Parenteral composition, method for preparing and using composition

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to pharmaceutical industry, namely to an immunomodulatory composition for injection into a mammal. The immunomodulatory composition for injection into a mammal containing a hydrolysate prepared by acid and/or enzymatic hydrolysis of one or more bioresources specified in a group consisting of bivalve molluscs, annelids, leeches, and water taken in certain proportions. A method for preparing the immunomodulatory composition for injection into a mammal. A method of treating a pathological condition in a mammal in need thereof involving the injections of the immunomodulatory composition into the above mammal. Using the composition for normalising metabolism into the mammal in need thereof.

EFFECT: composition enables extending the range of products with immunomodulatory activity for injections.

19 cl, 7 tbl, 7 ex

 

The technical FIELD

The present invention relates to the field of veterinary medicine. In particular, the present invention relates to compositions for parenteral administration based hydrolysate derived from natural biological resources, the method of obtaining this composition, method of treatment and/or prevention of pathological conditions in need of this mammal using the above compositions and different applications of the specified compositions for therapeutic and/or prophylactic, and other purposes. More specifically, the present invention relates to a composition having immunomodulatory properties based hydrolysate derived from biological resources, for parenteral administration to an organism in need of a mammal.

According to the present description of the proposed composition for parenteral administration based hydrolysate derived from biological resources. This composition contains a wide range of essential amino acids, melanoidins, acting as antioxidants, regulatory peptides, saturated and unsaturated fatty acids, vitamins and trace elements in a balanced natural ratio with a number of positive properties used in therapeutic and/or prophylactic purposes in veterinary medicine and medicine. In particular, the author of this image is the shadow was unexpected was detected, the composition for parenteral administration containing 1-10% aqueous solution of the hydrolyzate obtained from the above biological raw materials for parenteral administration, has considerable advantages in comparison with the compositions known from the prior art. In addition, a number of effects inherent in the present compositions when administered parenterally, is not observed in the case of oral administration of pure hydrolysate, or observed, but after a longer period of time. On the other hand, the present composition also has a number of advantages compared with the known parenteral forms. It was found that parenteral composition method disclosed in the present description, is safe and does not cause allergic reactions and other undesirable side effects.

The LEVEL of TECHNOLOGY

The prior art a variety of drugs and biologically active additives, representing protein hydrolysates obtained from raw materials of animal origin, in particular seafood, such as mussels, fish, mammal meat, etc. These protein hydrolysates, as a rule, are supplements intended for oral use individually or in combination with other drugs, and also as a food Supplement.

From German patent DE 409339 known biologically active pharmaceutical natural product of nature, which contains an acid hydrolysate of the molluscs of the genus Mytilus, containing amino acids, melanoidins and traces elements of edible mussels (Mytilus edulis) and the black sea mussels (Mytilus galloprovincialis). Additionally, the invention includes different ways to use this product. In particular, the specified product is used as a prophylactic, therapeutic radiopharmaceutical agent for humans and animals, has antipyretic effect, stimulates the production of blood especially during radio - and chemotherapy.

Also the prior art product - MIGS (MIG-PL®, liquid for oral administration), which represents the solution obtained by hydrolysis of the black sea mussels, used as nutritional supplements.

Well-known dietary Supplement restorative actions with similar properties MIGS-K, obtained from the meat of the mussels in the White sea - MYDEL. Therapeutic technique in combination with traditional therapies or as a standalone drug MYDEL recommended for immunodeficiency various etiologies.

One of the drawbacks of oral hydrolysates obtained from meat of mussels is relatively low speed achieve the desired effect in their daily admission.

The prior art also known composition, the region is non immunomodulatory effects, obtained from raw materials of animal origin, other than mussels.

In the patent of the Russian Federation 2221456 proposed universal biologically active substance (BAS), which is a protein hydrolysate obtained by acid hydrolysis and subsequent neutralization of fish meat, protein-rich waste products from animals or fish, filtration to obtain a hydrolyzate and the precipitate, followed by drying the hydrolysate. The patent States that the stated substance use as a food additive for animals, the basics of veterinary medicine, oral nutrition, perfumes and cosmetics, dairy products, confectionary products and other

In addition to the specified source various products and compositions on the basis of the hydrolysates obtained from meat, fish, known from EP 1653981, US 2009111747, US 2011124570.

In the patent of the Russian Federation 2402320 proposed product containing various amino acids, salts and trace elements, in the form of an aqueous solution, and a method of prevention and correction of pathological conditions in animals, consisting in injecting a specified drug in the body of the animal with the preventive purpose 2 times a week for months at doses of 1.5-2.0 ml per 10 kg of body weight, with the purpose of treatment at a dose of 3.0-5.0 ml per 10 kg of body weight 2 times a day for 3-5 days, with intoxication due to poisoning manufactured the definition and/or food poisons - in a ten-fold therapeutic dose. In particular, by subcutaneous or intramuscular injection of a specified drug provides a more effective prevention and treatment of diseases of different etiologies. However, obtaining the specified synthetic drug is a time-consuming and energy-consuming process by which they develop optimal qualitative and quantitative compositions and methods preparation of aqueous solutions of amino acids, vitamins, micro - and macroelements, glucose, normalizerbase metabolic processes in the animal body.

The prior art is also known for a variety of drugs for injection, such as, for example, Minton. Due to the complex three homeopathic components entering into its composition, Minton stimulates the phagocytic activity and the barrier function of the lymph nodes, skin, mucous membranes and other organs and tissues. Minton is an immunomodulator used in diseases of bacterial and viral etiology - plague distemper, feline panleukopenia, feline parvovirus enteritis, viral hepatitis, rhinotracheitis, etc. Minton administered daily, the course of treatment lasts from a few days (viral enteritis) up to 2-3 weeks (viral hepatitis).

All of the above technical solutions have a relatively low eff what aktivnosti in therapeutic and preventive purposes. A positive effect in patients on oral drugs derived from living aquatic resources, usually occurs over a long period of time provided it is continuous. Oral forms are used as biologically active additives to food, while the quantity of a drug during the entire course of treatment varies and may reach tens of milliliters to several liters. On the other hand, is known for its injectable form does not have a broad spectrum of activity, possibly due to the fact that their compositions either poorly balanced, as, for example, in the case of artificially produced compositions, or depending on natural origin, does not contain the entire set of connections that are necessary and sufficient to achieve rapid preventive or therapeutic effects in humans and animals, such as fish hydrolysates.

Thus, in the present description of the disclosed composition having various therapeutic and preventive properties to achieve the desired effect in a relatively short period of time regardless of ability and/or desire of the subject to food intake and the ability and the possibility of ingestion of the drug and its absorption through the gastrointestinal tract. This composition m which may be particularly useful in the field of veterinary medicine, where in many cases the animal is suffering from a pathological condition or needs in the means of prevention of certain diseases, refuses a meal containing oral drug, because of the taste characteristics of the latter. This composition also allows to solve this problem.

It was unexpectedly discovered that the properties disclosed in the present description of the composition include together with immunomodulating activity, phagocytic activity, hepatoprotective activity, adaptogenic activity, Antiherpes virus effect activity, detoxification activity, antiviral activity, antibacterial activity, antialgae and wound-healing properties. While specified therapeutic effect can be achieved after the first few techniques of composition. In some cases, the positive dynamics of treatment is observed with the first dose of the proposed compositions for parenteral administration.

Also in the present description disclosed a method of obtaining the above-mentioned composition. This method is simple in technical execution, and its implementation does not require special production capacity and depth of professional skills that have a positive impact on its economic feasibility.

The specified composition for parenteral administration note the following in the treatment and/or prevention of pathological conditions in need of this mammal. That is, according to another aspect of the present description of the disclosed method of treatment and/or prevention of pathological conditions of various etiologies in in need thereof of a subject using the above composition. This method includes the best mode of reception of the above composition, ensuring the achievement of positive preventive and/or therapeutic effect, in which regardless of the individual characteristics of the subject did not reveal any side effects including a single case of individual intolerance mammal a composition.

Thus, in the present description discloses immunomodulating composition for parenteral administration containing a hydrolysate derived from biological raw materials of animal origin, methods of obtaining the specified compositions, methods of treatment and/or prevention of pathological conditions in need of this mammal using the above composition and various applications of the indicated composition.

SHORT DESCRIPTION

According to the first aspect of the proposed composition for parenteral administration to a mammal containing from 1 to 10 wt.% the hydrolysate derived from biological resources, and water.

The term "biological resources" should be understood as any raw materials of animal or vegetable origin. For example,as biological resources can be used by animals and/or plants marine or terrestrial origin.

