Stable fat emulsion (versions), method for preparing it, emulsifying agent and methods for stabilising prostaglandin and fat drops

FIELD: medicine, pharmaceutics.

SUBSTANCE: stable fat emulsion contains prostaglandin as an active ingredient and phospholipids containing phosphatidylcholine and phosphatidyl glycerol in mass ratio 85:15 to 99.7:0.3. The fat emulsion under the invention and its active ingredient (prostaglandin) possess physical and chemical stability thereby increasing shelf life to approximately two years, and/or extended range of storage temperature to 10°C as compared with a commercially available fat prostaglandin emulsion.

EFFECT: fat emulsion under the invention enables satisfactory effectiveness even in the introduction of a low amount.

25 cl, 10 tbl, 12 ex

 

The technical field to which the invention relates.

The present invention relates to a fat emulsion of prostaglandin, which exhibits excellent resistance and contains the active ingredient with excellent resistance to a method for producing and stabilizing the fat emulsion of prostaglandin and emulsifying agent used for stabilization.

The level of technology

Prostaglandin is a physiologically active substance, synthesized from the essential fatty acids containing from 3 to 5 double bonds. Prostaglandin is involved in regulation of inflammation, pain and swelling in functions regulation of blood pressure, cardiac activity and activity of the gastrointestinal tract, in the regulation of the secretion of digestive enzyme in the regulation of renal function, blood coagulation, platelet aggregation and allergic reactions in neuronal transmission in the development of various hormones, etc.

As far as getting prostaglandin E1 (PGE1), developed the fat emulsion for intravenous injection (see, for example, JP-B-1-57094, JP-B-1-57096 and JP-A-2001-10958). Some of these emulsions are commercially available.

However, commercially available fat emulsion PGE1 should be stored at low temperature (for example, at 5°C or below) in the dark.

The period at which guaranteed the quality of these fat emulsions PGE1, cor is current (for example, 5°C × 1 year) even if the fat emulsion PGE1 stored at low temperature. The reason for this is the chemical instability of PGE1 (active ingredient). Since the fat emulsion PGE1 should be protected from freezing during storage, it is necessary to lead a strict temperature control during storage of fat emulsions at 5°C. As commercially available fat emulsion PGE1 should be kept under strict temperature control, there is the difficulty of quality control fat emulsions PGE1 during storage. Have been made various attempts to improve the resistance of PGE1 (see, for example, JP-B-8-18989 and JP-A-4-338333). However, these attempts have not led to significant results. In some documents disclosed measures to improve the stability of fat emulsions (see, for example, JP-T-2005-500366 and WO 2004/52354). However, these documents do not mention the fat emulsion of prostaglandin.

In view of this situation, there remains a need in the development of the fat emulsion of prostaglandin, which exhibits excellent resistance and contains the active ingredient (prostaglandin) with excellent resistance.

Disclosure of inventions

The invention provides a fat emulsion of prostaglandin, which contains a prostaglandin as an active ingredient and exhibits excellent resistance of the active ingredient and is great for the th stability of the emulsion, the method of obtaining and stabilizing the fat emulsion of prostaglandin and emulsifying agent used for stabilization.

The fatty emulsion according to one aspect of the invention, contains a prostaglandin as an active ingredient, with the fat emulsion comprises a phospholipid that contains phosphatidylcholine (PC) and phosphatidylglycerol (FG) in the ratio (PC:PG) of 85:15 up to 99.7:0.3 to.

In the above fat emulsion, the ratio of PC to PG (PC:PG) may be from 95:5 up to 99.7:0.3 to. In this case, the ratio of PC to PG (PC:PG) may range from 97:3 to 99.5:0.5 in.

The above fat emulsion essentially may not contain phosphatidylethanolamine (pea).

In the above fat emulsion FG may include linear saturated or unsaturated residue of a fatty acid with 12-18 carbon atoms.

In the above fat emulsion FG can occur from egg yolk.

The above fat emulsion can essentially do not contain free higher fatty acid or its salt.

The above fat emulsion can essentially do not contain free oleic acid.

The above fat emulsion may contain free higher fatty acid or its salt in an amount of 0.015 wt. parts or less, relative to 1 mass. part of the phospholipid. The above fat emulsion can in order to keep the free higher fatty acid or its salt in an amount of 0.15 wt. parts or less, relative to 1 mass. part of the phospholipid. In this case, the free higher fatty acid may be free oleic acid.

In the above-mentioned fatty emulsion of the total content of PF and PG phospholipid can be as high as 95% or more.

The above fat emulsion can be used for intravenous injection.

In the above-mentioned fatty emulsion of the prostaglandin may be prostaglandin E1 or its derivative.

In the above-mentioned fatty emulsion of the prostaglandin may be prostaglandin E1.

In this case, the fat emulsion average particle diameter may be 300 nm or less, and the residual content of prostaglandin E1 after storage at 40°C for seven days - 70% or more.

In the fat emulsion average particle diameter may be 300 nm or less, and the residual content of prostaglandin E1 after storage at 40°C for seven days - 80% or more.

In the fat emulsion average particle diameter may be 300 nm or less, and the residual content of prostaglandin E1 after storage at 40°C for seven days - 85% or more.

In the fat emulsion average particle diameter may be 300 nm or less, and the residual content of prostaglandin E1 after storage at 20°C for two months - 80% or more.

In the fat emulsion of the average diameter of the particles may be 300 nm or less, and the residual content of prostaglandin E1 after storage at 5°C for one year is 80% or more.

The method of obtaining the fat emulsion according to one aspect of the invention includes preparing a fat emulsion containing prostaglandin as an active ingredient and a phospholipid that contains phosphatidylcholine (PC) and phosphatidylglycerol (FG) in the ratio (PC:PG) of 85:15 up to 99.7:0.3 to.

In the above method of obtaining the fat emulsion pH can be set at the level of 4 to 7.

In the above method of obtaining the fat emulsion pH can be set at the level of 4.5 to 6.5.

The method of stabilization of prostaglandin according to one aspect of the invention includes the use of phospholipid ratio piatevshchina (PF) to phosphatidylglycerol (FG) (PC:PG) of 85:15 up to 99.7:0.3, and essentially does not contain phosphatidylethanolamine (pea) in the fat emulsion, which contains a prostaglandin as an active ingredient.

Emulsifying agent, according to an additional aspect of the invention, contains a phospholipid, in which the ratio of phosphatidylcholine (PC) to phosphatidylglycerol (FG) (FG:FG) is from 85:15 up to 99.7:0.3, and essentially does not contain phosphatidylethanolamine (pea), emulsifying agent is used for the fat emulsion, which contains a prostaglandin as an active ingredient.

p> Because of the above fatty emulsion comprises a phospholipid that contains phosphatidylcholine (PC) and phosphatidylglycerol (FG) in the ratio (PC:PG) of 85:15 up to 99.7:0.3, and fat emulsion and active ingredient (prostaglandin) show excellent resistance. Therefore, the shelf life can be increased and/or the temperature range storage can be expanded in comparison with commercially available fat emulsion prostaglandin. For example, the period (5°C × 1 year), during which ensures the quality of the fat emulsion PGE1 may be extended to approximately two years, or temperature range of storage can be expanded up to 10°C (see results of examples described later). This facilitates the control of quality during storage. As the drops of fat in the above fat emulsions have a small and uniform diameter, the drug can effectively accumulate in the affected area. Therefore, the fatty emulsion exhibits excellent efficiency even with the introduction of a small amount.

The implementation of the invention

Below are the fatty emulsion, which comprises a prostaglandin as an active ingredient (hereinafter referred to in this document as "the fat emulsion prostaglandin or fat emulsion") according to one variant of implementation, its preparation and stabilization and emulgirujushchih the agent, used for stabilization. In the following implementation options under the sign "%" mean "mass%", and the term "part(s)" mean "part(s) by weight".

1. Fat emulsion

Fat emulsion prostaglandin according to this variant implementation is an emulsified product (emulsion of the type oil-in-water"), which includes prostaglandin (active ingredient), phospholipid (emulsifying agent), base oil and water. In particular, the fatty emulsion of prostaglandin according to this variant implementation is an emulsified product, in which droplets of fat dispersed in water. In every drop of fat prostaglandin and base oil is mainly engaged in the membrane, which contains a phospholipid. The content of each component in the fat emulsion of prostaglandin according to this variant implementation is as follows. In particular, the content of prostaglandin is from 0.2 to 100 μg/ml, the content of the base oil is 5-50% relative to the total amount of the fat emulsion and the concentration of phospholipid - 1-50% relative to the total amount of base oil.

Fat emulsion prostaglandin according to this variant implementation can also include higher fatty acid, an isotonic agent, an antioxidant agent, regulating the pH, if necessary

the pH of the fat emulsion according to this variant implementation is preferably from 4 to 7, and more preferably from 4.5 to 6.5. If the pH of the fat emulsion according to this variant implementation is less than 4, the stability of the emulsion may be reduced. If the pH of the fat emulsion according to this variant implementation of more than 7, the stability of the active ingredient may be reduced.

