composition compared to compositions in which only triterpene acid is present as the active ingredient.

Compositions containing encapsulated triterpenic acid: betulinic acid, ursolic acid or their derivatives in the form of salts and esters, triterpene alcohol - betulin and additionally interferon, or sanguirytrin, or lysozyme used in the prevention of viral infections, are not described.

Thus, it is an object of the present invention to provide a composition comprising encapsulated triterpene acid: betulinic acid, ursolic acid or their derivatives in the form of salts and esters, and optionally interferon or sanguirytrin or lysozyme, which can be used to prevent viral infections caused by DNA and RNA-containing viruses, such as influenza viruses, oncoviruses, herpes, shingles.

The present invention provides a composition containing encapsulated triterpenic acid as an active ingredient: betulinic acid, ursolic acid or their derivatives in the form of salts of 0.5 wt.% And an additional 0.001 wt.% Interferon, or 5.0 wt.% Sanguiryrin, or 5.0 wt.% Lysozyme and acceptable carriers and excipients.

Most preferred is a composition containing encapsulated betulinic acid as the active ingredient and, in addition, sanguirythrin and acceptable carriers and excipients, in the following ratio, wt.%:

betulinic acid 0.5 β-cyclodextrin 5,0 emulsifier 0.5 preservative 1,0 sanguirythrin 5,0 polymer carrier 1,0 hydrophobic base 80.5 pH regulator 1,0 demineralized water 5.5

The active ingredient of the claimed pharmaceutical composition is betulinic acid or its derivatives: betulin, stearyl betulinate, ammonium or potassium or sodium betulinate, and lysozyme. Betulin, betulinic acid and its salts are used as an antiviral agent for external and local use. Betulinic acid is active against DNA and RNA-containing viruses, including various strains of Herpes simplex, Varicella zoster viruses, human papilloma viruses, cytomegaloviruses. The antiviral effect is apparently associated with the induction of the formation of interferon. Interrupts the replication of viruses in the early stages, causes the virion to exit the capsid, thereby preventing its penetration into the cells. This is due to selective dose-dependent inhibition of phosphorylating kinase P. It interacts with the structures of the virus, changing various phases of the viral cycle, which is accompanied by irreversible inactivation of viral particles (which are in a free state outside the cells), blocking the introduction of active viral particles through the cell membrane into the cell, as well as impaired ability viruses to the synthesis of new structural components. Inhibits viruses in concentrations that are non-toxic to normally functioning cells. Virus strains resistant to acyclovir and iodouridine are highly sensitive to betulinic acid. It also has anti-inflammatory, analgesic and tissue regeneration-enhancing effects both in the early manifestations of viral infection and in ulcer forms. (Fujioka T. Compounds and methods of use to treat HIV infections. J. Nat. Prod. 1994. Vol. 57. P.243-247).

Thus, a multidirectional antiviral effect is carried out: interruption of virus replication in the early stages, stimulation of the release of the virion from the capsid, induction of interferon formation, as well as suppression of secondary microflora and inflammation in the affected area and acceleration of epithelization.

To prepare the composition using substances obtained from crushed plant materials by extraction, followed by purification and drying and, if necessary, etherification or hydrolysis.

It was unexpectedly found that triterpene acids and their derivatives: betulinic acid, ursolic acid, triterpene alcohol - betulin and beta-cyclodextrin can be included in a stable composition that has the ability to penetrate the nuclei of infected cells and mitigate side effects from the endocrine system when using triterpene acids.

The composition of the present invention is usually made by conventional methods using solid or liquid acceptable carriers or excipients selected from emulsifiers, dispersants, preservatives, flavorings, pH adjusters, polymer carriers and other excipients that are suitable for the preparation of compositions for topical use. The composition of the present invention can be prepared in the form of an ointment on various bases: hydrophilic, hydrophobic and combined.

Additional advantages of this composition are a two-fold reduction in the amount of active substance, reduction of side effects, as well as cheaper product, increased stability, multidirectional antiviral effect, ease of use, and ease of dosing.

The claimed composition has stability with respect to the components included in its composition during long-term storage, which is achieved by immobilization of the components on the media contained in the specified composition. As the indicated carriers, any lecithins, cyclodextrins, fullerenes and polymers capable of binding to the components to form component-carrier complexes can be used. Examples of such polymeric carriers are gums, polyvinylpyrrolidone, carrageenans, sodium alginate, chitosan polyethylene glycol ether, sodium carboxymethyl cellulose, polyacrylate, cyclodextrins, fullerenes, lecithins.

