3,3'-diindolylmethane (metindol)-base vaginal suppository

FIELD: medicine, pharmaceutical industry, pharmacy.

SUBSTANCE: invention relates to suppository used in treatment of genital condylomae and neoplastic diseases of reproductive organs. Agent for treatment of genital condylomae and neoplastic diseases of reproductive organs as a suppository comprises metindol (3,3'-diindolylmethane) as an active component, lipophilic base containing solid confectionary fat, polyvinylpyrrolidone and butylhydroxyanisole taken in the definite ratio. Proposed agent promotes to effective treatment of genital condylomae and neoplastic diseases of reproductive organs.

EFFECT: valuable medicinal properties of suppository.

2 cl, 4 dwg, 1 tbl, 5 ex

 

The invention relates to the field of medicine and pharmaceutical industry, namely concerns the means for the treatment of genital warts and neoplastic diseases of the reproductive organs.

Human papillomavirus infection (RSI) is the most common infectious disease, sexually transmitted diseases. According to statistics, more than 50% of sexually active population during the life of the infected with human papilloma virus (HPV). Biological and medical interest in papillomaviruses because they belong to epochality, can cause benign hyperplasia, and some of their types is to initiate the development of malignant tumors of the anogenital area: cancer of the cervix, vulva, vagina, etc. are the Result of infection with HPV is considered as cervical intraepithelial neoplasia (CIN)related to pre-cancerous conditions. HPV DNA high degree of carcinogenic risk (mainly 16 and 18 types) found in 50-80% of the samples of moderate and severe dysplasia of the squamous epithelium of the cervix and in 90% of invasive cancer.

Cancer of the cervix (cervical cancer) is the second highest incidence among cancers in women. Every year in the world there are nearly half a million new cases of this disease and 270,000 caused their deaths. Standarts the integration of the incidence of and mortality from cervical cancer is 16.2 and 9 per 100,000 population, respectively. While approximately 52% of all cases of cervical cancer occur in Asia. In Russia from cervical cancer (4,8% of all malignant tumors in women) annually kills more than 6 Tyson (women aged 20 to 40 years cervical cancer is the leading cause of death), the incidence rate is approximately twice as high.

Not least in importance medical problem are benign manifestations of the MBC, which include exophytic and endophytic lesions of various parts of the epithelium of the vulva, vagina and cervix, as well as the surrounding skin and mucous membranes, including the anus. Exophytic forms (genital warts) include diverse in appearance and size, pointed, papillary and papaloapan education, often complicated by secondary bacterial infection and accompanied not only psychological, but also physical suffering of the patient.

Frequency of genital condylomas (genital warts), which is a consequence of infection benign HPV types, according to the RF Ministry of health (2001) is 26 per 100 000 population. Endophytic forms (subclinical RSI) represent different morphological changes of the squamous epithelium without external growths.

Genital warts are also a cause of return is Atego respiratory papillomatosis in children due to vertical transmission of the virus from an infected mother to child (Minkin G.N., Manukhin IB, Frank G.A., Precancer, Moscow, "Airbrush-Media, 2001).

The main methods of treatment of genital warts and pre-cancerous neoplastic diseases of the reproductive organs, is caused by HPV, are currently the chemical destruction, surgical excision or ablation. These methods are often accompanied by complications, require the use of expensive medical equipment and are not always effective. Thus the huge number of young women with ambiguous clinical picture vaginal condylomata, not subject to destructive treatment, are carriers of the infection and pose a risk to sexual partners and unborn child.

During infection of epithelial tissues papillomavirus uses a set of mechanisms, subordinating their interests livelihoods infected cells. It is known that HPV DNA encodes the synthesis of two proteins E6 and E7 induces the transition of differentiated cells in S-phase of the cell cycle. At the stage of active reproduction of the virus genes E6 and E7 are regulated protein E2 gene product is a repressor of transcription of these genes. Therefore, while the virus is in the episomal state, they shall have a place of benign processes of proliferation of the infected tissue. A key event in the malignant transformation of cells is the integration of the virus into the genome of the host cell, which is accompanied by a deletion of the E2 gene.

Control of the cell cycle and differentiation through oncogenic proteins E6 and E7, inactivating key proteins-regulators of the proliferative activity of cells, proapoptotic protein p53 and retinoblastoma protein (pRB). Found that E7 protein capable of forming a stable complex with the protein pRB, causing its degradation, which leads to the release of transcription factor E2F, which stimulates the transcription of genes required for DNA replication in S-phase of the cell cycle. In addition, E7 affect the activity of a number of other proteins-regulators of the cell cycle, such as the a - and E-cycline, dk2-kinase and inhibitors of cyclin-dependent kinases P21 and P27.

