RU2542420C1 - Drug substance of polyprenylphosphates and beta-sitosterol and method for preparing it - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: invention represents a method for preparing a drug substance of polyprenylphosphates and beta-sitosterol consisting in pre-mixing polyprenylphosphates and sorbitol in a mortar, a size of which fits the total volume of the mixed substances. Between a wall and/or a bottom of the mortar and the introduced polyprenylphosphates, there is a layer of sorbitol; the blended mixture of the polyprenylphosphates in sorbitol is homogenised. Sorbitol, a dry mixture of polyprenylphosphates in sorbitol in a ratio of 1:8 and beta-sitosterol are added into a homogeniser, and pulsed homogenisation is performed for 10 minutes. The homogenisation is performed at a rotation rate of 500-700 rpm for 4 minutes, 1000-1200 rpm for 2 minutes and then 500-600 rpm for 4 minutes; the rotation rate variation requires a pause of 20 minutes. Each pause of the process is followed by the intensive agitation of a mixture cup; the prepared powder is sieved at a mesh size of 20 mcm; if a sieving weight makes less than 0.1% of the weight of the loaded ingredients; the homogenisation process is terminated.

EFFECT: preparing the drug substance with the maximum effective concentration of the active substances uniformly weight-distributed and higher bioavailability.

3 cl, 2 ex, 1 tbl, 7 dwg

Description

The invention relates to medicines and methods for their preparation. The drug is intended for the prevention and treatment of diseases of the heart and blood vessels, as well as dyslipidemia.

The closest analogue is a biologically active additive (BAA) and its production method (RU 2427284, publ. 08.27.2011).

However, the concentration of active components in the body obtained using this dietary supplement is significantly lower than therapeutic. In addition, the cooking technology described in patent RU 2427284, does not allow to achieve a uniform distribution of active substances in the tablet. As a result, the effectiveness of the use of dietary supplements decreases, and a pronounced therapeutic effect can be achieved only with prolonged use of the drug.

Various methods are used to mix active pharmaceutical substances and excipients (excipients) having different physicochemical properties: pre-dissolving and applying to the excipient, mixing molten APIs and substances, caramelization, etc.

The analysis shows that none of the above methods is suitable in our case, because there are limitations on the solubility and decomposition temperature of the active pharmaceutical substance sodium polyprenyl phosphate and sorbite excipient (filler).

Dry blending of polyprenyl phosphates and sorbitol is proposed.

            The technical result is to obtain a stable tablet form with the most effective concentration of active components uniformly distributed throughout the tablet mass and increased bioavailability of the ingredients.

The technical result is achieved by a method for producing a medicine based on polyprenyl phosphates and beta-sitosterol, which is characterized by rubbing polyprenyl phosphate and sorbitol in a mortar, the size of which corresponds to the total volume of the mixed substances, to obtain a dry mixture of polyprenyl phosphate and sorbitol, after which sorbitol, beta are homogenized - sitosterol and a dry mixture of polyprenyl phosphate and sorbitol, while homogenization is carried out for 10 minutes in a pulsed mode.

The mixing process consists of two main steps

Pre-mixing is carried out by grinding polyprenyl phosphates in sorbitol.

Homogenization of the resulting mixture of polyprenyl phosphates in sorbitol using a homogenizer.

Preliminary mixing of polyprenyl phosphates and sorbitol is carried out in a mortar, the size of which corresponds to the total volume of the mixed substances.

The optimal ratio for mixing by grinding: 20 g of sorbitol per 10 g of polyprenylphosphates.

To prepare the process, 20 g of sorbitol are poured into the mortar, the granule size is 80-120 mesh. Polyprenyl phosphates, with a total weight of 10 g, are administered in small portions of 1-2 g. After each administration, the polyprenyl phosphates are triturated by mixing with sorbitol to obtain a dry mixture. The grinding time should not be too long, because prolonged grinding leads to aggregation (sticking) of the crushed particles.

During dispersion, polyprenyl phosphates can be compressed and adhere to the walls of the mortar, so care should be taken to ensure that there is a sorbitol layer between the wall or bottom of the mortar and the introduced polyprenyl phosphates.

