Lipid raft modulation

FIELD: medicine.

SUBSTANCE: group of inventions refers to versions of applying an antisecretory protein which corresponds the amino acid sequence SEQ ID NO:6, its homologue, and/or a fragment containing the amino acid sequence SEQ ID NO:4 showing equivalent activity, and/or its pharmaceutically active salt for preparing a pharmaceutical composition and/or nutritional care for treatment and/or prevention a dysfunction, e.g. a pathological function, lipid raft, receptor and/or small pit hypo- or hyperfunction. The lipid raft, receptor and/or small pit dysfunction can be induced by or cause a number of the other conditions, such as vascular and pulmonary dysfunctions and/or endocrine disorders, e.g. diabetes and related disorders.

EFFECT: group of inventions enables controlling intracellular transport and cell product release, as well as normalising tissue component distribution in various diseases.

13 cl, 12 ex

 

The technical field to which the invention relates

The present invention relates to the use of antisecretory proteins, their derivatives, homologues and/or fragments with equivalent functional activity, and/or their pharmaceutically active salts, to pharmaceutical compositions for the treatment and/or prevention of structural disorganization and dysfunction of lipid rafts and/or kahweol in cell membranes, and binding proteins, receptors, neurotransmitters, ion channels, water channels, cytoskeleton and systems G-proteins associated with lipid rafts and caveolae, including the uptake and release of connections. The pharmaceutical composition of the present invention is used to control and/or improve the structure, distribution and numerous functions of lipid rafts, receptors and/or kahweol in the membranes. Examples of such positive effects can be neutralized pathological features, such as Hypo - or hyperthyroidism, repair, and/or normalization of lipid rafts, receptors and/or kahweol structurally and functionally improved survival and/or recovery for diseases, injuries, recovery processes, and other dysfunctions. In addition, the present invention relates to the use of specified pharmaceutical composition for contact is aerovane intracellular transport and release of cellular products and also to normalize the distribution of tissue components.

The prior art of the present invention

Lipid rafts are dynamic, heterogeneous microdomain forming a specific area in the cell membranes rich in cholesterol and glycolipids, especially sphingolipids, such as GM1 (Ross & Pawlina, 2006; Pollard &Earnshaw, 2002). One of the options for their detection in vitro is getting insoluble residue after extraction with the detergent Triton X-100, used in diluted form at low temperature. This membrane fraction that is resistant to the action of detergent, was rebuilt as a strip of low density allocated using gradient flotation. The prevalence of lipid rafts is reduced by reducing the number or disruption of cholesterol in cell membranes. Lipid rafts are closely related and connected to the cytoskeleton and are involved in cell polarization. They form a specific microdomain prevailing in the membranes of cells and tissues at any age, for example, in embryos, fetuses, and young, adults and elderly individuals.

Lipid rafts are rich in proteins, essential for cohesion and signal transmission, such as receptors, binding proteins of cells, ion carriers and complexes of ion channels, including, for example, aquap is Rina, as well as receptors chemokines, receptors, neurotransmitters, receptors, hormones and receptors for growth factors. These proteins and protein complexes interact with intracellular systems G-proteins, which transmit the signal received, for example, receptors in the cytoplasm and the cell nucleus (Dermine et al., 2001; Ross & Pawlina, 2006; Pollard &Earnshaw, 2002; Helms & Zurzolo, 2004; Chini & Parenti, 2004; Head et al., 2006; Mahmutefendic et al., 2007). It was shown that the distribution and concentration are key to controlling the activity of ion cells, Ca2+closely associated with lipid rafts and caveolae. Additional proteins, such as connexin, CD38, CD19, Thy-1 and CD59, saldarini in lipid rafts, usually through proteins and receptors associated with glycosylphosphatidylinositol (GPI), which allow them to interact with cellular functions. Sphingolipids, an example of which is ganglioside GM1, are the target of cholera enterotoxin and is widely represented in lipid rafts, as their characteristic. In addition, neurotransmitter receptors and other components of synapses and neural processes, as well as the receptors of growth factors belong to a large number of proteins that are prevalent in lipid rafts, for example, in such a highly specialized cells like neurons. Calcium channels and carriers play an important role in the regulation of cell function and interactions largely present in lipid rafts (A Spät, 2007). As an example we can cite the fact that the destruction of lipid rafts is difficult or even eliminates the ability of cells to skip the moving waves of Ca2+in cells. Moreover, the bursts of calcium ions significantly affect key functions of normal cells and abnormal cells, for example, cell division, cell survival and cell death.

In addition, an assumption was made that lipid rafts play a key role in intracellular transfer protein activity of receptors and lipids. Signals and the impact of all these receiving and transmitting proteins are transferred into the cells via G-coupled proteins, enzyme systems or part of the cytoskeleton (Triantafilou & Triantafilou, 2004). For example, it is known that the physical state of lipid rafts promotes localization and limited mobility of many proteins, which facilitates functional education relevant signaling complexes.

Moreover, it is assumed that lipid rafts have the ability to regulate the activation, signaling and alterations in the cytoskeleton, which makes them extremely important for the mechanisms that control cell localiza, including directional migration, as well as to under what the actual content of the shape and size of cells and the associated transport. The cytoskeleton, in addition, is important for intercellular transfer of cell components and for the perception of dynamic and static load on the cell.

Lipid rafts, thus, are dynamic structures, typically with a diameter of about 5-50 nm, with a considerable range of variation. There are many ways to determine lipid rafts, for example, immunohistochemical or immunochemical detection of GM1, which has a very high affinity to cholerea enterotoxin. Another way of detecting and identifying their location is the selection of cell membranes after cell disruption and subsequent allocation resistant to detergent fraction at a certain temperature, as described in the available manuals for cell biology. For immunohistochemical identification of lipid rafts can be used flotillin, caveolin is the essential component and reggie. Of these proteins flotillin can further contact with the lipid droplets. Technology visualization of lipid rafts complementary atomic force microscopy and similar methods. The effect on the cells of cyclodextrin and its variants, causing the removal of cholesterol from membranes, represents an alternative way to detect the existence of lipid rafts.

Caveola form a special type of lipid RA is tov, because they are dynamic structures, characterized by a focal increase in cholesterol and sphingolipids in the membrane, and the transmission of signals from the environment into the inner part of the cells, as well as a connection to the cytoskeleton. Unlike other lipid rafts, size kahweol more, and, as a rule, they have the form of dimples or spacelane in the form of a bulb in cell membranes and appearance of bubbles (Kurzchalia & Parton, 1999). Their size is usually about 0.1 μm, but varies considerably. Caveola common, for example, in cardiac and smooth muscle cells, endothelial cells, macrophages and adipocytes, i.e. in almost all mammalian cells, although with significant changes in concentration.

It has been described that lipid rafts and caveola contain a system of generation of NO (nitric oxide) and act on it. In addition to the transmission of signals inside the cell and outside caveola participate in the transfer liquid and chemicals into and out of cells, endocytosis and regulation, transport, outflow and preservation of fatty acids and cholesterol in the cells and their environment (Pohl et al., 2004; Rajendran et al., 2007). Components kahweol and lipid rafts, in addition, involved in the processing of the protein precursor of β-amyloid (βAPP) and β-amyloid (Aβ), a protein associated primarily with Alzheimer's disease, but also with other neurodegenerative the illnesses and injuries of the nervous system (Graham & Lantos, 2002).

It is known that tumor cells contain lipid rafts, and Kaveri. It is therefore possible, for example, to weaken the growth and migration of tumor cells through destruction or disintegration in various degrees of data structures (Marquez, D. C. et al., 2006; Freeman et al., 2007).

The importance of lipid rafts and kahweol was further investigated in genetically modified animals. Knocked out mouse deficient in caveolin is the essential component, suffer dilatational cardiomyopathy and pulmonary hypertension (Mathew et al., 2004). Moreover, lipid rafts and caveola associated with transport systems, stimulated by insulin, such as transport of steroid hormones.

It was concluded that lipid rafts and caveola, forming a highly dynamic structure are closely related, but in some aspects differ, mainly for distribution in mammalian cells, and perform many important functions. The methods available to date, allow you to destroy or reduce lipid rafts and caveola, but the means of restoring and/or normalization of the structure, distribution, concentration and/or function of lipid rafts and signal and transferring the weight proteins, components of lipid rafts, is not known.

Antisecretory protein is a protein weight 41 kDa, which, as originally described is ANO, provides protection against diarrhea and inflammation of the intestine (review presents Lange and Lόnnroth, 2001). Antisecretory protein was Siqueiros, and its cDNA cloned. Apparently, a similar activity manifests mainly peptide located between positions 35 and 50 in sequence antisecretory protein. Immunochemical and immunohistochemical studies showed that the antisecretory protein is present and can also be synthesized by most tissues and organs in the body. Were described synthetic peptides containing Antidiarrhoeal sequence (WO 97/08202; WO 05/030246). Before that, it was described that the antisecretory factors normalize the abnormal transport of liquid and/or inflammatory reactions, for example, in the intestine and gorodno plexus of the Central nervous system after the introduction of the cholera enterotoxin (WO 97/08202). Thus, in WO 97/08202 an assumption was made that the use of natural antisecretory factors in nutrition and feeding is effective for the treatment of edema, diarrhea, dehydration and inflammation. In WO 98/21978 described the use of products with enzymatic activity for producing food products, which induce the formation of an antisecretory protein. In WO 00/038535, it also describes the food, enriched antisecretory proteins is as such.

It was also shown that antisecretory protein and its fragments improve the recovery of the nervous tissue and proliferation, apoptosis, differentiation and/or migration of stem cells and precursor cells and cells derived from them, in the treatment of conditions associated with loss and/or increased cell (WO 05/030246).

Antisecretory factors (AF), particularly proteins and peptides, as described in detail in WO 97/08202, effective for the elimination of hypersecretory conditions and diseases of the gut such as diarrhoea. Other examples related to the impact of AF in relation hypersecretory conditions are, for example, inflammatory bowel disease, brain swelling, glaucoma, increased intracranial pressure syndrome Miniera and mastitis. Also considered was the use of AF for the treatment of glaucoma (WO 97/08202).

Recently it was discovered that the structure, prevalence, distribution and function of lipid rafts, which were changed as a result of the impairment of cell function, excessive or abnormal exposure, infection, or toxic compounds, or drugs, can be controlled and even to normalize due to certain specific proteins and related compounds.

Surprisingly, the inventors were able to prove that the protein antisecretory factor (AF) and its derived peptides, n is the sample, AF-16, AF-8, can affect the structure, prevalence, distribution and/or function of lipid rafts, receptors and/or kahweol that were changed due to abnormal cell function, excessive or improper exposure, infection or toxic substances or drugs, cells, tissues and/or organs, and thus, for the first time it became possible to control and/or even to normalize the function of lipid rafts and kahweol or associated.

The essence of the present invention

The present invention relates to the use of pharmaceutical compositions containing the antisecretory protein, its homologue, derivative and/or fragment, having antisecretory and/or equivalent in functionality and/or a similar activity, or pharmaceutically active salt, to obtain a pharmaceutical composition for the treatment and/or prevention of dysfunction of lipid rafts, receptors and/or kahweol in cell membranes, such as inadequate, insufficient, Hypo - or Hyper-function.

The present invention also relates to use of pharmaceutical compositions containing the antisecretory protein, its homologue, derivative and/or fragment with equivalent and/or similar activity, or pharmaceutically active salt, to obtain a pharmaceutical composition d is I the treatment and/or prevention of various conditions, associated with dysfunction of lipid rafts, receptors and/or kahweol, such as any condition selected from the group consisting of vascular dysfunction, cardiovascular dysfunction, pulmonary dysfunction, hyperplasia and/or hypertrophy of cells and tissues, cardiovascular dysfunction, cardiomyopathy and pulmonary hypertension, formation of scar tissue, reactive formation of excessive tissue and diabetes, such as diabetes I and/or type II.