According to one implementation variant bioresources selected from the group including bivalves, crustaceans, annelids, leeches. These biological resources allow us to obtain a hydrolysate having a balanced chemical composition, including, amongst others, a wide range of amino acids, including essential amino acids, saturated and unsaturated fatty acids, melanoidins, carbohydrates, micro - and macro-elements, on the basis of which can be obtained composition to achieve the desired therapeutic effect when administered parenterally in need of such treatment or prophylaxis of a mammal.

This composition may contain a hydrolysate obtained from one or more kinds of raw materials. In particular, the composition may contain a hydrolysate derived from bivalve shellfish and/or crustaceans and/or annelids, and/or leeches.

According to one implementation variant bivalves include the mussels {Anodonta), the thick-shelled river mussels (Unio), oysters (Ostreidae), mussels (Mytilidae}, tridacna (Tridacna), oyster (Pinctada}, scallops (Pectinidae), ship worms (Teredinidae), guidec (Panopea abrupta), Icelandic cyprine (Arctica Islandica) and other species of shellfish.

According to one implementation variant crustaceans include krill, shrimps, crabs, lobsters, langoustines, crayfish,lobster and other species. Preferably crustaceans include krill (Meganyctiphanes norvegica) and/or North Atlantic lobster (Homarus americanus).

According to another implementation variant bioresource is a marine red sea urchin (Strongylocentrotus franciscanus).

All of the above biological resources allow you to obtain a composition that has a range of preventive and therapeutic properties. While bivalves and crustaceans are common commercial products and therefore are also suitable biological resources for obtaining this composition from an economic point of view. The composition of the hydrolysate obtained from marine species may slightly vary depending on location and external environmental conditions of their habitat, however, the basic properties are in General similar. Annelid worms and leeches preferably grown in a changing production environment that provides strong consistency of the composition obtained from using them. However, in the absence of this possibility, the latter may also be obtained from natural sources in their habitat.

The term "hydrolysate derived from biological resources" in the context of the present description should be understood liquid solution obtained by the hydrolysis of biological resources.

According to one variant Rea is Itachi of the present invention, the amount of the hydrolysate in the composition is from 2 to 8 wt.%, preferably from 3 to 7 wt.%, more preferably from 4 to 6%, most preferably about 5 wt.%. According to another implementation variant, the number of the hydrolyzate can be from 1 to 4 wt.%, from 1 to 3 wt.%, from 1 to 2 wt.%, from 6 to 9 wt.%, from 7 to 9 wt.% and from 8 to 9 wt.%.

Typically, the amount of the hydrolyzate does not exceed 10 wt.% composition, because otherwise the composition will not be suitable for certain types of parenteral administration, in particular the injection, due to the presence of high concentration of the contained salt. On the other hand, the mass fraction of the hydrolysate, as a rule, is not less than 1 wt.% to provide the composition with a sufficient number of components for the occurrence of the desired effect in the treatment and/or prevention of various conditions in need of the subject.

The term "in need (treatment or prevention) mammal" should be understood as any animal of the class mammal or human, in need of treatment and/or prevention of pathological conditions, diseases, disorders, etc.

According to one implementation variant, the mammal is an animal or human. Preferably the animal is an agricultural animal selected from the group including cow, Buffalo, Yak, deer, pig, goat, is the Supreme ecclesiastical authority, rabbit, horse, donkey, camel, Lama, sable, Fox, mink, ferret, etc, or the animal can be a pet, such as dog, cat, rat, hamster, Guinea pig, etc. According to another implementation variant, the animal may be an animal living in the wild.

According to one implementation variant "parenteral administration" includes any method of introducing prevent leakage of the proposed composition in the organism of a mammal in need through the gastrointestinal tract.

According to one implementation variant, the composition is a composition for the injection. The specified injection can be carried out intravenous (IV), intradermally, subcutaneously (s/C), intramuscularly (I/m) or nutricote. The author of the present invention it has been unexpectedly discovered that injecting the above composition significantly enhances therapeutic and prophylactic effects achieved in comparison with receiving undiluted hydrolysate oral method, and in addition, significantly accelerates time to achieve such effects. In addition, parenteral reception of the specified composition has managed to achieve some effects that were not identified during oral undiluted hydrolysate, extracting the aqueous meat mussels, as well as injectable forms known from the prior art. It should also be noted that, in accordance with clinical studies on animals indicated composition had no side effects and do not cause allergic reactions when its systematic parenteral administration.

According to another implementation variant composition used for outdoor applications. In particular, the composition may be an ointment, paste, liniment, cream, lotion and equivalent dosage form, or be part of these funds for external use. The composition may be part of the cutaneous applications, including to provide a transdermal therapeutic system with prolonged action. In particular, the described composition can be applied on the body with thin skin needy mammal. In addition, the composition for external use can be used in the treatment of open wounds, burns, frostbite, bruises, dislocations, fractures and other cutaneous and subcutaneous lesions.

According to another variant of realization of the specified composition is characterized by the mass fractions of amino nitrogen component of at least 0.01 wt.%. Preferably, the mass fraction of amine nitrogen is from 0.01 to 0, 5 wt.%. More predpochtite the flax mass fraction of amino nitrogen is from 0.05 to 0.5 wt.%, most preferably from 0.02 to 0.1 wt.%. The value of the mass fraction of amine nitrogen characterizes the degree of completion of the hydrolysis of life and is the factor that allows us to assess the completeness of this process. The above description serves as an indirect indicator of the availability of the necessary components in the final composition for parenteral administration using biological resources in accordance with the present description.

According to another variant implementation of the proposed composition has a pH of 4 to 7, preferably 5 to 7, most preferably from 5.5 to 6.5. The pH in the above range indicates a sufficient degree of neutralization of the hydrolysate, and the possibility of applying the specified compositions for therapeutic and/or prophylactic purposes for mammals.

According to another variant implementation of the proposed composition has a mass fraction of solids of from 0.5 to 5 wt.%, preferably from 1 to 3 wt.%, most preferably 1.2 to 1.8 wt.%.

According to another implementation variant, the composition contains a wide range of amino acids depending on the type of biological resources used for it. In particular, the composition according to the present invention comprises amino acids selected from the group including taurine, asparaginase acid, Trevi is, serine, glutamic acid, sarcosine, glycine, alanine, valine, cystine, methionine, cystathionine, isoleucine, leucine, tyrosine, phenylalanine, β-alanine, γ-aminoadamantane acid, γ-aminomethane acid, ornithine, lysine, histidine, carnosine, arginine, hydroxyproline, Proline. Perhaps this composition also contains other amino acids.

According to another variant of implementation of the present composition includes at least one or more indispensable for the human body amino acids selected from the group comprising valine, isoleucine, leucine, lysine, methionine, threonine, tryptophan, phenylalanine, arginine and histidine.

According to one implementation variant, the composition comprises the following amino acid in a mass ratio relative to the total number of amino acids in the specified composition:

taurine from 1 to 10%

aspartic acid is from 1 to 30%

glutamic acid, from 1 to 30%

glycine from 1 to 20%

alanine from 1 to 10%

leucine from 1 to 15%

phenylalanine from 1 to 15%

lysine from 1 to 10%

arginine from 1 to 20%

Proline from 1 to 50%

serine from 1 to 10%

histidine from 1 to 10%

threonine is from 0.5 to 10%

valine from 0.1 to 10%

methionine from 0.1 to 10%

isoleucine from 1 to 10%

According to one implementation variant, the same composition may, but not necessarily, at the same time to have a mass fraction of dry substances, pH, m is sowwy share amine nitrogen and the qualitative and quantitative content of amino acids, as specified above.

According to one implementation variant of the invention disclosed in the present description, the composition is characterized in that

mass fraction of amino nitrogen is from 0.01 to 0.5 wt.%, and/or

the pH value is from 4 to 7, and/or

mass fraction of dry substances is from 0.5 to 2 wt.%

In addition to the amino acid composition disclosed in the present description, contains saturated and unsaturated fatty acid selected from the group comprising acids with long hydrocarbon chains of from 12 to 25 carbon atoms. In particular, the composition preferably contains one or more fatty acids selected from the group comprising tridesilon acid C13:0, myristic acid C14:0, tetradecanoyl acid C14:1, pentadecanoic acid C15:0, palmitic acid C16:0, palmitoleic acid C16:1, hexadecanoyl acid C16:2, hexadecanediol acid C16:5, heptadecanoic acid C17:0, heptadecenoic acid C17:1, heptadiene acid C17:2, stearinovuyu acid C18:0, oleic acid C18:1 linoleic acid C18:2, linolenic acid C18:3, octadecatetraenoic acid C18:4, Aksenova acid C20:1, eicosadienoic acid C20:2, eicosatrienoic acid C20:3, arachidonic acid C20:4, eykozapentaenovuyu acid C20:5, heptacosanoic acid C21.-5, docosanol acid C20:0, dokozagexaenovu the acid 22:1, docosanol acid C22:2, docosatetraenoic acid C22:4, docosahexaenoic acid C22:6, triazinetrione acid, C23:4, triazapentadiene acid, C23:5, tetracosanoic acid C24:1.

