Lyophilised formulation containing influenza vaccine, and method for preparing it

FIELD: medicine, pharmaceutics.

SUBSTANCE: group of inventions refers to medicine, namely to a lyophilised formulation containing an influenza vaccine, an aqueous solution prepared by lyophilisation and containing (i) he influenza vaccine, (ii) a hydrophobic amino acid, and (iii) arginine and an acid addition salt thereof; it also concerns a method for preparing the lyophilised formulation containing the influenza vaccine to be desalted.

EFFECT: group of inventions provides preparing the lyophilised formulation wherein the influenza vaccine exhibits improved stability.

15 cl, 3 tbl

 

The technical field to which the invention relates.

The present invention relates to liofilizirovannam drugs, containing influenza vaccine. More specifically, the present invention relates to liofilizirovannom the drug, in which the influenza vaccine has increased stability. The present invention also relates to a method for producing a lyophilized form.

Prior art

Influenza is a disease caused by influenza viruses that infect the respiratory organs. In General, an individual infected with influenza virus, the virus after a latent period of about 1 to 2 days of fever to 38º or higher, which is accompanied by General symptoms such as headache, malaise, and joint and muscle pain. Followed by respiratory symptoms such as cough and sputum, but the condition usually improves within weeks. However, when influenza infection affects such individuals as the elderly, young children, pregnant women, patients with chronic diseases of the respiratory or cardiovascular system, diabetes or patients with chronic renal failure may develop severe and sometimes fatal complications, such as pneumonia and bronchitis. In addition, the flu, to whom that causes serious health problems, is also vysokozaraznoy, and for a short period of time become infected many people, leading to enormous social and economic losses.

The introduction of the flu vaccine is the most effective way to prevent health damage caused flu-like infections, and reducing social and economic losses. Drugs influenza vaccine, such as liquid preparations for use in injection and frozen preparations for use in the form of nasal drops, have been known in the past; however, so far not produced dry preparations with satisfactory stability (see, for example, non-patent document 1).

When an influenza-like vaccines distributed in the form of liquid preparations, they should always be maintained at low temperatures during distribution and storage to prevent inactivation of the vaccine; that is, the necessary cold chain impacts. In addition, when vaccines are distributed in the form of frozen preparations, you need to keep vaccines in a frozen state during distribution and storage to maintain the stability of drugs. Thus, flu-like vaccines made in the form of liquid or frozen preparations require perfect temperature control during transport and storage (see non-patent documents is 2), complicating the distribution of these drugs in the areas lack electricity or maintenance activity of flu vaccines without the presence of vehicles, providing the conditions of low temperature.

To overcome such disadvantages of liquid or frozen preparations would effectively distribute flu-like vaccine in the form of dry preparations. Although recently it has been proposed a technique related to liofilizirovannam drugs flu vaccines, in which an influenza-like vaccines are manufactured in the form of drugs with the addition of lactose or trehalose, not specified anywhere sustainability of such drugs (see non-patent document 3).

Like other vaccines, flu-like vaccine wysokonapieciowe to heat and is known to lose activity at high temperatures or at temperatures below their freezing point (see non-patent document 2). Even if flu-like vaccines simply dried or lyophilized, and their activity is problematic decreases with time during production or storage. Haven't found a method to overcome this problem, and to date not reported successful examples of implementation of dry preparations of flu vaccines.

Disclosure of inventions

The problem addressed by the invention

The present invention was POPs the ANO with the above methods of the prior art. The present invention is the provision of lyophilized, in which the influenza vaccine shows significantly better resistance.

Tools for problem solving

The inventor has conducted extensive research to overcome the above problems and found that the dried product, in which the influenza vaccine shows significantly better resistance can be obtained by lyophilization of an aqueous solution, which meets the following conditions (a) to (C) upon receipt of a lyophilized preparation containing influenza vaccine:

(A) on (i) the influenza vaccine, (ii) a hydrophobic amino acid, and (iii) arginine and its acid additive salt;

(C) the proportion of the component (iii) is from 20 to 85% by mass. relative to the total amount of the resulting lyophilized drug; and

(C) the pH is brought to 8 to 10 by regulating the relationship of arginine to the acid additive salts, which form component (iii).

The present invention was carried out by making additional improvements on the basis of these data.

In summary, the present invention provides a lyophilized preparation containing influenza vaccine as set forth below.

Paragraph 1. Freeze-dried preparation containing grupposo the vaccine, obtained by lyophilization of an aqueous solution containing (i) influenza vaccine, (ii) a hydrophobic amino acid, and (iii) arginine or its acid additive salt; and the ratio of component (iii) to the total number of lyophilized ranges from 20 to 85 wt. -%; and the ratio of arginine to its acid salt additive is in such a range that the pH of the aqueous solution is from 8 to 10.

Paragraph 2. Dried product according to paragraph 1, where the influenza vaccine is subjected to a desalting process.

Paragraph 3. Dried product according to paragraph 1, where (ii) a hydrophobic amino acid is phenylalanine or a combination of phenylalanine, at least one of the valine, leucine and isoleucine.

Paragraph 4. Dried product according to paragraph 1, where the proportion of (ii) hydrophobic amino acids ranges from 14 to 75% of the mass. the total quantity of a lyophilized drug.

Paragraph 5. Dried product according to paragraph 1, where (iii) arginine and its acid additive salt are respectively arginine and its hydrochloride.

Paragraph 6. Dried product according to paragraph 1, where the proportion of acid additive salts of arginine is from 1 to 20 parts by mass relative to 1 part by weight of arginine.

Paragraph 7. Dried product according to paragraph 1, where the total amount is in components (i)-(iii) is from 80 to 100% of the mass. relative to the total quantity of a lyophilized drug.

Paragraph 8. Dried product according to paragraph 1, which is soluble to obtain injection solution before use.

Paragraph 9. Dried product according to paragraph 1, which is a pharmaceutical preparation for transpulmonary administration.

Paragraph 10. Dried product according to paragraph 1, which is a pharmaceutical preparation for intranasal administration.