Also the present invention relates to the use of pharmaceutical compositions containing the antisecretory protein, its homologue, derivative and/or fragment that has equivalent activity, or pharmaceutically active salt, to obtain a pharmaceutical composition for the treatment and/or prevention of various conditions associated with dysfunction of lipid rafts and/or kahweol under which specified the pharmaceutical composition will effectively influence, for example, the component transfer across cell barriers, restoration of tissues and organs, reactive formation of excessive tissue and/or regeneration of the epithelial cell surface.

In another aspect, the present invention relates to the use of pharmaceutical compositions containing the antisecretory protein, its homologue, derivative and/or fragment possessing the th equivalent functional activity, or pharmaceutically active salt to obtain a pharmaceutical composition for the treatment and/or prevention of various conditions associated with dysfunction of lipid rafts and/or kahweol, wybranych from the group consisting of Alzheimer's disease, and other neurodegenerative disorders and injuries of the nervous system.

In a preferred embodiment, the specified antisecretory protein consists of a sequence of the following formula:

X1-V-C-X2-X3-K-X4-R-X5

where X1 represents I, amino acids 1-35 of the sequence SEQ ID NO:6, or is absent, X2 represents H, R or K, X3 is a, S or L, X4 is a T or A, X5 is an amino acids 43-46, 43-51, 43-80 or 43-163 sequence SEQ ID NO:6 or missing.

In addition, the present invention relates to a method of treatment and/or prevention of diseases associated with dysfunction of lipid rafts, receptors and/or kahweol, such as described above, and the method includes the administration to a mammal, optionally, a therapeutically effective amount of a pharmaceutical composition containing an antisecretory protein, its homologue, derivative and/or fragment that has equivalent activity, and/or pharmaceutically active salt.

The present invention also relates to various doses and with whom persons introduction, suitable for the planned treatment goals, and also taking into account age, sex, condition of the patient, etc.

In addition, this pharmaceutical composition may, of course, contain two or more antisecretory protein, and, in addition, contain a pharmaceutically acceptable excipient. The pharmaceutical composition of the present invention is intended for intraocular, intranasal, oral administration, topical application, subcutaneous and/or systemic injections and may, for example, be obtained for use as a spray, aerosol or nebulizer or atomizer. Upon receipt of a composition for systemic injections in the blood of the specified composition preferably get in a dosage of from 0.1 μg to 10 mg per dose per kg body weight per day, for example, at a dosage of from 0.1 μg to 1 mg per dose per kg body weight per day, more preferably 1-500 μg per admission per kg of body weight per day, for example, 1-50 μg per admission per kg of body weight per day. This introduction can be done either as a single dose or as multiple daily injections.

Generally, the present invention relates to the use of antisecretory protein, homologue, derivative and/or fragment that has equivalent activity, or its pharmaceutically active salts, to pharmaceutical compositions for the treatment and/isopropylacetate different States, associated with dysfunction of lipid rafts and/or kahweol. In the preferred embodiment, this composition can be used to control and normalize the structure, distribution and functions of kahweol and lipid rafts, i.e. specialized microdomains, for example, to restore them structurally and functionally improve the survival and recovery for diseases, injuries, recovery processes, and other violations. In addition, the present invention allows the control of intracellular transport and release of cellular products, as well as to normalize the distribution of tissue components in various diseases and/or control the formation of reactive cells and tissues, as well as reduce the formation of scar tissue, including pathological connection of tissues and organs. Moreover, the composition can also be used for possible treatment effects of toxic substances and control their long-term effects. In a preferred embodiment, such a condition selected from the group consisting of trauma, intoxication, infection, malformation, degeneration and other disorders or diseases of cells, tissues and organs in the body of a mammal.

Not having a goal to limit the scope of the present invention to a specific theory, the mod is but to assume, the composition of the present invention, containing antisecretory protein, its homologue, derivative and/or fragment that has equivalent activity, and/or pharmaceutically active salt, can be influenced by effective impact on the normalization of the structure, distribution and/or function of kahweol and/or lipid rafts in cell membranes.

A description of the shape

Figure 1 shows the potential difference of the cell membrane in cells with defective GABA receptor. Added histamine or AF-16, and measured the impact of these compounds on the potential of the cell membrane, measured in I/I0.

Definitions and abbreviations

Reduction

KD: blood pressure; CSF: cerebrospinal fluid; CNS: Central nervous system, i.e. brain and spinal cord; DIR: pressure interstitial fluid; LH: pulmonary hypertension; FSB: phosphate-saline buffer; AF: antisecretory factor, AF-16: peptide consisting of amino acids VCHSKTRSNPENNVGL; oktapeptid IVCHSKTR; separated VCHSKTR; Hexapeptide CHSKTR; Pentapeptide HSKTR.

Definitions

Proteins are biological macromolecules composed of amino acid residues joined together by peptide bonds. Proteins in the form of linear polymers of amino acids are also called polypeptides. Usually proteins consist of 50-800 the amino acid is the shaft residues and, therefore, have a molecular weight in the range of from about 6000 to about several hundred thousand daltons or more. Small proteins called peptides or oligopeptides. In the context of the present invention, the terms "protein" and "peptide" may be used interchangeably.

"Pharmaceutical composition" in the context of the present invention refers to a composition comprising a therapeutically active amount of antisecretory protein, optionally in combination with pharmaceutically active excipients, such as a carrier or filler. Specified pharmaceutical composition to get the appropriate way of introduction, which can be different depending on the condition of the patient, as well as other factors, such as age or preferred choice. Pharmaceutical composition containing antisecretory protein, serves as a delivery system of the drug. The pharmaceutical composition after administration ensures the supply of the active substance in the human body or animal. Specified pharmaceutical composition may be in the form of, for example, tablets, pills, lozenges, capsules, rectal pills, gels, solutions, etc. but they are not limited.

The term "pharmaceutically active salt" means a salt antisecretory protein, which may represent Liu is th its salt, on the basis of the so-called sequence of Hofmeister. Other examples of pharmaceutically active salts include triptorelin, acetate and lisinoril, but the present invention is not limited.

The term antisecretory" means in this context inhibiting or reducing secretion, especially putting secrets. Therefore, the term antisecretory protein" refers to a protein capable of inhibiting or reducing secretion in the body.

"Medical food" in the context of the present invention refers to food that was received together with the composition with antisecretory protein. Specified meals can be any suitable food in liquid or solid form, such as liquid or powder, or any other suitable food product. Examples of such products can be found in WO 0038535. The specified component may also cause the engagement, education and the release of antisecretory protein.

In the context of the present invention "antisecretory protein" or its homolog, derivative or fragment may be used interchangeably with the term "antisecretory factors" or "protein-antisecretory factor", as defined in the patent WO 97/08202, and indicate the antisecretory protein or peptide, or a homologue, derivative and/or fragment, having antisecretory and/or the functional equivalent and/or similar activity. Thus, it should be understood that "antisecretory factor", "protein-antisecretory factor", "antisecretory peptide", "antisecretory slice or antisecretory protein" in the context of the present invention can also designate their derivative, homologue or fragment. All of these terms in the context of the present invention can be used interchangeably. Moreover, in the context of the present invention, the term "antisecretory factor" may be abbreviated as "AF". Antisecretory protein in the context of the present invention also refers to a protein with antisecretory properties, as defined earlier in WO97/08202 and WO 00/38535. Antisecretory factors were also described, for example, in WO 05/030246. Also the term antisecretory factor mean egg yolks, enriched antisecretory factors, as described in SE 900028-2 and WO 00/38535 and additionally described below.

"Inhaler" in the context of the present invention refers to a medical device that delivers liquid medication in the form of an aerosol in the respiratory tract. Compressors "inhaler" drive air through the tube in the medical Cup, filled with liquid medication. The force of the air breaks up the liquid into fine particles in an aerosol, which can be deeply inhaled into the respiratory tract.

The term "aerosol" in the context of the present subramanyaashtakam gaseous suspension of fine solid or liquid particles.

Detailed description of the invention

There is a need for new drugs intended for the pharmacological treatment of diseases associated with dysfunction of lipid rafts and/or kahweol, because at present no adequate therapy. Antisecretory proteins have an effective impact, as evidenced by the examples below.

The present invention relates to the control and regulation of the structure and function of lipid rafts and kahweol related structures, are widely represented in the cell membranes. Under lipid rafts understand membrane microdomain rich in cholesterol, which are proteins, for example, sphingolipids, such as GM1, which is important for transport ion binding cell growth, signaling and associate with the cytoskeleton. Caveola are vesicular or structure in the shape of the bulb, are presented in the cells, for example, cardio-pulmonary and vascular systems, including endothelial cells, smooth muscle cells, epithelial cells, fibroblasts and cardiomyocytes (Chan and Ye, 2007; Petersen et al., 2007), formed by one or more clusters of components of the lipid raft. Other proteins associated either individually or in combinations, are flotillin, caveolin is the essential component and Reggie (reggie); there are several variations of each.

The present invention relates to the use of pharmaceutical compositions containing the antisecretory protein, its homologue, derivative and/or fragment that has equivalent activity, or pharmaceutically active salt, for the manufacture of pharmaceutical compositions and/or therapeutic food for the treatment and/or prevention of dysfunction of lipid rafts and kahweol in cell membranes. The present invention also relates to treatment and/or prevention of various conditions associated with dysfunction of lipid rafts and kahweol, such as the component transfer through cellular barriers, restoration of tissues and organs, hyperplasia and/or hypertrophy of cells and tissues, cardiovascular function, the formation of scar tissue, reactive formation of excessive tissue and the restoration and regeneration of the epithelial cell surface.

Also the present invention relates to a method of treatment and/or normalization dysfunction of lipid rafts and kahweol, as described above, and the method includes the administration to a mammal, optionally, a therapeutically effective amount of the pharmaceutical composition and/or therapeutic diets containing antisecretory protein or a derivative, homolog, or fragment that has equivalent activity is, or pharmaceutically active salt.

The present invention also relates to various doses and ways to use, suitable for the intended purpose, and also taking into account age, sex, condition of the patient, and the like.

Treatment in accordance with the present invention is probably most effective for patients with an increased risk of developing or suffering from dysfunction of lipid rafts and kahweol, or from the capture or release of pathogenic substances. Moreover, such a treatment is effective also in other States, which differ pathological metabolism of cells and extracellular matrix components.

Found evidence that the antisecretory proteins and peptides that are likely to affect lipid rafts in cell membranes is certainly unexpected for specialists in this field. There are a large number of domains with an average size much smaller than microns, called lipid rafts, characterized by a high concentration of cholesterol and sphingomyelin. Lipid rafts contain a variety of integral and peripheral membrane proteins involved in the mass transfer and cell signaling. Such signaling platforms floating in the membrane and are essential elements for the respective functions, such as receptors, the fact that the market mate, the effector enzymes and compounds and substrates, whereby it is possible to obtain the transfer of certain ions, molecules and signals. These domains also interact with, for example, cytoskeleton and, in addition, affect the composition and metabolism of interstitial fluid and its pressure. Moreover, lipid rafts are associated with the metabolism of kahweol, as well as release and internalization, for example, viruses. In cell membrane formation occurs in clusters of receptors of growth factors, receptors of inflammatory signal, ion channels and carriers in lipid rafts, which undergo dynamic changes associated with the function, prevailing at the moment.

The use of antisecretory proteins and peptides (AF) is not limited to tissues, organs and anatomical structures described in the examples, but includes additional symptoms and diseases characterized by dysfunction, pathological function, Hypo - or hyperfunction of lipid rafts and/or kahweol.