According to another implementation variant specified composition contains trace elements such as potassium, calcium, magnesium, iron, zinc, copper, cadmium, manganese, Nickel, chromium, selenium, iodine.

Additionally, this composition may contain known in the art of biologically active additives known in veterinary medicine and medicine and drugs for the prevention and treatment of pathological conditions, diseases and disorders in humans and animals, macro - and micronutrients, amino acids, melanoidins, carbohydrates, peptides, vitamins. Preferably to this song add salt required to obtain physiological solution suitable for parenteral administration. According to one implementation variant of the invention specified composition before the introduction mixed with glucose.

According to the second aspect of the present description, a method for receiving immunomodulatory compositions for parenteral administration in need of the prevention or the treatment of the mammal. In particular, the method for obtaining the composition for parenteral administration to a mammal, including:

(a) the enzyme is active hydrolysis and/or acid hydrolysis of bioresources, selected from the group including bivalves, crustaceans, annelids, leeches with the first solution;

(b) filtering the first solution to obtain a second solution;

(c) mixing the second solution with water to form a composition with a mass content of the second solution is from 1 to 10 wt.%.

According to one implementation variant stage (a) includes enzymatic and acid hydrolysis. As the enzyme can be used enzymes of animal, plant, or microbial origin. In particular, these enzymes without restrictions include praemegaceros, protegrin, it, trypsin, pepsin, and other interchangeable enzymes. The amount of enzyme, pH, temperature and other conditions pick empirically depending on the type of enzyme and raw materials subjected to enzymatic hydrolysis. The pH value during enzymatic hydrolysis is from 1 to 12, more preferably from 1 to 10, most preferably from 1 to 9. The temperature during the enzymatic hydrolysis of support in the range of from 20 to 80°C., more preferably from 30 to 70°C., most preferably from 30 to 60°C. the Enzyme is added in the form of dry powder in the raw material with stirring. The specified stage does not require additional consumption of fresh water, and can be held in places fishery of the criminal code of the related raw materials. The enzymatic hydrolysis is carried out in a period of from 10 minutes to 5 hours, preferably from 20 minutes to 3 hours, most preferably from 30 minutes to 1 hour.

In that case, if the specified biological raw material is a bivalve mollusks, the enzymatic hydrolysis may be necessary to loosen the locking muscles shellfish and, thereby, separating the soft tissue of mussels from shells, as well as for the primary hydrolysis soft tissues of clams. After enzymatic hydrolysis mollusk shells are separated from the soft tissue using filters, such as a vacuum filter or a pressure filter, and/or vibrating screen. After separation of the soft tissue conduct subsequent acid hydrolysis or further enzymatic hydrolysis with the use of one or more enzymes.

Alternate leaf and shell bivalves can be separated mechanically by hand and/or using methods that exclude the use of enzymes, for example, by using hot steam. Also bivalve molluscs can be fragmented and subjected to acid hydrolysis without the use of enzymes.

Raw materials other than bivalves, such as crustaceans, in particular, krill, as well as leeches, annelids, can also be subjected to enzymatic hydrolysis.

According to one implementation variant by acid hydrolysis using an acid number of from 5 to 20 wt.%, preferably from 5 to 15 wt.%, more preferably from 5 to 10 wt.% from the quantity of processed raw materials. The specified amount of acid, as a rule, sufficient to ensure the completeness of the process of hydrolysis. However, the amount of acid may be selected empirically, taking into account the views of life and the depth of the passage of hydrolysis.

According to one implementation variant, the temperature acid hydrolysis set in the range from 80 to 120°C., preferably from 90 to 110°C., most preferably from 95 to 105°C. In particular, acid hydrolysis Prov is car Ried out at the boiling solution.

Acid hydrolysis is carried out in enameled or glass reactor equipped with a thermometer or thermocouple for temperature control and mixing device.

Alternative raw materials can not be subjected to acid hydrolysis, when such is not necessary. In particular, the completeness of hydrolysis can be achieved only through enzymatic hydrolysis. In the case of acid hydrolysis was carried out, at the next stage, carry out the first stage of neutralization solution, comprising adding a neutralizing agent in the first solution obtained by acid hydrolysis, in a quantity sufficient to bring the pH of the solution to values of 4-7, preferably from 5 to 7, most preferably from 5.5 to 6.5. As the neutralizing agent used agents, forming water and a neutral salt, such as alkali, such as NaOH and/or KOH, in a solid aggregate state or in solution, as well as basic salts and their solutions. In particular, as salt can be used carbonates and/or bicarbonates of sodium or potassium. Preferably the neutralizing agent is a solid sodium hydroxide.

Next, carry out the filtering of the received first solution with the second solution.

According to one implementation variant before filtrowanie the m first solution is performed in its assertion at a temperature of from 1 to 25°C, preferably from 4 to 15°C, most preferably from 4 to 6°C for at least 10 days, preferably from 10 to 20 hours, most preferably from 10 to 30 days. The specified stage provides a more complete precipitation of the insoluble components from the first solution and their separation at a later stage, thereby increasing the stability of the final composition for parenteral administration.

After deposition of sediment and its affiliates receive the second solution. Stage filtering the first solution with the second solution is performed using a mechanical filter, preferably a vacuum filters. The specified stage can be divided into several stages and enable filtering hot first solution after completion of the enzymatic and/or acid hydrolysis, and one or more re-stages filtration of the cooled first solution with the second solution. Preferably stage filtration is carried out after the stage of settling the first solution in the conditions specified above.

At the next stage, carry out mixing the second solution with water to form a composition with a mass content of the second solution is 1 to 10 wt.%, preferably from 2 to 8 wt.%, or from 3 to 7 wt.%, more preferably from 4 to 6%, most preferably about 5 wt.%. Solenoide one implementation variant, the mass content of the second solution may be from 1 to 4 wt.%, from 1 to 3 wt.%, from 1 to 2 wt.%, from 6 to 9 wt.%, from 7 to 9 wt.% and from 8 to 9 wt.%. Water is distilled water or water for injection. The specified weight percent of the second solution in the composition corresponds to such number, in which the resulting composition is suitable for parenteral administration, particularly intravenous administration. According to one implementation variant the result of the composition is isotonic to the blood of a mammal in need of prevention or treatment using the compositions. Before applying this composition may be mixed with other ingredients, such as vitamins, carbohydrates, additional amino acids, micro - and macronutrients, glucose, etc.

According to another implementation variant of the composition after adding distilled water and/or water for injection, sterilized and filled into containers for storage. As reservoirs for storage of the composition can be used ampoules, vials, bottles, bags, etc. a Method of obtaining carried out in aseptic conditions in compliance with all requirements stipulated by the state Pharmacopoeia to the specific forms for parenteral administration. Capacity with the obtained composition is stored at a temperature of from 0 to 25°C, preferably from 4 to 6°C. shelf Life is oppozitsii while maintaining appropriate conditions may be not less than 2 years. Preferably with all of the above conditions, the composition remains stable and suitable for use for 5 years or more.

According to a third aspect of the specified composition used in the treatment and/or prevention of pathological conditions in need of this mammal, wherein the specified composition is administered parenterally to a needy person.

According to another implementation variant in the present description, a method for treatment and/or prevention of pathological conditions in need of in a mammal comprising injecting a specified composition in need of such treatment or prophylaxis of a mammal.

Specified parenteral administration includes any kind of introduction, the ingress of specified composition in the body of a mammal through the organs of the gastrointestinal tract. In particular, parenteral administration is an injection selected from the group including subcutaneous injection, intradermal injection, intramuscular injection, intravenous injection or intraosseous injection. Specified parenteral administration can also be an external application of the above composition.

According to one variant, i.e. monitoring) reference and specified composition is administered in an amount of from 0.05 ml/kg 10 ml/kg of mass of a specified mammal from 1 to 5 times a day for 1 to 50 hours. More preferably, the composition is administered in an amount of from 0.1 to 1 ml/kg for 1-15 days.

According to another implementation variant specified composition is administered once in the amount of from 0.05 ml/kg 10 ml/kg of mass of a specified mammal every 1-5 days in the amount of from 0.05 to 10 ml/kg over 1 to 50 days. More preferably, the composition is administered in an amount of from 0.1 to 1 ml/kg for 1-15 days.

According to another implementation variant composition maybe used 1 time in 45-120 days during the year in the amount of from 0.05 ml/kg 10 ml/kg of mass of a specified mammal. More preferably, the composition is administered in an amount of from 0.1 to 1 ml/kg 1 time in 90 days.

