RU2533231C1 - Supramolecular complex, possessing anti-inflammatory and angioprotective activity and method of obtaining thereof - Google Patents

Abstract

FIELD: chemistry.

SUBSTANCE: supramolecular complex, possessing an anti-inflammatory and angioprotective activity, which includes dihydroquercetin, arabinogalactan and water with a specified content of components. The method of obtaining the supramolecular complex includes mixing arabinoglactan with water to complete dissolution, then the addition of dihydroquercetin, heating the solution, mixing, with the following drying of the obtained solution by a method of spraying, under specified conditions.

EFFECT: increase of dihydroquercetin water solubility.

2 cl, 1 dwg, 5 tbl, 8 ex

Description

The group of inventions relates to the pharmaceutical industry, relates to the production of a water-soluble complex with increased pharmacological activity, consisting of a drug which is poorly soluble in water, dihydroquercetin and a water-soluble complexing agent.

The range of applications of dihydroquercetin (DHQ) is quite wide, it is used in the manufacture of various products, including in the pharmaceutical industry for the production of dietary supplements and medicines, as well as in the food industry as an antioxidant. Its disadvantage is the low bioavailability due to poor water solubility. Due to the widespread use of DHQ in the food, medical, and cosmetic industries, the task of obtaining a water-soluble complex of dihydroquercetin with high pharmacological activity and the development of improved methods for its preparation remains relevant.

It is known that in order to obtain water-soluble forms of drugs, various water-soluble natural oligo- and polysaccharides, for example dextran, inulin, maltodextrin and cyclodextrin, are added to the water-insoluble drug, which form water-soluble complexes with the target drug and, accordingly, increase its bioavailability.

Known water-soluble composition, including a sparingly soluble antibiotic in an amount of 20-60% and a natural polysaccharide selected from the group - dextran, inulin and maltodextrin, taken in a certain ratio. The known composition is obtained by mixing solutions of antibiotics and polysaccharides, followed by drying and isolation of a dry adduct, in which antibiotics with polysaccharides are linked by non-covalent and non-ionic bonds (US patent 6821959 B1, op. 23.11.2004).

A water-soluble composition is known comprising a sparingly soluble drug substance and cyclodextrin, preferably beta-cyclodextrin. As a sparingly soluble drug substance, the composition contains a non-steroidal anti-inflammatory drug (paracetamol, ibuprofen, ketoprofen, flufenamic and mefenamic acids, etc.), a steroid, prostaglandin, prostacyclin, barbiturate, sulfonamide, cardiac glycoside. Water-soluble intermolecular complexes of lipophilic organic compounds are formed in solution by intercalating their molecules into this cavity (J. Szejtli, Industrial Applications of cyclodextrins. - In Inclusion Compounds, v. 3. Ed. Atwood JL, Davies JE, Mcnicob DD, Academic Press, NY., 1984, p. 313-390).

A known complex based on dihydroquercetin, which is an aqueous solution of dihydroquercetin in molecularly encapsulated form in the form of a water-soluble associate. To obtain this complex, dihydroquercetin is homogenized in a melt of a surfactant, preferably Cremophor RH 40, or in a common ethanol solvent with a surfactant to dihydroquercetin ratio of 5: 1 in the temperature range of 40-70 ° C, followed by stirring with water and concentration by evaporation water (patent RU 2406496 C1, op. 20.12.2010). The invention improves the solubility and bioavailability of dihydroquercetin.

A disadvantage of the known complex is the low solubility of dihydroquercetin, as well as a complex and lengthy method for its preparation.

A known pharmaceutical composition comprising a sparingly soluble drug and arabinogalactan (AG), taken in a ratio of 1: (5-20) by weight, which is obtained by machining the mixture, preferably in ball mills, in particular planetary (patent RU 2337710 C2, op. November 10, 2008).

However, the method of obtaining a water-soluble composition requires the use of shock-abrasion and other intense mechanical stresses, including pressure and shear deformations, which leads to the destruction of the crystal structure of dihydroquercetin.

The closest to the claimed complex based on dihydroquercetin and the method for its preparation - the prototype, is a solid nanocomposite for the delivery of biologically active substances containing 0.1-15% of the active component, the nanocomposition of which contains taxifolin, 40-95% of the polymer, which the nanocomposition contains arabinogalactan, or polyethylene glycols, or polyvinylpyrrolidones, or polyvinyls of various molecular weights, 0-56% of a water-soluble component selected from a number of possible excipients: kollidon VA64 (co vinyl pyrrolidone and vinyl acetate example), kollidon 90F (high molecular weight polyvinyl pyrrolidone with M.M. 1,000,000 to 1,500,000), ludipress (modified lactose), powdered sugar, isomaltose, 0-6% hydrophobic or inert polymer in order to achieve controlled release of the drug substance, where As a hydrophobic component, the nanocomposite contains compritol 888 ATO (a composition of mono-, di- and triglycerides of behenic acid), and as an inert component - kollidon SR. (a mixture of polymers of polyvinylpyrrolidone and polyvinyl acetate).