Antisecretory proteins, peptides, derivatives and homologues have the ability to control and even to normalize the function of lipid rafts and proteins involved in the transport of nutrients and cell signaling. A very wide range of applied effective dosing schemes indicates that the risk of side effects is unexpected complications is minimal. Thus, the method used for monitoring and control of the structures and functions associated with lipid rafts and caveolae, makes possible the treatment of excessive effects on cells and tissues, and also allows to treat the patient with a broad range of doses, taking into account the individual reactions and the severity of the disease and/or discomfort.

The pharmaceutical composition of the present invention may, in accordance with one aspect be administered topically, locally, in situ, oral, nasal, subcutaneous and/or systemically through the blood vessels or through the respiratory tract.

Antisecretory factors represent a natural class of proteins of an organism. Protein-antisecretory factor person is a protein weight 41 kDa, containing 382 amino acids, when released from the pituitary gland. The active site in relation to the favorable impact on the normalization of lipid rafts and/or controlling Cavea of the present invention can be found in the website of the protein, adjacent to the N-end of the protein, located from 1 to 163 amino acid sequence SEQ ID NO:6 or a fragment of this section.

The authors of the present invention showed that the antisecretory factor to some extent homologous protein S5a, also called Rpn 10, which forms the subunit of the component, which is widely distributed in the all cells, the 26 S proteasome, more specifically in 19 S/PA 700 cap. In the present invention antisecretory proteins is defined as the class of homologous proteins with the same functional properties. Proteasome perform many of the functions associated with the degradation of excess protein, as well as short-lived unnecessary, denaturirovannykh, properly Packed and with other pathological proteins. Moreover, antisecretory factor/S5a/Rpn10 is involved in the distribution and transport of cellular components, especially protein.

All homologues, derivatives and fragments of antisecretory proteins and/or peptides of the present invention possess similar biological activity, and can be effective for use in obtaining medicines for the treatment and/or prevention of disorders in lipid rafts and/or caveolin, as well as in the treatment of conditions associated with dysfunction in lipid rafts and/or caveolin. Homologues, derivatives and fragments, in the context of the present invention contain at least 4 amino acids of natural antisecretory protein, which can additionally be modified by changing one or more amino acids, to optimize the biological activity of antisecretory factor in the treatment and/or prevention of conditions related to the present from which retenu.

Fragment antisecretory protein usually contains peptide/amino acid sequence or its fragment in the product in which the protein content is more than 90%, for example 95%, 96%, 97%, 98% or 99% of the peptide and/or their fragments of the present invention.

In addition, any amino acid sequence at least 70% identical to, for example, at least 72%, 75%, 77%, 80%, 82%, 85%, 87%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence of antisecretory protein, peptide, homologue, derivative and/or fragment of the present invention, is considered as part of the scope of the present invention. In the context of the present invention, the terms homologous and identical are used interchangeably, i.e. amino acid sequence having a degree of identity with another amino acid sequence has the same level of gomologichnosti with the specified amino acid sequence.

Under derived in the context of the present invention means a protein that has equivalent activity and/or functional equivalent activity in relation to the antisecretory factor, as defined in the present description, which is obtained from another substance either directly or by modification or partial replacement is possible, in which one or more amino acids are substituted by another amino acid, this amino acid may be modified or appear unnatural amino acid. For example, derivatives of antisecretory factor of the present invention may contain N-terminal and/or C-terminal protective group. One example of the N-terminal protective group includes acetyl. One example of a C-terminal protective groups include amide.

Under proteins, homologues, derivatives, peptides and/or fragments with amino acid sequence that is at least, for example, 95% identical to a reference amino acid sequence, it is understood that amino acid sequence, for example, a peptide that is identical to the reference sequence except that the amino acid sequence may include up to 5 point mutations per each 100 amino acids of the reference amino acid sequence. In other words, to obtain a polypeptide with amino acid sequence that is at least 95% identical to a reference amino acid sequence, up to 5% of the amino acids of the reference sequence may be deleted or replaced by other amino acids, or amino acids up to 5% of all amino acids of a reference sequence can be embedded in the reference of th is sequence. These mutations of the control sequences can be amino - or carboxy-terminal positions of the reference amino acid sequence or anywhere between those terminal positions, scattered, either individually among the amino acids of the reference sequence or in one or more continuous bands in the reference sequence.

In the present invention for determining the identity is the most suitable program for the local algorithm. Local algorithm (such as the algorithm of Smith-Waterman) compare the subsequence of one sequence with a subsequence of the second sequence and find the combination of subsequences and compare these subsequences that gives the highest overall level of similarity. The internal gaps, if present, impose a fine. Local algorithms work well when comparing two multi-domain proteins with a common domain or at least a common binding site.

How to determine identity and similarity are systematized in public programs. Preferred methods for determining the identity and similarity of two sequences using computer programs include, but are not limited to, the GCG software package (Devereux, J. et al. (194)) BLASTP, BLASTN, and FASTA (Altschul, S. F. et al. (1990)). The program BLASTX publicly available from NCBI and other sources (BLAST Manual, Altschul, S. F. et al., Altschul, S. F. et al. (1990)). Each program sequence analysis is a table of scoring by default and penalties for omissions by default. Typically, a molecular biologist will use the default parameter set used by a computer program.

Antisecretory protein or peptide or a homologue, derivative and/or fragment that has equivalent activity as defined in the present description may contain 4 or more amino acids, as, for example, 5-16 amino acids, such as, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20 amino acids or more. In other preferred embodiments, the implementation of the antisecretory factor consists of 42, 43, 45, 46, 51, 80, 128, 129 or 163 amino acids. In preferred embodiments, the implementation of the antisecretory factor consists of 5, 6, 7, 8, or 16 amino acids.

In another preferred embodiment, antisecretory protein or peptide or a homologue, derivative or fragment that has equivalent activity in accordance with the present invention, consists of a sequence of the following formula:

X1-V-C-X2-X3-K-X4-R-X5

where X1 represents I, amino acids 1-35 of the sequence SEQ ID NO:6, or it is absent, X2 represents H, R or K, X3 pre is is a S or L, X4 represents T or A, X5 is an amino acids 43-46, 43-51, 43-80 or 43-163 sequence SEQ ID NO:6 or is missing.

Antisecretory factor according to the present invention can be obtained in vivo or in vitro, for example, synthesized by recombinant means, synthetically and/or chemically and/or obtained from a natural source antisecretory factors, for example, from the pituitary gland of the pig or bird eggs. After receiving antisecretory factors can be subjected to further processing, such as chemical or enzymatic cleavage into smaller fragments with antisecretory activity or by amino acid modifications. Currently when cleaning it is impossible to get antisecretory factor in its pure form. However, it is possible to obtain biologically active protein antisecretory factor by recombinant means or synthetically, as described in WO 97/08202 and WO 05/030246. In WO 97/08202 also described the preparation of biologically active fragments of this protein size 7-80 amino acids.

Antisecretory factor of the present invention may, additionally, contain N-terminal and/or C-terminal protective group. One example of the N-terminal protective group includes acetyl. One example of a C-terminal protective groups include amide.

In the preferred embodiment, this image is etenia antisecretory factor selected from posledovatelnosti SEQ ID NO:6, i.e. VCHSKTRSNPENNVGL (SEQ ID NO:1, in the context of the present invention is also called AF-16), IVCHSKTR (SEQ ID NO:2), VCHSKTR (SEQ ID NO:3), CHSKTR (SEQ ID NO:4), HSKTR (SEQ ID NO:5) or the amino acid sequence of antisecretory protein in accordance with SEQ ID NO:6, using the conventional one-letter symbols of amino acids. Sequence SEQ ID NO:1, 2, and 3 are described, for example, in WO 05/030246. As defined in the attached list of sequences, some of the amino acids in the above defined sequences can be replaced by other amino acids. Below in this paragraph the position of a certain amino acid in a particular amino acid sequence is evaluated from left, closest to the N-end amino acid is designated as being in the position 1 in the specified sequence. Any replacement amino acids, as defined below, may be implemented independently of any other substitute amino acids in this sequence. In the sequence of SEQ ID NO:1 C in position 2 can be replaced by S, H in position 3 can be replaced by R or K, S at position 4 can be replaced by L and/or T in position 6 can be replaced by A. In the sequence SEQ ID NO:2 C at position 3 can be replaced by S, H in position 4 can be replaced by R or K, S at position 5 can be replaced by L and/or T in position 7 can be replaced by A. In the sequence SEQ ID NO:3 C in position 2 can be replaced by S,H in position 3 can be replaced by R or K, S in position 4 can be replaced by L and/or T in position 6 can be replaced by A. In the sequence SEQ ID NO:4, C in position 1 can be replaced by S, H in position 2 can be replaced by R or K, S at position 3 can be replaced by L and/or T in position 5 can be replaced by A. In the sequence SEQ ID NO:5 H in position 1 can be replaced by R or K, S in position 2 can be replaced by L and/or T in position 4 replace A.

The present invention also relates to the combination of two or more of any fragments of the sequence SEQ ID NO:1-6.

In one embodiment of the present invention the pharmaceutical composition in accordance with the present invention also provides pharmaceutically acceptable excipient. The choice of pharmaceutically acceptable excipient and its optimal concentration for use in accordance with the present invention can be easily carried out by a specialist by experiments. Pharmaceutically acceptable excipients for use in the present invention include solvents, buffering agents, preservatives, chelating agents, antioxidants and stabilizers, emulsifying agents, suspendresume substances and/or diluents. The pharmaceutical composition of the present invention can be obtained in accordance with conventional pharmaceutical practice, for example in the under "Remington: The science and practice of pharmacy", 21st edition, ISBN 0-7817-4673-6 or "Encyclopedia of pharmaceutical technology", 2nd edition, ed. Swarbrick J., ISBN: 0-8247-2152-7. Pharmaceutically acceptable excipient is a substance that is essentially harmless to the individual, which must be entered in the composition. This excipient, as a rule, corresponds to the requirements of the national health authorities. Official Pharmacopoeia, such as, for example, the British Pharmacopoeia, USP and European Pharmacopoeia, set standards for pharmaceutically acceptable excipients.

Below is an overview of suitable compositions for optional use in pharmaceutical compositions in accordance with the present invention. The review is based on a particular way of introduction. However, it should be understood that in those cases where the pharmaceutically acceptable excipient can be used in different dosage forms or compositions, the use of certain pharmaceutically acceptable excipient not limited to a specific dosage form or a specific function excipient. It should be emphasized that the invention is not limited to the use specified in further compositions.

Parenteral compositions. For systemic use of the composition according to the present invention can include conventional non-toxic pharmaceutically acceptable carriers, excepi the options including microspheres and liposomes.

The composition for use according to the present invention may include all types of solid, semi-solid and liquid compositions.

Pharmaceutically acceptable excipients may include solvents, buffering agents, preservatives, chelating agents, antioxidants and stabilizers, emulsifying agents, suspendresume funds and/or diluents. Examples of various substances are given below.

An example of the various funds

Examples of solvents include, but are not limited to, water, alcohols, blood, plasma, cerebrospinal fluid, ascitic fluid and lymph.

Examples of buffering agents include, but are not limited to, citric acid, acetic acid, tartaric acid, lactic acid, phosphoric acid, bicarbonates, phosphates, diethylamid etc.

Examples of chelating funds include, but are not limited to, EDTA and citric acid.

Examples of antioxidants include, but are not limited to, bottled hydroxyanisol (BHA), ascorbic acid and its derivatives, tocopherol and its derivatives, cysteine, and mixtures thereof.

Examples of diluents and dezintegriruetsja funds include, but are not limited to, lactose, sucrose, mdex™, calcium phosphates, calcium carbonates, calcium sulphate, mannitol, starches and microcrystalline cellulose.