In addition, the mode of administration of the composition can be installed depending on the type of pathological condition, being in need of a mammal or a specific destination specified composition.

According to one implementation variant specified pathological condition, treatment and/or prevention of occurrence of which is a liver disease of various etiologies, steatosis, intoxication, skin disease, disease of the hair and/or scalp, dermatitis, violation of the integrity of the skin due to injury or burn, obstructive lung disease, pneumonia, acute respiratory virus is th infection (ARI), influenza virus, circovirus infection, acidosis, herpes, stress, weakness in the postoperative condition, inflammation of the mucous membrane of the intestine, osteodystrophy, like status, osteoarthritis, degenerative disc disease, atherosclerosis and other diseases.

According to another implementation variant specified composition suitable for fast and effective immunity, normalize metabolism, in particular, calcipotriol metabolism, weight gain, if necessary, and its reduction in case of excess weight, increase the protein synthetic function of the liver, as a means of promoting the healing of wounds and burns, to contain the growth of benign and malignant tumors tools that stimulate the growth of hair, means to bring the farm animals in the state of estrus and improve reproductive function in animals.

According to one implementation variant specified composition used in any of poisoning, especially the poisoning of exogenous and endogenous nature, for example. Thus, the specified composition uses as a tool for detoxification. In this case, the composition is administered in an amount of from 0.1 to 10 ml/kg of the mammal in need at the same time. In most cases, requiring m is Capitalia gets rid of the symptoms of intoxication with the introduction of the dosage from 0.1 to 0.5 ml/kg With the resumption of symptoms of poisoning composition is administered in the same dosage from 1 to 5 times.

According to another implementation variant specified composition is used to improve the immune status in need of a mammal. In other words, the present composition is an immunomodulatory composition. In particular, the above composition when applied to the General weakness of the body due to various diseases, and also in those cases when the body of a mammal weakened, for example, as the result of previous treatment with antibiotics and/or drugs that weaken the immune system. For example, the specified composition is administered to the needy in the restoration of the mammal in the postoperative period or the period following chemotherapy, or in the period after pregnancy, during breast feeding offspring, etc. as a preventive measure specified composition is administered during epidemic infectious diseases such as influenza, SARS and other diseases.

According to another implementation variant specified composition is used for normalization of metabolism in need of this mammal. In addition to the core indicators, normalization of metabolism indirectly estimate the increase or reduction of body weight in a mammal in need of such increase or decrease the drop, respectively, increase the hair or wool on, to improve mobility and overall activity at the age of animals and other figures.

According to another implementation variant specified composition is used for normalization calcipotriol metabolism in need of this mammal. This normalization calcipotriol sharing judged by the content of calcium and phosphorus in the body of a mammal.

According to another implementation variant specified composition is used to bring the animal into a state of estrus, and overall improvements to its reproductive function. In particular, it was found that the intramuscular injection of agricultural animals from 1 to 3 doses of the present compositions in amounts of 0.005 to 1 ml/kg, preferably from 0.01 to 0.1 ml/kg, more preferably from 0.01 to 0.05 ml/kg weight of the animal during the day before insemination allows to achieve the above effect. In particular, the achievement of this effect was observed with intramuscular administration of the composition to the cows in the amount of from 0.01 to 0.05 ml/kg twice a day.

According to another implementation variant specified composition is used to reduce stress in need of this mammal. In particular, experimental animal models have shown that this composition exhibits a number of adaptogenic/antistress the new properties. For example, as shown, the composition of improved survival and increased weight gain of piglets in the post-Natal period of their development, and also gave positive results on model transport stress in horses.

According to another implementation variant specified composition is applied as a wound healing agent in need of this mammal. These wounds may be due to different physical damage, in particular damage to the integrity of skin and coat, as well as various diseases, such as dermatitis, psoriasis, herpes and other lesions of the skin of various etiologies.

According to another implementation variant specified composition is used as a growth-promoting means in a mammal. In particular, the growth-promoting action of the specified composition was assessed by total weight gain of farm animals receiving the specified composition, in comparison with control animals not receiving the specified composition or receiving only the hydrolysate for oral administration.

These and other advantages of the present invention will be more fully demonstrated in the following section a detailed description of specific examples of its implementation.

PODROBNEE DESCRIPTION

<> A composition for parenteral administration

Example 1

100 kg ciprini Icelandic (Arctica Islandica) were placed in a fermenter with constant mechanical stirring fired heating apparatus of the hot water to 42°C. the fermenter was added 1 kg of protocolin. Exercised control pH, the corresponding neutral environment for the reading of a pH meter.

Enzymatic hydrolysis was carried out for 40 minutes with constant stirring, after which the valves of shells were separated using roar. To the resulting solution was added concentrated hydrochloric acid and spent acid hydrolysis for 16 hours in enameled apparatus with a stirrer at a temperature of 100-105°C. after hydrolysis of the resulting solution under vacuum pumped into the catalytic Converter, cooled using a water circuit and neutralized with dry alkali to the value of pH in the range 4-6 with constant cooling. The neutralized hydrolysate was held out for 15 days at an ambient temperature of 20°C. Then made the filter on the vacuum unit for separating the resulting precipitate from the receipt does not contain solids solution of the hydrolyzate. To the obtained solution of the hydrolyzate with a mass of about 10 kg with stirring was added the required amount of distilled water ("the ode for injection") to produce a solution with a concentration of hydrolysate 5% by weight. Before filling the containers with the composition for the final application, conducted heat sterilization of the composition and, if necessary, subsequent filtration of the solution. Capacity with the finished composition was stored at a temperature of 4-6°C out of direct sunlight. The results of chemical analysis of the obtained compositions are presented below.

Mass fraction of amino nitrogen to 0.08%

Mass fraction of dry substances - 1.5 wt.% PH of 5.7

Trace elements (qualitative part):
Cali
Calci
3. Magnesium
4. Iron
5. Zinc
6. Copper
Cadmia
8. Manganese
9. Nickel
10. Chrome
11. Selenium
12. Iodine

Amino acid composition (including irreplaceable)Fatty acids (lipids)
1. Taurine1. Traditiona 13:0
2. The phosphoethanolamine2. Myristic C14:0
3. Asparagina the3. Tetradecene C14:1
4. Threonine4. Pentadecanol C15:0
5. Serine5. Palmitic C16:0
6. Glutamic the6. Palmitoleic C16:1
7. Sarcosin7. Hexadecanoate C16:2
8. Glycine8. Hexadecapole C16:5
9. Alanine9. Heptadecanoyl C17:0
10. Valine10. Heptadecanoyl C17:1
11. Cystine11. Heptadienal C17:2
12. Methionine12. Steo the new C18:0
13. Cystathionine13. Oleic C18:1
14. Isoleucine14. Linoleic C18:2
15. Leucine15. Linolenic C18:3
16. Tyrosine16. Octadecatetraenoic C18:4
17. Phenylalanine17. Aksenova C20:1
18. β-alanine18. Eicosadienoic C20:2
19. γ-aminoadamantane19. Eicosatrienoic C20:3
20. γ-aminomalonate20. Arachidonic C20:4
21. Ethanolamine21. Eicosapentaenoic (C20:5
22. Ornithine22. Heptanophenone C21:5
23. Lysine23. Docosanoate With C20:0
24. Histidine24. Dokozagexaenova C22:1
25. Carnosine25. Docosanoate C22:2
26. Arginine26. Docosatetraenoic C22:4
27. Hydroxyproline27. Docosahexaenoic C22:6
28. Proline28. Treasuretrove C23:4
29. Triazapentadiene C23:5
30. Tetracosanoate C24:1

Example 2

50 kg of krill (Meganyctiphanes norvegica) were placed in a reactor under stirring, and added concentrated hydrochloric acid. Acid hydrolysis for 24 hours in enameled apparatus with a stirrer at a temperature of 100-105°C. after hydrolysis of the resulting solution under vacuum pumped into the catalytic Converter, cooled using a water circuit and neutralized with dry alkali to the value of pH in the range 4-6 with constant cooling. The neutralized hydrolysate was survived for 20 days at an ambient temperature of 20°C. Then made the filter on the vacuum unit for separating the resulting precipitate from the receipt does not contain solids solution of the hydrolyzate. To the resulting solution was hydrolyzed under stirring was added the required amount of distilled water ("water for injection") to produce a solution with a concentration of hydrolysate 5% by weight. Before filling the containers with composities end use, conducted thermal sterilization of the composition and, if necessary, subsequent filtration of the solution. Capacity with the finished composition was stored at a temperature of 4-6°C, out of direct sunlight. The results of chemical analysis of the obtained compositions are presented below.