It is known that in the pathogenesis of viral infection, resulting in increased proliferative activity of infected cells of a benign nature (and sometimes with their malignant degeneration), plays a significant role in the endocrine system, in particular the level and activity of female sex hormones - estrogens (journal of dermatology and venereology, 2000, No. 6, p.20-23). The relative constancy of cell proliferative activity in estrogen-sensitive tissues is controlled with the aid of the through special metabolic mechanisms, converts estradiol (the most active estrogen) in biologically active derivatives. The main pool of endogenous estrogen utilized by localized in the liver microsomal cytochrome P-450 catalyzing the formation of its hydroxy derivatives, which facilitates their solubility and subsequent excretion from the body via the kidneys and bile ducts. Enzyme system cytochrome P-450 provides conversion of estradiol in two main metabolite: 16α-hydroxyestrone (16α -ONE) and 2-hydroxyestrone (2-ONE) (figure 1).

16α-ONE belongs to the category of "aggressive" hormones that cause long-term carcinogenic effect (Proc. Natl. Acad. Sci. USA, 1982, v.79, p.3047-3051). It is shown that this effect is due to the formation of strong covalent bonds 16α-ONE, which is an agonist of estrogen, with nuclear estrogen receptors. 2-ONE has moderate functions and, unlike the 16α-ONE, on the contrary, normalizes cell growth. When the level of 2-ONE tends to tumor cell death and prevention of their further education.

The study of the functions of these two metabolites has allowed to identify a clear relationship between the level 16α-ONE and the risk of developing tumors in estrogen-dependent tissues and conclude that the ratio of 2-ONE to 16α-ONE is at the same time the University is realnum biomarker and reliable diagnostic criterion for determining the risk and prognosis of estrogen-dependent tumors.

Long ago it was noted that tissue changes in the cervical canal, caused by HPV, mainly localized in estrogen-sensitive areas. Moreover, it was found that where there is an active expression of HPV proteins, a high level synthesis (16α-ONE), comparable to those in cancer cells of the breast.

It should be emphasized that in normal epithelial cells of the cervix are not able to provide the conversion of estradiol to 16α-hydroxyestrone. Thus, the active reproduction of HPV induces the formation of "aggressive" metabolite in infected cells (Kiselev V.I., Kiselev, I. the human papilloma Virus in cervical cancer. SPb. - M.: the rose of the world, 2003).

At the same time, it has been shown that HPV-infected keratinocytes show proliferative activity in vitro, but with no tumor phenotype microscopic examination. Adding to the culture medium exogenous 16α-ONE transforms cells in a typical cancer (Int. J. Cancer, 1991, v.49, p.867-869).

Thus, the vicious circle, in which the virus through education "aggressive" forms of estradiol creates favorable conditions for the development of tumors by stimulating the synthesis of oncoprotein E7. In turn, the oncoprotein E7, on the one hand, activates the mechanisms of pathological proliferation, as with the other blocks antiviral immunological protection (figure 2).

In sum, we can conclude that infection of epithelial cells HPV is a necessary but insufficient factor for malignancy. For the formation of irreversible alterations are necessary: 1) induction of metabolic mechanisms of conversion of estradiol in 16α-ONE playing a key role in cancer transformation of HPV-infected cells; 2) active expression of the oncogenes E6 and E7 virus-stimulated interaction of complex hormone-receptor promoter HPV DNA; 3) induction of multiple defects chromosomal DNA of the infected cell, completing the process of neoplastic transformation.

With the emergence of viral concept cervical carcinogenesis has fundamentally changed and approaches to the treatment of dysplastic cervical processes. Currently, surgical methods (due to the high percentage of relapses after their application) began to play a secondary role. First place went etiopathogenetic therapy with two main areas: 1) impact on the etiological factor is HPV; 2) blocking of the basic mechanisms of carcinogenesis.

Unfortunately, drugs that selectively affect HPV, does not currently exist. Most often for the treatment of HPV infections is AI used drugs interferon (IFN) - protein produced by cells of the immune system in response to the stimulation of viral antigens. However, in most cases, even long-term IFN-terpia does not lead to clinical improvement. It is shown that the resistance to IFN HPV-infected cervical cells is determined by an increased level of expression of the oncoprotein E7, intracellular inactivating factor regulation of IFN, which includes the transcription of genes coding for the synthesis of antiviral proteins.

For many years we are searching for natural and synthetic anticancer compounds that can stop the development of precancerous lesions of the cervix. Finally, these searches were unsuccessful. It was recently identified a chemical compound with anti-cancer properties - indole-3-carbinol (S) - phytonutrient, isoflavonoid contained in cruciferous vegetables. Numerous literature data indicate that long-term use of this compound prevents the development of tumors of the intestines, lungs, organs of the female reproductive system. S stimulates antitumor effect of many medicines, reduces the mutagenic activity of carcinogens.

Most studies on S, for its antitumor activity in the so-called estrogen-dependent tissues and organs (mammary gland, is endometry and cervix), characterized by cyclical changes in the level of cell proliferative activity. The uniqueness of the action of I3C is that he intervenes in both these ways, affecting their functioning and preventing thus cell proliferation. On the one hand, he has a strong anti-estrogenic effect, stimulating the formation antiproliferative 2-hydroxyestrone and thus improving the ratio of 2-ONE/16α-ONE in favor of the former, and on the other, prevents the phosphorylation of cytoplasmic proteins participating cascading transmission induced by EGF. Another important mechanism of the antitumor action of I3C is its ability to induce apoptosis - "programmed death" of tumor cells through a system of bax-bcl.