Each component of the fat emulsion according to this version of the implementation are described below.

1.1. Prostaglandin

The term "Prostaglandin" is the common name of the compounds, a basic skeleton which is postanova acid. The loop structure prostaglandins are classified into the following types: a, b, C, D, E, F, G, H, I, and the like. Prostaglandin is a physiologically active substance, which plays the role of alarm substance cells in relation to stimulation of smooth muscle, inflammation, allergies, secretion/aggregation/accumulation of cells in one location, to cell growth, neuronal transmission, etc. In particular, prostaglandin has a physiological activity, such as contractile activity of smooth muscles (e.g., muscles of the uterus and small intestine), anti-hypertensive activity, vasoconstrictor activity, antilipolytic activity, inhibition of gastric secretion, the effect on the Central nervo the system, the decrease in the adhesion of platelets, inhibition of platelet aggregation, inhibition of thrombogenesis, effect on epidermal growth and stimulation of keratinization depending on the type of prostaglandin.

Examples of prostaglandin included in the fat emulsion according to this variant implementation, are PGA1, PGB1, PGD2, PGE1, PGE2, PGF1-alpha, RR-alpha, PGI2 and their derivatives. Among them PGE1 and its derivatives are preferred. As derived prostaglandin preferred it difficult alkilany ether (see, for example, Japanese patent No. 2602964).

For example, PGE1 has vasodilator activity, protivogipertonicheskoe activity, activity to increase renal blood flow, natriuresis activity, activity to enhance the secretion of renin activity increased secretions eritropoyetina, activity inhibition of platelet aggregation, bronchodilator activity, activity to reduce uterine activity to accelerate the peristalsis of the intestine, the activity reduction of the longitudinal muscles of the stomach/intestinal activity on the relaxation of the circular muscle of the stomach/intestinal activity by inhibition of gastric secretion, protective effect to the gastric mucosa, immune activity, activity to suppress excretion of norepinephrine per feericheskih sympathetic nerve endings, etc. Therefore, according to this variant implementation of the fat emulsion, which contains PGE1, can be used to treat ulcers of the extremities and pain that occurs at rest due to chronic occlusion of the artery, treatment of skin ulcers caused by progressive systemic sclerosis or diffuse lupus erythematosus, cutaneous ulcers caused by diabetes, improve cognitive disorders caused by peripheral vascular disease, and restoration of disorders of the peripheral circulatory/nervous disorders/disorders of movement due to vibration, for the treatment of patent ductus arteriosus, resulting from a congenital heart defect that is dependent on the ductus arteriosus, and to improve resolution at the conducting pornografii through the superior mesenteric artery.

1.2. The phospholipid

The phospholipid used in the fat emulsion according to this variant implementation, includes phosphatidylcholine (PC) and phosphatidylglycerol (FG). The phospholipid may be in the form of a pharmaceutically acceptable salt, such as sodium salt, potassium salt or ammonium salt.

Examples of PF include egg yolk lecithin, soybean lecithin, lecithin, synthesized wholly or partly by a known method, etc. Among them egg yolk lecithin the two is to be preferred. In particular, purified egg yolk lecithin with a high content of phospholipid and highly purified egg yolk lecithin, which essentially does not contain FAA, are preferred.

The term "purified egg yolk lecithin" is defined in the Japanese Pharmaceutical Excipients, and the term "highly purified egg yolk lecithin" is defined in the Japanese Guide for Pharmaceutical Excipients (prepared for printing by the Japanese Council on Pharmaceutical Excipients). Purified egg yolk lecithin typically contains FAA in an amount of about 12-18%.

Preferably, the fat emulsion according to this variant implementation, essentially did not contain tea. Fat emulsion, which essentially does not contain tea, can be obtained by the use of purified egg yolk lecithin or lecithin obtained synthetically or semi-synthetic in the form of a phospholipid.

The fact that the phospholipid contains or essentially does not contain tea, you can check, for example, by the following method.

Namely phospholipids dissolved in a mixture of chloroform and methanol in the amount of 10% (weight to volume). On the lower part of the thin-layer plate of silica gel put 10 ál of a solution. The solution elute using a mixture of chloroform:methanol:water = 65:25:4), with subsequent drying. The village is e spraying ninhydrin solution is heated at 120°C for 10 minutes. The absence of staining in spots detected using a standard solution of PAA, which caused a similar fashion, determine that the phospholipid essentially does not contain tea.

FG can be chemically synthesized, extracted from plants or bacteria, or can be obtained through action of phospholipase D on lecithin, raw material is soybean or egg yolk, in the presence of glycerol by a known method (Biochemistry Experiments 3, Lipid Chemistry, str and 295, Tokyo Called Dojin, 1974). FG, as a rule, contains a linear or branched saturated or unsaturated residue of fatty acids with 12-22 carbon atoms. In the fat emulsion according to this variant implementation FY preferably contains a linear saturated or unsaturated residue of a fatty acid with 12-18 carbon atoms, and more preferably a linear saturated or unsaturated residue of fatty acids with 16-18 carbon atoms. When using FG with the remaining fatty acids with the number of carbon atoms within the above range (in particular, when using PF originating from egg yolk lecithin), chain length fatty acids PF almost equal to the chain length of the fatty acid FG, so it's more stable fat emulsion.

FG made by chemical synthesis, can represent, at the very measures which, one of the following substances:

dipalmitoylphosphatidylcholine, dioleoylphosphatidylcholine, dilaurylracglycerol, dimyristoylphosphatidylcholine, distearoylphosphatidylglycerol and palmitoyloleoylphosphatidylglycerol. In the case of natural FG, you can use FG with the balance of fatty acids that are specific for the substance, which is FG. Among the above-mentioned substances are preferred dipalmitoylphosphatidylcholine, FG, originating from soybean lecithin or FG originating from egg yolk lecithin, while FG originating from egg yolk lecithin is preferable from the viewpoint of stability of the active ingredient and biocompatibility.

In the fat emulsion according to this variant of implementation, the ratio of PC to PG (PC:PG) in the phospholipid is from 85:15 up to 99.7:0.3 to. When the ratio of PC to PG (PC:PG) is within the above range, stability of the active ingredient and the emulsion can be increased. If the number of FG is outside the above range, the resistance PGE1 deteriorates during storage.

The ratio of PC to PG (PC:PG) is preferably from 90:10 up to 99.7:0.3, and more preferably from 95:5 up to 99.7:0.3, and even more preferably from 97:3 to 99.5:0.5 in.

To improve the chemical stability of the active ingredient, the total content of PF and f is in the phospholipid should preferably be 95% or more, and more preferably 98% or more.

The phospholipid may include different phospholipid, in addition to the FG and FG (for example, at least one of the following connections:

sphingomyelin, phosphatidylinositol, phosphatidylglycerol, phosphatidylinositol, phosphatidylinositol, lysophospholipid or such compounds as Lisa-PF, Lisa-FG), in an amount not more than 5%. Natural phospholipid may include triglyceride, cholesterol and similar compounds in an amount of not more than 5%.

1.3. Base oil

Examples of the base oil used in the fat emulsion according to this variant implementation, include vegetable oils such as soybean oil, sesame oil, rapeseed oil, safflower oil, olive oil, castor oil, corn oil, cottonseed oil, rice oil, sunflower oil, grape seed oil and the oil of wheat germ, a triglyceride with a medium size chain (MCT) and the like, Vegetable oil preferably is a refined vegetable oil.

1.4. Higher fatty acid

In the fat emulsion according to this variant implementation of the free higher fatty acid plays the role of emulsifying additives.

More precisely, the stability of the fat emulsion according to this version of the implementation can be improved through the use of free higher fatty acid. Used in the same document, the term "free higher fatty acid" refers to a carboxylic acid. The term "salt free higher fatty acid", as used in this document, refers to pharmaceutically acceptable salts (e.g. alkali metal salts, such as sodium and potassium salts, and salts of alkaline-earth metals such as calcium salt) free higher fatty acid. The term "free higher fatty acid or its salt" excludes ester of fatty acid, which is produced from vegetable oils (base oils) and phospholipid.

The higher fatty acid is a linear or branched saturated or unsaturated fatty acid with 6 to 22, preferably 12 to 20) carbon atoms. Examples of the higher fatty acids include oleic acid, stearic acid, linoleic acid, palmitic acid, linolenic acid and myristic acid. Among them oleic acid is preferred.