In addition, the present invention also provides a method of obtaining a lyophilized preparation containing influenza vaccine as set forth below.

Paragraph 11. A method of obtaining a lyophilized preparation containing influenza vaccine, including:

the first stage for receiving an aqueous solution containing (i) influenza vaccine, (ii) a hydrophobic amino acid, and (iii) arginine and its acid additive salt; and the content of (i) influenza vaccine in aqueous solution equivalent to 20 to 80% of the mass. relative to the total amount received lyophilized; and pH of the aqueous solution is from 8 to 10; and

the second stage of freeze-drying the resulting aqueous solution.

Paragraph 12. The method according to paragraph 11, where (i) the influenza vaccine is subjected to a desalting process.

Paragraph 13. The method in accordance with sec the TSU-11, where (ii) a hydrophobic amino acid is phenylalanine or a combination of phenylalanine, at least one of the valine, leucine and isoleucine.

Paragraph 14. The method according to paragraph 11, where the content of (ii) hydrophobic amino acids in aqueous solution used in the first stage, the equivalent of 14 to 75% of the mass. relative to the total quantity of a lyophilized drug.

Paragraph 15. The method according to paragraph 11, where (iii) arginine and its acid additive salt are respectively arginine and its hydrochloride.

Paragraph 16. The method according to paragraph 11, where the proportion of the specified acid additive salts of arginine in aqueous solution used in the first stage is from 1 to 20 parts by mass relative to 1 part by weight of arginine.

Paragraph 17. The method according to paragraph 11, where the total content of components (i)-(iii) in aqueous solution used in the first stage, the equivalent of 80 to 100% of the mass. relative to the total amount received lyophilized form.

The effects of the invention

Dried product according to the invention can stably maintain the activity of the influenza vaccine during storage, thus facilitating the distribution and storage, compared with the previous preparations.

In addition, the dried product can be applied in the form of an injection solution, the solution is of the it solution for injection before use or in preparation for transpulmonary injection or intranasal, as he is; therefore, another advantage of the lyophilized drug is that it can be used with various types of injection.

Description of the preferred embodiments

In the description pH denotes a value measured at 25°C.

Dried product according to the invention contains (i) influenza vaccine, (ii) a hydrophobic amino acid, and (iii) arginine and its acid additive salt; in which is included a certain proportion of the component (iii), and the ratio of arginine to its acid salt additive comprising component (iii)is placed in a certain range.

Flu-like vaccines, appropriate as influenza vaccine used in the lyophilized preparation according to the invention (hereinafter sometimes referred to simply as "component (i)"), include subunit vaccines obtained after purification of viral particles grown in chicken embryos, and flu-like (hemagglutinin) vaccines, which are one type of split vaccines. Can also be used flu-like vaccines derived from tissue cultures. Alternatively, you can also use live vaccines derived from attenuated influenza virus, a component of the vaccine, derived from detoxificating influenza viruses by their immunogenicity of intact or whole viral particles, the floor is obtained from inactivated whole virus particles. Influenza vaccine for use in the present invention can be obtained from either or both of the strains a and b, but preferably can be obtained from the flu vaccine and a and b strains.

As the influenza vaccine for use in the invention can be used influenza vaccine received in accordance with a known method or a commercially available influenza vaccine.

When received the influenza vaccine or commercially available influenza vaccine is presented in the form of a solution or powder and contains salt, such as a buffer or preservative, the influenza vaccine is preferably subjected to pre-desalination process to remove salt. When influenza vaccine is exposed so prior to the desalting process, it is possible to more effectively improve the stability of influenza vaccine in a dried preparation according to the invention. Method of desalting influenza vaccine is not specifically limited, and its examples include ultrafiltration, precipitation, ion exchange, dialysis and the like. To prevent the reduction of the activity of influenza vaccine during the process of desalting desalting preferably is carried out using an aqueous solution, the pH of which is raised to the level of 8 to 10, and preferably from 8 to 9, the alkaline substance is m, such as sodium hydroxide, potassium hydroxide, arginine or the like. In particular, the solution with the same composition as the composition of the solution subjected to lyophilization in obtaining a lyophilized drug, with the exception of influenza vaccine, ideal as a solution for use in the desalting process.

The proportion of the component (i)used in the lyophilized preparation according to the invention may be determined in accordance with the method of application of lyophilized and the like. For example, the proportion of the component (i) relative to the total quantity of a lyophilized drug ranges from 0.3 to 60 wt. -%, preferably, from 1 to 50 wt. -%, preferably, from 2 to 40 wt. -%, more preferably, from 3 to 30 wt. -%, and particularly preferably from 3 to 20 wt. -%

Specific examples of hydrophilic amino acids contained in the lyophilized preparation according to the invention (hereinafter sometimes referred to simply as "component (ii)"), include the building blocks of protein, amino acids such as valine, leucine, isoleucine, phenylalanine and the like. In the invention of these hydrophobic amino acids may be used singly or in combination. In the invention, for example, phenylalanine, or a combination of phenylalanine, at least one of the valine, leucine and isoleucine can preferably be used in which the quality of hydrophobic amino acids.

The proportion of component (ii)used in the lyophilized preparation according to the invention may be determined in accordance with the type of component (ii), with the method of application of lyophilized and the like. For example, the proportion of component (ii) with respect to the total quantity of a lyophilized drug ranges from 14 to 75 wt. -%, preferably, from 20 to 75 wt. -%, preferably, from 20 to 70 wt. -%, more preferably, from 25 to 65 wt. -%, and particularly preferably from 30 to 60% of the mass.

Dried product according to the invention also contains arginine and its acid additive salt (hereinafter sometimes referred to simply as "component (iii)"). In other words, the dried product essentially contains a combination of arginine and its acid additive salt.