Cervical cancer associated with HPV, was also investigated as a potential target for therapy of I3C. First promising results were obtained in 1999 by a group of American researchers on the model of transgenic mice containing the genome integrated form of HPV-16. Previously it was found that the content of these mice on a diet, which includes 17-α-estradiol, leads to the development of cervical cancer (J.Cell. Biochem., 2000, 34 (Suppl.): 103-114). Further studies were able to reliably confirm the diversity of antitumor activities of I3C in HPV-transformed cell of the cervical epithelium. It was shown that in vitro and in vivo:

1) I3C reduces estradiol-dependent induction of oncogene E7, sharply reducing the level of expression of the oncoprotein E7 and preventing hormone-dependent proliferation of infected cells;

2) I3C normalizes the metabolism of estradiol in cells infected with HPV, preventing the formation of carcinogenic metabolite 16α-ONE, stimulating the expression of HPV oncogenes;

3) I3C induces apoptotic processes HPV-infected cells, causing selective cell death with tumor properties (J. Nutrit, 2001, 131, 3294-3302).

Antitumor activity of I3C as a means of prevention and treatment of cervical cancer was confirmed in a recent placebo-controlled clinical trials (Gynecol. Oncol, 78, 123-129, 2000). Patients with CIN II and III degree, daily taking 200-400 mg I3C (experimental group), there was complete regression of tumors in 47% of cases, whereas in the control group was not a single documented case of regression.

Discussing antitumor and antiviral activity of I3C, be aware that this compound is extremely unstable and readily undergoes oligomerization, which are amplified in an acidic environment. Thus the main oligomeric product of I3C is its dimeric form - 3,3'-diindolylmethane (DIM). As demonstrated by pharmacokinetic studies is Finance, under the influence of the acidic environment of the stomach adopted oral I3C almost instantly turns into DIM (Ameson DW, Hurwitz A, McMahon LM, Robaugh D: Presence of 3,3'-diindolylmethane in human plasma after oral administration of indole-3-carbinol (abstr.) Proc. Am. Assoc. Cancer Res, 40, 2833, 1999).

Experimentally proved that almost all multiple antitumor mechanisms induced by I3C in vitro and in vivo, typical for the DIM.

In a recent experimental studies have shown the ability to DIM to cause apoptosis of cervical HPV-infected keratinocytes in vitro. At one of the three investigated cell lines of cervical cancer DIM demonstrated several times greater efficiency than I3C (value (LD50) was 50-60 microns to DIM and 200 μm for I3C, respectively), but as well as its metabolic precursor (I3C), did not cause apoptotic changes in normal (untransformed) keratinocytes (J.Nutrit, 2001, 131, 3294-3302).

A few years ago using the method of microarray analysis, it was found that the similarity of antitumor and antiviral activity of I3C and DIM manifests and molecular (genetic) level. In cell lines of cervical cancer DIM, like S, constitutive altered the transcription of more than 100 genes, most of which directly or indirectly controls cell proliferation (apoptosis), as well as expre is this HPV oncogenes in virus-infected cells (J.Nutr. 132, 3314-3324, 2002).

In conclusion, it is necessary to mention another recently described, the most important property DIM - its immunomodulatory activity. It was shown that in vitro in tumor cells DIM stimulates IFNα-dependent signaling cascades by activating the expression of IFN receptorsαand other IFN-responsive regulatory protein (Cells. Mol. Pharmacol, 2006, 69, 430-439). Having in mind high antiviral and antitumor activity of IFNαand, this property can be of great clinical value when using DIM for the treatment of cervical dysplasia due to HPV.

Summarizing the above, we can conclude that the drug is based on the DIM must have high antitumor and antiviral activity against HPV-infected cells.

However, although indomethacin is more stable connection than indole-3-carbinol, he has low bioavailability, which may limit its use as a medicine.

To improve the solubility and bioavailability have been developed in different dosage forms DIM. However, they also have drawbacks. Therefore, the search for new, more effective compounds is still relevant.

Known vaginal suppositories DIM for the treatment of leishmaniasis, obtained by heating cetostearyl the first alcohol with an active substance, and with the addition of ceramides or their derivatives, with the subsequent introduction of the mixture with triglyceride base and molded candles (WO 2005/107747, 2005-11-17).

Known patent US 6689387, 10.02.2004 describing various forms of diindolylmethane for the treatment of mastalgia and endometriosis. Including notes that you can use as a suppository, with traditional binders and such media as triglycerides.

Known suppositories do not provide prolonged action of the active substance for treatment of human papillomavirus infection. In addition, the treatment of neoplastic disorders requires a considerable time. At the same time, triglyceride basis for long-term use cause ulceration of the mucous membranes.

As the closest analogue may be specified patent RU 2196568.