Mass fraction of amino nitrogen - 0,10%

Mass fraction of dry substances - 0.8 wt.%

pH is 5.5

No.Amino acidThe amino acid content, wt.%Fatty acid composition of lipidsTrace elements
1taurine0,020myristic acid C14:0Potassium
2aspartic acid0,042palmitic acid C16:0Calcium
3glutamic acid0,045palmitoleic acid C16:1Magnesium
4Glycine 0,030heptadiene acid C17:2Iron
5alanine0,017steinova acid C18:0Zinc
6cystine0,003oleic acid C18:1Copper
7leucine0,031linolenic acid C18:3Chrome
8tyrosine0,0063Aksenova acid C20:1Iodine
9phenylalanine0,041arachidonic acid C20:4
10gamma-aminobutyric acid0,002eicosapentaenoic acid C20:5
11Ethanolamine0,003 docosanoate acid C22:2
12lysine0,018docosahexaenoic acid C22:6
13arginine0,023treasuretrove acid C23:4
14Proline0,114triazapentadiene acid C23:5
15serine0,018tetracosanoic acid C24:1
16histidine0,008docosanoate acid C22:2
17threonine0,015
18valine0,012
19 methionine0,006
20isoleucine0,014

Toxicological studies of the composition on the basis of bivalves

The task of the first stage of the research of the composition for parenteral administration containing 5% aqueous solution of the hydrolyzate obtained in Example 1 (hereinafter the composition (A) were:

1. Holding toxic and biological research: determination of acute and subacute toxicity (including irritant and allergenic activity).

2. The definition of embryo tropic actions (embryotoxic and teratogenic).

3. Conducting experiments to identify possible sub-chronic toxicity in different species of animals.

Materials and methods research

The experiments were conducted on 115 white rats and 55 white mice using existing pharmacological, Toxicological, hematological, biochemical and immunological methods. Separate the methods and schemes of experiments on determination of toxicity to laboratory animals and other animal species presented during the presentation of experimental research.

The experiments were conducted twice with statistical clicks the processing of the obtained results.

The definition of acute and subacute toxicity (including irritant and allergenic activity)

Acute toxicity of the composition As determined by single intramuscular injection of therapeutic doses of 0.2 ml/head and doses increased in 2 and 4 times, that is, to 0.4; 0.8 ml/head, which was 1600 and 3200 mg/kg No visible deviations in the behavior of animals was not observed in rats willingly ate food and drank water. On Toxicological classification LD50more than 1000 mg/kg of the drug belongs to toxic substances. For the studied compositions of this figure is several times higher than the dose of 1000 mg/kg

When defining sub-chronic toxicity of the drug at a dose of 0.2 ml/head (or 800 mg/kg) was administered to rats for 10 consecutive days. As in the acute experience was not observed in the negative influence of composition on the animal organism.

Irritant effects were determined using the method episotomy applications. To determine a possible irritant, allergic drug action created two groups of white rats, weighing 250-270 g In each group of five animals. On shaved areas of the back (1,5×2 cm), hips (1×0.5 cm) and on the conjunctiva of the eyes of animals was applied for 20 days: the first group - composition for parenteral administration, the second group was 0.9%solution of sodium chloride.

For the experiment, which was cityval following indicators: appearance of hyperemia of the conjunctiva, the presence of edema of the skin and eyelids, inflammation, skin reactions (redness, rash) when using drugs. Observing the experimental laboratory animals, there were no allergic and local irritation of the composition. Skin reactions such as redness, rashes in the area shaved areas of the hips and back were observed.

The results of the action of the composition for parenteral administration, and isotonic sodium chloride solution on the mucous membrane of the eyes and shaved areas of the thighs and back of white rats are presented in Table 1.

Table 1
Indicators1st group2nd group
Skin rednessnono
Swelling of the skinnono
Condition ageunchangedunchanged
Conjunctivitisnono
Keratitis nono
Lacrimationnono
Painful reactionnono

From the presented data shows that the composition And has no local irritant, allergic action. The mucous membrane of the eye white rats, the state of age remained satisfactory, signs of inflammation, edema and hyperemia, lacrimation were absent, and there was no painful reaction after application of the solutions. The General condition of the white rats were satisfactory, they were mobile, willingly drank water and ate food.

As an additional indicator of irritants determined the permeability of the capillaries of the skin, using a sample Mack Clara-Aldrich. The sample was consisted of intradermal administration of experimental animals on day 15 of the experience of 0.2 ml of physiological solution in the area of application of the test drug and symmetrically located control plot. Fixed time resorption of salt blister on both sites. Studies have shown that drugs do not affect the permeability of the capillaries of the skin.

The definition of embryo tropic action of the composition (embriotoksicescoe and teratogenic)

Evaluation of embryotoxic and teratogenic action of the composition And were conducted according to the guidelines published Achtergekomen (1988).

Experiments to determine the possible embryotoxic action of the drug was carried out on 15 pregnant female rats weighing 150-180 g and 3 males first and second generation.

To the females, in the stage of estrus and proestrus, in the evening, was helping males, one male to four females. Detection of sperm in the vaginal smear the following morning considered the first day of pregnancy. The research began with the introduction of therapeutic composition in a dose of 0.2 ml on the head. The drug was administered intramuscularly in the period from 1 on the 17th day of pregnancy. The control animals at the same time injected intramuscularly isotonic solution of sodium chloride in the same dose.

Pregnancy was seen by examining vaginal smears females on the fourth or fifth day after fertilization and pregnancy, 10-11 th days of pregnancy and by weighing females at 1, 7, 14 and 20 days of pregnancy. On the 20th day of pregnancy, females were decapotable, counted the number of yellow bodies in pregnancy the ovaries and the number of seats implantation.

To determine the embryotoxic action of the composition And calculated preimplantation death of zygotes (the difference between the number of yellow bodies of pregnancy in the ovaries and the number of seats implantation in the uterus of the total number of yellow bodies), postimplantation embryos death (the difference between the number of seats implantation and the number of live fetuses in the uterus from the number of implantation) and total embryonic mortality (the difference between the number of yellow bodies of pregnancy and fresh fruits in the percentage of the number of yellow bodies in the ovaries).

With the introduction of the composition And pregnant rats at a dose of 0.2 ml per head within a specified time of embryogenesis and organogenesis (1-17 initial days of pregnancy) violations during pregnancy is not detected, the animals willingly drank water and ate food. No violations were found also in the study of internal organs by Wilson and the skeletal system by Dawson.

Main indicators of preimplantational the death of zygotes in the experimental group was $ 1,90, and in control - 2,3%, postimplantation embryos death - 1,92% and 2.0%, respectively, of total embryonic mortality was 3.3 and 3.46%, respectively, were close in performance, indicating the absence of embryotoxic action of the composition. Weight, dimensions, number of fruits and protoplanetny factor in the experimental and control groups had no statistical differences were within the limits of fluctuations of physiological norm. Thus, the average number of fetuses per female with the drug was a 8.9±0,06 and in control - 8,7±0,06, respectively, the mass - 2093,3±8,3 and 2010,2±5.1 mg and sizes of 2.7±,9 and 2.7±0,1 see

Visual inspection and microscopic examination of internal organs by Wilson from rats that were injected drug was not detected malformed fetuses (external and internal abnormalities were absent).

Microscopic examination of the bones of the skeleton of the fruit on Dawson found that the investigated composition during the entire period of embryogenesis did not cause abnormalities in the bone system of the fruit.

No abnormalities in the bone system of the fruit is confirmed by the identical mass of experimental and control animals, as well as protoplanetary coefficient (26,97 and 27,60 respectively).

Found that the composition And dose of 0.2 ml per head daily during the entire pregnancy (from 1 to 17 days) showed no embryotoxic or teratogenic action. Animals received daily administration of the drug and the results of the research material from them is the weight and size of the fruit, the condition of internal organs and skeletal system were identical (in a separate indicators - weight, fruit size and the size of the individual bones even more) compared with those from control animals, which was confirmed by the confidence level that was equal to or greater than 0,05 (P≥0,05).

Thus, the composition And does not cause second generation embryotoxic and teratogenic effects in laboratory animals.

Experiments conducted on cows, calves, pigs, weaned and dogs. The song, And was administered intramuscularly for 10 days in the following doses: cows - 10 ml/head; calves (live weight - 30-33 kg) 5 ml/head; pigs (live weight 9,3 kg) - 2 ml/head; dogs (live weight 35-40 kg) 5 ml/head. Daily observation of the animals was taken into account the overall clinical condition, appetite, possible negative effects in the behavior of animals. In blood taken at the beginning and end of the experience, and determined some hematological and immunobiochemical indicators. In piglets and calves was determined by the initial and final live weight of the body.