An acid additive salt of arginine for use in the invention are not specifically limited, while it is pharmacologically acceptable, and examples include inorganic acid salt additive, such as hydrochloride, nitrates and sulfates; and organic acid additive salts such as acetate. Arginine hydrochloride is indicated as a preferred example of an acid additive salt of arginine for use in the invention. In the invention an acid additive salt of arginine can be used alone or two or more the acid additive salts of arginine can be used in combination.

In the dried product according to the invention the total amount of component (iii) (i.e. the total number of arginine and its acid additive salt) is added in proportions of from 20 to 85% by mass. relative to the total quantity of a lyophilized drug. Such proportion of the component (iii) prevents the decrease of activity of influenza vaccine during storage, thereby providing the product with excellent stability. The proportion of component (iii) relative to the total quantity of a lyophilized drug is preferably from 20 to 80 wt. -%, preferably, from 20 to 75 wt. -%, more preferably, from 25 to 70 wt. -%, and particularly preferably from 30 to 65% of the mass.

Component (iii) satisfies any of these proportions and also is such that the ratio of arginine to its acid salt additive is chosen so that the pH of the aqueous solution subjected to lyophilization, ranged from 8 to 10. In other words, arginine is an alkaline and an acid additive salt of arginine is an acid so that the pH of the aqueous solution subjected to lyophilization, is adjusted within this range by adjusting the relationship of arginine to acid additive salts of arginine, provided that the amount used of the component (iii) is placed in any of the above ranges. By regulating the pH in the aqueous solution, subjected to lyophilization in the above range with the use of arginine and its acid salt additive can improve the stability during storage influenza vaccine. Preferably, the ratio of arginine to its acid salt additive is chosen so that the pH of the aqueous solution subjected to lyophilization, ranged from 8 to 9. Used in the present description, the term "aqueous solution subjected to freeze-drying" means an aqueous solution subjected to lyophilization, upon receipt of a lyophilized drug, i.e. an aqueous solution obtained by adding all the components of a lyophilized preparation according to the invention to treated water. The aqueous solution subjected to lyophilization, is adjusted to, for example, the total number of lyophilized (total number of all components) in 1 ml of an aqueous solution ranged from 0.2 to 20 mg, preferably from 0.4 to 20 mg, preferably, from 0.4 to 15 mg, more preferably from 0.6 to 12.5 mg, particularly preferably from 1 to 10 mg, although the invention is not specifically limited to these amounts.

The ratio of arginine to the acid additive salt in component (iii) is not specifically limited, until the pH of the aqueous solution subjected to lyophilization, can be within the above range, and it may be determined in accordance with the tipo is an acid additive salt, the type of the component (ii) and the like. In particular, the proportion of acid additive salts of arginine relative to 1 part by weight of arginine may, for example, be from 1 to 20 parts by weight, preferably from 1 to 15 parts by weight, preferably from 1.25 to 12.5 parts by weight, more preferably from 1.5 to 12.5 parts by weight, and particularly preferably from 2 to 10 parts by mass.

Dried product according to the invention may contain, in addition to the components (i)-(iii), hydrophilic amino acids except arginine (hereinafter sometimes referred to simply as "component (iv)"), in such a range not to impair the effects of the invention. Hydrophilic amino acid may be any amino acid with a hydrophilic side chain, regardless of whether it is a component of protein amino acid or not. Specific examples of hydrophilic amino acids include basic amino acids such as lysine, histidine and the like; neutral hydroxynicotinate, such as serine, threonine and the like; acidic amino acids such as aspartic acid, glutamic acid and the like; amino acids containing an amide group, such as asparagine, glutamine, and the like, glycine, alanine, cysteine, tyrosine and other amino acids. Preferred among these hydrophilic amino acids is Vlada alanine and glycine. Such hydrophilic amino acids may be used alone, or two or more of them may be used in combination.

When such hydrophilic amino acid found in dried preparation according to the invention, the proportion of hydrophilic amino acids is also not specifically limited; but, for example, the proportion of hydrophilic amino acids relative to the total quantity of a lyophilized drug may be from 5 to 50 wt. -%, preferably, from 5 to 40 wt. -%, preferably, from 5 to 30 wt. -%, more preferably, from 5 to 25 wt. -%, and particularly preferably from 10 to 25% of the mass.

In addition, in order to make the effect of stabilizing the influenza vaccine is more significant in the lyophilized preparation according to the invention, the total amount of components (i)-(iii)that does not include the component (iv), or the total amount of components (i)to(iv), which is included in component (iv)is from 80 to 100 wt. -%, preferably, from 85 to 100 wt. -%, preferably from 90 to 100 wt. -%, more preferably, 95 to 100 wt. -%, and particularly preferably 100% of the mass. relative to the total quantity of a lyophilized drug.

Dried product according to the invention may also contain the following components, while not impairing the effects of the invention: monosaccharides, such as glucose; disaccharides, such as sucrose, small the Tosa, lactose and trehalose; sugar alcohols, such as lures; oligosaccharides such as cyclodextrin; and polysaccharides, such as dextran 40 & pullulan; polyhydric alcohols such as polyethylene glycol; salts of fatty acids, such as capret sodium; human serum albumin; inorganic salts; gelatin; surfactants; buffers; etc. Surfactant can be anionic, cationic or non-ionic, as it is a surfactant commonly used in pharmaceutical products. In addition, freeze-dried preparation according to the invention optionally contains an adjuvant.

Dried product according to the invention is obtained by adding predetermined amounts of components (i)-(iii) to purified water, adding component (iv) and, optionally, other components and the effect on the mixture of the lyophilization process. More specifically, the dried product according to the invention can be obtained by the following first and second steps:

first stage: obtaining an aqueous solution containing: (i) influenza vaccine, (ii) a hydrophobic amino acid, and (iii) arginine and its acid additive salt, where the number (iii) arginine and its acid salt additive is equivalent 20-85% of the mass. relative to the total amount received lyophilized, and the pH of the leaves from 8 to 10; and

second stage: the lyophilization of an aqueous solution obtained in the first stage, to obtain a lyophilized form.