This decision concerns a pharmaceutical composition for the prevention and treatment of tumors associated with human papilloma virus, namely dysplasia and cervical cancer and laryngeal papillomatosis. The invention lies in the fact that the proposed arrangement on the basis of indole-3-carbinol as an active start, possibly in the form of vaginal suppositories, based witepsol. Since the active substance is not persistent, it may not provide the necessary effectively the TB treatment.

The present invention is the development of dosage forms for more effective treatment of genital condylomas and neoplastic diseases with no side effects, is stable when stored.

The problem is solved by the drug representing suppository vaginal ("Cervical"), containing as active substance indomethacin (3,3'-diindolylmethane - DIM), lipophilic base containing solid confectionery fat, polyvinylpyrrolidone and butylhydroxyanisole or equivalent in the following, wt%:

Indomethacin2,0-6,0
Polyvinylpyrrolidone0.5 to 1.2
Butylhydroxyanisole
or equivalent0,3-0,5
Lipophilic baserest

Indomethacin is contained in an amount of 0.045-0,110 g per dose.

As lipophilic bases, preferably using solid confectionery fat, such as tallow type "A" or "B", but it is also possible introduction of additional emulsifiers, esters of phthalic acid and alcohols, hydrogenated vegetable oils, waxes, paraffin, lanolin, cooking oil, cocoa butter.

It is known that solid cond the Terek fats are fine-grained structure, which is melted in a narrow interval, have good ductility and are easily mixed with many substances. Manufactured confectionery fats prepared at almadinah basis and on the basis of hydrogenated fat.

In addition, it was found that the use of this framework with PVP and antioxidant, almost had no undesirable effects on the mucous membrane, as in the case of foundations, consisting mainly of triglycerides, such as, witepsol, Astrium, widely used in pharmaceutical practice.

Butylhydroxyanisole and equivalent synthetic products related to the derivatives of phenol. Demonstrate effective antioxidant and is known preservatives. Their introduction into suppozitornyj mass gives additional effect to the affected mucous.

As polyvinylpyrrolidone preferably using PVP nizkomolekulyarnogo, for example, marks Kollidon CL, CL-M - molecular weight of 12600±2700 or Povidone - weighing 8000±2000.

The method of preparation of suppositories is that melt components lipophilic bases, part of it separately grind with the active ingredient to obtain a homogeneous suspension, cooled to the remaining part of the basis add polyvinylpyrrolidone and an antioxidant, and then the obtained suspension is stirred until Oder the underwater condition and molded candles by the method of splashing in the form of a given size.

The invention is illustrated in the following graphics:

Figure 1 reflects the metabolism of estradiol.

Figure 2 reflects the role of estrogen in the carcinogenesis of epithelial cells of the cervix with HPV.

Figure 3 shows the E7 gene transcription in CaSki cells in the presence of estradiol (Figa - 1 - before adding estradiol, 2 - incubation for 4 hours 3 - 24 hours, 4 to 48 hours) and in the presence of estradiol + Cervical (Figb - 1 - before adding estradiol + Cervical, 2 - incubation for 4 hours 3 - 24 hours, 4 to 48 hours).

Figure 4 shows the induction of apoptosis in the presence of Cervione (Figa - cells visible condensation of chromatin, indicating cell death).

The invention is illustrated by the following examples.

Example 1

Vaginal suppositories containing

diindolylmethane0.05 g
3,3'-Diindolylmethane0.05 g
Tallow type "A"2,02 g
Kollidon CL0.02 g
Butylhydroxyanisole0.01 g

TOTAL-2.1 g

Example 2

Vaginal suppositories containing
diindolylmethane1.0 g
,3'-Diindolylmethane 0.1 g
Tallow type "A"1,97 g
Kollidon CL0.02 g
equivalent0.01 g

TOTAL-2.1 g

Example 3

Materials and methods.

Experimental model. As the model used a cell line CaSki cervical carcinoma, containing multiple copies of integrated HPV-16 genome-th type, cell line s (not containing HPV DNA) and cultured epithelial cells derived from cervical region in patients with a diagnosis of HPV-induced dysplasia".

Cells were maintained in medium RPMI-1640 with the addition of 10% fetal bovine serum. Candles in example 1 was dissolved in a hydroalcoholic solution containing 20% ethanol, and added to the cells so that the final concentration is included in its composition 3,3'-diindolylmethane was 50 μm.

The allocation of RNA. The cell suspension was mixed with 0.5 ml of lyse solution containing 4M guanidinylation, 0,2% SDS, 25mM sodium citrate (P7,0), suspensively on the Vortex for 10 s and literally in an ice bath for 15 minutes

After that the tube was made of 50 μl of 2M sodium acetate (pH 4.0) and 500 ál of phenol, the mixture was intensively shaken, centrifuged and selected aqueous phase. The procedure was repeated twice and then the aqueous phase was added an equal volume of isopropanol. After 2 hours incubation in the refrigerator at 20°the precipitate was collected, centrifuged and dissolved in 50 µl TE buffer.