A method for producing a nanocomposite containing taxifolin and arabinogalactan in a weight ratio of 1:10 includes the following steps: arabinogalactan (10 g) is dissolved in 10 ml of water; separately prepare a solution of taxifolin by dissolving the latter (1 g) in 10 ml of water with stirring and by heating in a thermostated vessel until taxifolin is completely dissolved; in a vessel with a solution of arabinogalactan with stirring and thermostating add a solution of taxifolin to obtain a homogeneous medium followed by lyophilization of the product (patent RU 2351352 C2, op. 10.04.2009).

The disadvantages of the known solid nanocomposite are the complexity of the composition, the use of a large number of imported hard-to-reach components, the high cost of the target product, as well as the length and complexity of its production (the need to use expensive freeze drying).

The objective of the group of inventions is to create a complex based on dihydroquercetin with increased solubility in water and a method for its preparation.

Effect: increasing the water solubility of dihydroquercetin in the complex and increasing its pharmacological activity, reducing the duration of the method of obtaining the complex.

The problem is solved by the claimed composition of the complex and the method of its production.

The complex includes dihydroquercetin, arabinogalactan and water, with the following components, wt.%:

dihydroquercetin 1.0-22.8 arabinogalactan 75.2-96.8 water 2.0-5.0

The method of obtaining the complex consists in mixing arabinogalactan with water until completely dissolved, then adding dihydroquercetin, heating the solution to 40-45 ° C, stirring for 0.5-1 hours, followed by drying of the resulting solution by spraying. For this, arabinogalactan is dissolved in a container containing deionized water until it is completely dissolved, then dihydroquercetin is added to the solution in the ratio dihydroquercetin: arabinogalactan equal to 1: (3-100), preferably 1: (50-100) by weight, respectively, the solution is heated to 40-45 ° C, stirred for 0.5-1 hours, then the resulting solution was dried by spraying.

To dry the complex, spray dryers with a centrifugal spray (type SRC) equipped with a blade (flat working surface) high-speed disk or spray dryers with a spray nozzle (type SRF) equipped with pneumatic or mechanical (high pressure) nozzles can be used.

Arabinogalactan is a water-soluble polysaccharide with m.m. 9-18 kD, the main chain of which consists of galactose units, the side chains of arabinose and galactose units. This structural feature contributes to the formation of strong intermolecular complexes of drugs, the molecules of which are most likely bound by intermolecular hydrogen bonds in the space formed by the side chains. Given the conformational mobility of arabinogalactan macromolecules, this space can vary, contributing to the formation of intermolecular complexes with a wide range of substances. In addition, it is possible to form complexes of different stoichiometric composition, when one molecule of arabinogalactan can bind to several molecules of other organic compounds - drugs, allowing you to change the ratio of these components over a wide range. This is the advantage of arabinogalactan as a complexing agent compared to cyclodextrins commonly used for these purposes.

In the same way, it is possible that dihydroquercetin molecules interact with the complexing agent arabinogalactan. The dihydroquercetin molecule can be introduced between the long polysaccharide chains of arabinogalactan, forming a supramolecular complex, which has a higher solubility in water compared to poorly soluble dihydroquercetin. As a result, while maintaining the therapeutic effect of dihydroquercetin, it is possible to increase its solubility and effectiveness by tens of times.

The defining differences of the proposed complex and the method of its production in comparison with the prototype are:

1. The claimed supramolecular complex after drying contains dihydroquercetin and arabinogalactan taken in the ratio dihydroquercetin: arabinogalactan equal to 1: (3.3-96.8) by weight, respectively, as well as water, which allows to increase the solubility of DHQ in water by 11.8 -22.5 times and increase its pharmacological activity.

2. A method of producing a complex based on dihydroquercetin involves mixing arabinogalactan with water until completely dissolved, then adding dihydroquercetin in a ratio of dihydroquercetin: arabinogalactan equal to 1: (3-100), preferably 1: (50-100) by weight, respectively, heating the solution to 40-45 ° C, stirring for 0.5-1 hours, followed by drying the resulting solution by spraying, which reduces the duration of the method while maintaining the high quality of the target product.

