RU2743010C1 - Method of producing vitamin d3 nanocapsules - Google Patents

Abstract

FIELD: chemistry.

SUBSTANCE: invention relates to a method for producing nanocapsules vitamin D₃ in the medium Tween-80, at a weight ratio of core:shell 1:1, characterized in that successively in a beaker with heated purified water were added an oil concentrate of vitamin D₃, 2-hydroxy-β-cyclodextrin, emulsion (pre-melted) wax and Tween-80, then the system is stirred using an ultrahomogenizer at rate of 2000 rpm for 5 minutes, further, a magnetic stirrer with heating (exposure time 5 minutes) is used, then the obtained nanoemulsion is filtered out, dried on a lyophilic drier and powder of nanocapsules is obtained.

EFFECT: technical result is a novel method of producing nanocapsules of vitamin D₃ in a tween-80 medium, with a weight ratio of core to shell of 1:1.

1 cl, 3 ex, 3 dwg

Description

The invention relates to nanotechnology, namely the production of 7 nanocapsules from nanoemulsions used as carriers of active substances in pharmaceutical compositions and can be practically applied in the pharmaceutical industry and veterinary medicine.

Known for the following patents close to this invention.

In US Pat. RU 2 494 729 С2 Liposomal pharmaceutical preparation and method of its manufacture, published 10.10.2013 Russian Federation, describes a liposomal preparation that may also contain additional excipients, in particular excipients for further changing the characteristics of the liposome surface in order to endow the liposome with greater functionality in organism. Such auxiliary substances include, for example, lipids and the like, modified with hydrophilic polymers. As a phospholipid bilayer, substances with a melting point higher than body temperature are used, so that the phase shift temperature of the liposome is higher than body temperature. At the phase shift temperature, the fluidity of the membrane will increase, and the drug encapsulated in the liposome will have the maximum elimination rate. Thus, membrane fluidity has a direct effect on liposome stability.

In this invention will be used surfactant tween-80 (polysorbate-80) with a boiling point of 55-60 ° C, which dissolves in the oil phase.

In patent RU 2 680 096 C2 Liposomal preparations, published on February 15, 2019, Russian Federation, the production of a liposome is limited exclusively to mixing the lipid with an aqueous solution. These types of liposomes exist in the lowest energy state in which a lipid can exist when in aqueous solution, and the reproducibility of this liposomal preparation is not a problem. Above critical concentrations (about 20% w / v), non-liposomal structures will begin to form in an aqueous solution. These liposomes exist in their lowest energy state and are thermodynamically stable, self-forming liposomes.

In this invention, the concentration of the lipid component will not exceed 5% w / v in aqueous solution, which will make the system thermodynamically stable.

In US Pat. RU 2491917 C2 Nanoemulsion, published 09/10/2013, describes a technology for producing a nanoemulsion, which contains an aqueous component and a carrier comprising a lipophilic component and a surfactant. The way to obtain a nanoemulsion consists in mixing the components using a container and mixer, optimized to obtain a very quickly homogenizing mixture of components (in seconds), avoiding the formation of foam.

Thus, in addition to highly efficient homogenization, the emulsion preparation method of the present invention requires the use of high shear devices such as ultrasonic devices or high pressure homogenizers.

Known technology for producing nanoemulsions of the type water in oil with biologically active substances (RF Patent RU 2535022 C2 publ. 10.12.2014), which contains 35-80% of the hydrophobic phase, 1-15% of the hydrophilic phase, surfactant. A water-in-oil nanoemulsion prepared in a given proportion of phases has good storage stability.

There is also known a nanoemulsion with biologically active substances (RF Patent No. 2362544, A61K 9/10, A61K 9/107, publ. 2008), a transparent or slightly opalescent nanoemulsion of the water-in-oil type for oral, transdermal use for use in ophthalmic practice with biologically active compounds, characterized in that it contains 35-80% of a hydrophobic phase, 17-43% of a surfactant, 3-7% of a co-solvent and 1-15% of an aqueous phase. As a hydrophobic phase, mixtures of mono-, di- and triglycerides with mono- and diesters of saturated and unsaturated fatty acids are used, the surfactant is selected from the group of nonionic surfactants - sorbitans in a mixture with an auxiliary surfactant (from the group of polyhydroxyalkanes or monohydric alcohols).

In US Pat. 2359662 MPK A61K 009/56, A61J 003/07, B01J 013/02, A23L 001/00 published on June 27, 2009 The Russian Federation proposed a method for producing sodium chloride microcapsules using spray cooling in a Niro spray cooling tower under the following conditions: air inlet temperature 10 ° С, air temperature at the outlet 28 ° С, rotation speed of the spray drum 10000 rpm. The microcapsules of the invention have improved stability and provide controlled and / or prolonged release of the active ingredient. The disadvantages of the proposed method are the duration of the process and the use of special equipment, a set of certain conditions (air temperature at the inlet 10 ° C, air temperature at the outlet 28 ° C, the rotation speed of the spray drum 10000 rpm).