Analysis of transcription of the gene E7 HPV type 16. The study of the E7 gene transcription was performed Northern hybridization. For this, 10 μg of total RNA isolated from cell suspension was separated by electrophoresis in agarose gel containing formaldehyde, and transferred to nitrocellulose filters. The probe for hybridization was prepared by amplification of a fragment of E7 gene in the presence of labeled dTTP(p32). As matrix were used for amplification plasmid rtne containing the oncogene E7 HPV 16-th type.

Mass spectrometric (GC-MS) analysis of metabolites of estradiol. The samples were prepared from supernatants cells treated with estradiol and the claimed composition in concentrations of respectively 1 and 50 μm. In the supernatant (5 ml) was made deuterated estradiol as standard and was applied on the column With 18 Sep-Pak (Water Associates, Milford, MA).

The steroid elution was performed with a solution of methanol/ascorbic acid. The total volume of the eluate was brought to 0.5 ml of a soft heated at 50°under vacuum, and the remaining samples were dried in liquid nitrogen. Each sample was dissolved in 10 μl of pyridine and 50 μl of bis(trimethylsilyl) trifluoroacetamide (BSTFA) and left overnight at room temperature. 2 μl of each arr is SCA were analyzed on a gas mass spectrometer. The analysis results were expressed in ng/mg cell protein.

Test biotransformation of estrogens. Analysis of the biotransformation was carried out radiometric method based on the formation NO by proton transfer from specifically labeled estradiol as described in (Telang NT, Bradlow HL and Osborne MP "In vitro biotransformation of estradiol by explant cultures of murine mammary tissues" Breast Cancer Res. Treat. 1989, 13, 173-181).

Labeled estrogen was added to the cell monolayer in a solution of 0.1% propylene glycol. After 24 hours of incubation were counted radioactivity in the culture fluid. From the obtained values, subtract the background level of biotransformation in culture medium without cells was counted per mg protein, determined by the method of Lowry.

The experimental part.

The study expressive oncogenes of human papillomavirus in transformed cells when exposed to Indianola. The expression of the E7 gene of HPV-16 was investigated in cell lines CaSki, containing up to 500 copies of the genome of HPV-16 in the integrated state. In the culture medium was made by estradiol (final concentration 1 μm) and the composition of example 1 (final concentration 50 ám DIM), incubation was continued for 4, 24 and 48 hours. At the indicated time, cells were harvested, isolated total RNA was determined by the level of E7 gene expression using Northern hybridization. The results are shown in figure 3.

the present experiment investigated the level of E7 gene expression in CaSki cells when added to the culture medium of estradiol.

From the results shown in figure 3, it is seen that estradiol significantly induces the synthesis of mRNA of the gene E7, and this effect increases over time (figa), and the addition of the claimed composition is almost completely eliminates this effect (figb).

Study of the metabolism of estradiol in epithelial cells infected with HPV. As was established earlier, in cells transformed by HPV, high risk, there is a change in the metabolism of estradiol in the direction of education "aggressive" 16α-hydroxyestrone. This metabolite has carcinogenic properties and are able to form covalent complexes with estrogen receptors, causing prolonged hormone-dependent effects, including stimulating the expression of HPV oncogenes. We investigated the effects of Indianola on the metabolism of estradiol, in particular, education 16α-hydroxyestrone in cells transformed by HPV (see table).

Education 16α-hydroxyestrone in cells transformed by HPV type 16
No.The cell type% of 16α - hydroxyestrone in µg cell protein
EstradiolEstradiol + Cervical
1CaSki In The H (+) 16,0±0,12,2±0,5
2With 33 And HPV (-)0,08±0,030,08±0,08
3Cervical epithelium0,7±00,5±0,1
4The cervical epithelium of transformed zones12,6±0,52,1±0,3

Example 4

Study the induction of apoptosis of cells transformed by HPV

in the presence of the drug according to example 2.

We have investigated the ability of the drug to induce apoptosis of cells infected with human papilloma virus.

To quantify the survival of cells and the proportion of cells in apoteose, used a combination of two dyes ethidium bromide, penetrates cells with damaged membranes (i.e. dead and necrotic cells) and coloring their nucleus in bright orange, almost red, and acridine orange staining living cells in bright green color and allows you to observe the different phases of condensation of chromatin and blebbing membrane. In cells that are in late stages of apoptosis, observed a bright orange region of condensed chromatin in the nucleus, which distinguishes them from necrotic cells, which in this case are uniformly colored in orange Zwettl cells in apoptosis was calculated as a percentage of the total number.

Cells CaSki and 33 And incubated with the fluorescent dye Hoechst 33342 (Sigma) in culture medium at a concentration of 5 μl/ml at 37°30 min, then fixed in 2% paraformaldehyde at room temperature for 10 min, once washed PBS and analyzed using a fluorescent microscope Opton.

As the experiments showed, after 24-hour incubation of CaSki cells in culture medium containing the claimed drug (final concentration 50 ám DIM), in the field of view was observed more than 50% of apoptotic cells (see figa), whereas in cells lines 33 And under the same conditions, the percentage of cells in apoptosis was not more than 5% (figb).

Example 5. Study of the impact on genital warts.

We also conducted studies on patients diagnosed with genital genital warts".

