RU2430733C2 - Pharmaceutical composition for treating prostate diseases (versions) - Google Patents

Abstract

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to medicine, namely to peptide preparations for treating prostate diseases. The pharmaceutical composition (the first version) comprises 7-60 wt % of a bioregulatory peptide complex of bovine prostate containing water-soluble peptides 9 % more, and 40-93 wt % of a chelate zinc-arginine-glycin complex. The pharmaceutical composition in the form of suppositories (the second version) contains 0.015-0.130 g of the bioregulatory peptide complex of bovine prostate and 0.086-0.200 g of the chelate zinc-arginine-glycin complex as a bioactive compound in one fat-based suppository 2.7-2.9 g.

EFFECT: higher efficacy of the preparation, wider spectrum of its therapeutic action and minimised side effects.

3 cl, 5 ex, 3 tbl

Description

The claimed invention relates to medicine, in particular to peptide-based drugs, and in particular to pharmaceutical compositions containing an agent for treating diseases of the prostate gland (prostate). It can find application in the wide clinical and outpatient practice of treating these diseases.

Known agent for the treatment of prostate diseases, which is a preparation of a polypeptide nature, obtained by extraction from the prostate gland of cattle, followed by precipitation with acetone [RU No. 1417244, M. CL ⁵ . A61K 35/52, 1994]. The specified tool is a complex of bioregulatory peptides of the prostate gland of cattle with a content of water-soluble peptides of at least 20%. It is produced by the pharmaceutical industry under the brand names "Prostatilen" and "Samprost" in the form of lyophilized sterile powder.

The specified tool was used and is used to treat prostate diseases in the form of a solution for injection, for which the powder is dissolved in water or in an isotonic solution of sodium chloride, or in a 0.5% aqueous solution of novocaine. Injections of “Prostatilene” help to reduce prostate edema, reduce leukocyte infiltration of interstitial tissue and venous thrombosis [Auth. St. SU No. 1740004, M. cl. ⁵ . AK61K 35/48, 1992], in addition, have a tonic effect on the smooth muscles of the bladder [RU No. 2058780, M. cl ⁶ . A61K 35/48, 1996]. "Prostatilen" is also used as part of complex therapy for the treatment of prostate diseases [RU No. 2281101, M. cl. ⁷ . AK61K 31/495, 2006], for the treatment of chlamydial urethritis complicated by prostatitis [RU No.2003303069 / 14, M. cl. ⁷ . A61K 35/48, 2000], for the treatment of chronic pyelonephritis [RU No. 2104011, M. cl ^6. , A61K 35/48, 1998], etc.

The later developed method for producing a complex of bioregulatory peptides of the cattle prostate by ultrafiltration provides a preparation with a content of water-soluble peptides of at least 9% (most often 12-18%), but free of ballast protein fractions, fats, etc. It is also produced on an industrial scale under the brand name "Prostatilen" [FSP 42-0182-6850-05].

However, injections of "Prostatilen" are not effective enough in the treatment of chronic forms of prostate diseases. It is with chronic diseases of the prostate that most often encounter urologists and andrologists.

The technical result, to which the claimed invention is directed, is to increase the effectiveness of the pharmaceutical composition.

This result is achieved in that the pharmaceutical composition, comprising a complex of bioregulatory peptides of the prostate gland of cattle with a water-soluble peptide content of at least 9%, additionally contains a zinc-arginine-glycine chelate complex in the following ratio of components in the composition (% wt.):

complex of bioregulatory peptides cattle prostate gland - 7-60; zinc-arginine-glycine chelate complex - (first version of the pharmaceutical composition) 40-93.

The complex of bioregulatory peptides of the prostate gland of cattle ("Prostatilen") is produced by the industry in accordance with FSP No. 42-0182-6850-05. It is a lyophilized white (or yellowish) powder, readily soluble in water, and contains at least 9% (possibly at least 20%) of water-soluble peptides.

The chelate complex zinc-arginine-glycine was obtained by us as follows (example 1).