Target dried product can be obtained by exposure to an aqueous solution obtained in the first stage, the process of lyophilization. Therefore, in the first stage an aqueous solution, subjected to freeze-drying, it turns out that the ratio in the aqueous solution of the components other than the component (components)that were removed by lyophilization, is the same as the ratio of the components in the resulting product. In addition, the pH of the aqueous solution obtained in the first stage, brought by regulating the relationship of arginine to acid additive salts of arginine, as described above.

Himself dried product according to the invention can be a dried solid (freeze-dried cake)obtained by the process of freeze-drying, or may be a powder obtained from the dried solids.

The method of administration of a lyophilized preparation according to the invention are not specifically limited. For example, the drug may be dissolved in a diluent (solution for injection) before use and subcutaneously introduced in the form of injections. Dried product can also be dissolved in the diluent for the liquid mixture and put transpulmonary is Ino or intranasally in the form of a pharmaceutical preparation for injection through the lungs or in the form of nasal drops. In addition, the dried product can be produced in powder form by exposure to the air strike; therefore, when the dried product is used as a pharmaceutical preparation for injection through the lungs or in the form of nasal drops, he may be given transpulmonary or intranasally as he is, with use of the device for inhalation of a dry powder, which may be accompanied by air blow.

The method of administration of a lyophilized preparation can be defined depending on age and other parameters of the patient, but the drug may be injected, for example, in an amount equivalent to 3 to 300 μg influenza vaccine, once or twice with an interval of about 1 to 4 weeks.

When the dried product is introduced in the form as it is, without dilution in diluent before use, it is preferably placed in the container for each single dose (number of input at a time) with regard to ease of use. When lyophilized drug is dissolved in the diluent before use, it can be placed in the container for each single dose, or multiple doses of the drug can be placed together in one container.

Dried product contains a reduced amount of filler compared with drugs, is holding a flu-like vaccines, which was previously used in the form of injections. Therefore, one characteristic of the drug is that the amount of the drug in the container is low. In the present invention the quantity of a lyophilized drug, placed in the container is not specifically limited, but is, for example, from 0.1 to 10 mg, preferably from 0.15 to 8 mg, preferably, from 0.2 to 6 mg, more preferably from 0.25 to 5 mg, and particularly preferably from 0.3 to 5 mg

Examples

The present invention will be described in more detail below with reference to examples and the like, which are not intended to limit the invention.

Reference example 1: study of the conditions of desalination for flu vaccines

Investigated the conditions of the desalting process for solutions of flu IN the vaccine. In particular, influenza vaccine “Seiken” (product name, manufactured by Denka Seiken, Lot No. 308-A) was used as a solution ON influenza vaccines and a suitable amount of this solution was poured into each Vivaspin concentrator with a capacity of 4 ml (manufactured by Sartorius) and centrifuged at 3000 rpm for 30 minutes. Each of the solutions of the compositions shown in table 1, were then added to each Vivaspin concentrator and the mixture was centrifuged at 3000 rpm for 30 minutes. This procedure was repeated three times.

The activity of p is obtained desalted flu ON vaccines (size) was measured in accordance with the following method.

Measurement of flu activity IN vaccines

Receive 5% vol. suspension of chicken red blood cells

Canned chicken blood (supplied by Nippon Biotest Labo.) was placed in a test tube and centrifuged for 5 minutes at 900 g with the subsequent removal of the supernatant liquid and leukocyte layer. Then to erythrocytes in vitro was added to the diluent composition shown below (solution 1/200 mol/l sodium chloride, phosphate buffered (pH 7,2)). The mixture was mixed and subsequently centrifuged to remove supernatant. The procedure was repeated three times. Erythrocytes in vitro was aspirated with a pipette and placed in a container containing a diluent and mixed to obtain 5% vol. suspension of chicken erythrocytes.

The composition of the diluent

NaCl8.5 g
Na2HPO4·12H2O1,425 g
KH2PO4is 0.135 g
Purified water1000 ml

2. Dimension value

Each of the portions 50 μl of influenza vaccines with different factors dilution was added to the micropipette and then to it was added 50 μl of 5% vol. suspensions chicken ericr is Titov (method a two-stage cultivation). The samples in the microplate were thoroughly mixed and left to stand at room temperature for one hour, then was evaluated the highest dilution factor among the samples of each influenza vaccines when complete agglutination of erythrocytes as its value.

The results are shown in table 1. The results confirmed that there is a decrease in the activity of flu IN the vaccine subjected to desalting process solution with a pH of 7 or below; however, flu-like vaccines subjected to desalting process solution with a pH of 8 to 9, were able to maintain stable activity.

Table 1
Compositions and pH solutionsThe residual activity
The condition 10.5 mg Phe, added to 0.5 ml of purified water; pH brought NaOH to 3.012,5%
The condition 20.5 mg Phe, added to 0.5 ml of purified water; pH brought NaOH to 4.06,3%
Condition 30.5 mg Phe, added to 0.5 ml of purified water; pH brought NaOH to 5.712,5%
Condition 40.5 mg Phe, added to 0.5 ml of purified water; pH brought NaOH to 7.050%
Condition 5Without the addition of Phe in 0.5 ml of purified water; pH brought NaOH to 7.050%
The condition 60.5 mg Phe, added to 0.5 ml of purified water; pH brought NaOH to 8.0100%
Condition 7Without the addition of Phe in 0.5 ml of purified water; pH brought NaOH to 8.0100%
Condition 80.5 mg Phe, added to 0.5 ml of purified water; pH brought NaOH to 9.0100%
#"Residual activity" in table 1 indicates the percentage (%) values after the desalting process relative to the values before the desalting process.

The test example 1:% sustainability Assessment of flu vaccines

Influenza vaccine “Seiken” (product name, manufactured by Denka Seiken, Lot No. 308-A) was used as a solution ON influenza vaccine and 1 ml (equivalent to 90 mcg influenza vaccines) of this solution was poured in each Vivaspin concentrator with a capacity of 4 ml (produced by what company Sartorius) and centrifuged at 3000 rpm for 30 minutes. Each of the solutions of the compositions shown in table 2, was then added to each Vivaspin concentrator and the mixture was centrifuged at 3000 rpm for 30 minutes. This procedure was repeated three times. The final solutions obtained after concentration and desalting / buffer exchange), had the compositions shown in table 2, and their volume was brought to the original volume (1 ml). The glass bottle was filled with 0.5 ml of each of the obtained solutions and the solutions liofilizirovanny to obtain a lyophilized product.