Experiments on cows. Found that daily administration cows above composition at a dose of 10 ml/head does not have a negative effect on the external behavior of the animals that habitually ate forage diet and drinking water. Any negative factors in the behavior of animals were found. When the Hematology blood test also found no statistically significant differences in the content of erythrocytes, leukocytes and hemoglobin. Biochemical studies have shown some decrease in serum carotene and total protein, but these changes were unreliable. The content of calcium and phosphorus in the serum were within normal limits, changes in the indicators, the floor is built up and in the end experience is not marked. The positive moment of introduction of the composition was the possibility of earlier (7-10 days) fertilization 6 out of 10 experimental cows.

Experiments on calves. The composition was administered to calves immediately after birth. The negative aspects of their behavior were noticed. On the contrary, from 10 calves get sick with diarrhea only 3, while the average is about 67-75%. In the study of blood found that hematological indices were almost identical, both before and after the introduction of the composition. Of the biochemical parameters were observed trend of increasing total protein, although the difference was statistically insignificant. At the same time the age of one month experimental calves had higher live weight (51,3+2.1 kg, against 50.1+2.7 kg in the control calves) and among these animals is less than the recorded occurrence of bronchopneumonia.

Experiments on pigs. The piglets composition And were injected with for 10-13 days before weaning. Any deviations from the behavior of piglets was observed. Blood also has not been set changes from the initial and final data on the content of formed elements of blood, Sa, and the RV was only a trend of increasing total protein content, which subsequently showed higher live weight of 3.7±0.2%, compared with the control animals.

Experiments on dogs. Experiments conducted on the property is under the middle age group (5-7 years). Found that intramuscular injection of the composition And the dose of 5 ml/head for 10 days had no detrimental effect on behavioural responses of dogs. Animals willingly ate food and drank water. Hematological parameters of blood also did not differ from the beginning and end of the experiment.

The above studies on laboratory animals have confirmed the low toxicity of the composition And, including the absence of embryo tropic actions.

EXAMPLE 3. Immunostimulatory, adaptogenic, growth and wound healing action of the composition And

Materials and methods

The experiment was performed in 84 white rats and 36 white mice using existing drugs, hematological, biochemical and immunological methods. When studying the positive effects of the composition And on the organism of laboratory animals was taken into account some hematological and immunological indices in experimental and control animals. Hematological parameters were determined using existing methods in blood samples: the number of cells in 1 mm3counting in the camera Goryaeva. From immuno-biological indicators were considered: the blood levels of protein, glucose, phagocytosis and a number of other indicators that characterize the state of the organism. The amount of total protein in serum was determined by refraction IU the Odom. In the serum the amount of refraction depends mainly on the number of proteins.

The concentration of glucose was determined ortho-toluidine method, based on the staining compounds glucose ortho-toluidine in acetic acid solution, the intensity of which is proportional to the concentration of glucose.

The content of immunoglobulin G in the serum was determined according to the method of Mancini (1965) modified Lscolors et al. (1975). The method consists in the formation of a ring of precipitation as a result of interaction of a mixture of agar with anticorodal to the immunoglobulin and antigen test serum made in wells agar plates. The area of the precipitate, as the square of the diameter of the ring is directly proportional to the antibody concentration in the agar. However, the concentration of the test antigen and the area of the precipitate has a linear dependence.

The study of phagocytic activity based on the determination of the in vitro ability of neutrophils in the peripheral blood of the studied animals (production opsona-phagocytic reaction - ODF) to faviroute (absorb) the microbial cells. As the test culture for ODF is used Staphylococcus - Staph. albus.

The intensity of phagocytosis was determined by phagocytic activity (FA), phagocytic number (FC) and phagocytic index (PI).

Phagocytic activity is here (FA) - this percentage faguoqitirute of neutrophils to the total number counted. Phagocytic index (PI) is the number of absorbed microbial cells (M.K), in terms of a single neutrophil, of the total number of counted neutrophils. Phagocytic number (FC) - the number of microbial cells in terms of one active (faguoqitirute) neutrophil.

The experiments were conducted twice with the statistical processing of the results.

3.1. Definition immunostimulating action

Experimental white rats (three groups of 6 animals each) for 5 days were injected intramuscularly composition And dose of 0.2 ml/head. In the blood was determined by phagocytic index (PI), phagocytic activity (FA) and phagocytic number (FC), and immunological parameters immunoglobulins a and G (Table 2, 3). Paracetomol activity of the composition was compared with a known immunomodulator - eventnum. Part of Minton includes: Thuja D6, Vincetoxicum D4, Echinacea purpurea D4 and isotonic sodium chloride solution.

Table 2
Groups of animalsPhagocytic indexPhagocytic activityPhagocytic number
Composition And8,2±1,42 50,0±1,2419,6±1,47
Minton10,0±1,3460,0±1,4515,33±1,29
Control6,72±1,8733,33±1,398,0±1,64

Table 3
Groups of animalsImmunoglobulin aImmunoglobulin G
Composition And2,99±0,59of 8.06±1,57
Minton2,9±0,627,42±1,32
Control1,85±0,376,85±0,32

The results showed that the use of a composition contributed to the increase in phagocytic activity of neutrophils compared to the control group was slightly higher production quantities of immunoglobulin G compared with the action of the drug Minton and a control group of white rats.

Therefore, the composition And has a specific immunostimulating action on the organism of experimental animals and increases to mocny and humoral immunity indices.

3.2. Definition adaptogenic (anti-stress) properties

The experiment was carried out in 12 rats, 4 heads in each group. Stress in Wistar albino rats were generated using shuttel device that simulates transport (mechanical) stress. On shuttel apparatus was placed a box with the animals and kept them for 30-40 minutes while the machine is on. In the blood of white rats was determined the most informative indicators of the occurrence and during the stress response (stress mediators): glucose, total protein, and leukocyte numbers.

Pre for 3 days before stressaway animals of the 1st group was injected intramuscularly composition And dose of 0.2 ml of Animals of the 2nd group were injected with 2.5%solution of chlorpromazine, belonging to the group of neuroleptics, at a dose of 0.5 mg per head. Animals of the 3rd group (control) was administered intramuscularly isotonic solution of sodium chloride. The study of blood in rats to determine the above parameters was carried out after 1 and 24 h after stressaway.

According to laboratory tests, it was found that under stress glucose, leukocytes increases and decreases the total protein content.

For example, within 1 h after stressaway the concentration of glucose in a group with composition a was equal to 4.97±0.1 mmol/l, against 6,46±0.45 mmol/l in the group with chlorpromazine and 10.0±0,7 Mall in the control group. A day after the stress this had increased to 5,42±0.2 mmol/l, 6,25±0.3 mmol/l and 8,33±0.4 mmol/l, respectively. Results of laboratory analyses of blood, after 1 h and 24 h after stress are shown in tables 4 and 4' respectively.

Table 4
The blood 1 hour after stress
GroupGlucose mmol/lTotal protein, g/lLeukocytes, 103/µl
Composition Andequal to 4.97±0,167,5±1,25,6±1,7
Chlorpromazine6,46±0,4566,0±0,54,3±0,4
Isotonic NaCl10,0±0,766,0±0,95,8±0,52
Table 4'
The blood counts of white cover the 24 hours after stress
GroupGlucose, mmol/lTotal protein, g/lLeukocytes, 103/µl
Composition And5,42±0,272,7±2,35,6±0,1
Chlorpromazine6,25±0,372,4±1,66,1±0,3
Isotonic NaCl8,33±0,466,2±2,57,05±0,2

3.3. The study of the growth-promoting actions

The influence of composition And on the growth and development of laboratory animals was studied in 18 white mice, weighing 19-23, Created 3 groups of animals each with 6 goals. As the comparison drug used complex homeopathic medication for animals "Minton". All animals were weighed before and at the end of the experiment. At the end of the experiment was taken blood for clinical and immunobiochemical research. During the experiment, which lasted 15 days, the animals showed no signs of bespoke Ista, willingly ate food and drank water. The highest mobility was noted in animals of the experimental group, which was subcutaneously injected specified composition And dose of 0.1 ml per head. Animal comparison group was subcutaneously injected complex homeopathic medication for animals "Minton", in a dose of 0.1 ml per head. For the control group was used isotonic solution of sodium chloride, which was administered subcutaneously at a dose of 0.1 ml per head. The rate of injection was 3 days.

The best indicators of weight gain compared to control animals were obtained using the composition A. the Data presented in Table 5.