Example 1. The synthesis of the chelate complex zinc-arginine-glycine

In a 25 ml beaker, 1 g of arginine (accurately weighed) and 0.43 g of glycine (exact weighed) are dissolved in 5 ml of water. To the resulting solution at room temperature and with constant stirring on a magnetic stirrer, 1.63 g (accurately weighed) of zinc sulfate heptahydrate is added. At the end of the complete dissolution of zinc sulfate, 1.76 g (accurately weighed) of barium hydroxide is added to the solution. The resulting mixture was stirred vigorously for 1 hour at room temperature and then left to stand for 12 hours. The precipitate of barium sulfate is filtered through a paper filter, washing the precipitate several times with small portions of water. The resulting filtrate is freeze-dried. The result is a white crystalline powder of the zinc-arginine-glycine complex. Product yield: 70-80%.

Gross formula: C ₈ H ₁₇ N ₅ O ₄ Zn

Structural formula:

Molecular Weight: 312.63 g / mol

Physical properties: white crystalline powder, readily soluble in water and in dilute solutions of mineral acids.

The content of active components in the complex:

- Arginine - 55.40%

- Glycine - 23.69%

- Zinc - 20.91%

When this complex is dissolved in water, coordination bonds between nitrogen and zinc atoms break, with the formation of a positively charged structure due to the addition of hydrogen atoms to nitrogen atoms of amino groups:

The aqueous solution of the complex has a pH in the range of 8-10, which indicates the presence of free hydroxyl groups in the solution. If we assume that the dissociation of the complex proceeds to its complete destruction in solution with the formation of the zwitterions of the corresponding amino acids and the Zn ²⁺ cation, then inevitable formation of zinc hydroxide, which is slightly soluble in water, would occur, which would cause cloudiness of the complex solution and, in the long term, zinc hydroxide precipitation in sediment. This is not observed when maintaining an aqueous solution of zinc-arginine-glycine for 24 months.

The toxicity of the zinc-arginine-glycine complex was studied in accordance with the requirements of the Guidelines for the Experimental (Preclinical) Study of New Pharmacological Substances (2005), including acute toxicity with a single administration, subacute and chronic toxicity with prolonged administration of the substance. A study of acute toxicity was performed on 72 white outbred male mice; a study of subacute toxicity was performed on 42 white mongrel male rats; a study of chronic toxicity was performed on 76 male guinea pigs.

When studying acute toxicity, it was found that a single administration of the studied drug to animals in a dose exceeding the therapeutic recommended for clinical use by more than 5000 times does not cause toxic reactions, which indicates a large therapeutic breadth of the drug.

The study of subacute and chronic toxicity of the drug indicates the absence of side effects with prolonged use of the drug in doses exceeding the therapeutic by 100-1000 times. During the study, there was no significant effect of the studied drug on the morphological, biochemical parameters in the peripheral blood, as well as on ESR and erythrocyte resistance. Along with this, the content of total protein in blood serum was determined by the method of Lowry, potassium and sodium by plasma spectrophotometry. After the experiment was completed, a pathomorphological study of the brain and spinal cord, spinal ganglia, thyroid gland, parathyroid glands, adrenal glands, testes, pituitary gland, heart, lungs, aorta, liver, kidneys, bladder, pancreas, stomach, small intestine, colon, thymus, spleen, lymph nodes, bone marrow.

When assessing the general condition of animals, morphological and biochemical parameters of peripheral blood, the morphological state of internal organs, the state of the cardiovascular and respiratory systems, and liver and kidney function, pathological changes in the body were not detected.

The absence of the general toxic effect of the zinc-argonine-glycine complex allows us to recommend the pharmaceutical composition as an active principle for conducting in vivo tests for prostate diseases. The pharmacological effect of the claimed composition on an experimental model of chronic prostatitis was studied.

To obtain a pharmaceutical composition, Prostatilene lyophilized powder was mixed with zinc-arginine-glycine complex powder.