<> Examples of the linking means include, but are not limited to, sucrose, sorbitol, gum Arabic, sodium alginate, gelatin, chitosan, starches, cellulose, carboxymethylcellulose, methylcellulose, hydroxypropylcellulose, polyvinylpyrrolidone and polyethylene glycol.

The pharmaceutical composition of the present invention may in one context be used either topically or by peripheral intravenous infusion, or by intramuscular or subcutaneous injection to a patient, or buccal, pulmonary, nazalnam, dermal or oral route. In addition, the pharmaceutical composition can be entered through a surgically inserted shunt into the ventricle of the patient's brain.

In one of the embodiments of the pharmaceutical composition used in accordance with the present invention, a receive channel, local, intranasal, oral, subcutaneous and/or system administration. In a preferred embodiment, the composition of the present invention is administered in the form of suspension or, more preferably, in the form of a powder for inhalation spray, spray, inhaler, or nasal spray and/or in the respiratory tract.

The introduction of the powder containing antisecretory factors, gives additional advantages from the point of view of stability and is Osinovka. The pharmaceutical composition of the present invention can also be used topically, intraocular way, intranasally, orally, subcutaneously and/or enter systemically through the blood vessels. In a preferred embodiment, the pharmaceutical composition was prepared for intravenous, intramuscular, local, oral or nasal administration. Usually, when topically applied to the eye, the concentration in the composition of the present invention is from 1 μg to 1 mg per dose, preferably 50-250 μg, or as a single dose per day or repeating several times a day (multiple entry), but is not limited thus.

With the systemic administration of blood dosage is in the range from 0.1 μg to 10 mg per dose per kg body weight, such as from 0.1 μg to 1 mg per dose per kg body weight, preferably 1-500 μg/ kg body weight, more preferably 1-50 mg/ kg body weight, either as a single dose per day or repeated injections several times a day. When used in accordance with the present invention enriched antisecretory factors of egg yolk such a composition is preferably administered orally.

Accordingly, the present invention relates to the use of antisecretory protein or its homologue, derivative and/or fragment with equivalentofficially, and/or pharmaceutically active salts, to pharmaceutical compositions and/or medicinal food for the treatment and/or prevention of dysfunction of lipid rafts and/or kahweol. In one of the embodiments specified antisecretory protein consists of the sequence in accordance with the following formula:

X1-V-C-X2-X3-K-X4-R-X5

where X1 represents I, amino acids 1-35 of the sequence SEQ ID NO:6, or it is absent, X2 represents H, R or K, X3 is a, S or L, X4 is a T or A, X5 is an amino acids 43-46, 43-51, 43-80 or 43-163 sequence SEQ ID NO:6 or is missing. In another embodiment, the present invention relates to the use of antisecretory protein that contains the amino acid sequence of SEQ ID NO:1. In another embodiment, the present invention relates to the use of antisecretory protein that contains the amino acid sequence of SEQ ID NO:2. In yet another embodiment, the present invention relates to the use of antisecretory protein that contains the amino acid sequence of SEQ ID NO:3. In yet another embodiment, the present invention relates to the use of antisecretory protein that contains the amino acid sequence of SEQ ID NO:4. In another embodiment, the implement is placed, the invention relates to the use of antisecretory protein, which contains the amino acid sequence of SEQ ID NO:5.

In addition, in another embodiment, the invention relates to the use of antisecretory protein, which is a protein with amino acid sequence SEQ ID NO:6, or its homologue, derivative and/or fragment containing amino acids 38-42 sequence SEQ ID NO:6.

In yet another embodiment, the present invention relates to the use of pharmaceutical compositions as disclosed in the present description, which contains two or more antisecretory protein selected from the proteins of SEQ ID NO:1-6, and SEQ ID NO:6 or a homologue, derivative and/or fragment containing amino acids 38-42 sequence SEQ ID NO:6, or a sequence described by the General formula defined in the present description. These sequences are all equally preferred for use in the present invention.

In one of the embodiments of the present invention mentioned pharmaceutical composition additionally contains a pharmaceutically acceptable excipient. This excipient may be any preferred excipient selected as suitable for a particular purpose. Examples of excipients are disclosed in the present description.

In another embodiment, the present image is to be placed is specified pharmaceutical composition was prepared for intraocular, intranasal, oral, local, subcutaneous and/or system administration. The selected method of administration will vary depending on the condition of the patient, age and sex of the patient, etc.

In another embodiment, the pharmaceutical composition was prepared for introduction in the form of a spray, aerosol or via atomizer or nebulizer. In yet another embodiment, the present invention relates to pharmaceutical compositions and/or therapeutic feeding, which get for the introduction of systemically in the blood in a dose of from 0.1 μg to 10 mg per dose per kg body weight per day, such as, for example, from 0.1 μg to 1 mg per dose per kg body weight per day, preferably 1-500 μg per admission per kg of body weight per day, more preferably 1-50 μg per admission per kg of body weight per day. In another embodiment, the specified dose of 1-1000 μg per admission per kg of body weight per day, such as 1-100 μg per admission per kg of body weight per day. The amount of the pharmaceutical composition that is administered to the patient when the need arises, much will depend, of course, on the particular patient being treated, and is determined by the person skilled in the art, such as a practicing physician for each case. This introduction can be done or not in single dose, or as multiple daily applications.

At last the ies became obvious what lipid rafts, caveola and caveolin play a role in several pathological conditions in humans, such as, but not limited to, muscular dystrophy (e.g., poznakomitstsa muscular dystrophy), the imbalance of cellular cholesterol (e.g., Niemann-pick disease type C) and/or dysfunction of education amyloid peptide from a protein-amyloid precursor (APP) (for example, Alzheimer's disease). Thus, the present invention in one of the embodiments also relates to use of pharmaceutical compositions, as described in the present description, for the prevention and/or treatment of muscular dystrophy, imbalance of cellular cholesterol and/or dysfunction of education amyloid peptide from the protein precursor of amyloid.

Lipid rafts are a type of cell of the domain in which the lipid a specific chemical structure can be dynamically associated with each other, forming a platform that is essential for the ordering of membrane proteins and education carry weight and signaling complexes.

It is known that some cell receptors are found in lipid rafts and/or caveolin cell membrane, for example, it was shown that the rafts are essential for the regulation of GPCR receptors at all stages of their life cycle, i.e. on associtation path in the plasma membrane and n is acetosa way. Found that lipid rafts and/or caveola involved in the regulation of the stability of the receptor, regulating signaling and transport of any particular GPCR. In addition, you can control a fast metabolism receptors, protein ion channels and water channels and other transmitters of signals. Thus, in one embodiment implementing the present invention relates to the use of pharmaceutical compositions of the present invention for regulating the stability of the receptors, the regulation of signaling and/or transport of one or more receptors located in the cell membrane.

Many receptors there are no more than 3-5 hours. As a result, for permanent production of new receptor proteins necessary to produce the receptor proteins were safely and correctly aimed and mounted at the appropriate places in the cell membrane. The present invention discloses the possible use of pharmaceutical compositions in accordance with the present invention to improve and/or facilitate specified positioning of receptors in the cell membrane.

Moreover, recent discoveries indicate that the classical steroid hormone receptors are found in lipid rafts and/or caveolin in the cell membrane, and thus can function in these structures and on OSU. Examples of such receptors, but not limited to, are receptor tyrosine kinase, i.e. signaling pathways of EGFR and androgenous receptor, Akt1, IL-6, STAT3, estrogen receptor (ER). Thus, the present invention also relates to use of pharmaceutical compositions of the present invention for the treatment and/or prevention of conditions associated with dysfunction of the receptors of steroid hormones.

I believe that insulin resistance, defined as a reduced ability of cells or tissues to respond to physiological insulin is the primary disturbance in the pathophysiology of type II diabetes. It is known that in such resistance plays a major role TNFα, as viriato, and other cytokines and lymphokines. At the present time it is clear that the unique effects of hormone closely associated with compartmentalization of signaling molecules in lipid rafts in the cell membrane. Thus, it was shown that the corresponding function of lipid rafts plays a critical role for the compartmentalization of insulin signaling, for example, in adipocytes. The destruction of the structure and/or function of lipid raft in turn leads to inhibition and/or destruction of metabolic insulin signaling, probably at least partially due to the aberrant expression of glycosphingolipid the century The currently favored option of implementing the present invention thus relates to the use of pharmaceutical compositions containing the antisecretory protein or a homologue, derivative and/or fragment that has equivalent activity, and/or pharmaceutically active salt, for normalization of metabolic insulin signaling. The present invention therefore, in this embodiment, also relates to treatment and/or prophylaxis of diabetes, such as type II diabetes.

Lipid rafts and caveola act as equipped with the centers of the transmission signal. As the insulin receptor, and TC10 are located in lipid rafts. Erroneous hit TC10 in the domain of non-lipid raft prevents its activation by insulin and blocks the action of insulin. Numerous studies have demonstrated that lipid rafts act as organizing centers for insulin signaling in adipocytes. The activated insulin receptor, in particular, catalyzes the tyrosine phosphorylation of certain proteins in the lipid rafts, including caveolin is the essential component and CbI. Components of insulin signaling, including some or all of the insulin receptors, flotillin and TC10, constitutive located in lipid rafts.

Insulin stimulates the transport is lukosi in fat and muscle cells through a process regulated vesicular recycling, which ensures the transport of glucose Glut4 moves from intracellular sites to the plasma membrane. In unstimulated cells, Glut4 undergoes endocytosis in endosome and then distributed by specialized accumulating the vesicles that move to the plasma membrane after activation of the insulin receptor. Then vesicles couple and merge into certain areas of the membrane that leads to extracellular exposure of the carrier. Signaling cascade from the insulin receptor involves the tyrosine phosphorylation of several intracellular substrates and activation of the PI3-kinase pathway and activation of a G protein, which in turn is associated with a variety of effectors, including squirrels ectocyst Exo70.

Moreover, the complex of ectocyst, sogelease Exo70, Sec6 and Sec8, involved in the compartmentalization containing Glut4 vesicles in the domains of lipid rafts in the cell membrane, for example in adipocytes. Part of the complex of ectocyst are recruited G-proteins upon activation by insulin and are essential for insulinstimulated seizure of glucose in the cells. Moreover, it was recently shown that their focus on lipid rafts are necessary to capture the glucose and docking of Glut4 to the plasma membrane. Interestingly, this complex also necessary protein with bearing-domain catalystbed with the complex after it is moved in the lipid raft. Concentrations of ecosystem in lipid rafts create target sites for Glut4 vesicles, which is temporarily associated with these microdomains after stimulation of cells with insulin.

As shown in the experimental part of this application, the inventors were able to demonstrate that antisecretory factors have an effective impact on the development of symptoms in chemically induced diabetes. Detailed speaking, experimentally induced diabetes caused by exposure of streptozocin, was studied to determine whether treatment with AF-16 effectively influence it. In preferred currently embodiment, the present invention therefore relates to the use of pharmaceutical compositions containing the antisecretory protein or a homologue, derivative and/or fragment that has equivalent activity, and/or pharmaceutically active salt, for the treatment and/or prevention in a patient of a disease associated with diabetes and/or complications associated with diabetes, such as, for example, selected from diabetes I and II.

In another preferred embodiment of the present invention the pharmaceutical composition of the present invention is used to normalize the formation and/or function of lipid rafts in muscle cells, endotheli the selected cells, fat cells and/or red blood cells for the treatment and/or prevention of dysfunction Glut4 and/or accumulation of ecosystem in these cells. Therefore, the above mentioned pharmaceutical composition used in the present invention for the treatment and/or prevention of diabetes and/or complications associated with diabetes, such as, for example, selected from diabetes I and II.