Since the fat emulsion according to this variant implementation and its active ingredient have excellent resistance, the fat emulsion can essentially do not contain free fatty acid or its salt (for example, free oleic acid). When the fat emulsion contains a higher fatty acid or its salt, the amount of higher fatty acid or its salt is preferably 0.15 parts or less, and more preferably of 0.015 parts or IU the it per 1 part of the phospholipid. If the number of higher fatty acids exceeds 0.15 parts per 1 part of the phospholipid, the stability of the active ingredient can be reduced. If the amount of the higher fatty acid is of 0.015 parts or less, per 1 part of the phospholipid, it is possible to obtain an emulsion with excellent stability, while maintaining the stability of the active ingredient. Used in this document the wording "essentially does not contain a free higher fatty acid"means that the free higher fatty acid is not added intentionally, and eliminates the free higher fatty acid formed by the decomposition of vegetable oil (base oil) or phospholipid, and free fatty acid, which was even unintentionally.

The fact that the fat emulsion contains or essentially does not contain free oleic acid, can be checked, for example, by the following method.

Namely, to 1 ml of fat emulsion was added 1 ml of ethanol, 0.5 ml of diethyl ether and 0.5 ml of petroleum ether. Then the mixture is stirred. After centrifugation of the mixture collect the supernatant. Then solvent evaporated using nitrogen. The resulting oil phase was dissolved in a mixture of chloroform and methanol in the amount of 10% (weight to volume). On the lower part of tongolo the Noah plates of silica gel put 2 ál of a solution. The solution elute using a mixture of chloroform:methanol:water = 65:25:4), with subsequent drying. After spraying 50% (by weight) sulfuric acid solution is heated at 120°C for 30 minutes. The absence of staining in spots detected using a standard solution of oleic acid, which caused a similar fashion, determine that the fat emulsion, essentially, does not contain free oleic acid. Since the fat emulsion according to this variant implementation, essentially, does not contain fee and/or free higher fatty acid (e.g., free oleic acid), the number of components contained in the fat emulsion may be reduced. Moreover, the stability of the active ingredient can be further increased.

1.5. Other components

The fat emulsion according to this variant implementation may include isotonic agent (e.g., glycerol, glucose and sodium carbonate), an antioxidant (for example, ascorbic acid and its salts, benzoic acid, citric acid and its salts, dibutylaminoethanol, dibutylaminoethanol, alpha-tocopherol and D-sorbitol) and the agent regulating the pH (for example, sodium hydroxide, hydrochloric acid and phosphate), if necessary.

1.6. The way to obtain

The method of obtaining the fat emulsion of prostaglandin according to one version done by the means of the invention includes the preparation of fat emulsions using phospholipid, which includes phosphatidylcholine (PC) and phosphatidylglycerol (FG), the ratio PF cfg (PC:PG) is from 85:15 up to 99.7:0.3, and preferably from 90:10 up to 99.7:0.3, and more preferably from 95:5 up to 99.7:0.3 to, and even more preferably from 97:3 to 99.5:0.5 in. In particular, the fat emulsion is prepared by emulsification of prostaglandin (active ingredient), phospholipid (emulsifying agent), base oils and water.

FG and FG can be mixed in a carrier oil or water. After homogenization of the mixture of water or base oil can be added to the mixture for the preparation of the emulsion. Preferably homogenization FG and FG in the base oil.

For example, phosopholipid (FX and FY), prostaglandin (e.g., PGE1), additives (e.g., glycerol) and similar compounds are mixed in a certain number of base oils (e.g. soybean oil), and the mixture is homogenized using a homogenizer. After adding a certain quantity of water, the mixture is homogenized using a homogenizer to obtain an emulsion of the type oil-in-water". Thus can be obtained fat emulsion according to this variant implementation.

FG and FG can be added separately or simultaneously. You can use FG obtained by partial conversion of FG in FG under the action of the enzyme. You can use the phospholipid obtained by adding neo is imennogo or purified product, which contains FG or FG, so that the ratio of PC to PG (PC:PG) ranged from 85:15 up to 99.7:0.3 cleansing process FG or FG. When using PF originating from egg yolk, PF in the egg yolk can be turned into FG under the action of the enzyme with the aim of obtaining egg-yolk liquid, in which the ratio of PC to PG (PC:PG) is from 85:15 up to 99.7:0.3 to. After that, the phospholipid contained in the egg-yolk liquid, you can clean and use.

Additives such as a stabilizer, isotonic agent and the agent regulating the pH, you can add in the fat emulsion according to this variant implementation with the purpose of convenience. The resulting fat emulsion can be filtered or heated (for example, subjected to high-temperature heat treatment).

The method of obtaining the fat emulsion of prostaglandin according to this variant implementation may further include adjusting the pH of the fat emulsion is preferably in the range from 4 to 7 (more preferably from 4.5 to 6.5). If the pH of the fat emulsion according to this variant implementation is less than 4, the stability of the emulsion may be reduced. If the pH of the fat emulsion is greater than 7, the resistance of the active ingredient may be reduced.

The method of obtaining the fat emulsion of prostaglandin according to this variant implementation, for the ome, include the location of the fat emulsion, the pH of which is adjusted to the level of 4 to 7, in a sealed container and carrying out high-temperature treatment, the fat emulsion at a certain temperature for a certain period of time. The fat emulsion can be sterilized by high temperature heat treatment, while suppressing the decomposition of the active ingredient (prostaglandin). High-temperature heat treatment preferably is a thermal steam sterilization, high pressure or spray.

Thermal steam sterilization, high pressure or spraying is preferably carried out at 110-140°C. for 0.5-30 minutes. For example, heat treatment vapor pressure can be carried out at 127°C for one minute. Conditions - 127°C and 1 minute time period is consistent with the value F0 of about 9÷10, which results in the killing of microorganisms.

Fat emulsion obtained by the above method, and its active ingredient (prostaglandin) show excellent resistance. For example, the fatty emulsion is subjected to high temperature heat treatment has a residual content of prostaglandin at the level of 70% or more, preferably 80% or more, and preferably 85% or more, on a fat emulsion, not subjected to vyskot mperatures heat treatment.

1.7. Method of stabilizing and emulsifying agent

The method of stabilization of the fatty emulsion of prostaglandin according to one variant embodiment of the invention includes the use of phospholipid with a ratio of PC to PG (PC:PG) of 85:15 up to 99.7:0.3, and essentially does not include the use of FEA in the fat emulsion, which contains a prostaglandin as an active ingredient. Emulsifying agent according to one variant embodiment of the invention is used for the fat emulsion of prostaglandin, while emulsifying agent comprises a phospholipid with a ratio of PC to PG (PC:PG) of 85:15 up to 99.7:0.3, and essentially does not contain tea. Examples of such emulsifying agent is the above-mentioned highly purified egg yolk lecithin, which contains FG and FG in the above ratio.

1.8. Application and properties

Since the fat emulsion according to this variant implementation contains a phospholipid, which includes phosphatidylcholine (PC) and phosphatidylglycerol (FG) in the ratio of PC to PG (PC:PG) of 85:15 up to 99.7:0.3, then the stability of the active ingredient (prostaglandin) can be increased while maintaining the stability of the emulsion, even if the fat emulsion, essentially, does not contain free higher fatty acid such as oleic acid. Since the fat emulsion according to this variant implementation, is usesto, does not contain FAA, the resistance of the active ingredient (prostaglandin) can be further increased.

The fat emulsion according to this variant implementation can be used for intravenous injections. In this case, after adjusting the pH of the fat emulsion obtained according to this variant implementation, it is loaded into a sealed container such as an ampoule, a vial or pre-filled syringe container. After that, the fat emulsion is subjected to high temperature heat treatment or similar treatment.

The average diameter [particle] the fat emulsion according to this variant implementation is preferably 300 nm or less, and more preferably 150÷250 nm. If the average diameter exceeds 300 nm, the system fat emulsion may become unstable. If the average diameter is less than 150 nm, the level of accumulation in the inflamed site (in particular, on the inner wall of the peripheral vessels) or the number of macrophages decreased with the introduction of intravenous fat emulsion. In the fat emulsion may be insufficient physiological activity after administration.

The fat emulsion according to this variant implementation preferably has an average diameter of 300 nm or less, preferably prostaglandin PGE1 is, and the residual content is of PGE1 in the fat emulsion after storage at 40°C for seven days is 70% or more, preferably 80% or more, and more preferably 85% or more. The residual content of prostaglandin E1 in the fat emulsion after storage at 20°C for two months is 70% or more, preferably 80% or more, and more preferably 85% or more. The residual content of prostaglandin E1 in the fat emulsion after storage at 5°C for one year is 80% or more, preferably 85% or more, and more preferably 90% or more. In the invention, the storage test is a test to assess the resistance of the fat emulsion during long-term storage (see, for example, JP-A-4-338333). Storage test at 20°C for two months - this is a test to assess the resistance of the fat emulsion during prolonged storage under this option implementation (see, for example, the recommendations of the ICH Q1A).

Since the fat emulsion according to this variant implementation contains a phospholipid, which includes FX and FY in the ratio (PC:PG) of 85:15 up to 99.7:0.3, then PGE1 (active ingredient) fat emulsion has excellent stability, there is no need to limit other additives to the fat emulsion or in increasing the average diameter of the droplets of fat contained in the fat emulsion, or need to change particle size distribution of the droplets of fat contained in the fat Amul the FIC. As the drops of fat contained in the fat emulsion according to this variant implementation, have a small and uniform diameter, conventional in the art, the drug effectively accumulates in the affected area. Therefore, the fatty emulsion exhibits excellent efficiency even when introduced in a small amount.