Example 2. The effect of the claimed pharmaceutical composition on the course of experimental chronic prostatitis

The study was conducted on 44 white outbred male rats with an average body weight of 220 g. Bacterial prostatitis in rats was induced by transurethral administration of a suspension of enterotoxigenic E. coli K-2935 (01: K1: H-). After light ether anesthesia, the animals, after treatment of the external opening of the urethra with 70% alcohol, introduced a catheter greased with glycerol into the urethra, through which 0.2 ml of a suspension of bacteria (10 ⁹ CFU / ml) was installed with a syringe. Control animals were injected with a 0.2 ml saline solution through a catheter. 45 days after bacterial infection, rats with developed chronic prostatitis were injected subcutaneously daily with a solution of the pharmaceutical composition at a dose of 0.5 mg / kg per injection. The introduction of the drug was continued for 10 days. The control groups consisted of intact animals, animals with chronic prostatitis without treatment, and rats receiving 0.9% saline according to the same injection schedule.

All rats were euthanized by decapitation on the 70th day of the experiment. The animals underwent complete autopsy, carefully examined the testes and their appendages, the prostate gland, seminal vesicles, bulbourethral glands, the bladder, kidneys with ureters, and other internal organs. For histological examination, the ventral part of the prostate was fixed in 10% neutral formalin and, after standard wiring, through a series of solvents, it was poured into paraffin. Tissue sections 5-7 μm thick were stained with hematoxylin-eosin.

To compare the severity of prostatitis in different groups, the following 5-point scale was developed:

1 point - traces of prostatitis (small foci of fibrosis, interstitial lymphocytic and histiocytic infiltration);

2 points - foci of chronic prostatitis with inflammatory infiltration (mainly interstitial), occupying less than 1/4 of the gland section;

3 points - foci of prostatitis with large inflammatory infiltrates around the ducts, acini and blood vessels, with an accumulation of leukocytes in the glands of the glands, occupying 1 / 4-1 / 2 sections of the gland;

4 points - pronounced prostatitis, exciting 1 / 2-3 / 4 sections of the gland;

5 points - pronounced prostatitis with a lesion of more than 3/4 gland sections.

As a result of the study, it was found that the use of the pharmaceutical composition in 7 cases out of 12 contributed to the positive dynamics of the inflammatory process in the prostate gland. On histological examination, mainly signs of chronic inflammation were observed, capturing less than 1/2 section of the gland. However, in most ducts and acini inflammatory infiltrates were not detected. The average severity of prostatitis was 1.82 + 0.06, as can be seen from table 1.

Table 1.
Characterization of the severity of chronic bacterial prostatitis in rats Group of animals The number of rats in the group The severity of prostatitis in points Total With prostatitis To the rat in the group On a rat with prostatitis Intact rats 9 - - - Control 1 (E.coli) (without treatment) eleven 8 2.93 ± 0.14 4.05 Control 2 (E. coli + physical solution) 12 7 2.46 + 0.17 3.79 E. coli + claimed pharmaceutical composition 12 7 1.82 ± 0.06 * 2.81 * * P <0.05 compared with indicators in 1 and 2 control groups.

Thus, the use of the pharmaceutical composition parenterally contributed to the activation of local immunity reactions, increase the antimicrobial protection of prostate tissue, which was accompanied by a decrease in inflammatory reactions.

As an alternative to injectable solution as a dosage form, suppositories are used in the practice of treating prostate diseases in which Prostatilene is the active substance (substance). The suppository dosage form gives a quick pharmacological effect due to the rapid absorption of the substance by the mucous membrane of the rectum and the close relative position of the prostate (diseased organ) and the rectum (organ of drug administration).

Known means for the treatment of prostate diseases in the form of suppositories containing 0.04-0.06 g of a complex of bioregulatory peptides of the cattle prostate in the form of a powder containing at least 20% water-soluble peptides and 1.15-1.35 per suppository g basis - mono-, di- and triglycerides of natural saturated fatty acids with a carbon chain length of C ₁₂ -C ₁₈ , i.e. fats [RU No. 2112504, M. cl ⁶ . A61K 9/02, 1998]. The action of the obtained suppositories was studied experimentally in animals. Clinical data on the treatment of prostate diseases in humans are not given.

The agent according to RU No. 2112504 has an organotropic effect on the prostate, however, the level of this activity is not high due to the low content of Prostatilene in it.