The activity of the claimed composition is due to the activity of triterpene acid or its derivatives included in its composition. It was found that the composition containing betulin or triterpene acid 0.5 wt.%, Has stability during its long-term storage, therefore, the claimed composition can be used for the effective prevention of DNA and RNA viruses, such as influenza viruses, oncoviruses, herpes shingles.

The present invention is illustrated by the following examples.

EXAMPLE 1

The composition in the form of a gel with a hydrophilic base based on betulin has the following composition:

betulin 0.5 β-cyclodextrin 5,0 emulsifier 0.5 preservative 1,0 lysozyme 5,0 polymer carrier 1,0 pH regulator 1,0 demineralized water 86.00

The specified composition is prepared according to the following method. In demineralized water, β-cyclodextrin, betulin, lysozyme is dissolved, an emulsifier, preservative, a polymeric carrier, and a pH regulator are added. The resulting gel is stirred, vacuum and homogenized.

EXAMPLE 2

The composition in the form of a liniment based on sodium ursolate has the following composition:

ursolate sodium 0.001 hydrogenated lecithin 0.001 emulsifier 0.5 preservative 1,0 interferon 0.001 polymer carrier 1,0 pH regulator 1,0 demineralized water 96,497

The specified composition is prepared according to the following method. Ursolate sodium, interferon are dissolved in demineralized water, an emulsifier, a preservative, a polymeric carrier, hydrogenated lecithin, and a pH regulator are added. The resulting liniment is filtered.

EXAMPLE 3

The composition in the form of a cream with a hydrophobic base based on betulinic acid has the following composition:

betulinic acid stearyl ether 0.5 β-cyclodextrin 5,0 emulsifier 0.5 preservative 1,0 sanguirythrin 5,0 polymer carrier 1,0 hydrophobic base 80.5 pH regulator 1,0 demineralized water 5.5

The specified composition is prepared according to the following method. The suppository base is melted at a temperature of 40 ° C, β-cyclodextrin, betulinic acid stearyl ether, sanguirytrin are dissolved in demineralized water, an emulsifier, preservative, a polymeric carrier, and a pH regulator are added. The resulting mass is mixed, homogenized, cooled and packaged.

EXAMPLE 4

The composition in the form of a cream with a combined base based on ammonium betulinate has the following composition:

ammonium betulinate 0.001 hydrogenated lecithin 0.001 emulsifier 0.5 preservative 1,0 interferon 0.001 polymer carrier 1,0 pH regulator 1,0 demineralized water 96,497

The specified composition is prepared according to the following method. Ammonium betulinate, interferon are dissolved in demineralized water, an emulsifier, a preservative, a polymer carrier, hydrogenated lecithin, and a pH regulator are added. The resulting cream is filtered and packaged.

EXAMPLE 5

Assessment of the stability of betulin and triterpenic acid

To assess the stability of betulin and triterpenic acid, a comparative assessment was made of the conservation of betulin and triterpene acid in the composition (using gel and liniment as an example). The content of triterpenic acid was determined by non-aqueous titration, and betulin by HPLC.

The results are shown in the following table.

Table 1 Test samples Main ingredients Storage stability one 2 3 Gel number 1 According to Example 1 99% Betulin Content over 5 Years of Observation Liniment No. 2 According to Example 2 The content of triterpene acid 99% for 5 years of observation Gel number 3 According to Example 1 (without β-cyclodextrin) The content of betulin 70% for 5 years of observation Liniment No. 4 According to Example 2 (without hydrogenated lecithin) The content of triterpenic acid 68% for 5 years of observation

As can be seen from the table, the use of polymer carriers and stabilizers in the composition allows to obtain stable compositions with preservation of betulin and triterpene acid for 5 years of observation. Similar data were obtained using the compositions of EXAMPLES 3, 4.