As lumps of polyprenyl phosphates form, which when combined with sorbitol lose their ability to adhere to the walls of the mortar, 3-5 g of sorbitol is added and mixed to obtain a homogeneous mixture. The resulting mixture is sent to stage 2.

Homogenization of the resulting mixture of polyprenyl phosphates in sorbitol using a homogenizer.

The mixture obtained in stage 1 is homogenized using a universal homogenizer for solid samples of the Universal Mixer GK type.

To do this, the calculated amount of sorbitol (but not more than 1 kg) and a mixture of polyprenyl phosphates in sorbitol are poured into the homogenizer. The optimal ratio of polyprenyl phosphate: sorbitol is -1: 8 (sorbitol introduced in stage 1 is taken into account).

The ingredients are loaded into the homogenizer in the following order:

1st layer - sorbitol, 1/2 of the calculated amount

2nd layer - a mixture of polyprenols and sorbitol

3rd layer - sorbitol, the second half of the estimated amount.

Homogenization is carried out for 10 minutes in a pulsed mode. After each stop of the process - intensive shaking of the bowl with the mixture.

Time and speed of rotation:

4 minutes - 500-700 rpm, a break of 20 minutes.

2 minutes - 1000-1200 rpm, a break of 20 minutes.

4 minutes - 500-600 rpm - the end of the process.

           To verify the uniformity of the mixture, the resulting powder should be sieved through a sieve with a mesh size of 20 microns. When screenings appear on the surface of the screen, it is necessary to determine its weight and composition. If the dropout weight is less than 0.1% of the weight of the loaded ingredients, the homogenization process ends. If the screening weight is more than 0.2% of the weight of the loaded ingredients, and polyprenyl phosphate particles are visible in the screening, the homogenization process continues at a rotation speed of 1000-1200 rpm for one minute. After this, re-screening and determination of the weight of the screenings are made. If the dropout meets the conditions mentioned above, the process stops.

Visual assessment is made by looking at the surface of the obtained powder: with the naked eye at a distance of 25 cm, no separate particles, spangles or blotches of yellow or yellow color should be observed.

When preparing the mixture to obtain a tablet form of a drug based on polyprenylphosphates and beta-sitosterol, beta-sitosterol is also added along with sorbitol at the homogenization stage.

The optimized ratio of active pharmaceutical substances (polyprenyl phosphates and beta-sitosterol) and excipient (sorbitol) for high-quality tablet compression, elimination of sticking, and destruction of the 250 mg tablet form is 8 mg of polyprenyl phosphates and 10 mg of beta-sitosterol. The decrease in tablet weight by reducing the input of sorbitol does not allow direct tabletting of the tablet mass. At the same time, an increase in the mass of introduced substances> 10 mg for beta-sitosterol and, in particular,> 8 mg for polyprenyl phosphate leads to the destruction of the tablet form and sticking.

Thus, the maximum content of polyprenyl phosphates in one tablet is 8 mg, and beta-sitosterol is 10 mg.

           To determine the optimal content of beta-sitosterol in the preparation, 3 samples of the mixture are prepared containing the substance of beta-sitosterol (BSS) in an amount of: 0.0 mg (control); 6.0 mg (sample 1) and 10.0 mg (sample 2) and the filler is sorbitol.

All participating in the experiment animals cause severe hyperlipidemia by intraperitoneal administration of poloxamer 407 at a dose of 7.5 mg / mouse. The indicated dose of poloxamer is administered four times, every other day, in the morning (from 10 a.m. to 11 a.m.). To study the effect of drugs on cholesterol and triglycerides in the blood serum of hyperlipidemic mice, the following groups of experimental animals are formed (7 mice in each group):

1. The control group - mice with hyperlipidemia, which are intragastrically injected with 50 mg / mouse of food sorbitol in 0.5 ml of solvent (RPMI-1640 medium).

2. Experimental group I - mice with hyperlipidemia, which are administered intragastrically 50 mg / mouse of sample No. 1 in 0.5 ml of solvent.

3. Experimental group II - mice with hyperlipidemia, which are intragastrically injected with 50 mg / mouse of sample No. 2 in 0.5 ml of solvent.

All studied drugs are administered daily, starting from the day after the first administration of poloxamer, in the morning, through a gastric tube. Solutions and suspensions of preparations in a solvent are prepared immediately before administration to animals.