Table 5
The growth-promoting action of drugs
Groups of animalsWeight before experiment (g)Weight at the end of the experiment (g)Weight gain for 10 days. (g)Weight gain relative to control (%)
Composition And25,75±0,69232,0±0,756,25±0,62113,6
Minton24,75±0,56 30,75±0,566,0±0,56105,1
Control21,5±1,1927,0±1,45,5±1,34100,0

3.4. Study of wound healing action of the composition

Wound healing is three main phases: inflammation, regeneration and reorganization of the scar and epithelization. In any phase of healing is possible protracted course of the wound process with sluggish growth of granulations and delayed epithelialization.

Experiments on determination of wound healing action of the composition And was carried out on 6 rats, weighing 250 g, in each group of 4 animals. Previously shaved areas of the back 2×2 cm, nepilirovanny skin is cleaned, then surgical forceps took the skin and scalpel made an incision of 1 cm

On the course of healing of experimental wounds were judged on the following parameters:

1. Visual observation

the time of appearance of granulation in the wound

- closing the bottom of the wound granulation

- filling the granulation of the wound cavity

- the quality of the granulation

move epithelialization

- the state of the tissue around the wound.

2. Registration of changes in the area of wound

3. For the term of the final wound healing took time lapse of the scab

The experimental group during the former is eriment introduced above composition And intramuscularly at therapeutic dose of 0.2 ml per head, within 4 days. The control animals at the same time injected intramuscularly isotonic solution of sodium chloride in the same dose.

Granulation in the wound appeared on the 2nd day in both groups, closing the bottom of the wound and carry out granulation of the wound cavity in the experimental group, and control, noted on 5 days from the beginning of the experiment. During the experiment in the experimental group was a marked reduction in the area of the wound on day 4 - 0.3 cm in the control group the reduction in the area of the wounds in this period is not marked.

The deadline for final wound healing was in the experimental group was 6 days, and in control - 8.

The above research has established a number of positive pharmacological effects of the composition And such as immunostimulatory, adaptogenic, growth and wound healing effect. On the basis of the conducted research and obtained results were elaborated recommendations on the application of the composition And to increase the productivity and the natural resistance of the organism of animals.

The data obtained from clinical studies on laboratory animals have formed the basis for testing of farm and domestic animals, the results of which are given below.

EXAMPLE 4. The influence of composition on the performance of leukogram and bone metabolism, in the blood p is reset, located in the settlements

The study was performed within 2 weeks the piglets at the age of 45 and 65 days (n=15)contained in the conditions of the farm. We have formed 2 groups of animals: experimental group of animals, which is 1 time per day was administered intramuscularly composition And dose of 2 ml per head every other day for 14 days, and the control animals without specific treatment. In the group of animals were selected by the method of analogues.

Research results: the effect of composition on the metabolism of electrolytes and metabolism of bone tissue in the blood of piglets in the blood of pigs are presented in Table 6.

Table 6
№ p/ pIndexEd. MEAs.The control groupExperienced group
BeforeAfterToAfter
experimentexperiment
1calciummmol/l2,32±0,3 2,4±0,52,52±0,52,4±0,6
2phosphorusmmol/l1,83±0,31,78±0,51,76±0,31,84±0,27
3alkaline phosphataseIU/lof 87.8±7,886,2±5,560,9±2,739,5±1,2

From the research results we can conclude the following: in the control group of piglets in an imbalance in the ratio between calcium and phosphorus, as well as increased levels of alkaline phosphatase, which may indicate that (despite age-related changes) on inflammatory processes in the intestinal mucosa, juvenile osteodystrophy, like status.

After two weeks of using the drug in piglets of the experimental group in contrast to the control was noted positive dynamics: normal appetite, the animals were more active.

Dynamics of biochemical indices following: the level of alkaline phosphatase decreased by 35.1%, the level of calcium and phosphorus approached physiological but the moms in contrast to control animals group, at which the data changes of biochemical parameters was not observed.

Thus, the use of the above composition And helps to normalize calcipotriol exchange that allows us to recommend its use in treatment of osteo-articular pathologies.

Research results: the effect of composition on the performance of leukogram in the blood of piglets at weaning are shown in Table 7.

Table 7
IndexUnit.NormaThe control groupExperienced group
Before the start of the experimentAfter the experimentBefore the start of the experimentAfter the experiment
Leukocytesthousand/ál; 109/l8,7-or 37.910,9±1,215,1±2,111,05±1,616,1±2,5
Neutrophils Yu%0-2 0,15±0,020,17±0,010,14±0,020,17±0,12
Neutrophils P%2-41,9±0,041,01±0,0111,85±0,33,5±0,7
Neutrophils%40-4865,45±12,159,21±9,864,3±14,455,2±8,9
Eosinophils%1-30,8±0,010,9±0,0120,85±0,031,6±0,04
Basophils%0-10,7±0,0130,78±0,010,73±0,131,3±0,3
Monocytes%2-63,5±0,244,2±0,93,6±0,233,1±0,07
Lymphocytes %40-5027,5±3,132,73±3,628,53±4,737,13±4,9

The data obtained show that in piglets marked decrease in the level of lymphocytes, basophils, eosinophils, however, the level of segmented neutrophils increased, these changes may indicate a protective and adaptive reactions of the organism to stress. After two weeks of using the drug in piglets of the experimental group, in contrast to the control, was noted positive dynamics: normal appetite, animals became more active. Hematological parameters of blood started to come closer to physiological values in the experimental group, in contrast to the control. Thus, the composition And can be recommended for adjuvant therapy in the correction paleochannel stress in farm animals.

EXAMPLE 5. The study of the detoxification properties of the composition

We observed 17 animals, in which the diagnosis of acute piroplasmosis was confirmed by the presence of parasites Babesia in smears of peripheral blood. The status of all animals was regarded as serious: rectal temperature of 41.0°C, mucous membranes pale with yellowish tint, dark urine with blood, reducing appet is that including refusal of feed, gait disturbance, weakness, in a few cases, loss of consciousness.

As a specific therapy medication used Pyro-Stop (intramuscularly twice with an interval of 24 hours). In addition to the commonly used treatment (solution of ringer-Locke intravenous, 60 drops/min, 10-15 ml/kg 2 times a day, cardiac drugs) used the composition And 5-15 ml (depending on weight) intravenously (jet) 1 time per day. In especially severe cases, the drug was administered in the first 2-3 days intravenously 2 times a day, then moved on intramuscular injection of 1 time per day.

It was noted that the use of the above composition substantially increased the effectiveness of the treatment in comparison with traditional therapy. So, the General condition of all the sick animals were significantly improved after 3 to 5 hours after the first injection of the drug. Animals recovered rapidly appetite, decreased temperature, decreased heart rate. All dogs hemoglobin came to normal by the end of 2 weeks from the start of treatment.

Detoxification action of the above-mentioned composition due to its hepatoprotective properties, as evidenced by the results of the dynamic analysis of liver from infected animals. Even after a single intravenous injection of the composition mentioned normal is tion of ALT, ACT, albumin, GGT, globulin, total protein, prothrombin, and alkaline phosphatase, positive dynamics of the level of total bilirubin and indirect fractions of bilirubin, amylase.

Thus, the proposed in the present description, the composition has a strong detoxifying effect, in particular, due to the presence of hepatoprotective properties.

EXAMPLE 6. The influence of the composition And the musculoskeletal system in dogs

Composition And used as mono - and combination therapy of dogs with age-related changes in the musculoskeletal system. The group consisted of 5 dogs with normal and overweight. The main symptoms were apathy animals, lack of appetite, reluctance to walk, limping, different weights, different pain syndromes. Upon examination of the animals revealed stiffness in the joints, pain response to palpation in the region of the spine (thoracic and lumbar sections). On radiographs revealed varying degrees of osteoarthritis TBS, osteochondrosis.

Three Russian Borzoi at the age of 13 years old, weighing about 25 kg (norm) was administered intramuscularly in a dose of 5 ml of a composition containing 5% aqueous solution of hydrolysate derived from the meat of the bivalve mollusks, in the amount of 15 injection.

Registerer at the age of 12 years (female)suffering from overweight with painful the spine of the spine, the above composition was assigned to 4 ml intramuscularly every other day in the amount of 10 injections. English bulldog at the age of 10 years, suffering from obesity and lack of activity, the above composition was assigned as part of an integrated treatment: intramuscular injection of 4 ml a day in the amount of 10 injections, banjaran intravenous 2 ml of 1 time per week 4 injections, Kiesel 10 ml of 1 times a day for 2 months. In dogs before and after the drugs were taken clinical and biochemical blood tests. After treatment the condition of the animals was markedly different: all dogs recovered appetite, increased motor activity, decreased levels of alkaline phosphatase from elevated to normal, decreased activity of alanine transaminase (ALT), increased levels of urea within the norms that collectively speaks a positive effect of the composition And condition of the musculoskeletal system and liver function.

EXAMPLE 7. The treatment of pathological conditions in dogs

Below are the results of applying the composition in the treatment of various pathologies in dogs.