The residual content of PGE1 (%) in the fat emulsion after storage calculated by the following formula (1) based on the content of PGE1 in the fat emulsion immediately after high-temperature heat treatment and the content of PGE1 in the fat emulsion after storage, determined by high performance liquid chromatography (HPLC). Details of the method of determining the content of PGE1 in the fat emulsion by HPLC are shown in the examples described below.

The residual content of PGE1 (%) = (Content of PGE1 in the fat emulsion after storage / Content PGE1 in the fat emulsion after high-temperature heat treatment) × 100 (1)

1.9. Dosage form

The fat emulsion according to this variant implementation is preferably introduced into the human or other mammal besides humans, parenteral way. More preferably the fat emulsion for intravenous injection (including intravenous drip injection). For example, fatty em is lsiu according to this variant implementation, which contains a prostaglandin in a concentration of 0.2-100 μg/ml, can be administered by intravenous injection, either directly or after mixing with the infusion solution.

2. Examples

The invention is described below with examples. It should be noted that the invention is not limited to the following examples.

2.1. Method of evaluation

In the examples, the content of PGE1 in the fat emulsion is determined by the following methods, and the average diameter of the droplets of fat contained in the fat emulsion, measured by the following method.

2.1.1. The PGE1 content

The fat emulsion is pre-processed using the cartridge SEP-PAKC18(manufactured by Waters), and the content of PGE1 in the fat emulsion is determined by HPLC using post-column method.

Working conditions the following

Detector - UV absorptiometer (measured wavelength of 278 nm).

Column - octadecylsilane silica gel (inner diameter 4 mm, length 15 cm).

The temperature of the column - oloko 60°C.

The mobile phase is a mixture of phosphate buffer concentration 1/150 mol/l) and acetonitrile (3:1) (pH 6.3).

The flow rate of the mobile phase, 1 ml/min

(Reaction conditions after the exit from the column).

The reaction liquid is a solution of potassium hydroxide with a concentration of 1 mol/L.

A flow rate of 0.5 ml/min

Reaction round - Teflon (argovie name) tube (inner diameter - about 0.5 mm, the length is about 10 m).

The reaction temperature is about 60°C.

2.1.2. The average diameter of the droplets of fat

The average diameter of the droplets of fat contained in the fat emulsion, measured with a device for submicron measurement of particle size distribution N4PLUS (manufactured by Beckman Coulter, Inc.).

2.1.3. The detection is fed into the fat emulsion

To 1 ml of fat emulsion was added 1 ml of ethanol, 0.5 ml of diethyl ether and 0.5 ml of petroleum ether. Then the mixture is stirred. After centrifugation of the mixture collect the supernatant. Then solvent evaporated using nitrogen. The resulting oil phase was dissolved in a mixture of chloroform and methanol in the amount of 10% (weight to volume). On the lower part of the thin-layer plates of silica gel put 2 ál of a solution. The solution elute using a mixture of chloroform:methanol:water = 65:25:4), with subsequent drying. After spraying ninhydrin plate heated at 120°C for 30 minutes. The absence of staining in spots detected using a standard solution of PAA, which caused a similar fashion, determine that the fat emulsion essentially does not contain tea.

2.2. Example 1

50 g of soybean oil (base oil)8,82 g of lecithin egg yolk PC-98N" (manufactured by Q.P. Corporation, purity phosphatidylcholine: 98,8%, FAA not detected above the method detection FAA) (a phospholipid) and 0.18 g of dipalmitoylphosphatidylcholine (produced by NOF Corporation) was dispersed and homogenized, using a mixer-homogenizer. After adding 3.5 mg of prostaglandin E1, to the mixture add water for injection, which added 11,05 g of concentrated glycerin (described in the Japanese Pharmacopoeia) to obtain 500 g coarse emulsioning fluid. Rough emulsified liquid is passed through the homogenizer of Menton-Gaulin (manufactured by APV) fifteen times under pressure of 600 kgf/cm2to obtain the fat emulsion of example 1 (average particle diameter - 193 and 213 nm). Measure the average diameter of the droplets of fat contained in the fat emulsion, and determine the content of PGE1 in the fat emulsion.

The content of PGE1 in the fat emulsion (content of PGE1 in the fat emulsion immediately after preparation) are shown in Tables 1 and 2, and the average diameter of the droplets of fat contained in the fat emulsion (average diameter of the droplets of fat contained in the fat emulsion immediately after preparation), are shown in Tables 3 and 4.

The fat emulsion is filtered through a porous filter with a pore diameter of 0.45 µm. Then the pH of the fat emulsion was adjusted to 5.0, and 5.5 (two samples) or to 6.0 with an aqueous solution of sodium hydroxide. After pipetting the fat emulsion in 2 ml ampoule it is subjected to heat sterilization spray (i.e. high-temperature heat treatment at 110°C for 5 minutes. After opening the ampoule calculate the residual content of PGE1 in igrovoy emulsion (residual content of PGE1 in the fat emulsion immediately after heating) and measure the average diameter of the droplets of fat, contained in the fat emulsion (average diameter of the droplets of fat contained in the fat emulsion immediately after heating). The results are presented in Tables 1-4.

The residual content of PGE1 in the fat emulsion immediately after heating calculated by determining the content of PGE1 in the fat emulsion immediately after the high-temperature treatment above described method of quantitative determination of the content of PGE1 and calculate the ratio (%) content of PGE1 in the fat emulsion immediately after high-temperature heat treatment to the contents of PGE1 in the fat emulsion immediately after preparation.

Ampoule, subjected to high-temperature heat-treated, stored at 40°C for seven days. After opening the ampoule determine the content of PGE1 in the fat emulsion after storage at 40°C for seven days. The residual content of PGE1 in the fat emulsion after storage at 40°C for seven days are calculated according to the formula (1). Also measure the average diameter of the droplets of fat contained in the fat emulsion after storage at 40°C for seven days. The results are presented in Tables 1-4. FAA not detected in the fat emulsion of example 1.

In examples 1, 2 and 4-6 and comparative examples 2, 3 and 6 show the results for the case when the pH was reduced to 5.0. In the examples 1, 2, 4, 7, 9, 10 and 12 and comparative examples 1-4, 6 and 7 show the results for the case of, when the pH was brought to 5.5, and in examples 1, 3, 7, 8 and in comparative examples 3 and 5 show the results for the case when the pH was increased to 6.0 (see Tables 1-4).

In examples 1-12 and comparative examples 1-7, the number of farms is calculated by multiplying the number of phospholipid (lecithin egg yolk) purity (0,988=98,8%) FG.

In Tables 1 and 2 in the column "oleic acid (mass. parts)" is specified oleic acid content (mass. parts) relative to 1 mass. part of the phospholipid.

2.3. Example 2

The fat emulsion of example 2 (average particle diameter: 222 nm) are obtained in the same manner as in example 1 except that the amount of lecithin from egg yolk PC-98N change on 8,55 g, and the number of dipalmitoylphosphatidylcholine change by 0.45, the pH of the fat emulsion was adjusted to 5.0 or 5.5 with diluted hydrochloric acid.

The fat emulsion of example 2, examples 3-12 and comparative examples 1 to 7, described below, is subjected to high temperature heat treatment and stored at 40°C for seven days in the same manner as in example 1. Determine the residual content of PGE1 in each of the fat emulsion and the average diameter of the droplets of fat contained in the fat emulsion. The results are presented in Tables 1-4. FAA not detected in the fat emulsion of example 2.

2.4. Example 3

The fat emulsion of example 3 (average particle diameter: 229 nm) receive the same manner as in example 1, except that the amount of lecithin from egg yolk PC-98N change on the 7.65 g of 1.65 g FG made by processing the PF obtained by purification of egg yolk phospholipid by phospholipase D in the presence of glycerol (hereafter in this document referred to as "egg-yolk FG) is used instead of dipalmitoylphosphatidylcholine, and conditions of high-temperature heat treatment changed to 127°C and one minute. With regard to residues of fatty acids in egg-yolk FG, then calculated the number of residues of fatty acids with 16 carbon atoms is 35%and the number of residues of a fatty acid with 18 carbon atoms is 57%. the pH of the fat emulsion was adjusted to 6.0 with an aqueous solution of sodium hydroxide. FAA not detected in the fat emulsion of example 3. The residual content of active ingredient (PGE1) in the fat emulsion of example 3 immediately after heating, for example, lower its content in the fatty emulsions of examples 8 and 11, described below. Therefore we think that the increase in the number of FG adversely affect the stability of the active ingredient after heating.