Diagnosis was made on the basis of cytological, histological examination of biopsy specimens, detection of antibodies to HPV and detection of HPV DNA and oncoprotein E7.

Of the 25 patients genital warts in about 90% of the cases were associated with other urogenital infections. About 10% had previously been treated by electrocoagulation method.

15 patients were administered 2 times a day candles in example 2 and 10 patients candles in example 1 for 10 weeks - 12 weeks before genital warts? is the n. At the control examination after a year and a half of recurrence of genital warts in the main group, 3 patients noted a relapse. Assigned additional treatment with increasing doses.

Conclusions

Our studies allowed us to establish that the new dosage form Zervigon (vaginal suppositories containing substance indomethacin (3,3'-diindolylmethane - DIM), has a high specific antitumor activity against epithelial human cells infected with HPV, which is implemented through the following mechanisms:

1. Cervical removes estradiol-dependent induction of oncogene E7 HPV-16, thus inhibiting hormone-dependent proliferation of infected cells.

2. Cervical normalizes the metabolism of estradiol in cells infected with HPV, preventing the formation of carcinogenic metabolitethe hydroxyestrone, stimulating the expression of HPV oncogenes.

3. Cervical induces apoptotic processes HPV-infected cells, causing selective cell death with tumor properties.

There is every reason to believe that topically applied in the form of vaginal suppositories new drug has greater antitumor and antiviral activity against HPV-infected cervical cells comparedwith taken orally DIM (in the same final concentration) and its metabolic precursor - I3C into DIM in the gastrointestinal tract.

Thus, the drug can be recommended in the treatment of dysplastic cervical processes caused by human papilloma virus.

In addition, suppositories provide the necessary time at high bioavailability of the active substance, which allows to apply them in such intractable pathologies as genital condyloma.

1. Cure for genital warts and neoplastic diseases of the reproductive organs in the form of a vaginal suppository, characterized in that it contains as active substance indomethacin (3,3'-diindolylmethane), lipophilic base containing solid confectionery fat, polyvinylpyrrolidone and butylhydroxyanisole at the following content, wt.%:

Indomethacin2,0-6,0
Polyvinylpyrrolidone0.5 to 1.2
Butylhydroxyanisole
or equivalent0,3-0,5
Lipophilic baserest

2. The tool according to claim 1, in which the lipophilic bases contains solid fat type ' a'.



 

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3 ex

FIELD: organic chemistry, medicine, pharmacy.

SUBSTANCE: invention relates to derivatives of piperidine of the general formula (I): or their pharmaceutically acceptable salts or isomers wherein Q means nitrogen atom (N); X and Z are chosen independently from group consisting of -CH and N under condition that one or both groups among Q and Z mean N; R, R4, R5, R and R are chosen independently from group consisting of hydrogen atom (H) and (C1-C6)-alkyl; R1 means H, (C1-C6)-alkyl, R9-aryl-(C1-C6)-alkyl-, (C1-C6)-alkyl-SO2-, (C3-C6)-cycloalkyl-SO2-, fluoro-(C1-C6)-alkyl-SO2-, R9-aryl-SO2-, R9-heteroaryl-SO2-, -N(R22)(R23)-SO2-, (C1-C6)-alkyl-C(O)-, (C3-C6)-cycloalkyl-C(O)-, fluoro-(C1-C6)-alkyl-C(O)-, R9-aryl-C(O)-, CH3CH2-NH-C(O)- or R9-aryl-NH-C(O)-; R2 means H or (C1-C6)-alkyl, and R3 means H, (C1-C6)-alkyl, (C1-C6)-alkoxy-(C1-C6)-alkyl-, (C3-C10)-cycloalkyl-, (C3-C10)-cycloalkyl-(C1-C6)-alkyl-, R9-aryl, R9-aryl-(C1-C6)-alkyl- or R9-heteroaryl under condition that each X and X doesn't mean N, or R2 and R3 in common mean =NOR10. Proposed compounds can be used as selective CCR5 antagonists. Compounds are useful in HIV treatment. Also, invention describes a pharmaceutical composition based on compounds thereof and combination with antiviral or anti-inflammatory agent.

EFFECT: valuable medicinal properties of compounds and pharmaceutical composition.

16 cl, 4 tbl, 5 ex

FIELD: veterinary virology, biotechnology.

SUBSTANCE: the suggested vaccine contains an active substance and a target additive. As the active substance it contains the suspension of avirulent and purified antigenic material out of "KPR-96" strain of the virus of swine's reproductive-respiratory syndrome (SRRS), Arteriviridae type, Arterivirus genus, FGU collection "VGNKI" "KPR-96"-DEP in efficient quantity. The vaccine in question induces high level of antibodies against SRRS, provides reliable passive maternal immunity in youngsters up to the age of 25-30 d that enables to protect the youngsters against field virulent SRRS virus in earlier period of life.

EFFECT: higher efficiency.

7 cl, 3 dwg, 7 ex, 16 tbl

FIELD: veterinary medicine.