Example 5
Table 2
Soustava and pH of the solutions used in examples and comparative examples
Example 10.5 mg Phe, 0.5 mg of Arg-HCl and 0.075 mg Arg added to 0.5 ml of purified water; pH 8
Example 20.5 mg Phe, 0.3 mg of Arg-HCl and 0,050 mg Arg added to 0.5 ml of purified water; pH 8
Example 30.5 mg Phe, 0.2 mg Ile, 0.3 mg of Arg-HCl and 0,057 mg Arg added to 0.5 ml of purified water; pH 8
Example 40.5 mg Phe, 0.2 mg Val, 0.3 mg of Arg-HCl and to 0.055 mg Arg added to 0.5 ml of purified water; pH 8
0.5 mg Phe, 0.2 mg Leu, 0.3 mg of Arg-HCl and 0,052 mg Arg added to 0.5 ml of purified water; pH 8
Example 60.5 mg Phe, 0.2 mg Ala, 0.3 mg of Arg-HCl and 0,052 mg Arg added to 0.5 ml of purified water; pH 8
Example 70.5 mg Phe, 0.2 mg Gly, 0.3 mg of Arg-HCl and 0,052 mg Arg added to 0.5 ml of purified water; pH 8
Comparative example 10.5 mg Phe and an appropriate amount of NaOH added to 0.5 ml of purified water; pH 8
Comparative example 20.5 mg Phe and to 0.060 mg Arg added to 0.5 ml of purified water; pH 8
Comparative example 30.3 mg of Arg-HCl and 0.030 mg Arg added to 0.5 ml of purified water; pH 8
Comparative example 40.5 mg Phe, 0.5 mg His and 0.30 mg Arg added to 0.5 ml of purified water; pH 8
# the pH of the solutions used in examples 1-7 and Comparative examples 2-4, drove up to 8, using the specified number of components without the use of other regulators of pH. NaOH was added to the solution used in Comparative Example 1, so that the pH was 8 after adding the given number of comp is the component.

Each of the obtained lyophilized preparations was hermetically sealed in a glass bottle and stored for 4 weeks in a dark place at 25°C and a relative humidity of 60%. Immediately upon receipt and after 1, 2, 3 and 4 weeks after saving ON influenza vaccine each lyophilized preparation was carried out measurement values in accordance with the same method described in Reference example 1.

The results are shown in table 3. When using solutions containing phenylalanine whose pH was brought to 8 with only sodium hydroxide and arginine, it was impossible to prevent the reduction in the activity of flu IN the vaccine during storage (Comparative examples 1 and 2). Also in the case of using the solution, pH of which was brought to 8 by adding arginine and arginine hydrochloride without addition of phenylalanine, which is a hydrophobic amino acid, it was impossible to prevent the decline in activity ON influenza vaccine (Comparative example 3). In addition, in the case of using the solution, pH of which was brought to 8 by using a combination of phenylalanine, histidine, which is a basic amino acid, and arginine, it was impossible to prevent the decline in activity ON influenza vaccines

On the contrary, it was found that kogarashi for use in obtaining a lyophilized drug meets the conditions (1)-(3), below, you can steadily maintain the activity of influenza vaccines in the final dried product:

(1) includes a hydrophobic amino acid, arginine and acid additive salt of arginine; (2) the ratio of arginine to acid additive salts of arginine selected so that the pH of the solution is 8; (3) the total number of arginine to acid additive salts of arginine is chosen so that it was equal to or exceeded the target concentration in the dried drug.

Table 3
The compositions of lyophilised preparationsThe residual activity
Immediately
after receiving
After 1 weekAfter 2 weeksAfter 4 weeks
Example 14,0% vaccines
44,6% Phe, 51,4% Arg
and Arg-HCl
100%100%100%100%
Example 25,0% vaccines
55,9% Phe, 39,1% Arg
and Arg-HCl
100% -100%-
Example 34.1% a vaccine,
45.4% of Phe, 18,1% Ile, 32,4% Arg and Arg-HCl
100%-100%-
Example 44.1% a vaccine,
45.4% of Phe, 18,2%
Val, 32,3% Arg
and Arg-HCl
100%-100%-
Example 54.1% a vaccine, 45.6% of Phe, 18,2% Leu, 32.1% of Arg and Arg-HCl100%-100%-
Example 64.1% a vaccine, 45.6% of Phe, 18,2% Ala, 32.1% of Arg and Arg-HCl100%-100%-
Example 74.1% a vaccine, 45.6% of Phe, 18,2% Gly, 32.1% of Arg and Arg-HCl100%-100%-
Comparative example 1Approximately 8.3% of the vaccine, approximately 91.7% of Phe, a suitable quantity aOH 100%25%6,3%3,1%
Comparative example 27.4% vaccines, 82.7% of Phe, 9,9% Arg100%50%25%25%
Comparative example 37,8% FOR vaccines, 92,2% Arg and Arg-HCl100%-25%-
Comparative example 44,2% FOR vaccines, 46.5% of Phe, 46.5% of His, 2,8% Arg100%50%12,5%6,3%
# "Residual activity" in table 3 indicates the percentage (%) values of lyophilized after saving relative values of lyophilized immediately after receipt.
In table 3 unit % in the column "Composition of lyophilised compositions" means % by mass.
In table 3, " - " means "not measured"

1. Freeze-dried preparation containing influenza vaccine obtained by lyophilization of an aqueous solution containing (i influenza vaccine, (ii) a hydrophobic amino acid, and (iii) arginine and its acid additive salt;
moreover, the proportion of the component (iii) arginine and its acid salt additive relative to the total quantity of a lyophilized drug is from 20 to 85 wt.%; and the ratio of arginine to its acid salt additive is in such a range that the pH of the aqueous solution is from 8 to 10, with influenza vaccine subjected to desalting process.