2.5. Example 4

The fat emulsion of example 4 (average particle diameter: 226 nm) are obtained in the same manner as in example 1, except that soybean oil is added 0.6 g of oleic acid. the pH of the fat emulsion was adjusted to 5.0 diluted the Oh hydrochloric acid or up to 5.5 with an aqueous solution of sodium hydroxide. FAA not detected in the fat emulsion of example 4.

2.6. Example 5

The fat emulsion of example 4 (average particle diameter: 210 nm) are obtained in the same manner as in example 1, except that soybean oil is added 1.2 g of oleic acid. the pH of the fat emulsion is 5.0. FAA not detected in the fat emulsion of example 5.

2.7. Example 6

The fat emulsion of example 6 (average particle diameter: 230 nm) are obtained in the same manner as in example 2, except that soybean oil is added 1.2 g of oleic acid. the pH of the fat emulsion was adjusted to 5.0 with diluted hydrochloric acid. FAA not detected in the fat emulsion of example 6.

2.8. Example 7

The fat emulsion of example 7 (average particle diameter: 218 nm) are obtained in the same manner as in example 3, except that the amount of lecithin from egg yolk PC-98N change on 8,82 g, the number of egg-yolk FG is changed to 0.18 g, and conditions of high-temperature heat treatment is changed to 110°C. and five minutes. the pH of the fat emulsion was adjusted to 5.5 with diluted hydrochloric acid or up to 6.0 with an aqueous solution of sodium hydroxide. FAA not detected in the fat emulsion of example 7.

2.9. Example 8

The fat emulsion of example 8 (average particle diameter: 197 nm) are obtained in the same manner as in example 3, except that the amount of lecithin from egg yolk PC-98N change is and 8,91 g, the number of egg-yolk FY change 0.09, the pH of the fat emulsion was adjusted to 6.0 with an aqueous solution of sodium hydroxide. FAA not detected in the fat emulsion of example 8.

2.10. Example 9

The fat emulsion of example 9 (average particle diameter: 217 nm) are obtained in the same manner as in example 8, except that soybean oil is added 0.12 g of oleic acid. the pH of the fat emulsion was adjusted to 5.5 with an aqueous solution of sodium hydroxide. FAA not detected in the fat emulsion of example 9.

2.11. Example 10

The fat emulsion of example 10 (average particle diameter of 198 nm) are obtained in the same manner as in example 8, except that the amount of lecithin from egg yolk PC-98N change on 8,955 g, and the number of egg-yolk FY change on 0,045, the pH of the fat emulsion was adjusted to 5.5 with diluted hydrochloric acid. FAA not detected in the fat emulsion of example 10.

2.12. Example 11

The fat emulsion of example 11 (average particle diameter of 195 nm) are obtained in the same manner as in example 8, except that the amount of lecithin from egg yolk PC-98N change on 8,973 g, and the number of egg-yolk FY change on 0,027, the pH of the fat emulsion was adjusted to 6.0 with an aqueous solution of sodium hydroxide. FAA not detected in the fat emulsion of example 11. Since the average diameter of the particles in the fat emulsion of example 11 increases th the heating of about 50 nm, it is considered that thermal stability of the fat emulsion of example 11 somewhat smallish. However, not observed separation of oil phase and the stability of the active ingredient (PGE1) is excellent.

2.13. Example 12

The fat emulsion of example 12 (average particle diameter of 227 nm) are obtained in the same manner as in example 1, except that FG made by processing soybean phospholipid by phospholipase D in the presence of glycerol (hereafter in this document referred to as "soy FG"), is used instead of dipalmitoylphosphatidylcholine, and conditions of high-temperature heat treatment changed to 127°C and one minute. With regard to residues of fatty acids in soybean FG, then calculated the number of residues of fatty acids with 16 carbon atoms is 14%and the number of residues of a fatty acid with 18 carbon atoms is 78%. the pH of the fat emulsion was adjusted to 5.5 with diluted hydrochloric acid. FAA not detected in the fat emulsion of example 12.

2.14. Comparative example 1

The fat emulsion of comparative example 1 (average particle diameter: 214 nm) are obtained in the same manner as in Example 5, except that the amount of lecithin from egg yolk PC-98N" is changed to 9.0 g, and the number of dipalmitoylphosphatidylcholine is changed to 0, the pH of the fat emulsion was adjusted to 5.5 with an aqueous solution of hydroxide intothree is. FAA not detected in the fat emulsion of comparative example 1.

2.15. Comparative example 2

The fat emulsion of comparative example 2 (average particle diameter: 189 nm) are obtained in the same manner as in example 1, except that the amount of lecithin from egg yolk PC-98N" is changed to 9.0 g, and the number of dipalmitoylphosphatidylcholine is changed to 0, the pH of the fat emulsion was adjusted to 5.0 with diluted hydrochloric acid or up to 5.5 with an aqueous solution of sodium hydroxide. FAA not detected in the fat emulsion of comparative example 2.

2.16. Comparative example 3

The fat emulsion of comparative example 2 (average particle diameter: 216 nm) are obtained in the same manner as in comparative example 1 except that the conditions of high-temperature heat treatment changed to 127°C and one minute. the pH of the fat emulsion was adjusted to 5.0 with diluted hydrochloric acid or diluted to 5.0 with hydrochloric acid or up to 5.5 or 6.0 with an aqueous solution of sodium hydroxide. FAA not detected in the fat emulsion of comparative example 3.

2.17. Comparative example 4

The fat emulsion of comparative example 4 (average particle diameter: 221 nm) are obtained in the same manner as in example 12, except that instead of soy FG use phosphatidylserin (FS)prepared by processing the PF obtained by processing the of stolipinovo egg yolk, the phospholipase D in the presence of L-serine ("egg-yolk FS"). the pH of the fat emulsion was adjusted to 5.5 with an aqueous solution of sodium hydroxide. FAA not detected in the fat emulsion of comparative example 4.

2.18. Comparative example 5

The fat emulsion of comparative example 5 (average particle diameter: 192 nm) are obtained in the same manner as in example 8, except that the amount of lecithin from egg yolk PC-98N change on 8,991 g, and the number of egg-yolk FY change on 0,009, the pH of the fat emulsion was adjusted to 6.0 with an aqueous solution of sodium hydroxide. FAA not detected in the fat emulsion of comparative example 5. Since the fat emulsion of comparative example 5 is separated after heating, the definition of PGE1 and storage test is not performed.

2.19. Comparative example 6

The fat emulsion of comparative example 5 (average particle diameter: 195 nm) are obtained in the same manner as in comparative example 1 except that the amount of lecithin from egg yolk PC-98N" is changed to 7 g and add 2 g of egg yolk lecithin "PL-100M" (manufactured by Q.P. Corporation, the content of FAA: 15.8 percent). the pH of the fat emulsion was adjusted to 5.0 with diluted hydrochloric acid or up to 5.5 with an aqueous solution of sodium hydroxide. The content of FAA in phospholipid is 3.5% (i.e. FAA find in the fat emulsion of comparative example 6).

2.2. Comparative example 7

The fat emulsion of comparative example 7 (average particle diameter: 238 nm) are obtained in the same manner as in example 12, except that instead of soy FG use dipalmitoylphosphatidyl acid. the pH of the fat emulsion was adjusted to 5.5 with an aqueous solution of sodium hydroxide. FAA not detected in the fat emulsion of comparative example 7.

Table 2
Oleic acid (mass. part)FG:FGpHImmediately after cookingImmediately after heatingAfter storage at 40°C for 7 days
PGE1 (μg/ml) (a)PGE1 (μg/ml) (b)the residual content of PGE1 (%) (b/a)PGE1 (μg/ml) (C) (%) (C/b)the residual content of PGE1
Comparative
tion example 1
of 0.133100:05,57,20of 87.84,2166,7
Comparative
hydrated example 2
0100:05.06,34of 5.8992,95,5894,8
5,5of 5.84to 92.15,5995,8
Comparative
hydrated example 3
of 0.133100:05,07,125,95to 83.54,3673,4
5,55,7680,94,0770,6
6,077,53,6365,9
Comparative
hydrated example 4
0(FH:FC=95:5)5,56,114,7477,62,5654,0

Oleic acid (mass. part)FG:FGpHImmediately after cookingImmediately after heatingAfter storage at 40°C for 7 days
PGE1 (μg/ml) (a)PGE1 (μg/ml) (b)the residual content of PGE1 (%)
(b/a)
PGE1 (μg/ml) (C) (%) (C/b)the residual content of PGE1
Comparative
hydrated example 5
099,9:0,16,0--- --
Comparative
hydrated example 6
of 0.133(PF:FAA=96,5:3,5)5,07,105,6679,72,5244,5
5,55,6179,02,4443,5
Comparative
hydrated example 7
0(FH:FC=95:5)5,56,93is 4.9371,33,0060,7