SUBSTANCE: the vaccine suggested contains as antigens: the cell suspension of pure cultures of Escherichia coli, Salmonella typhimurium and Streptococcus fecalis obtained due to selecting the affected organs from dead nutrias out of local epizootic focus, preparing the suspension, inoculation onto differential-diagnostic media, isolating pure cultures of the agents of Escherichia coli, Salmonella typhimurium and Streptococcus fecalis. The obtained pure cultures of microorganisms should be separately grown in beef-extract broth with glucose up to the concentration of microbial cells being 4-5 bln/cu. cm to be then mixed at equal ratios. The vaccine, also, contains formalin, glucose and aluminum hydroxide at the following ratio of the components, weight%: cell suspension of Escherichia coli pure culture isolated from the affected organs out in dead nutrias out of local epizootic focus in nutritive medium at the titer being 4-5 bln microbial cells/cu. cm 18.0-21.0; cell suspension of Salmonella typhimurium pure culture isolated from the affected organs in dead nutrias out of local epizootic focus in nutritive medium at the titer being 4-5 bln microbial cells/cu. cm 18.0-21.0, cell suspension of Streptococcus pneumoniae pure culture isolated from the affected organs in dead nutrias out of local epizootic focus in nutritive medium at the titer being 4-5 bln microbial cells/cu/ cm 18.0-21.0 cell suspension of Streptococcus fecalis pure culture isolated from the affected organs in dead nutrias out of local epizootic focus in nutritive medium at the titer being 4-5 bln microbial cells/cu. cm 18.0-21.0, glucose 2.0-1.0; formalin 2.0-1.5; aluminum hydroxide - the rest. The vaccine in question is of high specificity, safety and immunogenicity.

EFFECT: higher efficiency.

5 ex, 1 tbl

Anti-viral agent // 2316320

FIELD: pharmaceutical industry, in particular agents for viral disease treatment including HIV infection (AIDS and HIV-associated diseases).

SUBSTANCE: claimed agent in form of suppository contains per one 2 g suppository Fullevir (fullerenopolyaminocaproic acid sodium salt) 20 mg as active ingredient and ancillary substances such as propylene glycol 200 mg and balance: Vitepsol.

EFFECT: effective agent having no adverse influence on peripheral blood and body systems.

FIELD: pharmaceutical industry, pharmacy.

SUBSTANCE: invention relates to producing interferon-containing suppositories. Interferon-containing suppository comprises a fatty base, emulsifier and human lyophilized interferon purified from low-molecular and high-molecular components of the definite antiviral activity. Mixture of mono-, di- and triglycerides and saturated fatty acids from coconut oil and palm oil of sort "Vitepsol" H-15 and W-35 taken in equal ratio are used as a fatty base. Distilled monoglycerides taken in the definite ratio are used as an emulsifier. Above describes suppositories retain specific activity and effectiveness in treatment of viral and bacterial diseases for a long time and show stability against mechanical effects.

EFFECT: improved and valuable properties of suppository.

1 tbl, 1 ex

FIELD: chemical-and pharmaceutical industry.

SUBSTANCE: claimed agent contains DNA sodium salt isolated from bovine testis and is useful in peroral application in form of powder, tablets, pellets, and capsules for treatment and prophylaxis of anemia, secondary immunodeficit conditions and as bracing agent. Agent may additionally contains biologically active components and filler.

EFFECT: effective and convenient agent having antianemic, immunomodulating and adaptogenic actions.

3 cl, 3 ex, 6 tbl

FIELD: medicine, chemical-pharmaceutical industry, homeopathy, gynecology.

SUBSTANCE: invention relates to creature of novel vaginal homeopathic suppository that can be used in treatment of female genitals. Invention proposes nine variants of homeopathic suppository containing sea-buckthorn oil, homeopathic oily extract, oils and homeopathic essences. Proposed suppositories possess the enhanced bioavailability based on addition of homeopathic oily extract in the prescription, improve regenerative function of organs and tissues and enhance effect of basic components that allows their using in homeopathy in treatment of fungal colpitis, vulvovaginitis, endometriosis and papillomas.

EFFECT: improved and valuable medicinal properties of suppository.

9 cl, 20 ex

FIELD: medicine, pharmaceutical industry.

SUBSTANCE: the present innovation deals with developing new medicinal preparations for treating urogenital diseases. The suggested preparation for treating prostatic diseases designed as a suppository that contains the complex of bioregulatory peptides of cattle prostate as a powder with the content of water-soluble peptides being 20%, not less and the foundation, moreover, antimicrobial preparation being the antibiotic or anti-viral preparation, or anti-protozoan preparation, or the mixture of antibiotic and anti-protozoan preparation taken at 1:1 ratio, or the mixture of anti-viral and anti-protozoan preparation taken at 1:1 ratio and the foundation at the following ratio of the components, g/suppository: complex of bioregulatory peptides of cattle prostate 0.05-0.4; antimicrobial preparation 0.7, not more; the foundation - sufficient quantity to obtain the suppository of 2.15-2.35 g weight. The suggested preparation in the form of suppository is of high level of organotropic activity and high level of antimicrobial action that enables to implement etiotropic and pathogenetic therapy of bacterial prostatitis. The innovation provides the action upon an agent and all pathological processes in the affected organ. The innovation enables to treat bacterial prostatic inflammation and that of functionally associated organs.