In US Pat. 2173140 IPC A61K 009/50, A61K 009/127 Russian Federation published on September 10, 2001, a method for producing organosilicon microcapsules using a rotary cavitation unit with high shear forces and powerful hydroacoustic phenomena of the sound and ultrasonic range for dispersion is proposed.

The disadvantage of this method is the use of special equipment - a rotary cavitation unit, which has an ultrasonic effect, which affects the formation of microcapsules and can cause side reactions due to the fact that ultrasound has a destructive effect on protein polymers, therefore the proposed method is applicable when work with polymers of synthetic origin.

The closest to our proposed method is the method proposed in US Pat. RU 2 703 269 C1 A method for producing vitamin B ₄ nanocapsules published on October 16, 2019, a method for producing nanocapsules by drying at room temperature a nanoemulsion containing hydrophilic and hydrophobic phases and, without fail, surfactants is proposed.

The disadvantage of this method of obtaining nanocapsules is the use of a representative of aromatic perfluorinated organofluorine compounds - fluorobenzene.

The disadvantages of the known nanoemulsions are low stability during storage, possible allergic reactions due to the presence of chemical components, as well as high cost and the need for strict control due to the use of raw materials of synthetic origin.

The technical problem is to obtain a stabilized nanoemulsion of vitamin D ₃ , followed by obtaining nanocapsules of vitamin D ₃ with a high biological potential.

The solution to the technical problem is achieved by the method of obtaining nanocapsules of vitamin D ₃ , characterized in that tween-80 is used as the shell of the nanocapsules, and vitamin D ₃ is used as the core when obtaining nanocapsules by mechanical fragmentation of the oil phase in the aqueous phase in the presence of a surfactant. Very small oil particle size is often achieved by at least one pass through a high pressure homogenizer or ultrasonic device.

A distinctive feature of the proposed method is the production of nanocapsules by a highly efficient physical method using an IKA T 18 digital ULTRA-TURRAX ultrahomogenizer and a shaft speed of 2000 rpm, exposure time 5 minutes.

The oil concentrate is preliminarily prepared according to the following technology: 86.5 g of C8 / C10 triglyceride and 1 g of vitamin E (tocopherol acetate) were introduced into a beaker covered with foil in a stream of nitrogen. With stirring without heating and under a stream of nitrogen, 12.5 g of vitamin D ₃ (40 million IU / g) was added. The mixture was stirred in a stream of nitrogen until complete dissolution and transferred in a stream of nitrogen into a dark glass jar, previously purged with nitrogen. Before closing the can, an additional nitrogen purge was made for 2 minutes.

The method of obtaining nanocapsules of vitamin D ₃ in a tween-80 medium is characterized by the fact that tween-80 is used as a shell, and vitamin D ₃ is used as a core, with a mass ratio of core: shell of 2: 1. Into a beaker with purified water heated to 50-60 ° C add 5 g of vitamin D ₃ oil concentrate prepared in advance according to the indicated technology, 3 g of 2-hydroxy-β-cyclodextrin, 2 g of emulsion wax (pre-melted) and Tween-80 5 d. The nanoemulsion is obtained by mixing the components at room temperature in a beaker using an IKA T 18 digital ULTRA-TURRAX ultrahomogenizer and a shaft speed of 2000 rpm, exposure time 5 minutes and 3 different exposure modes:

1) magnetic stirrer with heating MR Hei-Standard (Heldolph Instruments, Germany), exposure time 5 minutes;

2) centrifuge - High speed centrifuge (MECHANIKA PRECYZYJNA, Poland) 5000 rpm, exposure time 5 minutes;

3) ultrasonic bath UZV-7/100-MP-RELTEK UHL 4 TU 3444-005-26285789-2006, exposure time 5 minutes.

The resulting nanoemulsion is dried on a Zirbus VaCo2 freeze dryer under the following conditions.

At the first stage of freeze drying, nanoemulsion samples are frozen to a low temperature (-60 ° C) and placed in an open container in the working chamber of the device for 2 hours. The freeze drying chamber is connected to a low temperature compartment called a condenser, which is kept at a low temperature, while the temperature in the condenser is always lower than in the lyophilization chamber.

The second drying step is direct removal of ice or solvent crystals from the frozen solution. Why a high vacuum is created in the entire system. Due to the difference in the partial pressure of water vapor, the water in the condenser is frozen, i.e. water from the frozen solution, bypassing the liquid state, is frozen on the condenser coil. Removal of residual moisture within 1.5 hours.

The result of the proposed method is the production of vitamin D ₃ nanocapsules.

EXAMPLE 1. Obtaining nanocapsules D ₃ , the ratio of the core: shell 1: 1

5 g of vitamin D ₃ oil concentrate is added to purified water heated to 50 ° C in the presence of 5 g of tween-80, 3 g of 2-hydroxy-β-cyclodextrin, 2 g of emulsion wax (previously melted). The resulting emulsion was exposed to an IKA T 18 digital ULTRA-TURRAX ultrahomogenizer, mode 2000 rpm, duration 5 minutes and a magnetic stirrer with heating MR Hei-Standard (Heldolph Instruments, Germany), exposure time 5 min. Next, the nanoemulsion is dried on a Zirbus VaCo 2 freeze dryer according to the above technology.