EXAMPLE 5

Evaluation of the antimicrobial activity of the composition

The antimicrobial activity of the composition of EXAMPLE 1 was determined by serial dilution. As test microorganisms, cultures: Staphylococcus spp., Streptococcus spp., Enterococcus spp., Shigella spp., Escherichia spp., Salmonella spp., Proteus spp., Acinetobacter spp., Citrobacter spp., Pseudomonas spp. Serratia ., Klebsiella spp., Antracoides spp., Cryptococcus spp., Fungi of the genus Microsporum, Trichophyton, Nocardia, Aspergillus, Candida, Actinomycetes, protozoa: Entamoeba histolytica, Trichomonas vaginalis. A one-day culture of 1000 cells / ml was added to each tube. All cultures of microorganisms, except Microsporum, Trichophyton, Nocardia, Aspergillus, Candida, Actinomycetes, were grown in test tubes on a medium with meat and peptone broth at 37 ° C for 24 hours, and cultures of Microsporum, Trichophyton, Nocardia, Aspergillus, Candida, Actinomycetes C on agarized at 24 ° C for two days. Titration was started at a concentration of 10 mg / ml (1/10 dilution). As a control, a test tube was used, into which only nutrient medium and microorganism were introduced. The concentration in the last tube with no growth was taken as the minimum inhibitory concentration. To assess the bactericidal effect (minimal inhibitory concentration), from the test tubes with the lack of growth, they were reseeded into plates with meat and peptone broth. The cups were thermostated at 37 ° C for a day. Bactericidal action was noted in the absence of growth on a solid nutrient medium.

table 2 Name of culture MIC (minimum inhibitory concentration) mg / ml MPC (minimum inhibitory concentration) mg / ml Staphylococcus spp. 12.5 25 Streptococcus spp. 12.5 25 Enterococcus spp. 12.5 25 Shigella spp. 12.5 25 Escherichia spp. 12.5 25 Salmonella spp. 12.5 25 Proteus spp. 12.5 25 Acinetobacter spp. 12.5 25 Citrobacter spp. 12.5 25 Pseudomonas 12.5 25

spp. Serratia spp. 12.5 25 Klebsiella spp. 12.5 25 Antracoides spp. 12.5 25 Cryptococcus spp. 12.5 25 Microsporum 25 fifty Trichophyton 25 fifty Nocardia 25 fifty Aspergillus 25 fifty Candida 25 fifty Actinomycetes 25 fifty Entamoeba histolytica 25 fifty Trichomonas vaginalis 25 fifty

As can be seen from table 2, the minimum inhibitory concentration is: for Staphylococcus spp., Streptococcus spp., Enterococcus spp., Shigella spp., Escherichia spp., Salmonella spp., Proteus spp., Acinetobacter spp., Citrobacter spp., Pseudomonaspp ., Serratia spp., Kiebsieila spp., Antracoides spp., Cryptococcus spp. - 12.5 mg / ml; for Microsporum, Trichophyton, Nocardia, Aspergillus, Candida, Actinomycetes, Entamoeba histolytica, Trichomonas vaginalis - 25 mg / ml; the minimum inhibitory concentration is: for Staphylococcus spp., Streptococcus spp., Enterococcus spp., Shigella spp., Escherichia spp., Salmonella spp., Proteus spp., Acinetobacter spp., Citrobacter spp., Pseudomonas spp., Serratia Klebsiella spp., Antracoides spp., Cryptococcus spp. - 25 mg / ml; for Microsporum, Trichophyton, Nocardia, Aspergillus, Candida, Actinomycetes, Entamoeba histolytica, Trichomonas vaginalis - 50 mg / ml. Similar results were obtained in determining the antimicrobial activity of the compositions of EXAMPLES 2-4.

EXAMPLE 6

Evaluation of the effectiveness of the composition against herpes simplex viruses and Varicella zoster virus

To evaluate the effectiveness of the composition, an open comparative study of the composition of EXAMPLE 3 was conducted. The study involved 39 patients aged 18 to 45 years with clinical manifestations of herpes virus infection. By gender, the group was divided as follows: men - 11 people, women - 28 people. The duration of the disease ranged from 2 years to 12 years. At the same time, the overwhelming majority of patients had a mild and moderate course of herpes infection - 2-3 times a year - 14 people, 4-6 times a year - in 22 and 6-8 times a year in 3 patients.

Among women in the observed group, orofacial manifestations were noted in 18, and genital localization in 10 people. In the group of men, genital herpes and herpes zoster were registered in 7 patients, and in 4 patients there was a lesion in the face area. At the same time, in 7 people (men - 2, women - 5), conditionally pathogenic bacterial flora joined the manifestations of herpes with the development of complications in the form of strepto-staphyloderma.

In most patients, a small lesion area was recorded, ranging from 0.5 to 1.0 cm in 29 people, foci of 1.5 cm in 9 patients.

The comparison group included 11 patients with manifestations of herpes infection who received external therapy with Acyclovir cream. By gender, the comparison group was divided as follows: there were 7 men, 4 women. The manifestations of herpes in 4 men were in genital localization, in 7 men in the genital and lumbar region, in 1 woman herpes also had genital localization, and 3 women had orofacial herpetic eruptions. According to the severity of the course, 8 patients of the control group had a mild course, 3 of moderate severity.