The spray drying process is extremely fast (usually 15-30 seconds) and the particles of the composition in the zone of elevated temperatures have a saturated surface. Due to instant drying and low temperature of the sprayed particles of the complex, the dried product is of good quality, because there is no violation of the native properties of dihydroquercetin and arabinogalactan, which is of particular importance for the pharmaceutical industry.

In addition, since the mixing of substances is carried out in the liquid phase, preferably in deionized water, there is a uniform distribution of substances throughout the volume of the solvent, resulting in the homogeneity of the obtained complexes.

When using spray drying, the technological cycle of obtaining the finished dry complex of dihydroquercetin with arabinogalactan is significantly shortened and completely mechanized. In this case, crystallization, precipitation, filtration, centrifugation, drying, grinding, grinding of the finished product are excluded, which significantly reduces energy consumption.

The dried material during drying does not come into contact with the surfaces of the dryer until it dries. This simplifies the solution to the problem of corrosion and the choice of material for the drying chamber. In other drying methods, the wet product is in contact with metal surfaces.

The technical and economic indicators of the spray drying method can be significantly improved due to the intensification of the evaporation process in spray dryers. As practice has shown, when drying highly dispersed materials, the process can be significantly intensified, as a result of which the dimensions of the installation and the consumption of electricity and heat are reduced.

The present invention is illustrated by the following examples.

Example 1. Obtaining complexes based on dihydroquercetin

Arabinogalactan and dihydroquercetin, taken in mass ratios of 1: 3, 1: 5, 1:10, 1:20, 1:30, 1:50 and 1: 100, respectively, were added to containers containing deionized water, respectively, and the solution was heated to 40-45 ° C, was stirred for 0.5-1 hours and received aqueous solutions of the complexes DKV / AG, which were then subjected to physico-chemical methods of analysis.

As the dihydroquercetin used the commercial product "Lavitol (dihydroquercetin)", batch No. 700v from 08.23.11, TU 9325-001-70692152-07), and as an arabinogalactan, the commercial product "Lavitol-arabinogalactan", batch No. 44 from November 9, 2011, TU 9325-008-70692152-08). The results of the study of the composition of the obtained complexes, including water, in wt.%, Before the spray drying process, are presented in table 1.

From the data of table 1 it can be seen that to obtain the complex of the desired ratio, it is possible to vary the wt.% Of dihydroquercetin and the liquid phase at constant wt.% Of arabinogalactan.

Example 2. Obtaining a complex of dihydroquercetin with arabinogalactan in a ratio of 1: 3

The complex of dihydroquercetin with arabinogalactan in a ratio of 1: 3 by weight, respectively, was obtained as follows. An arabinogalactan powder weighing 9.00 kg was dissolved in a container containing 48.0 L deionized water. After complete dissolution of arabinogalactan, dihydroquercetin powder weighing 3.00 kg was added to the solution, the solution was heated to 40 ° C, stirring was carried out for 1 hour, then the resulting solution was spray dried.

The complex was dried by spraying in a dryer of the GLP-60 brand with a centrifugal spray (type СРЦ) equipped with a high-speed blade (flat working surface).

The time spent by the dried material in the dryer chamber is 1.5-2.0 seconds, the temperature of the supplied air at the inlet is 180-200 ° C, at the outlet 70-80 ° C.

The result was a water-soluble complex containing, wt.%:

dihydroquercetin 22.8 arabinogalactan 75,2 water 2.0

Example 3. Obtaining a complex of dihydroquercetin with arabinogalactan in a ratio of 1:20

A complex of dihydroquercetin with arabinogalactan in a ratio of 1:20 by weight, respectively, was obtained as follows. In a container containing deionized water with a volume of 50.55 L, arabinogalactan powder weighing 9.00 kg was dissolved. After complete dissolution of arabinogalactan, dihydroquercetin powder weighing 0.45 kg was added to the solution, the solution was heated to 45 ° C, stirring was carried out for 0.5 hours, then the resulting solution was dried by spraying as in example 2.

The result is a water-soluble complex containing, in wt.%:

dihydroquercetin 4.4 arabinogalactan 93.1 water 2,5

Example 4. Obtaining a complex of dihydroquercetin with arabinogalactan in a ratio of 1: 100

The complex of dihydroquercetin with arabinogalactan in a ratio of 1: 100 by weight, respectively, was obtained as follows. An arabinogalactan powder weighing 9.00 kg was dissolved in a container containing 50.91 L deionized water. After complete dissolution of arabinogalactan, dihydroquercetin powder weighing 0.09 kg was added to the solution, the solution was heated to 45 ° C, stirring was carried out for 0.5 hours, then the resulting solution was spray dried. The result is a water-soluble complex containing, in wt.%:

dihydroquercetin 1,0 arabinogalactan 96.8 water 2.2

In a similar manner, complexes of arabinogalactan with dihydroquercetin in the remaining ratios were obtained.