1. English bulldog

Age: 1 year 3 months.

Weight: 30 kg

The diagnosis of Demodicosis, urine acid diathesis, streptoderma: smitten the whole head, cheeks, forehead, between and behind the ears, all in purulent cork, get wet in the Aya eczema.

Appointed: General symptomatic treatment and Composition 3 ml intramuscularly once a day for 5 days.

The result in a week: the skin is dry, overgrown; swelling and itching no, no pus.

Clean, overgrown skin.

Mobiman

Age: 2 years 6 months.

Weight: 23 kg

Diagnosis: Severe thinness, despite the good food, the lack of worms and parasites. Dandruff on the skin bilirubin in the urine.

Appointed: Composition 5 ml/m once a day for 5 days.

The result, after a month: the increase of mass 2 kg, bilirubin normal, the hair is clean, shiny.

3. American bulldog

Age: 5 years

Weight: 60 kg

The Diagnosis Of Leptospirosis. Symptoms: purulent otitis, bloody diarrhea, vomiting.

Appointed: Dropper and symptomatic treatment, as well as Composition 5 ml/m once a day for 5 days.

The result: within 2 days stopped vomiting and diarrhea, one week after the start of treatment, clinical blood and urine in norm. After 4 months of recurrence was not observed.

4. Dalmatian

Age: 1 year 7 months old.

Weight: 22 kg

Diagnosis: Trichophytosis, complicated allergic dermatitis. Affected the entire back and sides.

Appointed: symptomatic treatment.

Treatment within one month no results.

Appointed: Composition 3 ml/m, 5 injections.

The result after two weeks: all symptoms disappeared, hair net

5. Yorkshire Terrier

Age: 1.5 months.

Weight:1 kg

Diagnosis: a Comprehensive re-inoculation was done a week ahead of schedule - after 4 days the temperature of 41.6°, bronchopneumonia, unilateral purulent expiration of the eyes and nose.

Appointed: symptomatic treatment and Composition 5:0 at 1 ml/m once a day for 5 days.

The result after two days: normalized temperature, absence of purulent effusions and rales in the lungs.

6. German shorthaired pointer

Age: 12 years old

Weight: 17 kg

Diagnosis: Chronic leptospirosis. Temperature of about 40° in two months, pain in the liver and kidneys, the depletion of the dye, the smell of skin, barely walking, lack of appetite. Traditional methods of treatment do not help.

Appointed: the first week - with large doses of antibiotics and symptomatic treatment. Clinical blood returned to normal, but the symptoms remained.

Appointed: Repeal of antibiotics, symptomatic treatment of liver and kidney and Composition 3 ml/m once a day for 5 days.

The result after two days: the temperature is normal for 3 weeks improved the condition of the liver and kidneys, disappeared dandruff, improved coat, appetite.

After two months, the relapse: infectious arthritis - two injections of the composition of 3 ml/m once a day.

All symptoms disappeared.

1. Immunomodulatory composition d is I injected the mammal, contains:
from 1 to 10 wt.% the hydrolysate obtained by acid and/or enzymatic hydrolysis of one or more biological resources selected from the group including bivalve molluscs, annelids, leeches; and
water - the rest.

2. Immunomodulatory composition according to claim 1, characterized in that the water is water for injection.

3. Immunomodulatory composition according to any one of claim 1 or 2, additionally containing glucose.

4. The composition according to claim 1, characterized in that the mammal is an animal or human.

5. The composition according to claim 1, characterized in that the biological resources selected from the group comprising of mussels (Anodonta), the thick-shelled river mussels (Unio), oysters (Ostreidae), mussels (Mytilidae), tridacna (Tridacna), oyster (Pinctada), scallops (Pectinidae), ship worms (Teredinidae), guidec (Panopea abrupta), Icelandic cyprine (Arctica Islandica), krill (Meganyctiphanes norvegica), North Atlantic lobster (Homarus americanus).

6. The composition according to claim 1, where the specified hydrolysate is characterized in such a way that:
mass fraction of amine nitrogen in the composition is from 0.01 to 6.5 wt.%, and/or
the pH of the composition is from 4 to 7, and/or
mass fraction of solids in the composition is from 0.5 to 2 wt.%

7. The composition according to claim 1, where the specified hydrolysate contains one or more amino acids selected from the group comprising Toure is, asparaginase acid, threonine, serine, glutamic acid, sarcosine, glycine, alanine, valine, cystine, methionine, cystathionine, isoleucine, leucine, tyrosine, phenylalanine, β-alanine, γ-aminoadamantane acid, γ-aminomethane acid, ornithine, lysine, histidine, carnosine, arginine, hydroxyproline, Proline.

8. The composition according to claim 7, containing the following amino acid in a mass ratio relative to the total number of amino acids in the specified composition:
taurine from 1 to 10%
aspartic acid is from 1 to 30%
glutamic acid is from 1 to 30%
glycine from 1 to 20%
alanine from 1 to 10%
leucine, from 1%to 15%
phenylalanine from 1 to 15%
lysine from 1 to 10%
arginine, from 1 to 20%
Proline, from 1 to 50%
serine from 1 to 10%
histidine from 1 to 10%
threonine is from 0.5 to 10%
valine from 0.1 to 10%
methionine from 0.1 to 10%
isoleucine from 1 to 10%.

9. The composition according to claim 1, where the specified hydrolysate contains one or more fatty acids selected from the group comprising tridesilon acid C13:0, myristic acid C14:0, tetradecanoyl acid C14:1, pentadecanoic acid C15:0, palmitic acid C16:0, palmitoleic acid C16:1, hexadecanoyl acid C16:2, hexadecanediol acid C16:5, heptadecanoic acid C17:0, heptadecenoic acid C17:1, heptadiene acid C17:2, stearinovuyu acid C18:0, oleic acid C18:1 linoleic acid C18:2, linoleic to the slot C18:3, octadecatetraenoic acid C18:4, Aksenova acid C20:1, eicosadienoic acid C20:2, eicosatrienoic acid C20:3, arachidonic acid C20:4, eykozapentaenovuyu acid C20:5, heptacosanoic acid C21:5, docosanol acid C20:0, dokozagexaenovu acid C22:1, docosanol acid C22:2, docosatetraenoic acid C22:4, docosahexaenoic acid C22:6, triazinetrione acid, C23:4, triazapentadiene acid, C23:5, tetracosanoic acid C24:1.

10. The method of obtaining immunomodulatory compositions for injecting the mammal according to any one of claims 1 to 9, including:
(a) enzymatic hydrolysis and/or acid hydrolysis of one or more biological resources selected from the group including bivalve molluscs, earthworms, leeches, with receipt of the first solution;
(b) filtering the first solution to obtain a second solution;
(c) mixing the second solution with water to form a composition with a mass content of the second solution in water from 1 to 10 wt.%.

11. The method according to claim 10, further comprising the first stage of neutralization of the solution with the base.

12. The method according to claim 10, further comprising sterilizing the resulting composition.

13. The method according to claim 10, further comprising settling the first solution for at least 10 days before stage (b).

14. A method of treating Pat the logical States in need thereof of a mammal, includes injecting the specified mammal a composition according to any one of claims 1 to 9, and
specified pathological condition selected from the group including
liver damage,
steatosis,
the intoxication of the organism,
skin disease,
disease of the hair and/or scalp,
dermatitis,
violation of the integrity of the skin due to injury or burn,
obstructive lung disease,
pneumonia
acute respiratory viral infection,
circovirus infection
acidosis,
stress,
inflammation of the mucous membrane of the intestine,
the fibrosa,
like the state,
arthrosis,
osteochondrosis,
atherosclerosis,
and
the specified composition is administered in an amount of from 0.05 ml/kg 10 ml/kg of mass of a specified mammal from 1 to 5 times a day for 1 to 50 hours.

15. The method according to 14, characterized in that the specified composition is administered once in the amount of from 0.05 ml/kg 10 ml/kg of mass of a specified mammal every 1-5 days in the amount of from 0.05 to 10 ml/kg over 1 to 50 days.

16. The method according to 14, characterized in that the injection is an introduction, selected from the group including subcutaneous injection, intradermal injection, intramuscular injection is whether intravenous injection.

17. The method according to 14, further comprising introducing one or more traditionally used remedies for the treatment or prevention of a specified pathological conditions.

18. The use of a composition according to any one of claims 1 to 9 for the normalization of metabolism in need of this mammal.

19. Use p, characterized in that the normalization of metabolism includes normalization calcipotriol metabolism in need of this mammal.



 

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16 cl, 4 tbl

FIELD: chemistry.

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22 cl, 5 dwg, 5 tbl, 7 ex

FIELD: medicine.

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2 cl, 5 ex

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