EFFECT: higher efficiency of therapy.

4 cl, 132 ex, 32 tbl

FIELD: chemical-pharmaceutical industry.

SUBSTANCE: it has been suggested to apply a solid composition for manufacturing a pharmaceutical tablet or a suppository, its melting point being 25°C or higher and it contains an uninterrupted lipid component that contains either one or more than one galactolipids, one or more glyceride ether of fatty acids, possibly one or several out of the following: water and mono-triatomic alcohol at the quantity being up to 15 weight% against the weight of composition, and one or more agent chosen among pharmacologically active agent. The method for manufacturing the composition mentioned includes mixing galactolipids and glyceride ethers of fatty acids followed by dissolving pharmacologically active agents in a liquid phase, cooling up to a solid state and forming a tablet or a filled capsule. Pharmaceutical tablet or suppository are depicted that include uninterrupted lipid phase possibly containing an inert nucleus and, also, food tablets or suppositories of similar composition that include food agents instead of pharmacologically active agent and, possibly, having got one or more submembranes consisting of food excipients. The innovation provides economy and increased comfort in usage.

EFFECT: higher efficiency.

42 cl, 13 ex, 10 tbl

FIELD: microbiology, biotechnology, medicine, in particular production of living vaccines against viral infections.

SUBSTANCE: invention relates to living vaccines for rectal administration against viral infections based on attenuated recombinant Salmonella strains bearing protective viral antigens. Suppository for immunoprophylaxis of viral infections contains (per one 2 g suppository, mass %): cell slurry of attenuated recombinant Salmonella strains transformed with pGEX-2T-TBI, pcDNA-TCI, or pKHBc bearing gene of protective viral antigens, mixed with suppository base in amount of 106-109 living cells 1 %; fatty base 94 %; and emulsifier T2 5 %. As fatty solid cooking oil (94 %); or hydrolyzed cotton oil (94 %); or mixture of cooking oil (60 %), Cocoa fat (24 %), and solid wax (10 %).

EFFECT: agent for inducing of humoral and cell immune response to respective viral antigen.

8 tbl, 11 ex, 3 dwg

FIELD: medicine, gynecology, proctology, pharmacy.

SUBSTANCE: invention relates to medicaments made as vaginal and rectal suppository used in treatment of proctological, non-inflammatory and inflammatory diseases of female pelvis organs, among them complicated with suppurative-inflammatory processes. Invention relates to an agent used in treatment of gynecological and proctological diseases it is made as a suppository and comprises 2-ethyl-6-methyl-3-oxypyridine succinate as an active component and a base chosen from group involving polyethylene oxide-1500, polyethylene oxide-400, cacao butter and hydrogenated fats. Invention provides prolonged release of a curative preparation in mucosa and soft tissues and provides its effect for about 12 h. The procedure is suitable and easy in performance and can be carried out by patients independently under ambulatory conditions.

EFFECT: valuable medicinal and pharmaceutical properties of agent.

3 cl, 2 tbl, 2 ex

FIELD: pharmaceutical industry, in particular agent for prophylaxis and treatment of rheumatoid arthritis and osteoarthritis.

SUBSTANCE: disclosed are suppository having mass of 2-2.5 g containing (g): diclofenac 0.0225-0.0525; glucosamine hydrochloride 0.175-0.525; lecithin 0.057-0.063; and balance: solid fat.

EFFECT: agent for prophylaxis and treatment of arthritis, degenerative joint and backbone diseases with increased anti-inflammatory action and reduced ulcerogenic action.

1 tbl, 3 ex

FIELD: medicine, pharmacy.

SUBSTANCE: invention relates to the development of a novel vegetable-base agent that can be used in treatment of inflammatory diseases of small pelvis organs and diseases of rectum. Agent used in prophylaxis and treatment of diseases of small pelvis organs and diseases of rectum comprises long turmeric (Curcuma longa L.) rhizome dried extract standardized by curcumin, anesthetic agent and pharmaceutically acceptable excipients. Agent is made as a suppository and comprises benzocaine as an anesthetic agent and vitepsol and cacao butter as a suppository base. Method for preparing agent used in prophylaxis and treatment of diseases of small pelvis organs and diseases of rectum involves mixing long turmeric dried extract standardized by curcumin with anesthetic agent and a pharmaceutically acceptable excipient. Method for prophylaxis and treatment of diseases of small pelvis organs and diseases of rectum involves using the proposed agent.

EFFECT: improved preparing agent, valuable medicinal properties of agent.

9 cl, 3 ex

FIELD: medicine.

SUBSTANCE: method involves smearing 10% Indomethacin ointment over duodenal mucous membrane as 3 cm long thin layer outward from the wound immediately after duodenotomy.

EFFECT: suppressing inflammatory phenomena in duodenal wall; preventing pancreatic exocrine secretion regulation disorder; preventing intraductal hypertension of pancreas.

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