Example 5. The study of the composition of the obtained complexes

The identification and quantitative content of dihydroquercetin in the complex was determined by HPLC using a Milichrom A-02 device (Ekonova CJSC, Novosibirsk), 290 nm, gradient mode, acetonitrile: water was used as an eluent, caffeine was used as an internal standard, as an external standard - a standard sample of dihydroquercetin company Sigma-Aldrich. Data processing was performed using the "Multichrom" software.

Chromatograms of a standard sample of DKV (a) and a complex of DKV: AG (1: 3) (b) using the internal standard of caffeine are shown in the drawing. The peak of the dihydroquercetin included in the composition in the chromatogram corresponds to the peak of the standard sample.

Each complex obtained in the above ratios was analyzed by chromatographic method under similar conditions.

The AH content in the composition was determined by the photometric method by reaction with an anthron in an acidic medium. The obtained data on the composition of the obtained water-soluble complexes are shown in table 2.

From table 2 it follows that when a complex is formed by spray drying, no new chemical bonds are formed, the chemicals do not change in composition, and only intermolecular bonds between the DHQ and AG molecules increase, which increase the solubility of DHQ in water.

Example 6. The study of the water solubility of complexes of dihydroquercetin with arabinogalactan

An experiment was conducted to study the water solubility of the obtained complexes. For this, the samples were dissolved in 100 ml of distilled water at a temperature of 20 ° C on a magnetic stirrer (400 rpm). Dissolution was carried out until the added samples stopped dissolving. The concentration of DHQ in the solution was determined by the photometric method.

From table 3 it is seen that with increasing AG content in the complex, the solubility of DHQ increases.

Example 7. The study of the anti-inflammatory activity of complexes of dihydroquercetin with arabinogalactan

Anti-inflammatory activity was investigated in a model of acute exudative inflammation in mice caused by subplantation of 0.04 ml of a 2% formalin solution in the left hind paw. The studied complexes were administered intragastrically in water purified 1 hour before the introduction of formalin. A group of control animals received an equivolume amount of purified water. 3 hours after the injection, the mass of the left and right hind legs was measured and the severity of edema was judged by the difference between them.

The obtained data on the effect of a single intragastric administration of the studied complexes on the increase in the mass of the left foot in relation to the right (Δ,%) are presented in table 4.

From the data of table 4 it is seen that the complex of dihydroquercetin with arabinogalactan in the ratio of (1: 3), (1: 5) and (1:10) showed greater anti-inflammatory activity than pure dihydroquercetin.

Example 8. The study of angioprotective activity of complexes of dihydroquercetin with arabinogalactan

The study of angioprotective activity was carried out by reaction to xylene, which was administered to rats in an amount of 0.02 ml intradermally in the depilated region of the abdomen 10 minutes after intravenous injection of 2 ml / kg of 1% Evans blue solution. The criterion of vascular permeability was the time (s) between the introduction of xylene and the appearance of the first signs of skin color. The test substances were administered intragastrically for 7 days in purified water. A group of control animals received an equivolume amount of purified water. The last injection of substances was carried out 1 hour before the injection of a solution of blue Evans. Data on the effect of course intragastric administration of complexes on capillary permeability in rats are shown in table 5.

From the data of table 5 it follows that the studied complexes of dihydroquercetin with arabinogalactan have pronounced angioprotective properties.

The invention allows to increase the solubility of dihydroquercetin in water to 11.8-22.5 times, to improve its bioavailability by increasing the absorbability of the complex, as a result of which it becomes possible to reduce the dose of the drug dihydroquercetin while maintaining its pharmacological activity.

The invention also allows to significantly reduce the duration of the method of obtaining the complex while maintaining the high quality of the target product.

Claims (2)

1. Supramolecular complex with anti-inflammatory and angioprotective activity, including dihydroquercetin, arabinogalactan and water, with the following components, wt.%:
dihydroquercetin 1.0-22.8 arabinogalactan 75.2-96.8 water 2.0-5.0 2. The method of producing the supramolecular complex according to claim 1, comprising mixing arabinogalactan with water until completely dissolved, then adding dihydroquercetin, heating the solution to 40-45 ° C, stirring for 0.5-1 hours, followed by drying of the resulting solution by spraying .

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