2. Dried product according to claim 1, where (ii) a hydrophobic amino acid is a phenylalanine and, optionally, included at least one component selected from valine, leucine and isoleucine.

3. Dried product according to claim 1, where the proportion of (ii) hydrophobic amino acids ranges from 14 to 75 wt.% of the total number of lyophilized.

4. Dried product according to claim 1, where (iii) arginine and its acid additive salt are respectively arginine and its hydrochloride.

5. Dried product according to claim 1, where the proportion of acid additive salts of arginine is from 1 to 20 parts by mass relative to 1 part by weight of arginine.

6. Dried product according to claim 1, where the total of components (i)-(iii) is from 80 to 100 wt.% relative to the total quantity of a lyophilized drug.

7. Dried product according to claim 1 which is soluble to obtain injection solution before use.

8. Dried product according to claim 1, which is a pharmaceutical preparation for transpulmonary administration.

9. Dried product according to claim 1, which is a pharmaceutical preparation for intranasal administration.

10. A method of obtaining a lyophilized preparation containing influenza vaccine, including:
the first stage for receiving an aqueous solution containing (i) influenza vaccine, (ii) a hydrophobic amino acid and (ii) arginine and its acid additive salt; and the content of (iii) arginine and its acid additive salts in aqueous solution equivalent to 20 to 85 wt.% relative to the total amount received lyophilized; and pH of the aqueous solution is from 8 to 10; and
the second stage of freeze-drying the resulting aqueous solution;
when influenza vaccine is subjected to desalting process.

11. The method according to claim 10, where (ii) a hydrophobic amino acid is a phenylalanine and, optionally, included at least one component selected from valine, leucine and isoleucine.

12. The method according to claim 10, where the content of (ii) hydrophobic amino acids in aqueous solution used in the first stage, the equivalent of 14 to 75 wt.% relative to the total quantity of a lyophilized drug.

13. The method according to claim 10, where (iii) arginine and its acid additive salt Ave is astavliaut respectively arginine and its hydrochloride.

14. The method according to claim 10, where the proportion of the specified acid additive salts of arginine in aqueous solution used in the first stage is from 1 to 20 parts by mass relative to 1 part by weight of arginine.

15. The method according to claim 10, where the total content of components (i)-(iii) in aqueous solution used in the first stage, the equivalent of 80 to 100 wt.% relative to the total amount received lyophilized form.



 

Same patents:

FIELD: biotechnologies.

SUBSTANCE: method to produce a flu virus is proposed, according to which hens are injected with a vaccine against flu, eggs are collected from vaccinated hens, the process of embryogenesis is initiated, eggs with a developing embryo are infected, introducing a flu virus into an allantoic cavity, infected eggs with developing embryos are incubated under temperature and moisture, which provide for virus replication, and the allantoic liquid is collected, which contains a virus. Also application of the flu virus and eggs produced by this method is proposed.

EFFECT: method improvement.

26 cl, 9 tbl, 5 ex

FIELD: biotechnologies.

SUBSTANCE: methods are described to prevent a virus flu infection and detection of efficiency of a flu virus vaccine using a molecule of hemagglutinin (HA) of a flu virus A of subtype H5, immunogenicity of which is increased by replacement of amino acids in the HA sequence.

EFFECT: replacement of specific remains in HA, such as introduction of asparagin into the position 223 in HA H5, makes it possible to increase sensitivity of hemagglutination moderation reaction as a result of variation of receptor specificity or ability to binding of antibody-antigen.

24 cl, 7 tbl, 7 ex

FIELD: medicine.

SUBSTANCE: invention refers to pharmaceutical industry, namely an antiviral agent. The soluble melanine antiviral agent prepared by extraction of Inonotus obliquus basidium fungus and possessing antiviral activity on viruses of influenza, type 2 herpes simplex, immunodeficiency (HIV-1) and variolovaccine.

EFFECT: agent possess a wide spectrum of antiviral action.

6 cl, 4 tbl, 13 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention relates to medicine and deals with combined medication for elimination of catarrhal diseases and flu, which contains phenylephrine hydrochloride, N-acetyl-L-hydroxyproline and cetirizine hydrochloride.

EFFECT: invention ensures suppression of inflammatory pain, which corresponds to pharmacological profile of medication as anti-inflammatory and pain-killing remedy.

1 cl, 2 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to pharmaceutics and medicine and concerns a HIV-1 and influenza A virus reproduction inhibitor representing 2-hydroxyimino-3-oxolup-20(29)-en-28-oic acid and possessing high antiviral activity.

EFFECT: preparing the agent with high antiviral activity.

1 tbl, 2 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: present invention refers to antiviral preparations produced of herbal raw materials, i.e. cinnamon, leechee and peanut. According to the present invention, there are presented a composition and a method for preparing said composition containing pentamerous, trimerous and tetramerous flavonoid procyanidine.

EFFECT: composition improves an immune response, and as it is found, it is applicable for treating and controlling HIV infection and AIDS, as well as for preventing, treating and controlling influenza virus and infection.

25 cl, 5 tbl, 14 ex, 5 dwg

FIELD: medicine.

SUBSTANCE: bacterial Lactobacillus casei CNNM 1-1518 strain is used for preparing an oral composition for increasing humoral immunity induced in influenza vaccination, preferentially in patients aged 65 years and older. The composition is found in the form of an enzyme milk product, in the form of a foodstuff or a food additive, and with the content Lactobacillus casei CNNM 1-1518, preferentially 1x108 to 1x109 CFU/ml.

EFFECT: intensified humoral response induced by influenza vaccination.