EXAMPLE 2. Obtaining nanocapsules D ₃ , ratio core: shell 2: 1

5 g of vitamin D ₃ oil concentrate is added to purified water heated to 50 ° C in the presence of 5 g of tween-80, 3 g of 2-hydroxy-β-cyclodextrin, 2 g of emulsion wax (previously melted). The resulting emulsion was exposed to an IKA T 18 digital ULTRA-TURRAX ultrahomogenizer, mode 2000 rpm, duration 5 minutes and a High speed centrifuge (MECHANIKA PRECYZYJNA, Poland) 5000 rpm, exposure time 5 minutes. Then the nanoemulsion is dried in a freeze dryer Zirbus VaCo 2 using the above technology.

EXAMPLE 3. Obtaining nanocapsules D ₃ , core: shell ratio 3: 1

5 g of vitamin D ₃ oil concentrate is added to purified water heated to 50 ° C in the presence of 5 g of tween-80, 3 g of 2-hydroxy-β-cyclodextrin, 2 g of emulsion wax (previously melted). The resulting emulsion was exposed to an ultrahomogenizer IKA T 18 digital ULTRA-TURRAX, mode 2000 rpm, duration 5 minutes and an ultrasonic bath UZV-7/100-MP-RELTEC UHL 4 TU 3444-005-26285789-2006, exposure time 5 minutes. Next, the nanoemulsion is dried on a Zirbus VaCo 2 freeze dryer according to the above technology.

Figure 1 shows a histogram of the distribution of the average hydrodynamic radius in the analyzed sample after exposure to a magnetic stirrer with heating MR Hei-Standard (Heldolph Instruments, Germany), exposure time 5 min.

Figure 2 shows a histogram of the distribution of the average hydrodynamic radius in the analyzed sample after exposure to a centrifuge - High speed centrifuge (MECHANIKA PRECYZYJNA, Poland) 5000 rpm, exposure time 5 minutes.

Figure 3 shows the histogram of the distribution of the average hydrodynamic radius in the analyzed sample after exposure to the ultrasonic bath UZV-7/100-MP-RELTEC UHL 4 TU 3444-005-26285789-2006, exposure time 5 minutes.

Before determining the particle size, each sample was diluted 2 times with distilled water, the particle size was determined by photon correlation spectroscopy on a Photocor Complex setup (manufactured by OOO Antek-97, Russia). Computer data processing was carried out using the DynaLS software.

Claim

The method of obtaining nanocapsules of vitamin D ₃ in a tween-80 medium is characterized by the fact that tween-80 is used as a shell, and vitamin D ₃ is used as a core, with a mass ratio of core: shell 1: 1, or 2: 1, or 3: one. Into a beaker with purified water heated to 50 ° C add 5 g of vitamin D ₃ oil concentrate prepared in advance according to the indicated technology, 3 g of 2-hydroxy-β-cyclodextrin, 2 g of emulsion wax (pre-melted) and Tween-80 5 g. The nanoemulsion is obtained by mixing the components at room temperature with stirring in a beaker using an ultrahomogenizer and a shaft speed of 2000 rpm, exposure time 5 minutes and 3 different exposure modes:

1) stirring with a magnetic stirrer with heating MR, exposure time 5 minutes;

2) centrifugation at 5000 rpm, exposure time 5 minutes;

3) sounding in an ultrasonic bath: the power of the ultrasound source is 100 W, the exposure time is 5 minutes.

The resulting nanoemulsion is dried in a freeze dryer under the following conditions.

At the first stage of freeze drying, nanoemulsion samples are frozen to a low temperature (-60 ° C) and placed in an open container in the working chamber of the device for 2 hours. The freeze drying chamber is connected to a low temperature compartment called a condenser, which is kept at a low temperature, while the temperature in the condenser is always lower than in the lyophilization chamber.

The second drying step is direct removal of ice or solvent crystals from the frozen solution. Why a high vacuum is created in the entire system. Due to the difference in the partial pressure of water vapor, the water in the condenser is frozen, i.e. water from the frozen solution, bypassing the liquid state, is frozen on the condenser coil. Removal of residual moisture within 1.5 hours.

The result of the proposed method is the production of vitamin D ₃ nanocapsules.

Claims (1)

A method of obtaining nanocapsules of vitamin D ₃ in a tween-80 medium, characterized by the fact that tween-80 is used as a shell, and vitamin D ₃ is used as a core, with a mass ratio of core: shell of 1: 1, characterized in that the chemical a glass with heated purified water, an oil concentrate of vitamin D ₃ , 2-hydroxy-β-cyclodextrin, emulsion wax (pre-melted) and tween-80 are added, then the system is stirred using an ultrahomogenizer at a speed of 2000 rpm for 5 minutes, then used a magnetic stirrer with heating (exposure time 5 minutes), then the resulting nanoemulsion is filtered off, dried in a freeze dryer and nanocapsule powder is obtained.

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