Caveola are dynamic structures that can develop in the plasma membrane, forming cytoplasmic carriers participating in mediated by receptors capture solutions in the cell, and transcytosis are activated through the cell. Although caveola found in many types of cells, they are especially common in adipocytes, where they can be combined in a ring structure (socket Cavea), often associated with actin filaments. They take part in the capture of fatty acids, as well as in the transport of fatty acids and/or the binding of fatty acids by the cells. Moreover described that Kaveri and caveolin is the essential component-1 participate in the transport of cholesterol to the cell surface is regulated by the level of cholesterol in the cell. Thus, the present invention in another embodiment also relates to the use of pharmaceutical compositions containing the antisecretory protein or a homologue, derivative and/or fragment with ek is Valentey activity and/or pharmaceutically active salt for the treatment and/or prevention in a patient of a disease associated with dysfunction of the capture of fatty acids, transport of fatty acids, binding of fatty acids and/or cholesterol.

Dysfunction kahweol associated with some human diseases. For example, caveolin is the essential component-1 (CAV1) -null cells exhibit increased proliferation, and lack of CAV accelerates tumor development. In some forms of breast cancer is negative regulation of CAV1, and was discovered a number of sporadic mutations CAV1 in samples of breast cancer person, that is, in particular, in conjunction with positive estrogen receptor alpha. CAV3, isoform of caveolin is the essential component characteristic of the muscles, also closely associated with the disease. Many CAV3 mutations are associated with a number of muscle disorders in humans.

In another, equally preferred embodiment, the pharmaceutical composition in accordance with the present invention is used to normalize the formation and/or function of kahweol to counteract and/or stabilize high blood pressure of the patient when the need arises. As shown in the experimental part, applying to the mammal a pharmaceutical composition in accordance with the present invention can reduce the slojnenia, caused by hypertension, both small and large circle circulation of a mammal. Thus, the present invention can be used for the treatment and/or prevention of myocardial infarction, high blood pressure, lung disease, belascoaran and/or traumatic injury of the vascular system, such as heart and lung, as well as hormonal dysfunction and dysregulation.

In another aspect, the present invention relates to a method of treatment and/or prevention of dysfunction of lipid rafts and/or kahweol in mammals when the need arises, and this method includes the introduction of an effective amount of a pharmaceutical composition containing an antisecretory protein or a homologue, derivative and/or fragment having equivalent functional activity, and/or pharmaceutically active salt. In one embodiment, the present invention relates to a method, wherein said antisecretory protein consists of the sequence in accordance with the following formula X1-V-C-X2-X3-K-X4-R-X5, where X1 represents I, amino acids 1-35 of the sequence SEQ ID NO:6, or is absent, X2 represents H, R or K, X3 is a, S or L, X4 is a T or A, X5 is an amino acids 43-46, 43-51, 43-80 or 43-163 sequence is eljnosti SEQ ID NO:6 or missing. In another embodiment, the present invention relates to a method, wherein said antisecretory protein contains the amino acid sequence of SEQ ID NO:1. In yet another embodiment, the present invention relates to a method, wherein said antisecretory protein contains the amino acid sequence of SEQ ID NO:2. In yet another embodiment, the present invention relates to a method, wherein said antisecretory protein contains the amino acid sequence of SEQ ID NO:3. In addition, the present invention relates to a method, wherein said antisecretory protein contains the amino acid sequence of SEQ ID NO:4. In yet another embodiment, the present invention relates to a method, wherein said antisecretory protein contains the amino acid sequence of SEQ ID NO:5. In yet another embodiment, the invention relates to a method, wherein said antisecretory protein is a protein with amino acid sequence SEQ ID NO:6 or a homologue, derivative and/or fragment containing amino acids 38-42 of SEQ ID NO:6. In one embodiment, the present invention relates to a method in which the specified pharmaceutical composition comprises two or more antisecretory protein selected from the proteins of SEQ ID NO:1-, and SEQ ID NO:6 or a homologue, derivative and/or fragment containing amino acids 38-42 sequence of SEQ ID NO:6 or the sequence described General formula in the present description. In one embodiment, the mentioned pharmaceutical composition is for intraocular, intranasal, oral, topical, subcutaneous and/or system administration. In yet another embodiment, the specified pharmaceutical composition and/or health food form for administration as a spray, aerosol or via atomizer or nebulizer. A variant of implementation of the present invention encompasses the way in which the pharmaceutical composition is for administration systemically in the blood at a dosage of from 0.1 μg to 10 mg per dose per kg body weight per day, such as, for example, from 0.1 μg to 1 mg per dose per kg body weight per day, preferably 1-500 μg per admission per kg of body weight per day, more preferably 1-50 μg per admission per kg of body weight per day. In one embodiment, the specified method of the specified introduction is carried out or as a single dose, or as multiple daily applications. The present invention also relates to a method of treatment and/or prevention of dysfunction of lipid rafts and/or kahweol in mammals when the need arises, and this method includes the maintenance of an effective amount of the pharmaceutical composition, containing antisecretory protein or a homologue, derivative and/or fragment having equivalent functional activity, and/or pharmaceutically active salt.

In one preferred embodiment, the present invention also relates to a method of treatment and/or prevention of dysfunction of lipid rafts and/or kahweol in mammals when the need arises, and this method includes the introduction of an effective amount of a pharmaceutical composition containing an antisecretory protein or a homologue, derivative and/or fragment that has equivalent activity, and/or pharmaceutically active salt.

It must be clear that the method comprising introducing an effective amount of the pharmaceutical composition to the mammal when the need arises and/or other medical use of pharmaceutical compositions containing the antisecretory protein or a homologue, derivative and/or fragment that has equivalent activity, and/or pharmaceutically active salt, in accordance with the present invention, directed to all of the States described in the present description, associated with dysfunction of lipid rafts and/or kahweol.

Experimental part

Example 1

Immature and adult rats, with weight the body 150-300 g of either sex, treated experimentally to cause pulmonary hypertension and reactive changes in the lungs. Pulmonary hypertension (LH) is defined as the presence of pressure in excess of normal pulmonary arterial pressure, ~20 mm RT. Art., typically in the range of 30-40 mm RT. Art. It is a progressive disease with high prevalence and mortality in humans and animals. Experimental models make possible the study of the triggering mechanisms. A separate application to the rat monocrotaline is a recognized method of inducing LH (Cf. Mathew et al., 2004). This plant alkaloid is activated in the liver to pyrrole metabolites with a half-life of only a few seconds, and therefore mainly affects the endothelium of pulmonary arteries, causing destruction of endothelial cells and the permeability of the pulmonary vessels. Then in the next few days should be a significant stimulation of DNA synthesis and hypertrophy of endothelial cells of the pulmonary arteries. I believe that these endothelial cells secrete growth factors and stimulating migration, which contribute to the migration and reactive changes of the adjacent smooth muscle cells. Within two weeks the right ventricle was hypertrophied due to the increased workload caused by the induced crotaline the changes in the pulmonary vascular system.

The group of rats Sprag-Dole made one intraperitoneally injection crotaline (60 mg/kg body weight; Sigma). Every second animal was implanted subcutaneously on the back of osmotic minimalis Alzet (type 2001; filling volume ~235 μl; sweep rate ~1 mm/h; pre-launched; filling solution contains 20 mg/ml AF-16, dissolved in the FSB, with the addition of 15% ethanol). The pump, therefore, gave ~20 μg of AF-16 per hour for at least 10 days. In one experiment rats with pump Alzet 2001 received implantation of one intramuscular injection of 2 mg AF-16. For comparison, each of the second rat when injected Catalina implanted pump Alzet 2001, filled with the filler, the FSB with 15% ethanol. Additional groups of rats with a suitable weight carried only implanted pumps, filled or AF-16, or filler, but have not received Catalina.

After 18 days, rats were anestesiology and determined their weight. Those that received crotalin were thin and small for his age, were less active and had a hard fur and had less weight compared with untreated normal control. Those that received crotalin and which was poured AF using minnesoto, looked healthy and had almost the same weight and appearance as the control. The pressure in the right ventricle were determined using wholesale the miniature fiber measuring system (Samba System 3200 & Samba Preclin sensor 420; Samba Sensors AB, V. Frölunda, Sweden)inserted through the right jugular vein. The mean pulmonary arterial blood pressure was approximately 20 mm RT. Art. in rats, which were injected filler or introduced AF-16. In contrast, rats that were administered crotalin (one intraperitoneally injection; 60 mg/kg body weight; crotalin dissolved in the FSB) and the filler, imported by Alzet pump, had for 18 days pulmonary arterial pressure above 30 mm RT. Art. and hypertrophy of the right ventricle, as determined by comparing its full weight full weight of the left ventricle and with the weight of the respective structures untreated normal rats. The introduction of the AF-16 rats, which was introduced crotalin, return the blood pressure in the right ventricle and pulmonary arterial blood pressure close to normal, and there was no significant hypertrophy of the right ventricle compared to the normal untreated rats. These results are reproducible, that whenever performed in repetition. It is known that compounds derived from Catalina, cause the destruction of, for example, protein caveolin is the essential component that leads to disorganization of kahweol and lipid rafts, which effects lead to changes in signaling of endothelial cells in this model LH (Mathew R, et al., 2004).

We believe that the use of AF-16 stun the Alo reconstruction of the pulmonary vascular system and, consequently, the development of pulmonary disorders and hypertrophy in the right half of the heart, which in other cases have been reported after use Catalina. Induced disruption of lipid rafts and kahweol in the endothelium and smooth muscle cells of blood vessels and subsequent reactive changes were thus reduced by using AF-16 and has not resulted in the expected vascular disorders, no ventricular hypertrophy right half of the heart.

Example 2

In the second experiment investigated whether the introduction of the AF-16 in the dosage, which, as shown above, stops the development of pulmonary hypertension, the healing process of damaged arteries.

Shot adult rats carried osmotic Meninas (Alzet type 2001; filling volume ~235 μl; ~1 mm/h; pre-launched; filling solution contains 10 mg/ml AF-16, dissolved in the FSB, with the addition of 15% ethanol), implanted subcutaneously on the back. In addition, 1 mg AF-16 was administered via intramuscular injection immediately after implantation. After this, the skin on the right side of groin was shaved, cut and highlighted the right iliac and common iliac artery. About artery installed clip Piana and closed it three times, each time for 15 seconds and then removed. A special effort was made not to puncture the vessel or any adjacent structures. Check the whether the patency of the femoral and iliac arteries and veins, to ensure proper blood circulation. Then the wound edges lined and stitched. In parallel with other rats pinched right femoral and iliac artery, but they have introduced filler, the FSB with 15% ethanol, instead of the active material.

After 10 days, the animals were once again exposed to anesthesia and injected 1 ml/kg body weight of a mixture of 2% Evans blue and 3% albumin (bovine serum albumin: Sigma), dissolved in FSB. After 15 minutes, the animal was fixed by transcardial perfusion buffer formalin solution after the initial washing of the FSB, to which was added heparin. The right and left iliac and femoral arteries were carefully separated and fastened with their ends immersed in a solution of formalin in the FSB. After 1 hour, the vessel longitudinally cut with sharp scissors and investigated the prevalence blue staining of the surface of the lumen of the arterial wall. The surface of the lumen of intact arteries showed no staining. On the contrary, those who have the right iliac and femoral artery were injured and treated filler, showed a continuous, clear, intense blue staining throughout the damaged surface of the vessel, with fairly sharp edges. Injured iliac and femoral arteries of the third group of animals treated with AF-16 after damage to the bore stains and intermittent irregular areas, which were painted blue, separated, seemingly unpainted, in the damaged part of the vessel. As far as can be judged on appearance, more than half the field in the damaged area left unpainted treated with AF-16. This means that AF-16 contributes to the restoration of the original dinadiawan inner wall of the vessel so that it becomes covered inside cells. Moreover, the observed clot was less visible compared to vessels that have been damaged in animals treated filler. Light microscopy damaged areas showed that the number of leukocytes, platelets, macrophages and mast cells in the damaged tissue was lower after treatment with AF-16 in comparison with the number after treatment with filler. Moreover, mononuclear cells attached to the surface was less. Were found smooth muscle cells infiltrated into adluminal layer, but to a lesser extent after treatment with AF-16 in comparison with the processing of filler. Endothelial cells, fibroblasts and smooth muscle cells in normal conditions characterized by a large number of kahweol and vesicles on and in their plasma membrane. The operation of these surface structures is disturbed during and after mechanical injury in the blood vessel and stop the is to a considerable extent in this state for at least 10 days i.e. the period of time studied in the described experiments. However, in animals that were injected AF-16, there is a marked tendency to normalization, compared to the state prevailing at those who were injected filler. It is known that the signal transmission for the above cell types to a large extent mediated by lipid rafts and caveolae. You can, therefore, conclude that treatment with AF-16 reduces inflammatory and reactive changes of the damaged blood vessels and normalizes to a considerable extent the structure and function of cells in these areas. AF-16 exerts its beneficial effects by affecting lipid rafts and caveola.