Table 3
pHImmediately after cookingImmediately after heatingAfter storage at 40°C for 7 days
Average particle diameter (nm) The standard deviation (nm)Average particle diameter (nm)The standard deviation (nm)Average particle diameter (nm)The standard deviation (nm)
Example 15,0193862118220066
5,52148821388
5,5213322116321272
6,02127821382
Example 25,02226821880225 78
5,52195522085
Example 36,0229562267023074
Example 45,0226862258822950
5,52277422580

PHImmediately after cookingImmediately after heatingAfter storage at 40°C for 7 days
Average particle diameter (nm)The standard deviation (nm)Average particle diameter (the m) The standard deviation (nm)Average particle diameter (nm)The standard deviation (nm)
Example 55,0210852117121766
Example 65,0230742236023268
Example 75,5218862196821580
6,02217321378
Example 86,0197811986019682
Example 95.5217772217121174
Example 105,5198581915618164
Example 116,0195672395323472
Example 125,5227932338522674

Table 4
pHImmediately after cookingImmediately after heatingAfter storage pri°C for 7 days
Average particle diameter (nm)The standard deviation (im)Average particle diameter (nm)The standard deviation (nm)Average particle diameter (nm)The standard deviation (nm)
Comparative
tion example 1
5,5214332194721963
Comparative
hydrated example 2
5,018969731315423195
After heating stripes
5,5After heating stripes, and measuring ranovitsa impossible 413185
Comparative
hydrated example 3
5,021678419189427179
5,53117634099
6,02277522667
Comparative
hydrated example 4
5,5221292257721980

pHImmediately after cookingImmediately after heatingAfter storage of n and 40°C for 7 days
Average particle diameter (nm)The standard deviation (nm)Average particle diameter (nm)The standard deviation (nm)Average particle diameter (nm)The standard deviation (nm)
Comparative
hydrated example 5
6,019278753354--
After heating stripes
Comparative
hydrated example 6
5,0195701917619772
5,51876818671
Comparative
hydrated example 7
5,5238 532396523866

In Tables 1 and 2 FG:FG" refers to the ratio of PC to PG phospholipid used in each of the fat emulsion.

As follows from the results presented in Tables 1-4, since the fat emulsions of examples 1-12 contain a phospholipid that contains FG and FG in the ratio (PC:PG) of 85:15 up to 99.7:0.3, and the residual content of PGE1 after storage at 40°C for seven days is 70% or more (80% or more, 85% or more, depending on conditions). Moreover, the average diameter of the droplets of fat is 300 nm or less, the average diameter of fat droplets is small immediately after cooking, immediately after heating and after storage at 40°C for seven days. Therefore, the fatty emulsions of examples 1-12 exhibit excellent resistance and excellent resistance manifests its active ingredient (PGE1).

As for the fat emulsion of comparative example 1, the residual content of PGE1 in the fat emulsion after storage at 40°C for seven days significantly lower its content in the fat emulsion immediately after preparation. Regarding the fat emulsion of comparative example 2, the average diameter of the droplets of fat immediately after heating and after storage at 40°C for seven days PR is more than the average diameter of the droplets of fat immediately after cooking. Moreover, the average diameter of the droplets of fat immediately after heating and after storage at 40°C for seven days significantly more change in the average diameter of the droplets of fat immediately after cooking, and there is a stratification of the drug. Regarding the fat emulsion of comparative example 3, the diameter of the droplets of fat increases immediately after heating at pH 5.0 and 5.5, and the residual content of PGE1 after storage at 40°C for seven days reduced at pH 6.0. In particular, since the ratio of PC to PG (PC:PG) in the phospholipid beyond the range of 85:15-99,7:0,3 in the fatty emulsions of comparative examples 1-3 and 5, the stability of the active ingredient (PGE1) and/or the stability of the emulsion worse. In comparative example 4, in which instead use FG FS, the stability of the emulsion is stored immediately after heating and after storage at 40°C for seven days. However, the residual content of PGE1 after storage at 40°C for seven days is much lower.

In comparative example 7, in which instead use FG FC, the stability of the emulsion is stored immediately after heating and after storage at 40°C for seven days. However, the residual content of PGE1 after storage at 40°C for seven days is much lower. I.e. when using phosphatidylserine (PS) or phosphatidic acid (FC) instead of FG resistance is mulshi saved but PGE1 disappears quickly.

In comparative example 6 stability of the emulsion increases with the introduction of tea in the phospholipid. However, PGE1 disappears faster in comparison with comparative examples 1-3.

2.21. The results after storage at 20°C for two months.

Fat emulsions (pH 5.5) of examples 1 and 7 and comparative example 1, stored at 20°C for two months. In Tables 5 and 6 shows the residual content of PGE1 and the average particle diameter after storage. The residual content of PGE1 after storage at 20°C for two months calculated according to the formula (1).

As follows from the results presented in Tables 5 and 6, the active ingredient (PGE1) in the fat emulsion of example 1 has a higher stability compared to the active ingredient in the fat emulsion of comparative example 1 even after storage at 20°C for two months.

Table 5
Oleic acid (%)FG:FGPHImmediately after cookingImmediately after heatingAfter storage at 20°C for two months
PGE1 (μg/ml) (a)PGE1 (μg/ml) (b)the residual content of PGE1 (%) (b/a)PGE1 (μg/ml) (C) residualthe PGE1 content (%) (c/b)
When
measures 1
098:25,55,69lower than the 5.3794,3equal to 4.9792,5
When
measures 7
098:25,56,646,1392,35,59to 91.1
Comparison
the preserver
tion example 1
of 0.133100:05,57,206,32of 87.84,3669,1

Table 6
Oleic acid (%)SRAS is after cooking Immediately after heatingAfter storage at 20°C for two months
Average particle diameter (nm)The standard deviation (nm)Average particle diameter (nm)The standard deviation (nm)Average particle diameter (nm)The standard deviation (nm)
When
measures 1
5,5213322116321179
When
measures 7
5,5218862196822436
Comparison
the preserver
tion when
measures 1
5,5214332194722867

2.22. The results after storage at 5°C for six months the s

Fat emulsions (pH 5.5) of examples 1 and 7 and comparative example 1 stored at 5°C for six months. In Tables 7 and 8 shows the residual content of PGE1 and the average particle diameter after storage. The residual content of PGE1 after storage at 5°C for six months calculated according to the formula (1).

As follows from the results presented in Tables 7 and 8, the active ingredient (PGE1) in the fatty emulsions of examples 1 and 7 has a higher stability compared to the active ingredient in the fat emulsion of comparative example 1 even after storage at 5°C for six months.

Table 7
Oleic acid (%)FG:FGPHImmediately after cookingImmediately after heatingAfter storage at 5°C for six months
PGE1 (μg/ml) (a)PGE1 (μg/ml) (b)the residual content of PGE1 (%) (b/a)PGE1 (μg/ml) (C)the residual content of PGE1 (%) (C/b)
Example 1098:25,55,69lower than the 5.3794,3are 5.3699,8
Example 7098:25,56,646,1392,35,9997,6
Comparative example 1of 0.133100:05,57,206,32of 87.85,3284,2

Table 8
pHImmediately after cookingImmediately after heatingAfter storage at 5°C for six months
Average particle diameter (nm)Standard on klonnie (nm) Average particle diameter (nm)The standard deviation (nm)Average particle diameter (nm)The standard deviation (nm)
When
measures 1
5,5213322116322787
When
measures 7
5,5218862196823664
Comparison
the preserver
tion example 1
5,5214332194723152

2.23. The results after storage at 5°C for one year

Fat emulsions (pH 5.5) of examples 1 and 7 and comparative example 1 stored at 5°C for one year. In Tables 9 and 10 shows the residual content of PGE1 and the average particle diameter after storage. The residual content of PGE1 after storage at 5°C for one year R is schityvat by the formula (1).

As follows from the results presented in Tables 7 and 8, the active ingredient (PGE1) in the fatty emulsions of examples 1 and 7 has a higher stability compared to the active ingredient in the fat emulsion of comparative example 1 even after storage at 5°C for one year.

Table 9
Oleic acid (%)FG:FGpHImmediately after cookingImmediately after heatingAfter storage at 5°C for one year
PGE1 (μg/ml) (a)PGE1 (μg/ml) (b)the residual content of PGE1 (%) (b/a)PGE1 (μg/ml) (C)the residual content of PGE1 (%) (c/b)
When
measures 1
098:25,55,69lower than the 5.3794,35,1395,6
When
Mar
098:25,56,646,1392,3the ceiling of 5.6091,4
Comparison
the preserver
tion when
measures 1
of 0.133100:05,57,206,32of 87.84,7174,5

5,5
Table 10
pHImmediately after cookingImmediately after heatingAfter storage at 5°C for one year
Average particle diameter (nm)The standard deviation (nm)Average particle diameter (nm)The standard deviation (nm)Average particle diameter (nm)The standard deviation (nm)
When
measures 1
213322116320578
When
measures 7
5,5218862196821874
Compare
AUX in
measures 1
5,5214332194721165

1. Stable fat emulsion comprising a prostaglandin as an active ingredient and phospholipids containing phosphatidylcholine and phosphatidylglycerol mass ratio of from 85:15 up to 99.7:0.3 to.

