Process for producing fine-grained biologically active materials

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention relates to pharmaceutical industry and represents a method for producing fine-grained biologically active materials containing active substances, characterised by the fact that the liquid with biologically active substances is dispersed to the microdrop state in a layer of dry fine-grained inert hydrophobic aerosil in the proportion 10:1.5 to 10:6, to form thereby liquid microdrops surrounded by hydrophobic aerosil particles and powdered which, if necessary, are dried by a technologically acceptable method.

EFFECT: invention provides increasing dispersion of biologically active materials.

11 ex, 2 dwg

 

The invention relates to medicine, biotechnology and pharmaceutical industry and relates to a method of obtaining fine-grained materials containing biologically active ingredients.

A known method of producing microcapsules containing living microorganisms, in which the freeze-dried culture of microorganisms dispersed during the cooling 30-50% aqueous solution of polyvinylpyrrolidone with a molecular weight of 15,000 in the ratio from 1:10 to 1:15 by weight with stirring to a suspension containing individual particles of the desired size, to which is added the solution of cross-linking agent is 10-30% aqueous solution of tannin, the mixture is stirred for 30-180 minutes, then cured microcapsules are collected by filtration, washed and dried (patent RU 2220716 CI, AC 9/54, 10.01.2004).

Known dry probiotic preparation and method thereof, involving the receipt of liquid biomass by mixing native culture of lactic acid bacteria with a protein-carbohydrate complex, contact the dehydration of the obtained liquid biomass moisture capacity ion exchange resin KB-4P-2 with a particle size of from 1 to 800 microns, pre-treated with a mixture of anhydrous lactose and hydrophobic Aerosil (patent RU 2268926 C2, C12N 1/20, AS 9/12, F26B 5/16, 10.03.2005).

Known capsules, comprising the microorganism, the matrix component and m is Nisha least one encapsulated material in which the matrix component and the encapsulated material does not originate from a microorganism, and the encapsulated material includes at least one functional agent, which is characterized by the calculated value of the distribution coefficient in a mixture of octanol-water clogP less than 3 and the method of preparation of capsules containing the stage of the preparation of aqueous fluid comprising at least a microorganism and water, adding the encapsulated material comprising functional agent having a clogP 1 or higher, mixing, shaking or mixing the aqueous liquid and encapsulated material, adding a matrix component, drying components and, optionally, granulating the dried slurry to obtain capsules (application EN 2007104041 AND A 1/22, 10.08.2008).

You know the concentration of the Mare's milk as a preparation for oral administration, dried biologically inert, fine matrix, which is used highly dispersed silicon dioxide (application EN 2004134353 AND AC 9/70, 10.06.2005).

A method of obtaining dry bacterial drugs, according to which fine dry powder of silicon dioxide is added to a broth culture of microorganisms in the ratio 1:2 and stirred until a homogeneous density mass which is dried in a thermostat at 27-32°C or in air and then dispersed for two hours until finely dispersed state (patent RU 2104299 C1, C12N 1/04, 10.02.1998).

The main disadvantage of this method is the inability to provide a high dispersion of the biologically active materials containing active substances.

The basis of the claimed invention is the task of improving the dispersion of the biologically active materials containing active substances.

The problem is solved by the fact that the liquid bioactive active substances dispersed to drip status in the layer of dry finely dispersed inert hydrophobic disconnector with nano particles and, if necessary, dried technologically acceptable method.

As a result of our research for the first time it is shown that the dispersion liquid in a layer of fine-grained hydrophobic disconnector with nano-particles formed of a material called us drip powder. It represents the droplets of fluid, surrounded by the hydrophobic particles of the disconnector is in the form of powder (Figure 1) and has all its powder properties, such as flowability (flow), bulk density, consobrina, shear strength, but has no lipomastia, hygroscopicity and wettability.

The principle of education and drip powder based on the ability of drops of liquid, covered with hydrophobic understand what setelem, not to join in collisions. Figure 2 shows the difference drops of 1% solution of CuSO44 mm diameter, spreading on the surface (left), and a drop of this liquid is ideal spherical shape, covered with a layer of hydrophobic Aerosil R972, not wetting the surface and preserves the form (on the right needle hinder the descent of the droplets).