A rectal agent for the treatment of chronic prostatitis occurring against the background of immunodeficiency is also known, including one suppository weighing 2.0-2.2 g 4-6 mg of the concentrate of the extract of the prostate gland of cattle, as well as a concentrate of liquid purified α-interferon with activity 10 -40 thousand ME or recombinant interferon with an activity of 250-500 thousand ME [RU No. 2160114, M. cl ⁶ . A61K 35/48, 2000]. Gelatin, glycerin and sodium carbonate — bicarbonate buffer — are used as the base. The concentrate of the extract of the prostate gland of cattle is an intermediate product of the production of dry injectable "Prostatilene". In clinical trials, the agent according to RU No. 2160114 showed a higher efficiency in the treatment of chronic prostatitis than therapy with prostatilene injections.

However, side effects are possible: fever, headaches, arthralgia. The action of interferon is aimed mainly at antiviral protection. In addition, interferon is unstable in time in this composition.

Closest to the claimed tool in terms of essential features is a tool for the treatment of prostate diseases, made in the form of a suppository, containing on a suppository weighing 2.15-2.35 g of 0.05-0.40 g of a complex of bioregulatory peptides of the prostate gland of cattle and up to 0.7 g of antimicrobial agents [RU No. 2304979, M. cl. ⁷ . A61K 38/02, 2007]. As an antimicrobial agent, the suppository may include antibiotics from the group of fluoroquinolones or from the group of penicillins, or from the group of cephalosporins, or from the group of tetracyclines, or from the group of sulfanilamides; ribavirin or lamivudine were used as an antiviral agent; suppositories may also include an antiprotozoal agent such as metronidazole, ornidazole and tinidazole. Perhaps a combination of these funds in one suppository. The effectiveness of suppositories was studied on a model of prostatitis caused by the action of various bacteria in white male rats.

However, when using suppositories according to RU No. 2304979 in a person, usual complications are possible caused by a rather high content of antimicrobial agents, such as the development of antibiotic resistance, allergic reactions on the rectal mucosa, and also a decrease in the level of immunity usually caused by antibiotics.

The technical result achieved in the claimed invention is to increase the effectiveness of the drug and minimize side effects.

The specified technical result is achieved in that the pharmaceutical composition for treating prostate diseases in the form of suppositories, comprising a complex of bioregulatory peptides of the cattle prostate in the form of a powder with a content of water-soluble peptides of at least 9%, a bioactive compound and a fat base, as a bioactive compound contains zinc-arginine-glycine chelate complex in the following ratio of components in grams per suppository weighing 2.7-2.9 g:

complex of bioregulatory peptides cattle prostate gland - 0.015-0.130; zinc-arginine-glycine chelate complex - 0,086-0,200 fat base - (second embodiment of the pharmaceutical composition) rest.

A pharmaceutical composition in the form of a suppository may also further comprise 0.04-0.06 g of dimethyl sulfoxide per suppository.

The complex of bioregulatory peptides of the prostate gland of cattle obtained in accordance with FSP No. 42-0182-6850-05.

Zinc-arginine-glycine chelate complex was obtained by us as described in example 1.

As a base, for example, cocoa butter, edible fats, a mixture of mono-, di- and triglycerides of fatty acids, known as Vitepsol, and other fatty bases permitted in the Pharmacopoeia can be used.

Suppositories are prepared as follows.

Example 3. The technology of manufacturing suppositories

1. Getting suppository mass.

1.1. Getting premix.

Premix is prepared in a heated jacket homogenizer.

To prepare the premix, a sufficient amount of suppository base (Witepsol / Witepsol / W35) is placed in the homogenizer and melted.

Then pre-weighed components are placed: “Prostatilen” substance, zinc-arginine-glycine complex, possibly dimethyl sulfoxide. Homogenization of the premix is carried out for 20-30 minutes. At the end of the process, the quality of the premix is checked - the mass should be uniform.

1.2. Mix premix with base

The suppository base (Witepsol / Witepsol / W35) is placed in a reactor and melted. Then the premix is added to the base and mixed for 1 hour. Suppository mass should be homogeneous without visible inclusions.