Blood samples are taken from animals from the tail veins (the tip of the tail is cut into mice and 20-30 μl of blood is taken into a plastic capillary), three times a day, with the first sample taken one day after the second injection of the poloxamer (that is, after two injections of the studied drug) .

The data obtained are presented in FIG. 1. Sample No. 2, containing the highest possible dose of BSS, significantly lowers cholesterol.

            FIG. 1. Shows the effect of beta-sitosterol on serum cholesterol in hyperlipidemic mice.

The specific properties of the resulting mixture can be illustrated by the following examples.

Example 1

Prepare 6 samples of a dosage form containing 10 mg BSS and different doses of polyprenyl phosphates (PPF).

Table 1 shows the numbers of the samples and the corresponding BSS content in the drug.

Table 1. Samples of the mixture and their corresponding dosage forms.

All participating in the experiment animals cause severe hyperlipidemia by intraperitoneal administration of poloxamer 407 at a dose of 7.5 mg / mouse. The indicated dose of poloxamer is administered four times, every other day, in the morning (from 10 a.m. to 11 a.m.). To study the effect of drugs on cholesterol and triglycerides in the blood serum of hyperlipidemic mice, the following groups of experimental animals are formed (7 mice in each group):

            1. Control group I - mice with hyperlipidemia, which are administered intragastrically 50 mg / mouse of sample No. 1 in 0.5 ml of solvent.

2. Experimental group I - mice with hyperlipidemia, which are intragastrically injected with 50 mg / mouse of sample No. 2 in 0.5 ml of solvent.

3. Experimental group II - mice with hyperlipidemia, which are administered intragastrically 50 mg / mouse of sample No. 3 in 0.5 ml of solvent.

4. Experimental group III - mice with hyperlipidemia, which are administered intragastrically 50 mg / mouse of sample No. 4 in 0.5 ml of solvent.

5. Experimental group IV - mice with hyperlipidemia, which are administered intragastrically 50 mg / mouse of sample No. 5 in 0.5 ml of solvent.

6. Experimental group V - mice with hyperlipidemia, which are intragastrically injected with 50 mg / mouse of sample No. 6 in 0.5 ml of solvent.

All studied drugs are administered daily, starting from the day after the first administration of poloxamer, in the morning, through a gastric tube. Solutions and suspensions of preparations in a solvent are prepared immediately before administration to animals.

Blood samples are taken from animals from the tail veins (the tip of the tail is cut into mice and 20-30 μl of blood is taken into a plastic capillary), three times a day, with the first sample taken one day after the second injection of the poloxamer (that is, after two injections of the studied drug) .

The data obtained are presented in FIG. 2. The addition of PPF dose-dependently enhances the effect of BSS and leads to an additional decrease in cholesterol, with the maximum effect being observed at the maximum possible PPF content in the tablet (8 mg).

FIG. 2. Shows the effect of polyprenyl phosphate on serum cholesterol.

Thus, the maximum therapeutic effect, determined by the level of lowering cholesterol, is achieved with a maximum content of polyprenyl phosphates (8 mg), and beta-sitosterol (10 mg) in one tablet.

           Example 2

Based on the mixture described above containing 8 mg of polyprenylphosphates and 10 mg of beta-sitosterol, tablets weighing 250 mg with sorbitol as a filler are prepared as a finished dosage form (GLF).

In mice of hybrids of the first generation (C57Bl / 6xDBA / 2) F ₁ or mice C57 B1 / 6 females, at the age of 2 months, pronounced hyperlipidemia is induced by intraperitoneal administration of poloxamer 407 at a dose of 7.5 mg / mouse. The indicated dose of poloxamer is administered two to four times, every other day, in the morning (from 10 a.m. to 11 a.m.). The following groups of experimental animals are formed:

1. Control group I - mice with hyperlipidemia, which are injected intragastrically with a solvent (RPMI-1640 medium - 0.5 ml) - placebo group.

2. Control group II - mice with hyperlipidemia.

3. Experimental group III - mice with hyperlipidemia, which are orally administered 0.2 mg / mouse sodium polyprenyl phosphate in 0.5 ml of solvent. (6.25 mg GLF).

4. Experimental group IV - mice with hyperlipidemia, which are orally administered with 0.8 mg / mouse of sodium polyprenyl phosphate in 0.5 ml of solvent (25 mg of GLF).