Example 3

In experiment investigated whether the introduction of the AF-16 in the healing process of damaged skin and cartilage, applying ear damage in rats by freezing-thawing as a model system.

Shot adult rats carried osmotic Meninas (Alzet type 2001; filling volume ~235 μl; ~1 mm/h; pre-launched; filling solution contains 20 mg/ml AF-16, dissolved in the FSB, with the addition of 15% ethanol), implanted subcutaneously on the back. In addition, 1 mg AF-16 was administered via intramuscular injection immediately after implantation of the pump. After that has been damaged by freezing-thawing, clenching his ear standard is mainly using clip Peana, chilled in liquid nitrogen. For comparison, other rats were simultaneously treated in the same way that they were introduced only a filler, the FSB with 15% ethanol, but not the peptide. In addition, for control used in normal rats, which are not subjected to any damage by freezing and thawing, as intact rats with implanted pumps with AF-16.

Rats treated with AF-16 in 2 weeks after damage by freezing, showed less swelling and inflammation compared with the injured, but treated filler. On both sides along the edges of the damaged elastic cartilage in the ear formed a new hyaline cartilage. In rats, which were injected AF-16, 2 weeks less than necrosis in the remaining elastic cartilage, less education surrounding hyaline cartilage, less edema and infiltration of inflammatory cells. The amount of collagen and density of inflammatory cells decreased in treated with AF-16 in comparison with the treated filler. Application to intact rats as filler, and AF-16 has not changed structures in the ear.

We can conclude that the outcome of the healing process in relation to damaged skin and underlying cartilage was more successful and resulted in the formation of smaller amounts of scar tissue, if the animals received AF-16. Povrezhdeniya freezing much stronger decreases the number of kahweol in cells in the damaged areas in the treated filler ears than observed in the treated AF-16.

Example 4

Adult rabbits were studied from the point of view, does the introduction of the AF-16 in the healing process of damaged skin and periosteum.

Shot adult new Zealand rabbits albinos (females, the body weight of 2.5-2.8 kg) to surgically open the bones of the skull stuck helmet, made of glass fiber reinforced plastics. The rabbit was injected intravenously injection or AF-16, or filler in the marginal ear vein. Dose of AF-16 was up to 5 mg per kg of body weight twice a day. The skin wound of the skull vault and sites of injection in the ear examined macroscopically through the week. Then the samples from both areas were cut and prepared by fixing, fill, production cuts and coloring to a study by light microscopy. The use of AF-16 led to the spread of a smaller swelling in healing the skin, which was less inflamed and less eye-catching, hypertrophic scar tissue, observable in the corresponding samples of animals treated filler. Light microscopy confirmed that in samples of animals who have introduced the AF-16, fewer inflammatory cells and less collagen than those who were injected filler. Moreover, in samples of animals who have introduced the AF-16, blood vessels look more Mature.

For the, turn, that AF-16 improves the healing of deep skin wounds, reducing significant otherwise the inflammation and spread of edema and excessive scar tissue. It was found that the introduction of the AF returns a reduced concentration of kahweol to a more normal than observed in treated filler.

Example 5

Adult rats were studied in relation to the fact that it affects the healing of experimentally induced gastric RAS introduction AF-16.

Shot rats carried osmotic Meninas (Alzet type 2001; filling volume ~235 μl; ~1 mm/h; pre-launched; filling solution contains 20 mg/ml AF-16, dissolved in the FSB, with the addition of 15% ethanol), implanted subcutaneously on the back. In addition, 2 mg AF-16 was administered via intramuscular injection immediately after implantation of the pump. After that shaved belly and surgically opened the abdominal wall. Was found in the stomach and opened his underparts, with great attention was paid to the fact, not to give various neighboring organs and structures to dry or to receive mechanical damage. Ventral side of glandularia (distal) part of the stomach is then treated with 80% acetic acid contained in a glass tube for 60 seconds. Then the glass tube was quickly removed and open serous membrane was washed a large amount of the FSB. Then the abdomen was sutured. The pair is parallel to the other rats were treated in the same way, but in the pump they had a filler, and they received an injection with an equal volume of filler. A week later the rats once again anestesiology, and they discovered and surgically opened belly. Investigated the distribution of adhesions and ascitic fluid as the healing of stomach ulcers. Less fluid in the abdominal cavity were those of rats that had received AF-16, compared with those who received the filler. The number of adhesions between the greater omentum and the stomach, intestines, spleen and liver, as well as the wounds of the abdominal wall, was less significant and less concentration and size in those who received AF-16. Light microscopy confirmed the macroscopic impression that the wall of the stomach was less swollen. The epithelial coating on the inner surface of the stomach was more complete and extensive and it seemed better ordered after application of AF-16. In addition, after application of AF-16 was less infiltration of inflammatory cells in the gastric wall. We can conclude that AF-16 improves the healing of wounds in the stomach wall. Moreover, decreased distribution of ascitic fluid, as the number and risk of adhesions of the abdominal cavity.

Example 6

In another experiment investigated whether the introduction of the AF-16 biointegration implanted in the body for foreign material.

Shot adult rats was carrying twins who I osmotic Meninas (Alzet type 2001; filling volume ~235 μl; ~1 mm/h; pre-launched; filling solution contains 10 mg or 20 mg/ml AF-16, dissolved in the FSB, with the addition of 15% ethanol), implanted subcutaneously on the back. In addition, 2 mg AF-16 was administered via intramuscular injection immediately after implantation. The implants consisted of thin membranes (1×2 cm)suture material and sponge (0,2×0,5×1 cm; designed for augmentation), made of degradable poliuretanoviy (Artelon®, received as gift from Artimplant, V. Frölunda, Sweden). In parallel to other rats implanted the same materials in the same areas directly under the muscle fascia on their back. The implantation procedure was carried out as carefully as possible to avoid excessive bleeding and tissue damage. Moreover, the sponge chitosan and chitosan membranes (Medicarb AB, Bromma, Sweden) was implanted as described above, rats that were injected or AF-16, or filler.

When checking after 10 days it was found that biointegrated implants was excellent in those animals that received AF-16, both at the macroscopic evaluation, and evaluation by light microscopy of thin colored stripes. Along the edges of the implants was less inflammatory cells and macrophages, and it was found that fibroblasts infiltrated in implantirovannoi sponge extracted from the stomach is s, treated with AF-16. In addition, healing of skin wounds in rats, which were injected AF-16, was more complete with less reactive changes compared to those who were injected filler. It is known that the signal transmission, which leads to reactive tissue changes, includes signal transfer through lipid rafts and caveola cells in the damaged tissue.

Example 7

Experiments were carried out to investigate whether the introduction of the AF-16 in ischemic reaction in the internal organ of the body.

Shot adult rats carried osmotic Meninas (Alzet type 2001; filling volume ~235 μl; ~1 mm/h; pre-launched; filling solution contains 20 mg/ml AF-16, dissolved in the FSB, with the addition of 15% ethanol), implanted subcutaneously on the back. In addition, 2 mg AF-16 was administered via intramuscular injection immediately after implantation of the pumps. Ischemia was induced unilaterally in the left kidney by impeding blood flow through the renal artery for 40 minutes followed by recycling. The right kidney was surgically removed at commit time. Then the wound was closed, and animals were given pain medication, but did not receive any additional treatment. In parallel, other rats that received the filler, but not the peptide was subjected to ischemia in the left kidney and right kidney were removed. Repatriable shot again or 4, or after 7 days. The kidney was separated, examined, measured and weighed and fixed in buffered formalin solution for further study using light microscopy.

Experiments have shown that ischemic and reactive changes in the kidney were the most significant in the proximal tubules. In those who were treated with AF-16, was observed reduced hemorrhage and necrosis.

Example 8

Experiments were carried out with neoplastic tumors to explore, it affects their growth in the use of AF-16.

Shot adult rats carried osmotic Meninas (Alzet type 2001; filling volume ~235 μl; ~1 mm/h; pre-launched; filling solution contains 20 mg/ml AF-16, dissolved in the FSB, with the addition of 15% ethanol), implanted subcutaneously on the back. In addition, 2 mg AF-16 was administered via intramuscular injection immediately after implantation of the pumps. The pumps were replaced with a new one once a week. Different tumors induced chemically or transplanted, studied in relation to the effect of the introduction of the AF-16, compared with the use of filler.

Those rats that received a carcinogenic chemical, developed smaller tumors of the mammary gland in the lower concentration after application of AF-16, compared with the use of filler. Reactive and inflammatory changes were also less than the visible. Were also reviewed additional experimental tumors, when using AF-16 was achieved beneficial effects compared to the filler.

Example 9

Experiments were carried out on rats with experimentally induced diabetes caused by exposure of streptozocin studied in relation to the fact, it affects them use AF-16.

Shot adult rats carried osmotic Meninas (Alzet type 2001; filling volume ~235 μl; ~1 mm/h; pre-launched; filling solution contains 20 mg/ml AF-16, dissolved in the FSB, with the addition of 15% ethanol), implanted subcutaneously on the back. For comparison to other rats were injected filler, but not a peptide. One dose of streptozocin (Sigma) were injected with rats and observed the appearance of glucose in the urine, measured using a commercial indicator strips, and the appearance of increased volume of urine.

The use of AF-16 reduced, in accordance with our preliminary results, the loss of glucose in urine and decreased urine volume. The level of glucose in the blood were lower after AF-16.

Thus, AF-16 exerts a beneficial effect on the development of symptoms in chemically induced diabetes mellitus.

Example 10

Experiments were carried out with AF, administered to people suffering from diabetes type II. Study b is lo blind to the physician and the patients. All patients had diabetes mellitus type II, and frequency of diseases other than diabetes, they all have been thoroughly researched in the University hospital in Lund in Sweden. Ethical permission was obtained from the regional Committee. One group x subjects gave AF, whereas x subjects in the second group for comparison was administered the same number of control material. After 12 weeks were collected and analyzed blood samples.

The observed result was that HbA1c levels declined significantly (p≥0.05), and 0.2 units of, for those who have introduced the AF compared to those who entered the inactive solution. This means that the level of glucose in the blood of subjects treated with AF, remained at a lower level and better controlled compared with the parameters from those who were in the comparison group (placebo). Thus, a beneficial effect in relation to the outcome of diabetes mellitus type II achieved for those subjects who received AF.

Example 11

Oscillating changes in the concentration of calcium ion constitute a very important part of the signal device in many types of cells, such as neurons and astrocytes, which are dominant in the brain, spinal cord and retina. Moreover, it has been described that endocrine cells, such as pancreatic β-cells, are the pulse of the regulating insulin secretion. Also it is shown that pancreatic endocrine cells are associated ion changes when, for example, the seizure of glucose. Suppose that the weak violation of the oscillations of calcium ions in astroglial cells violate the regulation, for example, extracellular glutamate, which in itself can lead to local microglial activation with production of proinflammatory cytokines, swelling of astrocytes and, consequently swelling, reduced extracellular space and eventually to the destruction of the brain. In this state, a decrease as the release of glutamate and neuronal transmission. Presumably, such a reduced transmission and impaired activity in the brain is correlated, it is likely that dysfunction of the nervous system and abnormal neurological activity and activity.