2. The emulsion according to claim 1, in which the mass ratio of phosphatidylcholine and phosphatidylglycerol is from 95:5 up to 99.7:0.3 to.

3. The emulsion according to claim 2, in which the mass ratio of phosphatidylcholine and phosphatidylglycerol is from 97:3 to 99.5:0.5 in.

4. The emulsion according to claim 1, in which the phospholipids, essentially free from phosphatidylethanolamine.

5. The emulsion according to claim 1, in which phosphatidylglycerol contains linear naydennyye unsaturated residue of a fatty acid with 12-18 carbon atoms.

6. The emulsion according to claim 1, in which phosphatidylglycerol comes from the egg yolk.

7. The emulsion according to claim 1, which is essentially free of higher fatty acid or a salt thereof.

8. The emulsion according to claim 7, which is essentially free of free oleic acid.

9. The emulsion according to claim 1, which contains a free higher fatty acid or its salt in an amount of 0.15 wt. including about 1 wt. including phospholipids.

10. The emulsion according to claim 9, in which the free higher fatty acid is oleic acid.

11. The emulsion according to claim 1, in which the total content of PF and PG phospholipids is at least 95%.

12. The emulsion according to claim 1 in which the prostaglandin is a prostaglandin E1 or complex alkilany ether.

13. The emulsion according to item 12, which as prostaglandin contains prostaglandin E1 and has an average particle diameter of at most 300 nm, and the residual content of prostaglandin E1 after storage at 40°C for seven days is at least 70%.

14. The emulsion according to item 12, which as prostaglandin contains prostaglandin E1 and has an average particle diameter of at most 300 nm, and the residual content of prostaglandin E1 after storage at 40°C for seven days is at least 80%.

15. The emulsion according to item 12, which as prostaglandin contains prostaglandin E1 and has an average diameter is the Tr of particles of at most 300 nm, in which the residual content of prostaglandin E1 after storage at 40°C for seven days is at least 85%.

16. The emulsion according to item 12, which as prostaglandin contains prostaglandin E1 and has an average particle diameter of at most 300 nm, and the residual content of prostaglandin E1 after storage at 20°C for two months is at least 80%.

17. The emulsion according to item 12, which as prostaglandin contains prostaglandin E1 and has an average particle diameter of at most 300 nm, and the residual content of prostaglandin E1 after storage at 5°C for one year is at least 80%.

18. Stable fat emulsion comprising a prostaglandin as an active ingredient and essentially free from phosphatidylethanolamine and free oleic acid phospholipids containing phosphatidylcholine and phosphatidylglycerol mass ratio of from 85:15 up to 99.7:0.3 to.

19. Stable fat emulsion comprising a prostaglandin as an active ingredient, essentially free from phosphatidylethanolamine phospholipids containing phosphatidylcholine and phosphatidylglycerol mass ratio of from 85:15 up to 99.7:0.3 and free higher fatty acid or its salt in an amount of 0.15 wt. including about 1 wt. including the above-mentioned phospholipid.

20. The way the LM is new emulsion according to any one of claims 1 to 19, includes preparation of fat emulsions, which comprises a prostaglandin as an active ingredient and phospholipids containing phosphatidylcholine and phosphatidylglycerol mass ratio of from 85:15 up to 99.7:0.3 to.

21. The method of claim 20, in which the pH of the fat emulsion regulate at the level of 4 to 7.

22. The method of stabilization of prostaglandin fat emulsion comprising a prostaglandin as an active ingredient, which is used essentially free from phosphatidylethanolamine phospholipids containing phosphatidylcholine and phosphatidylglycerol mass ratio of from 85:15 up to 99.7:0.3 to.

23. The method of stabilization of fat droplets in the fat emulsion comprising a prostaglandin as an active ingredient, which is used essentially free from phosphatidylethanolamine phospholipids containing phosphatidylcholine and phosphatidylglycerol mass ratio of from 85:15 up to 99.7:0.3 to.

24. Emulsifying agent for the fat emulsion comprising essentially free from phosphatidylethanolamine mixture of phospholipids containing phosphatidylcholine and phosphatidylglycerol mass ratio of from 85:15 up to 99.7:0.3, and when this fat emulsion contains a prostaglandin as an active ingredient.

25. The use of fat emulsion comprising a prostaglandin as an active ingredi the NTA and phospholipids, containing phosphatidylcholine and phosphatidylglycerol mass ratio of from 85:15 up to 99.7:0.3, and as a drug for intravenous administration to a human or other mammal.



 

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2 dwg, 7 tbl, 4 ex

FIELD: chemistry.

SUBSTANCE: present invention relates to dihydropyrazolone derivatives or of formula (I), where R1 denotes a heteroaryl group of formulae given below, where * denotes the linkage point with the dihydropyrazolone ring, A in each individual occurrence denotes C-R4 or N, wherein at most two ring members A represent N at the same time, E denotes O or S, R2, R3 and R4 are as defined in the claim. The invention also relates to a method of producing said compounds.

EFFECT: compounds of formula (I) inhibit HIF-propylhydroxylase activity and can be used to treat and/or prevent diseases, as well as for producing medicaments for treating and/or preventing diseases, particularly cardiovascular and haematologic diseases, kidney diseases, and for promoting the healing of wounds.

10 cl, 10 tbl, 178 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to medicine, namely new organic compounds, namely N,N'-substituted piperazines of general formula (I), wherein R1, R2: linear or branched alkoxy (C1÷C4), CH3C(=O)O; n=1-5; m=0-3; Z: C=O, SO2; X:C(=NH)NH2, C(=NH)NHC(-NH)NH2, G is low-molecular organic or mineral acid, sodium, potassium, ammonium cations or water influences the haemostasis system, showing antiagregant, anticoagulant and vasodilator properties, and to a method for preparing N,N'-substituted piperazines of formula 1 by reaction of N-substituted piperazines of general formula wherein R1, R2; linear or branched alkoxy (C1÷C4), CH3C(=O)O; n=1-5; m=0-3; Z: C=O, SO2; and 1H-pyrazole-1-carboxamidine, dicyane diamide and their salts in organic solvents or water at temperature 10-50°C in the presence of bases.

EFFECT: new substance are promising for prevention and treatment of the disturbed haemostasis system.

12 cl, 10 tbl, 14 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to medicine, namely new organic compounds, namely N,N'-substituted piperazines of general formula (I), wherein R1, R2: linear or branched alkoxy (C1÷C4), CH3C(=O)O; n=1-5; m=0-3; Z: C=O, SO2; X:C(=NH)NH2, C(=NH)NHC(-NH)NH2, G is low-molecular organic or mineral acid, sodium, potassium, ammonium cations or water influences the haemostasis system, showing antiagregant, anticoagulant and vasodilator properties, and to a method for preparing N,N'-substituted piperazines of formula 1 by reaction of N-substituted piperazines of general formula wherein R1, R2; linear or branched alkoxy (C1÷C4), CH3C(=O)O; n=1-5; m=0-3; Z: C=O, SO2; and 1H-pyrazole-1-carboxamidine, dicyane diamide and their salts in organic solvents or water at temperature 10-50°C in the presence of bases.

EFFECT: new substance are promising for prevention and treatment of the disturbed haemostasis system.

12 cl, 10 tbl, 14 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention relates to chemical-pharmaceutical industry, medicine, pharmacology and represents medication for prevention and treatment of atherosclerotic lesion of blood vessels, as well as pre- and thrombotic states, which possesses hypolipidemic and anticoagulant and antithrombotic activity, and is made in dosed drug form in form of coated tablets, consisting of core, which contains active substance and auxiliary substances, characterised by the fact that core contains substance of sulfated arabinogalactan potassium salt as active ingredient, and ludipress, aerosol and sodium stearate as auxiliary substances, components in medication being in definite ratio in wt %.

EFFECT: invention ensures efficient treatment and prevention of atherosclerotic lesion of blood vessels, as well as pre- and thrombotic states.

4 cl, 5 tbl

FIELD: medicine.

SUBSTANCE: invention refers to biotechnology, namely plasmid DNA pCID-PROC for expression of recombinant human protein C, cell lines of Chinese hamster ovary DG-CID-PROC-1 and a method for producing recombinant human protein C. The presented invention may be used for producing recombinant human protein C. Plasmid DNA pCID-PROC for expression of recombinant human protein C contains a sequence of full-length complementary DNA of a human protein C gene presented in List of Sequences as the sequence SEQ ID NO: 1. Plasmid DNA pCID-PROC is characterised by a physical map presented on dwg. 1. A cell line of Chinese hamster ovary DG-CID-PROC-1 is produced by transformation of the cell line of Chinese hamster ovary DG-44 by said expression plasmid DNA pCID-PROC. The method for producing recombinant human protein C involves culture of said cell line DG-CID-PROC-1 in a nutrient medium and recovery of produced target protein from a culture fluid.