5. Experimental group V — mice with hyperlipidemia, which were orally administered with 1.6 mg / mouse of sodium polyprenyl phosphate in 0.5 ml of solvent (50 mg GLF).

6. Experimental group VI - mice with hyperlipidemia, which are orally administered with 2.4 mg / mouse of sodium polyprenyl phosphate in 0.5 ml of solvent (75 mg of GLF).

7. Experimental group VII - mice with hyperlipidemia, which are orally administered with 3.2 mg / mouse of sodium polyprenyl phosphate in 0.5 ml of solvent (100 mg GLF).

All studied drugs are administered daily, starting from the day after the first administration of poloxamer, in the morning. Solutions and suspensions of preparations in a solvent are prepared immediately before administration to animals.

The number of animals in the group is formed on the basis of the need for triple blood sampling. Blood samples are taken from animals from the group after decapitation, three times, every other day, while the first sample is taken one day after the second injection of poloxamer (that is, after two injections of the test drug).

Determination of total cholesterol, triglycerides, LDL cholesterol, HDL cholesterol is carried out on an Olympus 640 biochemistry analyzer (Japan) using Olympus commercial kits (Cat. No. OSR6516, OSR6133, OSR6283, OSR6587, respectively).

In group I mice, cholesterol and triglyceride indices (Fig. 3) did not significantly (P> 0.05) differ from the corresponding indices of group II mice.

FIG. 3 shows a comparison of groups I (placebo) and II (control) in terms of cholesterol and triglycerides.

Comparison of groups of animals III-VII that received different GLF with the control group II allows us to determine the effective dose by the studied parameters.

FIG. 4 shows the effect of GLF on blood cholesterol in hyperlipidemic mice.

The blood cholesterol level of hyperlipidemic animals (Fig. 4) is most reduced in group IV after administration of 25 mg of GLF (0.8 mg of sodium polyprenyl phosphate), which corresponds to 1.25 g / kg. Doses of 75 and 100 mg did not significantly affect cholesterol levels.

A similar picture is observed when examining the level of triglycerides in the blood (Fig. 5): the maximum effect is observed with the introduction of 25 mg of the drug. Doses of 50-75 mg have a slightly smaller but significant effect (P <0.05) of a decrease in triglycerides.

FIG. Figure 5 shows the effect of GLF on the level of triglycerides in the blood of hyperlipidemic mice.

FIG. 6 - the effect of GLF on the level of HDL in the blood of hyperlipidemic mice.

FIG. 7 - the effect of GLF on the level of LDL in the blood of hyperlipidemic mice.

A significant effect of GLF on the level of HDL (Fig. 6) and LDL (Fig. 7) is observed in a wider range of doses - from 6.25 to 100 mg. At the same time, there is an increase in the level of HDL and a decrease in LDL.

Claims (3)

1. A method of obtaining a medicine based on polyprenyl phosphates and beta-sitosterol, which consists in pre-mixing polyprenyl phosphates and sorbitol in a mortar, the size of which corresponds to the total volume of the substances to be mixed, while there is a sorbitol layer between the wall and / or bottom of the mortar and the introduced polyprenyl phosphates obtained by mixing, the mixture of polyprenyl phosphates in sorbitol is homogenized, while sorbitol is added to the homogenizer, a dry mixture of polyprenyl phosphates in sorbitol in a ratio of 1: 8 and beta-sitosterol, and homogenization is performed for 10 minutes in a pulsed mode, and homogenization is carried out at a rotation speed of 500-700 rpm for 4 minutes, 1000-1200 rpm for 2 minutes and then at a speed of 500-600 rpm for 4 minutes, while changing the rotation speed, take a break of 20 minutes, and after each stop of the process, the bowl with the mixture is intensively shaken, after which the resulting powder is sieved through a sieve with a mesh size of 20 μm and if the screening mass is less than 0.1% by weight loaded ingredients, ol The homogenization process is completed. 2. A drug based on polyprenyl phosphates and beta-sitosterol for the treatment of hyperlipidemia, obtained by the method according to p. 1. 3. The drug according to claim 2, characterized in that it contains 8 mg of polyprenyl phosphates and 10 mg of beta-sitosterol.

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