Experiments were carried out to illustrate the oscillations of ions of calcium in astrocytes in the nervous tissue. Oscillations cause using the stimulator, such as histamine and/or monoaminergic transmitter, and determine the control activity with specific proteins, such as AF. The local concentration of calcium ions is determined using compounds, calcium-binding, the fluorescence of which varies quantitatively and qualitatively with the concentration of vnutri mocnych calcium ions. Used fluorescent microscopy and confocal scanning microscopy of astrocytes and neurons, mainly grown in vitro. Then used electrophysiological methods. To the cells was added a certain concentration of AF, in order thereby to determine the impact on system ionic carriers located in the lipid rafts in cells.

In addition, various types of muscle cells and connective tissue cells similarly analyzed for the distribution, frequency and amplitude of oscillations of calcium ions in the cytoplasm immediately before and after adding AF and related compounds.

As you can see on a typical figure 1, the observation of oscillations of calcium in the cell, tissue and/or body really allows you to observe the dysfunctions such signaling processes and assist in the normalization. Through this it becomes possible treatment of disorders and diseases.

Example 12

Aquaporins are a family of proteins that are integrated into the membrane and expressed in all living cells and organisms. The main function of aquaporins is to control the flow of water into and out of cells, i.e. between the cytoplasm and the extracellular environment. Each type of cell, tissue and organ has its own specific set of aquaporins with features u is adelene characteristics. Functional aquaporins mainly accumulate in lipid rafts and caveolin, whereas subunit and dissociatively aquaporins complexes can be observed outside of these membrane components.

Experiments on the brain and spinal cord showed that the expression and distribution of aquaporin, mainly aquaporin-1 and aquaporin-4, changes under the influence of ischemia or deformation and mechanical stress. Moreover, infection of the brain and spinal cord, such as encephalitis, the pattern and intensity of these two brain changes, as shown by immunohistochemistry and immunochemistry. The interaction between, for example, neurons and supporting glial cells and vascular structures, apparently, is confusing and complex, but important.

Conducted experiments, showing in more detail the results of injuries, infections and other pathological conditions. Courses of treatment with AF proteins, peptides, derivatives and homologues was administered topically and systemically, took in a single dose, or in multiple applications, or permanently, i.e. until the end of life of a subject of which they treat. The results characterize quantitatively and qualitatively is the impact on the structure and functions of different types of aquaporins in the body. Experiments demonstrate AF have a major impact on controlling and even normalization of the distribution, distribution and activity of aquaporins and complexes of ion channels, of which the latter interact with the first. Thus, on the basis of the achieved results can be new and important ways of treatment.

References

1. The use of antisecretory protein, which corresponds to the amino acid sequence of SEQ ID NO:6, its homologue or fragment contains the amino acid sequence of SEQ ID NO:4 that have equivalent activity, and/or pharmaceutically active salts, to pharmaceutical compositions
a. for the treatment and/or prevention of dysfunction of lipid rafts, receptors and/or kahweol in cell membranes, where this dysfunction is selected from the group consisting of vascular dysfunction, cardiovascular dysfunction, pulmonary dysfunction, diabetes, complications associated with diabetes, formation of scar tissue, repair of tissues and/or organs, reactive formation of excessive tissue repair and/or regeneration of the epithelial cell coverage, hyperplasia and/or hypertrophy of the cells and/or tissue, cardiomyopathy, pulmonary disorders and tumors and their complications, or
b. to improve biointegration implanted foreign material to the mammal.

2. Use p., where specified antisecretory protein selected from one or more protein containing the amino acid sequence of SEQ ID NO:1-6.

3. The use according to claim 1, where the specified pharmaceutical composition comprises two or more antisecretory protein.

4. The use according to claim 1, where the dysfunction is selected from the group consisting of abnormal, inadequate, Hypo - or hyperthyroidism.

5. The use according to claim 1, where the specified pharmaceutical composition further comprises a pharmaceutically acceptable excipient.

6. The use according to any one of claims 1 to 5, where specified pharmaceutical composition was prepared for intraocular, intranasal, oral administration, local application, subcutaneous and/or system administration.

7. The use according to any one of claims 1 to 5, where specified pharmaceutical composition was prepared for introduction in the form of a spray, aerosol, nebulizer or by spray.

8. The use according to any one of claims 1 to 5, where the pharmaceutical composition and/or health food get for systemic injections into the blood at a dosage of from 0.1 μg to 10 mg per dose per kg body weight per day, preferably 1-1000 μg per admission per kg of body weight per day.

9. The use of claim 8, where the specified introduction is carried out or as a single dose, or as multiple daily doses.

10. The use according to claim 6, where the specified pharmaceutical what kind of composition was prepared for administration in the form of spray, aerosol nebulizer or by spray.

11. The use according to claim 7, where the specified introduction is carried out or as a single dose, or as multiple daily doses.

12. The use of antisecretory protein, homologue and/or fragment that has equivalent activity, and/or pharmaceutically active salts, for the treatment and/or prevention of dysfunction of lipid rafts, receptors and/or kahweol in cell membranes, where this dysfunction is selected from the group consisting of vascular dysfunction, cardiovascular dysfunction, pulmonary dysfunction, diabetes, complications associated with diabetes, formation of scar tissue, repair of tissues and organs, reactive formation of excessive tissue repair and/or regeneration of the epithelial cell coverage, hyperplasia and/or hypertrophy of cells and/or tissue, cardiomyopathy, pulmonary disorders and tumors and their complications, or to improve biointegration implanted foreign material to a mammal,
where specified antisecretory protein consists of a sequence of the following formula
X1-V-C-X2-X3-K-X4-R-X5,
where X1 represents I, amino acids 1-35 of the sequence SEQ ID NO:6, or is absent, x2 represents H, R or K, X3 is a, S or L, X4 is T or A, X5 before the hat is amino acids 43-46, 43-51, 43-80 or 43-163 sequence SEQ ID NO:6 or missing.



 

Same patents:

FIELD: medicine.

SUBSTANCE: tripeptides Ile-Pro-Pro (IPP) and/or Val-Pro-Pro (VPP) are applicable for preparing a product removing arterial stiffness and for preparing a product improving vascular elasticity. They are made of casein or casein-containing starting material by fermentation with the strain Lactobacillus helveticus DSM 13137 or Lactobacillus helveticus DSM 17754 and recovery by concentration and nanofiltration. The product contains a ferment composition containing the casein-recovered peptides Ile-Pro-Pro and/or Val-Pro-Pro, and living lactic acid bacteria. For the purpose of removing arterial stiffness, the product containing Ile-Pro-Pro and/or Val-Pro-Pro is introduced into an individual. For the purpose of improving vascular elasticity, the product containing Ile-Pro-Pro and/or Val-Pro-Pro is introduced into an individual with using said biologically active peptides.

EFFECT: invention promotes prevention and normalisation of endothelial dysfunction, improves vascular elasticity and removes arterial stiffness.

12 cl, 3 tbl, 5 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention relates to medicine, namely pharmacology in cardiology, and may be used for preparing herbal teas for treating arterial hypertension. Herbal tea for treating adolescents suffering sympathicotonic vasomotor dyscrasia and degree 1 and 2 arterial hypertension contains dry ground herbal raw material. quinquelobate motherwort herb, red haw, melissa herb, Baikal woundwort herb, Baikal skullcap roots in certain proportions. The tea is presented in the form of an aqueous infusion.

EFFECT: herbal tea is effective in treating arterial hypertension in patients of various age groups including adolescent patients.

2 cl

FIELD: chemistry.

SUBSTANCE: disclosed are dodecapeptides of general formula X-Arg(NGY)-Pro-Arg-Leu-Ser-His-Lys-Gly-Pro-Nle-Pro-Phe-Z, where X=CH3, Y=H, Z-OH for compound (II), X=CH3, Y=H, Z=NH2 for compound (III), X=H, Y=NO2, Z=NH2 for compound (IV), having cardioprotective properties.

EFFECT: disclosed peptides can be used as cardioprotective agents in cardiology for cardiovascular disease therapy.

2 tbl, 4 ex, 1 dwg

FIELD: chemistry.

SUBSTANCE: present invention relates to novel 6-substituted isoquinolone derivatives of formula

or , or stereoisomeric forms and/or pharmaceutically acceptable salts thereof, where R2 denotes H or (C1-C6)alkyl; R3, R4 and R5 denote H; R6 and R6' independently denote H, (C1-C8)alkyl, (C1-C6)alkylene-R', (C1-C6)alkylene-C(O)O-(C1-C6)alkyl, C(O)-(C1-C6)alkylene-R', or R6 and R6', together with a N atom with which they are bonded form a (C5-C6)heterocyclyl group in which one or more carbon atoms can be substituted with 1, 2 or 3 nitrogen atoms, 1 or 2 oxygen atoms or a combination of different heteroatoms; R7 denotes H, halogen, (C1-C6)alkyl; R8 denotes H; n equals 1; m equals 1, 2, 3, 4 or 5, and L denotes O or O-(C1-C6)alkylene; where, R' denotes (C3-C8)cycloalkyl, (C5-C10)heterocyclyl, (C6-C10)aryl; where in residues R6 and R6' alkyl or alkylene can optionally be substituted one or more times with COOH groups; and where in residues R6 and R6' (C6-C10)aryl and (C5-C10)heterocyclyl are unsubstituted or substituted one or more times with suitable groups independently selected from a group comprising CONH2 and (C1-C6)alkyl; where if m equals 3, R6 cannot denote H; where if m equals 3 and R6 denotes (C1-C8)alkyl, then the alkyl is substituted once or more times, preferably one to three times, with a COOH group. The invention also relates to use of the compound of formula (I) and a medicinal agent based on the disclosed compounds.

EFFECT: novel isoquinolone derivatives which inhibit Rho-kinase and/or Rho-kinase mediated phosphorylation of the myosin light-chain phosphate.

31 cl, 6 tbl

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to medicine, more specifically to a new chemical compound, a 3-(2,2,2-trimethylhydrazinium)propionate-5-nicotinate hydroxide 3-(2,2,2-trimethylhydrazinium)potassium propionate derivative, (CH3)3N+HCH2CH2COOKRCOO- wherein exhibiting antiischemic activity.

EFFECT: what is produced and described is the new compound which may be effective as an agent showing antiischemic activity.

1 cl, 1 ex, 1 tbl

FIELD: chemistry.

SUBSTANCE: invention relates to medicine, specifically a novel chemical compound - a 3-(2,2,2-trimethylhydrazinium) propionate derivative - potassium glycinate 3-(2,2,2-trimethylhydrazinium) propionate (CH3)3N+NHCH2CH2COOKRCOO-, where , having antiischemic activity.

EFFECT: obtaining a compound with antiischemic activity.

1 cl, 1 ex, 1 tbl

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to pharmaceutics. A pharmaceutical granule is spherical or spheroidal and has volume density 0.6-1.3 g/ml and disintegration 0.5-5 minutes. Active pharmaceutical ingredients are preparations of Traditional Chinese Medicine, herbal preparations or their extracts with a nucleus made of an extract of Traditional Chinese Medicine or herbal preparations, and a pharmaceutically acceptable carrier. The content of the pharmaceutically acceptable carrier with respect to total granule weight makes 10-60 wt %. The granule diameter can be equal to 700-1500 mcm. The granule nucleus diameter can be equal to 200-750 mcm. The granule can be also coated with a layer of 2-5 wt % with respect of total granule weight. A method for making the granules consists in introducing initial granules into a layer of a fluidised material as an excipient; the active pharmaceutical ingredients are prepared in the form of a suspension or a solution of the viscosity controlled within 6.0-9.8 MPa·s with using a viscosity-control agent; then they are sprayed over a surface of the initial granule for making a finished granule.