EFFECT: invention provides higher production performance of the protein C expression system and simplified recovery, activation and purification of recombinant activated human protein C.

3 cl, 3 dwg, 2 ex

FIELD: chemistry.

SUBSTANCE: invention relates to biotechnology and specifically to obtaining factor VII (FVII) and factor Vila (FVIIa) albumin linked polypeptides, and can be used in medicine. A polypeptide, which is a FVII or FVIIa polypeptide is obtained in a recombinant manner, said peptide being linked with albumins through a glycerine-serine peptide linker of a special structure, which separates part associated with FVII or FVIIa from the albumin part, wherein the FVII or FVIIa polypeptide lies on the N-end of the fused protein. The linked polypeptide or vector structure, which contains its coding nucleic acid, is used as a medicinal agent for treating or preventing blood-clotting disorders.

EFFECT: invention enables to obtain a protein with FVII or FVIIa biological activity and longer functional half-time in plasma in vitro compared to non-linked FVII or FVIIa.

12 cl, 4 dwg, 6 tbl, 6 ex

FIELD: medicine.

SUBSTANCE: invention relates to medicine, namely to surgery, and can be applied for sealing interintestinal anastomosis. For this purpose before immersion into abdominal cavity and suturing middle wound, application of 30-40 g of dry lyophilised cryoprecipitate is performed on serous cover of small intestine on entire circumference of interintestinal anastomosis. After that, either 2-3 ml of 5% calcium chloride solution or 4-5 ml of sterile thrombin 15 U NIH/ml, dissolved in 5% solution of aminocaproic acid, are added drop by drop into said cryoprecipitate. 60 seconds after formation of gel-like fibrin film re-application of the first or the second two-component composition is carried out.

EFFECT: method ensures increase of strength and biological impermeability of intestinal sutures at the background of extensive peritonitis due to efficient fixation of fibrin, which does not produce damaging impact on tissues, is capable of quick formation of fibrin film, is homologous and safe, penetrates zone of suture, filling holes from needle punctures, making it possible to close anastomosis on circumference evenly.

4 dwg, 4 ex

FIELD: medicine.

SUBSTANCE: group of inventions refers to medicine, namely traumatology and surgery, and may be used for bleeding control. That is ensured by the use of a composition, as well as an adhesive material containing gelatin and transglutaminase wherein their relation is sufficient for bleeding control in a wound, and wherein said gelatin is not exposed to irreversible gel-formation and forms a solution with transglutaminase at temperature below natural one of a gel-sol junction of standard animal gelatin prepared from an animal source or representing recombinant gelatin or their combination.

EFFECT: use of the given inventions enables reducing time of bleeding control in the wound.

22 cl, 15 dwg, 12 tbl, 15 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: in formula (1): R1 means haloalkyl containing 1-6 fluorine atoms; R2 means C1-C6alkyl or halogen; R3 means -L-NR4R5, -X-NR-C(O)R8 or -X-NR-C(O)NR4R5 wherein L means -X-C(O), -(CR2)j, -O(CR2)1-4 or and X means (CR2)j or [C(R)(CR2OR)]; R4 and R5 independently mean H, C1-C6alkyl, halogen-substituted C1-C6alkyl, hydroxy group-substituted C1-C6alkyl, or (CR2)k-R6; R8 independently means (CR2)k-R6 or C1-C6alkyl, or halogen-substituted C1-C6alkyl; R7 means H; alternatively, R4 and R5 together with N atom in each NR4 R5 form a 4-7-member heterocyclic ring containing 1 -2 heteroatoms independently specified in N and O substituted by 0-3 groups R11; R11 means R8, (CR2)k-OR7, CO2R7, (CR2)k-C(O)-(CR2)k-R8, (CR2)kC(O)NR7R7 or (CR2)kS(O)1-2R8; each R means H or C1-C6alkyl; each k is equal to 0-6; and j and m are independently equal to 0-4; provided R1 does not mean trifluoromethoxygroup, provided R3 means C(O)NH2, C(O)NR12R13; wherein R12 and R13 together form piperazinyl; the values of the radical R6 are presented in the patent claim. The invention also refers to the pharmaceutical composition containing said compounds.

EFFECT: producing new 5-(4-(halogenalkoxy)phenyl)pyrimidin-2-amine derivatives showing c-kit, PDGFRα, PDGFRβ kinase inhibitory activity, optionally in the form of isomers or pharmaceutically acceptable salts.

12 cl, 77 ex, 1 tbl

FIELD: medicine.

SUBSTANCE: invention refers to medicine, particularly to applying a neutralising antibody to matrix metalloproteinase-10 (MMP-10) for preparing a drug applicable for antifibrinolytic therapy and treating bleedings and hemorrhagic complications of various aetiology: hyperfibrinolytic conditions caused by congenital pathologies, anticoagulant therapy, surgical procedures.

EFFECT: invention provides higher selectivity of inhibitors which block molecular mechanisms associated with specific matrix metalloproteinases that allows avoiding undesired events.

7 cl, 4 ex, 2 tbl

FIELD: chemistry.

SUBSTANCE: palladium-copper catalyst for homogeneous selective oxidation of thiols combines a functional binuclear thiolate bridge coordination compound of palladium (II) and a modifying thiolate complex of copper (I), having general formula [pd2II(µ-SR)2(NH3)4]·{Cu1k(SR)m} (I). SR is a residue of a thiolate ligand of glutathione or acetylcysteine, k ranges from 2 to 14, m≥3k. The invention also discloses a catalytic combination which stimulates proliferative activity, a pharmacological combination for enhancing therapeutic activity of a purine and(or) pyrimidine base or lithium ions, pharmaceutical compositions which stimulate proliferative activity, and for enhancing therapeutic activity of said bases or lithium ions, and methods for therapeutic treatment of a patient based on said catalyst.

EFFECT: catalyst has low toxicity and can be used in pharmacological solutions with medicinal agents for therapy of different duration where catalytic activity is needed in reactions for oxidative modification of thiols to disulphides in peptide molecules.

19 cl, 3 dwg, 11 tbl, 12 ex

FIELD: medicine.

SUBSTANCE: invention relates to medicine and deals with closed container, which contains composition of activated factor VII polypeptide in amount in range from more than 10 to 90 mg per 1 ml of container volume. Invention also relates to various methods of obtaining activated factor VII polypeptide in closed container; set, including such containers; and method of obtaining ready for application liquid aqueous pharmaceutical composition of activated factor VII polypeptide from the set.

EFFECT: invention ensures obtaining high FVII a concentrations.

31 cl, 27 ex, 1 dwg, 7 tbl

FIELD: medicine.

SUBSTANCE: invention relates to medicine, namely to surgery and disaster medicine, and can be used in case of necessity to stop hemorrhage caused by injury of parenchymatous organs in case of polytraumas. For this purpose at the stage of sorting out wounded people with symptoms of continuing intra-abdominal hemorrhage are detected, delivered to operation room, where in cause of taking anti-shock measures, intra-abdominal hemorrhage is diagnosed. After that, laparotomy is performed and source of hemorrhage is detected. On detected injuries of parenchymatous organs applied is hydrophilic powder two-component hemostatic preparation in form of mixture of synthetic zeolites of CaA and CaX types, synthesized in kaolin granules. CaA zeolites correspond to formula mCaO·nNa2O·2SiO2·Al2O3·0.5H2O in volume amount within the interval 70÷80 wt %. CaX type zeolites correspond to formula mCaO·nNa2O·2.5SiO2·Al2O3·0.5H2O in volume amount within the interval 30÷20 wt %. Crystal matrices of said zeolites contain calcium ions in quantity 8÷10 wt %. After that, gauze tampon is applied and pressed by hand for 5-7 min until stable hemostasis is achieved. Simultaneously detection of other injuries is continued by revision of abdominal cavity organs which results in determination of further patient treatment tactics.

EFFECT: method makes it possible to ensure efficient stable primary hemostasis in case of multiple hemorrhages.

FIELD: medicine, pharmaceutics.

SUBSTANCE: what is offered is an agent enhancing thrombocyte adhesion properties, representing cyclophilin A. It is shown that cyclophilin A not changing thrombocyte count increases collagen-induced thrombocyte aggregation without thrombin, and does not have an effect on ADP-induced and ristocetin-induced thrombocyte aggregation.

EFFECT: cyclophilin A provides an adhesive effect, and does not promotes development of thromboembolic complications.

3 tbl

FIELD: veterinary medicine.

SUBSTANCE: method includes injection of the preparation "FSH-super" and the preparation of prostaglandin F2α. On the day of artificial insemination the cows-donors are additionally injected with homeopathic preparation "Ovariovit" at the dose of 4.9-5.1 ml per head in a single dose.

EFFECT: method enables to obtain a greater number of embryos and to improve their quality.

5 tbl, 1 ex

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