As a disconnector is used Aerosil - organosilicon polymer of ultra-fine light white powder with the following unusual properties: high purity of 99.96%, a large specific surface area - 100-300 m2/g nano-sized particles - 7-40 Å (0,007-0,04 µm), mechanical strength, thermal and radiation stability [Senov P.L. Pharmaceutical chemistry. - M.: Medicine, 1978. - p.132-133]. Hydrophobic Aerosil (99%) is grafted on the surface of particles of silicon dioxide methyl - and dimethylsilicone groups. He is allowed by the State Pharmacopoeia [State Pharmacopoeia of the USSR. - 11th ed. - M.: Medicine, 1989. - Vol.2. - 400 C.] for use in medicinal preparations as excipients.

Because the disconnector has the nano-particles, so high dispersion drip powders is provided a method of dispersing fluid. For these purposes, can be used different equipment, hydraulic and pneumatic force the NCI, acoustic and piezoelectric atomizers. But we have selected the disk and electromagnetic dispersers as providing liquid droplets with the best dispersion.

The use of dispersion liquids containing active biologically active agents (solutions, suspensions or emulsions), allows to obtain highly dispersed drip powders based on them, which can be used immediately after preparation or dried technologically acceptable method. In the latter case, suitable biologically active dispersion liquid before mixing with protective environments.

The drip drying powders also can be done by various methods, for example by sublimation, sorption, sorption-contact, naturally. Criterion method of drying should serve as a required activity of the active substance in the resulting product.

In some cases, if required by the technology of preparation, after dehydration, the product is separated from the sorbent any, for example sieve method, resulting in obtaining a dry dispersible powder containing active substances.

The claimed method can be used for the preparation of highly dispersed materials containing immune, probiotic, analgesics is eticheskie, analiticheskie, antimicrobial, antiviral, protivodiabeticheskie, insecticidal active substances, but only of liquids that do not Deplete the hydrophobicity of fine disconnector. Therefore, the liquid containing organic solvents (such as alcohol, benzene, acetone)may be used for implementing the inventive method. Furthermore, materials of good quality can be obtained from solutions, suspensions and emulsions with a pH close to the neutral value of 7.0), with a ratio of liquid phase: the disconnector 10:1.5 to 10:6. The use of the disconnector in smaller proportions to the liquid phase than 1.5:10, leads to the fact that because of his lack of not all of the formed droplets of the liquid phase they are stabilized, gradually become larger, which consequently leads to the reduction of dispersion of the obtained materials. The use of the disconnector in large numbers with the liquid phase than 6:10, leads to the presence of a large number of materials that, at least not economically feasible.

According to the invention increase the dispersion of the biologically active materials containing active substances, provided that the liquid bioactive active substances dispersed to drip status in the layer of dry finely dispersed inert hydrophobic razor is Italia with nano particles and, if necessary, dried technologically acceptable method.

The inventive method for high dispersion of the biologically active material is new and are not described in literature.

The technical result of the claimed invention is to improve the dispersion of the biologically active materials containing active substances.

The invention is illustrated in the following examples, showing the increase in the dispersion of the biologically active materials containing active substances, when using the proposed method.

The content in the preparations of viable aerobic microorganisms: Francisella tularensis, Yersinia pestis, Serratia marcescens, Entherococcus faecium were identified by means of Pasteur-Koch on solid nutrient media. The content of viable anaerobic microorganisms Bifidobacterium bifidum was determined in a liquid nutrient medium by the method of limiting dilutions. The concentration of virus vaccine strain La Sota Newcastle disease was determined by cultivation in allantoine fluid of chicken embryos [Surin V.N., Belousov W., Fomin N. Veterinary Virology. - M.: Kolos, 1986]. Biological activity of antibodies preparations characterized protivoallergennoy activity (in the credits TPHA) [FS 42-3347-97]. The dispersion of the preparations was measured by a laser analyzer of grain Malvern Instruments)) S by the method developer. Sterilization of sorbents to combine the Menno dehydration was performed in a dry-heat Cabinet SUP-4 at a temperature of 120°C with exposure to steady-state thermal conditions for at least 2 hours.