10 dwg, 1 tbl

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to new substituted (3R,4R,5S)-5-amino-4-acylamino-3-(1-ethyl-propoxy)-cyclohex-1-ene carboxylic acids of general formula 1, their esters, pharmaceutically acceptable salts and/or hydrates possessing antiviral activity associated with inhibition of neuraminidase, particularly influenza virus neuraminidase. The invention also refers to pharmaceutical compositions and a based drug preparation, a method for inhibition of neuraminidase, and a method of treating influenza and pneumonia caused by influenza virus. In general formula 1 R1 represents hydrogen, C1-C7alkyl, C2-C7alkenyl or C2-C7alkinyl optionally substituted by C3-C6cycloalkyl, phenyl, pyridyl, C1-C3alkoxy; R2 represents hydrogen; -NH2; C2H5O-, substituted C1-C5alkyl specified in -CH2OH, -CH2-NH2, -CH=CH2, -CH=CHCl, -C=CH, -CH2F, -CHF2, -CF3, -CH2CF3, cyclo-C3H7, -CF2CF2CF2CF3, -CF2CF2CF2CHF2, -CH2CF2CF2CF2CF3.

EFFECT: compounds can find application in treating influenza virus and pneumonia caused by influenza virus.

10 cl, 3 tbl, 33 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to a new β-cyclodextrine clathrate complex (an inclusion compound) with 5-hydroxy-4-aminomethyl-1-cyclohexyl(or cycloheptyl)-3-alkoxycarbonylindole derivative: β-cyclodextrine 1:1 to 1:5, preferentially at the relation of 1:1 to 1:3 of general formula (I): wherein X means - hydrogen, chlorine, iodine, n=1 or 2, R3-C1-C3 alkyl, ALK means C1-C6 alkyl group, R1, R2 are independently specified in C1-C4-alkyl, preferentially methyl, or R1 and R2 together with a nitrogen atom (i.e. group - NR1R2) means the groups described by formulas: wherein Bn is benzyl, a Ph is phenyl with the molar ratio of 5-hydroxy-4-aminomethyl-1-cyclohexyl(or cycloheptyl)-3-alkoxycarbonylindole derivative: β-cyclodextrine 1:1 to 1:5, preferentially 1:1 to 1:3, especially preferentially in the relation of 1:2. The clathrate complex may represent nanoparticles of size not less than 100 nm. There are preferential clathrate complexes wherein 5-hydroxy-4-aminomethyl-1-cyclohexyl(or cycloheptyl)-3-alkoxycarbonylindole derivative represents 1-cyclohexyl-4-aminomethyl-5-hydroxy-2-methyl-1H-indole-3-carboxylic acid ethyl ester. The new clathrate complexes possess antiviral action and exhibit high activity versus influenza viruses. The invention also involves a pharmaceutical composition and a drug based on the clathrate complexes. Besides, the invention refers to liquid-phase and solid-phase synthesis of the clathrate complexes.

EFFECT: preparing the compounds which possess antiviral action and exhibit high activity versus influenza viruses.

20 cl, 2 ex, 2 tbl, 8 dwg

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to a pharmaceutical combination and to its use for treating an infection caused by influenza virus. The declared composition contains a pyrazine derivative of formula wherein R1 and R2 are identical or different, and each represents a hydrogen atom or a halogen atom; and R3 represent a hydrogen atom or a protective group for amino group or its salt, and, and a neuraminidase inhibitor. The neuraminidase inhibitor is specified in oseltamivir, zanamivir, peramivir or CS-8958.

EFFECT: invention provides preparing the combination which shows strong antiviral activity, smaller side effects and applicable for treating influenza.

12 cl, 8 tbl, 4 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: claimed invention relates to method of obtaining aripiprazole suspension. Method includes the following stages: (a) combining aripiprazole mass and carrier with formation of primary suspension; (b) first refining of primary suspension with obtaining secondary suspension using apparatus for refining with large shearing force, dispersant, in which shearing force is applied for material processing, or high-pressure homogeniser; and (c) second refining of secondary suspension with obtaining final sterile suspension by high-pressure homogeniser. Claimed composition also relates to methods of obtaining lyophilysed composition from aripiprazole suspension.

EFFECT: claimed invention provides possibility of obtaining aripiprazole suspension with the average size of particles from 1 to 10 mcm, without necessity to use special crystallisation technologies for obtaining initial aripiprazole mass and without application of vacuum mixing at the stage of combining aripiprazole mass and carrier.

33 cl, 13 tbl, 15 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: claimed invention relates to chemical-pharmaceutical composition and represents method of lyophilisation of particles, which contain frozen liquid and have contained in them pharmaceutical composition, including provision of heat-conducting container, which has bottom and side walls; filling container with a layer of particles, with the layer including multiple strata of particles and having aspect ratio mot less than 1; providing source of heat above upper stratum of particles, with source of heat having surface facing upper stratum of the layer, where said surface is characterised by emissivity factor equal, at least, 0.4; impact on container-filling particles with pressure lower than atmospheric pressure; heating at least container bottom and said surface to conduct heat to particles in order to ensure sublimation of frozen liquid at pressure lower than atmospheric pressure; ceasing conduction of heat to particles after sublimation of frozen liquid.

EFFECT: invention ensures uniform drying of multiple layer of particles with obtaining homogenous product.

11 cl, 4 ex, 2 dwg

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention relates to field of biotechnology and immunology. Claimed is medication for treatment and prevention of TNF-dependent disorder, which contains molecule of TNF-binding nanobody, lyoprotector, surface-active substance and buffer, method of such medication obtaining, method of lyophilised preparation reduction and method of analysis of medication-manufacturing process, as well as method and set for treatment or prevention of TNF-dependent disorder.

EFFECT: invention ensures obtaining stable pharmaceutical preparations of TNF-binding nanobodies and can be applied in therapy of TNF-dependent diseases.