2. Forming suppositories

2.1. Filling of blister strip packaging with suppository mass (packaging).

The preparation of the drug in blister strip packaging is carried out on a dosing machine. The suppository mass is transferred to the hopper of the metering machine. Packing suppository mass in the cells is carried out at a constant temperature and stirring.

Contour cell packaging filled with a suppository mass is wound into rolls using guide rollers and transferred to the operation “Suppository Cooling”.

2.2. Suppository cooling

Suppositories are cooled in a refrigerator at a temperature of 10-15 ° C for 1-2 hours.

The inventive pharmaceutical composition has passed clinical trials in the treatment of chronic prostatitis and prostate adenoma.

Example 4. The effectiveness of the claimed pharmaceutical composition in the form of suppositories in patients with chronic prostatitis

A study of the effectiveness of the clinical use of suppositories containing a pharmaceutical composition was conducted in 23 patients with chronic prostatitis aged 27-49 years.

The main manifestations of chronic prostatitis were aching pains of typical localization, various disorders of urination, disorders of the ability to commit sexual intercourse and signs of a neurotic condition.

All patients previously received traditional medicines of symptomatic and pathogenetic action, the use of which showed a short-term therapeutic effect, requiring an increase in the dose of drugs per treatment course and their long-term use. Suppositories were administered rectally at night before bedtime for 10-15 days, depending on the severity of the disease. The control group consisted of 11 similar patients who were prescribed traditional treatment.

The effectiveness of treatment with suppositories was evaluated based on the dynamics of complaints of patients, a general clinical study of blood and urine, a biochemical study of blood, a blood coagulogram before and after treatment. The degree of abdominal pressure during urination and the nature of the urine stream were expressed in points from 1 to 5 (1 - normal, 5 - the largest changes in the indicator). The maximum and average speeds and the time of urination, the time to reach the maximum speed of urination were determined, and the fluorometric index was evaluated. Along with the above studies, a prostate gland was palpated, a laboratory study of its secretion, a more in-depth study of the state of copulative function. An ultrasound examination of the prostate was also performed.

The main focus in assessing the results of treatment of chronic prostatitis was given to clinical criteria. At the end of treatment, the pain completely disappeared in 75.3% and significantly decreased in 24.7% of patients presenting the corresponding complaints. Of the number of patients suffering from impaired sexual function, 57.3% indicated its full recovery and 40.9% noted an improvement. The positive effect of suppositories was manifested both in relation to improving the quality of an erection, and strengthening the orgasm and eliminating its pain. An increase in the duration of sexual intercourse was also noted. By the end of treatment, 52.8% of patients reported restoration of libido.

Pollakiuria (increased urination) completely stopped disturbing 89.4% of patients. The need for nocturnal urination has disappeared. Stranguria (difficulty urinating) ceased to bother 81.6% of patients, 22.3% noted a marked increase in urine flow and relief of urination. The total information about the change in the act of urination in patients with chronic prostatitis after treatment with suppositories is presented in table 2.

Table 2.
The influence of the claimed pharmaceutical composition in the form of suppositories on the urodynamics of patients with chronic prostatitis Indicators Before treatment After treatment with suppositories Average urination rate (ml / s) 18.2 + 1.3 24.3 + 2.1 * Maximum urination rate (ml / s) 21.5 + 2.7 25.8 ± 3.5 Time to reach maximum urination rate (ml / s) 3.4 + 0.2 1.3 ± 0.1 * * - P <0.05 compared with the indicator before treatment.

Urofluograms recorded after treatment in patients with stage I and II chronic prostatitis showed restoration of the main parameters of urination to normal values. In stage III of the disease, this was prevented by a decrease in the elasticity of the neck of the bladder due to sclerotic changes in the tissue of the prostate gland, but they also showed a marked increase in the stream of urine. On palpation of the prostate gland through the rectum, a tendency to restore its size and consistency after treatment with suppositories was determined. At the same time, the compaction areas disappeared, and the study itself became painless. The observed decrease in the size of the prostate gland, regarded as a result of a decrease in edema of the interstitial tissue and indicating the elimination of the activity of the inflammatory process in it, was also confirmed by the results of ultrasound diagnostics. General clinical and biochemical blood tests performed in dynamics did not reveal any significant deviations from normal values. Side effects were not observed.