Example 1. Drip powder culture test to check air filters received, dispersive suspension of Serratia marcescens stuck-851 with a pH of 6.9 and a content of viable microorganisms 183×109CFU/ml in the layer of hydrophobic Aerosil AM-1-300 at a ratio of 10:4, in the electromagnetic disperser for 30 sec.

Check the dispersion obtained drip powder showed that the content of the target fraction of particles (10 μm) was 36% and in comparison with the product obtained by known manufacturing techniques, increased 1.2 times.

Obtained drip powder test culture with the content of viable microorganisms 130×109CFU/g was used directly (without drying) on purpose to check air filters on their compliance with safety and industrial hygiene.

Example 2. Drip powder tularemia vaccine received, dispersive suspension Francisella tularensis strain No. 33 NIIEG with a pH of 7.0 and a content of viable microorganisms 590×109CFU/ml in the layer of hydrophobic Aerosil AM-1-300 at a ratio of 10:3, in the electromagnetic disperser for 20 sec.

Drying drip powder vaccine for tularemia in the amount of 60 g with a content of viable microorganisms 408x109CFU/g assests is whether sorption-contact method, mix it with ion exchange resins KB-4P-2 residual moisture less than 1% in the ratio of 1:5.

After separation from the sorbent prepared the product was a dry powder beige color with a residual moisture content of 4.0%, a content of viable microorganisms 530×109CFU/g

The content of the target fraction of particles (up to 10 μm) in a final product compared to the product obtained by known manufacturing techniques (drying and subsequent grinding), increased by 1.5 times and amounted to 31%.

The viability of microorganisms during storage of the finished product within six months did not change and amounted to 100%.

Example 3. Drip powder plague vaccine was received, dispersive suspension Yersinia pestis strain EV, NIIEG with a pH of 7.1 and a content of viable microorganisms 315×109CFU/ml in the layer of hydrophobic Aerosil AM-1-300 at a ratio of 10:2,5, electromagnetic disperser for 25 C.

Drying drip powder plague vaccine in the amount of 50 g with a content of viable microorganisms 250×109CFU/g was carried out by sorption-contact method, mix it with ion exchange resins KB-4P-2 residual moisture less than 1% in the ratio of 1:6.

After separation from the sorbent prepared the product was a dry powder beige color with a residual moisture content of 4.1%, with the holding of viable microorganisms 405×10 9CFU/g

The content of the target fraction of particles (up to 10 μm) in a final product compared to the product obtained by known manufacturing techniques (drying and subsequent grinding)increased by 1.8 times and amounted to 42%.

The viability of microorganisms during storage of the finished product within six months did not change and amounted to 100%.

Example 4. Drip powder culture test to check air filters received, dispersive suspension of Serratia marcescens stuck-851 with a pH of 6.9 and a content of viable microorganisms 183×109CFU/ml in the layer of hydrophobic Aerosil AM-1-300 at a ratio of 10:4, in the electromagnetic disperser for 30 sec.

Drying drip powder culture test to check air filters in the amount of 65 g with a content of viable microorganisms 130×109CFU/g was carried out by sorption-contact method, mix it with ion exchange resins KB-4P-2 residual moisture less than 1% in the ratio of 1:4.

After separation from the sorbent prepared the product was a dry powder pink with a residual moisture content of 5.1%, a content of viable microorganisms 219×109CFU/g

The content of the target fraction of particles (up to 10 μm) in a final product compared to the product obtained by known techniques from the otopleniya (drying and subsequent grinding), increased 1.2 times and amounted to 36%.

The viability of microorganisms during storage of the finished product within six months did not change and amounted to 100%.

Example 5. Drip powder probiotic preparation was received, dispersive suspension Bifidobacterium bifidum STS with a pH of 7.0 and a content of viable microorganisms 2,5x109CFU/ml in the layer of hydrophobic Aerosil AM-1-300 at a ratio of 10:3, in the disk disperser for 15 sec.