Before treatment and 4 weeks after the end of therapy, all patients underwent the following studies:

- Survey and inspection.

- The study of material from the lesion using polymerase chain reaction to detect the human herpes simplex virus.

- The study of scrapings from the urogenital tract to detect chlamydia, myco and ureaplasmas (with damage to the urogenital tract.

- Testing blood serum for syphilis; HIV test after a pre-test interview.

- Clinical blood test

After obtaining informed consent for external treatment, all patients were prescribed a composition from EXAMPLE 3 4-6 applications per day, three patients with severe course were simultaneously treated with acyclic nucleoside preparations for 5 days. In the comparison group, patients received external therapy with Acyclovir cream 6 applications per day.

Treatment was completed by 31 patients. 8 patients after remission did not appear for follow-up. Most of the observed patients (28 people) noted a good tolerance to the drug. During the observation process, none of the patients showed allergic reactions to the composition from EXAMPLE 3. A positive effect from the use of the composition from EXAMPLE 3 was observed in all patients and was expressed in accelerated epithelialization of foci and the effect on the conditionally pathogenic bacterial flora with the resolution of infectious complications.

When examining the patients of the main group against the background of the treatment, a positive clinical effect was observed in the majority of those observed in the form of cessation of new rashes on average 3 ± 1.2 days, the beginning of epithelization on 6 ± 2.1 days, complete epithelization was observed to 11 ± 2.3 day. The timing of epithelization depended on the size of the initial focus and the severity of the course of herpetic infection. A positive therapeutic effect was also obtained in patients with bacterial complications of herpetic infection, which is probably due to the active components included in the composition. It is noteworthy that quite good results were obtained in patients with severe herpes infection. Moreover, the timing of epithelialization of foci in these patients was comparable with a group of patients with mild herpes.

In the group of patients where Acyclovir cream was used, similar results were obtained in terms of the termination of the appearance of new rashes (6 ± 1.4 days), in terms of the start of epithelialization (9 ± 2.3 days), while complete epithelization of the lesion foci occurred more slowly and amounted to 18 ± 2.1 days. A general clinical examination carried out before the start of treatment did not reveal abnormalities both in the main group and in the comparison group.

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

1. It was found that the use of the composition of EXAMPLE 3 is possible in the treatment of lesions caused by herpes virus type 1,2 and Varicella zoster virus.

2. The drug is well tolerated by patients.

3. Side effects in the area of application of the composition of EXAMPLE 3 in the form of allergic reactions during the study did not occur.

4. It is possible to use the composition of EXAMPLE 3 for bacterial complications of herpes infection, which is due to the presence in its composition of components with antimicrobial activity.

5. The use of the composition of EXAMPLE 3 promotes faster epithelialization of foci compared with the use of Acyclovir cream.

6. The composition of EXAMPLE 3 can also be used in combination therapy of patients with severe recurrent herpetic infection, while promoting faster epithelization.

7. Due to the fact that the active components act on the DNA of viruses, the cause of diseases caused by DNA-containing viruses, it can be concluded that the composition is effective in treating and preventing diseases caused by herpes viruses, herpes zoster, and human papillomas. Similar data were obtained when testing the compositions of EXAMPLES 1, 2, 4.

EXAMPLE 7

Evaluation of the effectiveness of the composition against oncoviruses by the example of treatment of melanoma

The pharmacological effectiveness of the obtained compositions from EXAMPLES 1-4 was studied on white mice, which subcutaneously inoculated the tumor with melanoma B 16. The animals were divided into 5 groups. The first control group consisted of animals that did not receive the drug. Animals from the second to fourth groups were applied topically three times a day: the second group — the composition according to EXAMPLE 1, the third group — the composition according to EXAMPLE 2, the fourth group — the composition according to EXAMPLE 3, the fifth group — the composition according to EXAMPLE 4. The preparations were started after 48 hours after inoculation of the tumor for 10 days. Tumor volume was measured on days 3, 7, 10, and 13 after the end of treatment. According to the measurement results, the inhibition of tumor growth (SRW) was calculated, which was calculated by the formula:

TPO (%) = (Vk-Vo) / Vk × 100,

where Vk and Vo are the average volume of tumors (mm 3) in the control and experimental groups, which was determined as the derivative of the sizes of three perpendicular diameters of the tumor node. The increase in life expectancy (UP) of the treated animals compared with the control was calculated by the formula:

YL% = (Mo-Mk) / Mk × 100,

where Mo and Mk are the average life expectancy (days) in the experimental and control groups of animals.