Due to the restoration of prostate function under the influence of suppositories, an improvement in the properties of its secretion was observed. This ensured an increase in the content of motile spermatozoa in the ejaculate by 21.4%. The decrease in the activity of the inflammatory process was confirmed by a decrease in the number of leukocytes in the ejaculate, prostate secretion and urine. At the same time, a decrease in the content of cells of desquamated epithelium in the test material was observed.

Example 5. The effectiveness of the claimed pharmaceutical composition in the form of suppositories in patients with prostate adenoma

Suppositories were used to treat 12 patients with stage I-II prostate adenoma aged 54-73 years. The characteristic symptoms of the disease in patients with prostate adenoma were persistent dysuria, not amenable to traditional methods of treatment, weakened stream of urine, decreased ability to perform sexual acts. All patients for various times received traditional medicines with symptomatic and pathogenetic effects.

Suppositories were administered at bedtime 20-30 days, depending on the clinical manifestations of the disease. The control group consisted of 9 similar patients who were prescribed traditional treatment. The effectiveness of treatment with suppositories was evaluated based on the dynamics of complaints of patients. The degree of abdominal pressure during urination and the nature of the urine stream were expressed in points from 1 to 5 (1 - normal, 5 - the largest changes in the indicator). The maximum and average speeds and the time of urination, the time to reach the maximum speed of urination were determined, and the fluorometric index was evaluated. An ultrasound examination of the prostate was performed. The dynamics of the results of the study in patients with prostate adenoma before and after the end of the course of treatment with suppositories is presented in table 3.

Table 3.
The influence of the claimed pharmaceutical composition in the form of suppositories on the state of urodynamics of patients with prostate adenoma Indicators Before treatment After treatment with suppositories Urinary retention time (min) 4,5 + 0,4 2.3 ± 0.1 * Urination count - in the daytime 7.9 ± 0.5 6.1 + 0.1 * - at night time 3.1 ± 0.2 2.7 + 0.1 The degree of abdominal pressure (points) 3.2 2.5 The nature of the urine stream (points) 3.6 2.1 * Average urination rate (ml / s) 10.6 ± 1.3 18.1 ± 1.4 * Maximum urination rate (ml / s) 15.8 + 2.1 20.4 ± 3.7 Time to reach maximum urination rate (ml / s) 6.8 + 0.2 4.3 + 0.1 * * - P <0.05 compared with the indicator before treatment

The condition of patients with prostate adenoma after treatment with suppositories was characterized by an improvement in subjective and objective indicators of urodynamics. In 48.5% of patients, increased libido was noted. Side effects were not observed.

Thus, the claimed pharmaceutical composition in the form of suppositories is effective in the treatment of inflammatory diseases of the prostate gland and dysuric disorders. Side effects were not observed.

Claims (3)

1. A pharmaceutical composition for the treatment of prostate diseases, comprising a complex of bioregulatory peptides of the prostate gland of cattle with a water-soluble peptide content of at least 9%, characterized in that it additionally contains a zinc-arginine-glycine chelate complex in the following ratio of components in the composition, wt .%:
complex of bioregulatory peptides cattle prostate 7-60 chelating complex zinc-arginine-glycine 40-93 2. A pharmaceutical composition for treating prostate diseases in the form of suppositories, comprising a complex of bioregulatory peptides of the cattle prostate in the form of a powder with a content of water-soluble peptides of at least 9%, a bioactive compound and a fat base, characterized in that the composition contains a bioactive compound chelating complex zinc-arginine-glycine in the following ratio of components, g per suppository weighing 2.7-2.9 g:
complex of bioregulatory peptides cattle prostate 0.015-0.130 chelating complex zinc-arginine-glycine 0,086-0,200 fat base rest 3. The rectal agent according to claim 2, characterized in that it additionally contains dimethyl sulfoxide in an amount of 0.04-0.06 g per suppository.