Drying drip powder probiotic preparation in the amount of 30 g with a content of viable microorganisms of 1.9×109CFU/g was carried out by sorption-contact method, by mixing it with a dispersed alumina residual moisture of 1% in the ratio of 1:8.

Ready the product was a dry powder beige color with a residual moisture content of 11.8%, a content of viable microorganisms of 1.6×108CFU/g

The median particle diameter of the finished product compared to the product obtained by known manufacturing techniques (drying and subsequent grinding), decreased 2.3 times and amounted to 24 microns.

The viability of microorganisms during storage of the finished product within six months did not change and amounted to 100%.

Example 6. Drip powder probiotic preparation was received, dispersive suspension Entherococus faecium with a pH of 7.2 and a content of viable microorganisms of 1.8×10 9CFU/ml in the layer of hydrophobic Aerosil AM-1-300 at a ratio of 10:3, in the electromagnetic disperser for 20 sec.

Drying drip powder probiotic preparation in the amount of 40 g with a content of viable microorganisms of 1.4×109CFU/g was carried out by sorption-contact method, mix it with ion exchange resins KB-4P-2 residual moisture less than 1% in the ratio of 1:7.

After separation from the sorbent prepared the product was a dry powder beige color with a residual moisture content of 4.1%, a content of viable microorganisms 1,3x×109CFU/g

The median particle diameter of the finished product compared to the product obtained by known manufacturing techniques (drying and subsequent grinding), decreased 1.4 times and amounted to 31 microns.

The viability of microorganisms during storage of the finished product within six months did not change and amounted to 100%.

Example 7. Drip powder immunobiological preparation was received, dispersive solution of immunoglobulins IgG, IgA, IgM with pH 7.0 and protivoallergennoy activity 1:640 in the credits TPHA in the layer of hydrophobic Aerosil R 972 at a ratio of 10:2, in the disk disperser for 20 sec.

Drying drip powder immunobiological preparation in the amount of 45 g of protivosemnye the heat activity 1:640 in the credits TPHA carried out sequentially aspirated and sorption-contact method, mix it with dispersed alumina residual moisture less than 1% in the ratio of 1:4.

Ready the product was a dry white powder with a residual moisture content of 4.5%, protivoallergennoy activity 1:320 in the credits TPHA.

The median particle diameter of the finished product compared to the product obtained by known manufacturing techniques (drying and subsequent grinding), decreased 1.4 times and amounted to 12 microns.

Protivogelmintny activity during storage of the finished product within six months did not change and amounted to 1:320 in the credits TPHA.

Example 8. Drip powder local anesthetic drug received, dispersive suspension of benzocaine concentration of the active substance 250 mg/ml and pH 6.5 in the layer of hydrophobic Aerosil R 972 at a ratio of 10:2,5, disk disperser for 20 sec.

Drying drip powder local anesthetic drug in the amount of 30 g with a content of active ingredient 200 mg/g was carried out by atmospheric dehydration at a temperature of 22°C.

Ready the product was a dry white powder with a residual moisture content of 3.9%, a content of active substance is 500 mg/g

The median particle diameter of the finished product compared to the product obtained by known manufacturing techniques (in susiana then crushed), decreased by 1.2 times and amounted to 16 microns.

Example 9. Drip powder complex biologic drug received, dispersive suspension of microorganisms Bifidobacterium bifidum STS in a mixture with a solution of immunoglobulins IgG, IgA, IgM with pH 7.0, protivoallergennoy activity 1:640 in the credits TPHA, the content of viable microorganisms 2×109CFU/ml in the layer of hydrophobic Aerosil AM-1-3 00 when the ratio suspension: Aerosil 10:3, in the disk disperser for 15 sec.

Drying drip powder complex immunobiological preparation in the amount of 35 g with protivoallergennoy activity 1:640 in the credits TPHA and content of viable microorganisms of 1.5×109CFU/g was carried out by sorption-contact method, by mixing it with a dispersed basic alumina residual moisture less than 1% in the ratio of 1:6.

Ready the product was a dry white powder with a residual moisture content of 12.2%, protivoallergennoy activity 1:320 in the credits TPHA and content of viable microorganisms of 1.2×108CFU/g

The median particle diameter of the finished product was 25 μm (preparation by known technology does not exist).