30 cl, 12 ex, 3 tbl, 32 dwg

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention relates to field of pharmaceutics. Lyophilised pharmaceutical composition contains IX factor, more than 90% of whose calcium-binding ability are preserved in storage within 6 months at 25°C and tregalose, taken in amount 0.5%-3% by volume. Method of preparing stable dry composition includes stages of mixing IX factor-containing solution with tregalose 0.5%-3% by volume with obtaining cryoprotective solution. Method of pharmaceutical composition lyophilisation includes stages of freezing pharmaceutical composition containing IX factor and tregalose by 0.5%-3% in volume at temperature -40°C or less, burning pharmaceutical composition at temperature approximately from -20°C to -35°C, reduction of pharmaceutical composition temperature to -40°C or less, drying pharmaceutical composition at the first stage of drying at temperature from 5°C to 20°C at lower pressure, drying pharmaceutical composition at the second stage of drying at temperature from 45°C to 55°C at lower pressure. Method of preparing IX factor-containing lyophilised composition ensures preservation of more than 90% of IX factor calcium-binding ability.

EFFECT: application of composition on the basis of tregalose in amount 0,5%-3% in volume ensures its stability for 6 months of storage at conditions of 25°C.

17 cl, 5 ex, 6 dwg

FIELD: medicine, pharmaceutics.

SUBSTANCE: method of obtaining drug-containing composition of polymeric micelles includes: dissolution of poorly dissolved in water drug and amphiphilic block copolymer in organic solvent; and addition to obtained mixture in organic solvent of water solution with formation of polymeric micelles. Method does not require carrying out separate operation of organic solvent removal before formation of micelles. Method is simple, reduces technological time and amenable to scaling.

EFFECT: in accordance with invention method polymeric micelles preserve their stability.

14 cl, 9 tbl, 6 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to chitosan carboxyalkylamide hydrogel and may be used for cosmetic and dermatological treatment of skin burns. Chitosan carboxyalkylamide hydrogel of pH close to that of skin and making 6.5 to 7.2 contains 40 to 90 mole % of the groups of N-carboxyalkylaminde D-glucosamine of formula (I) wherein n represents an integer 1 to 8, 60 to 10 mole % of the protic groups of D-glucosamine, and 5 to 15 mole % of the groups of N-acetyl-D-glucosamine. A method for preparing said hydrogel involves preparing an acid solution of chitosan of a degree of acetylation of 85 to 95%, providing a reaction of produced additive chitosan salt in an aqueous solution of diorganic acid and correcting pH of the prepared solution.

EFFECT: preparing the anhydrous product of chitosan carboxyalkylamide prepared by hydrogel dehydration.

12 cl, 1 dwg

FIELD: medicine, pharmaceutics.

SUBSTANCE: group of inventions refers to an non-aqueous pharmaceutical composition containing dehydrated active polypeptide containing 10-100 amino acids, and at least one semipolar proton organic solvent, with polypeptide dehydrated at pre-set pH differing by at least 1 pH unit from pi polypeptide in the aqueous solution and to a method for preparing the composition.

EFFECT: group of inventions provides improved stability of the non-aqueous composition of therapeutic peptide.

39 cl, 7 ex, 11 dwg

FIELD: medicine, pharmaceutics.

SUBSTANCE: method for preparing a composition for injections containing sodium cevtriaxone and sodium tazobactam involves the following stages: (a) suspension of raw materials, i.e. sodium cevtriaxone, sodium tazobactam, sterilised water for injections, mixed solution of ethyl acetate and isopropyl alcohol, and anhydrous ethanol in mass relation making 3-5:1:2:5:9, with volume relation of ethyl acetate to isopropyl alcohol making 1:2-4; (b) dissolution of sodium cevtriaxone and sodium tazobactam in sterilised water for injections with added activated hydrocarbon and filtration; (c) addition of the mixed solution of ethyl acetate and isopropyl alcohol to the filtrate and agitation of the mixture; addition of a seed crystal of sodium cevtriaxone to the solution for crystallisation initiation; and finally washing of the crystals in anhydrous ethanol and crystal drying; and (d) lyophilisation to form the composition for injections containing sodium cevtriaxone and sodium tazobactam.

EFFECT: composition is characterised by high uniformity, high degree of purity and safety of use.

6 cl, 2 tbl, 3 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: group of inventions relates to pharmaceutical composition, which contains parathyroid hormone (PTH), methionine, at least one SAS poloxamer and physiologically acceptable buffer preparation, which can be applied in animals and people, as well as to method of obtaining said composition and application.

EFFECT: group of inventions ensures composition solution stability and prolonged storage term.

19 cl, 11 ex, 22 tbl, 3 dwg

FIELD: medicine.

SUBSTANCE: invention refers to medicine. What is presented is a lyophilised preparation containing at least one nicotine - virus-like particle conjugate, and a stabiliser composition containing at least one nonreducing disaccharide and at least one non-ionic surfactant wherein the pre-lyophilisation pH value of the stabiliser composition makes 5.8 to 6.6, preferentially 6.0 to 6.4.

EFFECT: invention provides producing the effective lyophilised vaccinal preparation able to keep stability for a long period of time.

14 cl, 3 dwg, 4 tbl, 6 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to medicine, more specifically to an agent to be used for treating arterial hypertension and congestive cardiac failure. A pharmaceutical composition contains ramipril, a combination of sodium bicarbonate and arginine as a stabiliser, lactose monohydrate, microcrystalline cellulose, hypromellose, sodium croscarmellose, glidant and a lubricant and optionally a dying agent. A method for preparing the pharmaceutical composition involves moisturising of a mixture of ramipril, lactose monohydrate, microcrystalline cellulose, sodium bicarbonate, and optionally the dying agent in a solution of hypromellose and arginine, granulation, drying, dry granulation, addition of sodium croscarmellose, glidant and the lubricant, and tableting of the prepared mixture. The pharmaceutical composition in the form of a solid dosage form is characterised by decreased formation of all impurities, including ramiprilate and diketopiperazine ramipril, fast release of the active substance, high strength and storage stability of the quality characteristics guaranteed within more than 2 years of shelf life.

EFFECT: preparing the agent for treating arterial hypertension and congestive cardiac failure.

11 cl, 2 tbl, 12 ex

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