EFFECT: granules possess fast disintegration, contain a small amount of the carrier and contain a low single dose.

12 cl, 5 ex

FIELD: medicine.

SUBSTANCE: method implies introducing into a laboratory animal a phytococktail which contains mixed 70% alcoholate of snowdone rose, common licorice, eleuterococcus, horseheal taken in proportions 2:2:1:1, and dissolved in distilled water in proportions 1:50. The phytococktail is introduced in rats through probes in amount 1 ml per 150-200 g of animal's weight from 16.00 to 18.00 o'clock for 15 days.

EFFECT: higher vascular thromboresistance ensured by higher endothelial aggregation ability mediated by activation of antioxidant and anti-inflammatory effects of the vascular epithelium.

1 tbl

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to new compounds of general formula 1, or their pharmaceutically acceptable salts showing the properties of incretin secretagogues, preferentially the properties of a bile acid receptor TGR5 agonist. The compounds are applicable for treating metabolic diseases associated with glucose metabolism, such as diabetes, obesity, metabolic syndrome, etc. In formula 1 R1, R2 and R3 independently represent a cyclic system substitute specified in: hydrogen, C1-C3alkyl, halogen, a trifluoromethyl group, C1-C3alkoxy, a cyano group, a trifluoromethoxy group; an amino group substituted by C1-C3alkyl; or two radicals R3, found at carbon neighbours in a benzene ring, together with the benzene ring bound therewith form 3,4-methylene dioxyphenyl; R4 represents hydrogen, C1-C5alkyl, a carboxyl group, C1-C3alkoxycarbonyl or an amide group CONHR5; R5 is an optionally substituted by C1-C3alkyl, C5-C6cycloalkyl optionally substituted by phenyl, benzyl, pyridyl; X and Y represent two hydrogen atoms or an oxygen atom, provided Y=O, then X=2H, provided Y=2H, then X=O or X=Y=2H; the sign (N) shows the possibility of bioisosteric substitution of the benzene ring by the pyridine, pyrimidine, pyridazine, triazine or pyrazine ones.

EFFECT: preparing the pharmaceutical composition and the combined drugs with the use of the compounds of formula 1 or the based pharmaceutical composition and a protein kinase DPP-IV inhibitor specified in Vildagliptin or Sitagliptin, and/or an endogenous bile acid or mied bile acid secretagogues.

53 cl, 7 dwg, 8 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to compounds of formula I, as well as to their physiologically acceptable salts wherein: X means NH; R1 means (C1-C6)-alkyl; R2 means OH; R2' means H; R5' means (C1-C6)-alkylene-O-S(O)2-R6; R3, R3', R4, R4' and R5 independently mean H, OH, (C1-C6)-alkylene-O-S(O)p-R6, O-(CH2)m-phenyl; at least one of the radicals R3, R3', R4, R4' and R5 has the value -O-(CH2)m-phenyl; R6 means OH; m=1; p=2.

EFFECT: compounds can find application in medicine, eg as lipid-lowering agents.

FIELD: medicine.

SUBSTANCE: invention relates to medicine, namely to oncology and deals with method of treatment in case of urinary bladder cancer. Combined treatment with application of neoadjuvant system polychemical therapy by the following scheme: 1 day -5-fluoruracyl, 2 day - cyclophosphane+vinblastine, 14 day - methotrexate, 15 day - doxorubicin, 16 day -cisplatin, and adjuvant intrabladder chemical therapy with preparation doxorubicin during 2 years, on the first day after operation doxorubicin 40 mg is instilled, then 4 instillations of doxorubicin in dose 40 mg with 7 day interval, then doxorubicin 40 mg is instilled with 30 day interval during 11 months, after that during the second year instillations of doxorubicin in dose 20 mg are performed after 1 month.

EFFECT: invention ensures improvement of results of organ-preserving treatment by reduction of number of urinary bladder cancer recurrences.

1 ex

FIELD: medicine.

SUBSTANCE: there are offered: recovered polynucleotides and polypeptides for binding human EGFR as a part of an antibody, a vector and a host cell for antibody expression, a method for producing an anti-EGFR antibody and an antibody fragment, the anti-EGFR antibody and the antibody fragment. There are discussed a composition containing the anti-EGFR antibody or its fragment, as well as applying the antibody and its fragment for treating EGFR-associated disorders. Besides, there are described versions of glycosylation of the anti-EGFR antibody or its fragment.

EFFECT: invention can find further application in therapy of various EGFR-mediated diseases.

30 cl, 6 ex, 32 dwg, 39 tbl

FIELD: chemistry.

SUBSTANCE: invention relates to cytotoxic compounds of directed action and methods for therapeutic use thereof in treating neoplasm and other diseases.

EFFECT: high treatment efficiency.

9 cl, 12 tbl, 31 ex, 3 dwg

FIELD: medicine, pharmaceutics.

SUBSTANCE: present invention refers to {1-methyl-5-[2-(5-trifluoromethyl-1H-imidazol-2-yl)pyridin-4-yloxy]-1H-benzimidazol-2-yl}(4-trifluoromethylphenyl)amine salt. Also, the invention refers to a pharmaceutical composition based on the declared salt, to a method for preparing said pharmaceutical composition and to a method of treating cancer and/or angiogenesis based on the use of the declared salt.

EFFECT: there are prepared new {1-methyl-5-[2-(5-trifluoromethyl-1H-imidazol-2-yl)pyridine-4-yloxy]-1H-benzimidazol-2-yl}(4-trifluoromethylphenyl)amine salts showing effective biological properties.

42 cl, 12 dwg, 10 tbl, 7 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: present invention refers to new (2R)-2-phenylpropanamide derivatives of formula (I) or their pharmaceutically acceptable salts having an ability to inhibit CXCL1-induced chemiotaxis of human multiform nuclear leucocytes (PMNs), to a based pharmaceutical composition, to applying them for preparing a drug and a methods for making them. wherein R is specified from H, C1-C5-alkyl, C3-C6-cycloalkyl, thiazol substituted by trifluoromethyl, a residue of formula -CH2-CH2-O-(CH2-CH2O)nR", wherein R" represents H or C1-C5-alkyl, n is equal to an integer within 0 to 2, or R together with an NH-group whereto attached is a radical group of primary amides of natural amino acid such as (2S)-2-aminopropanamide; R' is specified in linear or branched C1-C5-alkyl, C3-C6-cycloalkyl, trifluoromethyl, phenyl, optionally substituted by a group specified in halogen, C1-C5-alkyl, C1-C5-alkoxy and trifluoromethyl; unsubstituted benzyl; thiophen.

EFFECT: preparing the new pharmaceutical composition.

9 cl, 1 tbl

FIELD: medicine.

SUBSTANCE: present invention offers a composition for treating tumours containing: (1) allogenic or xenogenic tumour cells; (2) a lysate of syngeneic tumour cell and (3) a pharmaceutically acceptable excipient wherein allogenic or xenogenic cells are tumour cells of the same or similar class as patient's tumour cells.

EFFECT: invention enables selecting a type of tumour cells/tissue for transplantation of tumour/cancer tissue/cell of one individual to another, enables avoiding rejection of the transplanted tissue/cell, and also promotes higher immunological alarm with respect to peptides being common for these tumour/cancer growths and other tumours having similar peptides.

8 cl, 1 ex, 9 dwg

FIELD: medicine.

SUBSTANCE: group of inventions refers to medicine, and concerns a combination of a proteasome inhibitor and a histone deacetylase inhibitor for tumour cell growth/proliferation inhibition. What is offered is a new combination wherein the proteasome inhibitor is Bortezomib, while the histone deacetylase inhibitor is R3 06465 . What is also presented is applying said combination for preparing a drug for tumour cell growth/proliferation inhibition, including haematologic cancer cells.

EFFECT: group of inventions provides the new combination with a therapeutic potential showing a synergetic effect of human haematologic cancer cell proliferation inhibition.

5 cl, 8 tbl, 7 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to chemical-pharmaceutical industry, particularly a technology of a new dosage form of 3,31-diindolylmethane. A method consists in mixing 3,31-diindolylmethane and a liquid fatty excipient and adding a wedding agent. In mixing, the prepared mixture is heated to 35-45°C to prepare a solution. Then the prepared solution is cooled to 21-23°C to fill gelatine capsules in the following proportions, g/capsule: 3,31-diindolylmethane - 0.01-0.1, the liquid fatty excipient - 0.05-0.1, the wedding agent 0.4-0.45. The prepared drug in the form of soft gelatine capsules containing 3,31-diindolylmethane in the form of the solution enables higher bioavailability of the active substance.

EFFECT: prepared drug is non-toxic, safe in prolonged introduction, has no local irritant action, no toxic effect on the immune and reproductive systems.

4 cl, 4 dwg, 7 tbl, 6 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: present invention refers to pharmaceutics and medicine, and concerns a combination showing IAP inhibitor and FLT3 inhibitor activity containing (S)-N-((S)-1-cyclohexyl-2-{(S)-2-[4-(4-fluorobenzoyl)thiazol-2-yl]pyrrolidin-1-yl}-2-oxoethyl)-2-methylaminopropionamide, the compound of formula

, a method of treating of acute myeloid leukemia, applying said combination for treating acute myeloid leukemia and a commercial packing containing said combination.

EFFECT: composition shows high efficacy.

9 cl, 16 dwg, 16 ex

FIELD: medicine.

SUBSTANCE: chimeric peptide VP-22_p16INK4a contains two sequences of amino acids one of which comprises a cycline kinase inhibitor in the form of an active fragment of p16INK4a as a therapeutic agent, while another one comprises the peptide VP22 of herpex simplex virus as a transport agent for transporting the cycline kinase inhibitor inside the target cells.

EFFECT: invention enables extending the range of application of said chimeric peptide.

4 dwg, 1 ex

FIELD: chemistry.

SUBSTANCE: invention relates to compounds of formula I , and pharmaceutically acceptable salts thereof, where L denotes O, S, or CH2; Y denotes N or CH; Z denotes CR3; G denotes CH; R1 denotes a heteroaryl ring of formula , where D1 denotes S, O; D2 denotes N or CR12; D3 denotes CR12; R2 denotes (C6-C10)-aryl; 5-9-member mono- or bicyclic heteroaryl with 1 or 2 heteroatoms independently selected from N or S; a saturated or partially saturated (C3-C7)-cycloalkyl; or a saturated 5-6-member heteocyclyl with 1 heteroatom selected from N, where said aryl, heteroaryl, cycloalkyl and heterocyclyl are optionally substituted with one or two groups independently selected from (C1-C6)-alkyl, F, Cl, Br, CF3, CN, NO2, OR6, C(-O)R6, C(=O)OR6, C(=O)NR6R7, saturated 6-member heterocyclyl with 2 heteroatoms independently selected from N or O, and S(O)2R6, and where said alkyl is optionally substituted with one -OR8 group; R3 denotes H; (C1-C6)-alkyl; (C2-C6)-alkenyl; Cl; Br; OR6; SR6; phenyl; or a 6-member heteroaryl with 1 heteroatom selected from N, where said alkyl and alkenyl are optionally substituted with one group selected from C(=O)OR8, -OR8, -NR8R9; or a saturated 6-member heterocyclyl with 1 heteroatom selected from N or O.

EFFECT: disclosed compounds are used in treating and preventing diseases mediated by insufficient level of glucokinase activity, such as sugar diabetes.

16 cl, 479 ex

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