Protivogelmintny activity and content of viable microorganisms during storage of the finished product for the of six months did not change and amounted to 100% from the original values in the finished product prior to storage.

Example 10. Drip powder aerosol drug received, dispersive solution of sodium chloride with a concentration of active ingredient 100 mg/ml and pH 5.5 in the layer of hydrophobic Aerosil AM-1-300 at a ratio of 10:1.5, electromagnetic disperser for 15 sec.

Drying drip powder aerosol formulation in the amount of 45 g with a content of active substance of 87 mg/g was carried out by sorption way above the layer of sorbent (macroporous silicagel IBCS) residual moisture of 1% at a temperature of 20°C.

Ready the product was a dry white powder with a residual moisture content of 5%, the content of active ingredient 200 mg/year

The median particle diameter of the finished product compared to a preparation obtained by known manufacturing techniques (drying and subsequent grinding), decreased to 1.25 times and amounted to 8 microns.

Example 11. Drip powder combined immune-antimicrobial drug received, dispersive suspension of microorganisms Bifidobacterium bifidum STS mixed with the antibiotic ooxacin with a pH of 6.0, a content of viable microorganisms 1,8x109CFU/ml and the concentration of antibiotic 30 mg/ml in a layer of hydrophobic Aerosil AM-1-300 when the ratio suspension: Aerosil 10:3, in the disk disperser for 20 sec.

Drying Mick is capelinha powder combined immune-antimicrobial drug with the content of viable microorganisms of 1.4×10 9CFU/g and the antibiotic concentration 23 mg/g was carried out by sorption-contact method, by mixing it with a dispersed basic alumina residual moisture less than 1% in the ratio of 1:6.

After separation from the sorbent prepared the product was a dry yellow powder with a residual moisture content of 5.1%, a content of viable microorganisms of 1.9×109CFU/g and the concentration of antibiotic 300 mg/g

The median particle diameter of the finished product was 30 μm (preparation by known technology does not exist).

The content of viable microorganisms during storage of the finished product within six months did not change and amounted to 100% from the original values in the finished product prior to storage.

Example 12. Drip powder viral vaccines received, dispersive suspension vaccine strain La Sota virus Newcastle disease with a pH of 7.0 and a content of viable viruses 10,4 lg EID50/ml, in the layer of hydrophobic Aerosil AM-1-3 00 at a ratio of 10:3, in the electromagnetic disperser for 20 sec.

Drying drip powder viral vaccines with the maintenance of viable viruses 10,3 lg EID50/g was carried out by sublimation.

Ready the product was a dry white powder with a residual moisture content of 4.0%, a content of viable virus is 10.2 lg EID 50/year

The content of the target fraction of particles (up to 10 μm) in a final product compared to the product obtained by known manufacturing techniques (drying and subsequent grinding)increased by 1.1 times and amounted to 77%.

The viability of viruses during storage of the finished product within six months did not change and amounted to 100%.

Example 13. Drip powder analgesic drug received, dispersive solution of paracetamol, with the concentration of the active substance 250 mg/ml and pH 6.5 in the layer of hydrophobic Aerosil AM-1-300 at a ratio of 10:6, in the electromagnetic disperser for 15 sec.

Drying drip powder analgesic drug in the amount of 35 g with a content of active substance 156 mg/g was carried out by atmospheric dehydration at a temperature of 22°C.

Ready the product was a dry white powder with a residual moisture content of 5%, a content of active substance is 500 mg/g

The median particle diameter of the finished product compared to the product obtained by known manufacturing techniques (drying and subsequent grinding), decreased by 3.9 times and amounted to 4.3 μm, and the content of fractions up to 5 µm increased 7.3 times and amounted to 56%.

The method of obtaining highly dispersed biologically active materials containing active the substances, characterized in that the liquid bioactive active substances dispersed to drip status in the layer of dry finely dispersed inert hydrophobic Aerosil in the ratio of 10:1.5 to 10:6, resulting in formation of droplets of liquid, surrounded by particles of hydrophobic Aerosil and having the form of powder, which, if necessary, dried technologically acceptable method.



 

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