Antitumor active drugs were considered to cause inhibition of tumor growth of 50% within 7 days after the end of treatment. The toxic effect of the drugs was judged by the number of dead animals before the death of mice in the control group. Statistical processing of the obtained data was carried out according to the Fisher-Student method. Differences in the compared values were considered significant at p = 0.05. The results of the evaluation of the antitumor activity of the compositions of EXAMPLES 1-4 with respect to the transplanted tumor of mice, melanoma B 16, were distributed as follows: it was found that immediately after treatment the tumor growth was inhibited in 2-5 groups compared with the control group of animals; the maximum effect was obtained with the composition of EXAMPLE 3 with three daily use for 10 days: 80% SRW immediately after treatment, 67% SRW for 7 days and the preservation of more than 50% SRW for 13 days. The compositions of EXAMPLES 1, 2, and 4 had a slightly lesser effect: 75% SRW immediately after treatment, 60% SRW for 7 days, and maintaining more than 50% SRW for 13 days. No deaths from toxicity were observed in any group of treated animals. Thus, based on the data obtained, it can be concluded that the antiviral effect of the compositions of EXAMPLES 1-4 in relation to oncoviruses that cause melanoma.

EXAMPLE 8

Evaluation of the effectiveness of the composition against influenza virus

To assess the effectiveness of the composition, a study was conducted of the composition of EXAMPLE 4. The study involved 50 patients aged 18 to 50 years with clinical manifestations of influenza. 25 patients were intranasally administered three times a day with the composition of EXAMPLE 4, 25 patients were given symptomatic treatment. In this study, the following clinical effects of the composition of EXAMPLE 4 were identified: acceleration of relief of fever was noted and prevalence of a critical type of temperature decrease in all age groups (on average in 53% of patients). This effect is explained by the antiviral effect of the composition and is confirmed by a reduction in the duration of detection of viral antigen in smears from the nasal mucosa. The duration of a runny nose and cough is significantly reduced. The marked clinical efficacy of the composition of EXAMPLE 4 was accompanied by a protective effect against subsequent diseases for 6 months. The number of episodes of influenza per patient observed was 3 times less in the treatment of the composition of EXAMPLE 4. An important advantage of therapy with the composition of EXAMPLE 4 was the prophylactic effect on the layering of the secondary bacterial flora: complications were significantly less common in those receiving the composition of EXAMPLE 4 (23, 7%) in contrast to the comparison group (45%) (p <0.05). Evaluation of the transformation of the form of rhinitis into purulent authentically confirmed the indicated effect - 7.9% during therapy with the composition of EXAMPLE 4 and 20% - comparison group.

Evaluation of the antiviral effect of the composition of EXAMPLE 4 was carried out on the basis of PCR diagnostics carried out on days 1-2 and 5-6 of the disease. In half of the patients receiving the composition from EXAMPLE 4, an etiological agent was not detected on the 5-6th day of the disease, while in the placebo group in a third of patients (p <0.05).

Similar data were obtained in the study of the compositions of EXAMPLES 1-3. Thus, based on the data obtained, it can be concluded that the antiviral effect of the compositions of EXAMPLES 1-4 against influenza virus.

Claims (3)

1. A composition having antiviral, antimicrobial activity, including betulin, β-cyclodextrin, emulsifier, preservative, lysozyme, polymer carrier, pH adjuster, demineralized water, in the following ratio, wt.%:
betulin 0.5 β-cyclodextrin 5,0 emulsifier 0.5 preservative 1,0 lysozyme 5,0 polymer carrier 1,0 pH regulator 1,0 demineralized water 86.00 2. A composition having antiviral, antimicrobial activity, including sodium ursolate or ammonium betulinate, hydrogenated lecithin, emulsifier, preservative, interferon, a polymer carrier, pH adjuster, demineralized water in the following ratio, wt.%:
ursolate sodium or ammonium betulinate 0.001 hydrogenated lecithin 0.001 emulsifier 0.5 preservative 1,0 interferon 0.001 polymer carrier 1,0 pH regulator 1,0 demineralized water 96,497 3. A composition having antiviral, antimicrobial activity, including betulinic acid, β-cyclodextrin, emulsifier, preservative, sanguirytrin, a polymeric carrier, a hydrophobic base, a pH regulator, demineralized water in the following ratio, wt.%:
betulinic acid 0.5 β-cyclodextrin 5,0 emulsifier 0.5 preservative 1,0 sanguirythrin 5,0 polymer carrier 1,0 hydrophobic base 80.5 pH regulator 1,0 demineralized water 5.5