Using alginate oligomers in biofilm control

FIELD: medicine, pharmaceutics.

SUBSTANCE: group of inventions refers to biotechnology. What is presented is an in vitro method for controlling a biofilm containing gram-negative bacteria, gram-positive bacteria or yeast, involving the contact of the above biofilm and an alginate oligomer. The alginate oligomer has an average molecular weight of less than 20000 Da and at least 80% of G residues, particularly α-L-guluronic acid. For the purpose of controlling the biofilm, including a biofilm infection, the above alginate oligomer can be also used as a part of a kit or a cleaning composition combined with other ingredients, as well as an abiotic surface coating.

EFFECT: above alginate oligomer can be used for preparing a therapeutic agent for using in treating or preventing the biofilm infection in an individual.

88 cl, 10 dwg, 7 tbl, 15 ex

 

The present invention relates to a method of combating biofilms. In particular, the present invention relates to the use of a specific class of alginates and, in particular, some alginate oligomers in combating biofilms on biotic and abiotic surfaces. Thus, offers both medical and non-medical applications and ways to combat biofilm infections or for combating biofilm formation on inanimate surfaces, for example for the purposes of disinfection and cleaning. The invention is based on the unexpected discovery that some of alginate oligomers can interact with the biofilm and to touch it.

In General terms biofilm is an aggregate or community of microorganisms surrounded by a matrix of extracellular polymers (also known in the art as glycocalyx). These extracellular polymers are polysaccharides, particularly polysaccharides produced by organisms, but they can also contain other biopolymers. Biofilm is normally attached to the surface, which may be inert or living, but in addition, it was observed that biofilms can be formed from microorganisms that are attached to each other or to any surface section. Thus, in General the m case, the biofilm is characterized as highly organized multicellular community of microorganisms, prisoners in the extracellular polymeric matrix, usually polysaccharide matrix, or surrounded by it, and usually in close contact with the surface or boundary surface. This pattern of growth is protective for microorganisms and makes difficult their removal or destruction (for example, as further discussed below, the durability or resistance to antimicrobial agents, or immune defenses of the host, or the mechanisms of purification). It is considered, according to the present invention that alginate oligomers can interact with the polymer matrix of the biofilm, and thus to weaken the biofilm. As further discussed below, biofilms cause significant commercial, industrial and medical problems, in terms of infections, infestations, contamination and damage, and thus, the present invention provides a significant advantage to enable or facilitate the control of such biofilms, including reduction or prevention of their formation, and their transfer to the state is more sensitive to the removal or reduction of, for example, are more sensitive to the action of antimicrobial agents (including disinfectants or antibiotics) or, in the case of infection, the immune response of the infected host. Thus, it can be enhanced the effectiveness of PR is tuomikoski as therapeutic agents, and non-therapeutic and including, in particular, antibiotics.

Biofilms are found throughout a wide variety of surfaces or interfaces (for example, the division surface water/solid and water/gas (e.g., water/air)), if there are conditions conducive to microbial colonization. Essentially, the biofilm can be formed wherever there are microorganisms and the surface boundary or surface, particularly the surface exposed to water or humidity, and biofilm currently recognized as the natural state of microbial growth on these surfaces or the surfaces of the partition. In fact, as noted above, biofilm is a complex and orderly arrangement of microbial colonies on the surface or on the surface of the partition, which can be, in particular, in the presence of water or humidity. The organization of such colonies is the result of the ability of microorganisms to produce an organized extracellular matrix, in which the embedded cells. Such a matrix is formed from biopolymers produced by microorganisms, and the basic polymer is a polysaccharide.

The microorganisms in the biofilm community are properties at the cellular level (phenotype)that do not have their planktonic (obognavshie) equivalents. Indeed, suppose that the microorganisms in the biofilm are very different from free-floating planktonic cells. Additional differences can also be observed at the community level, and they relate to the effects of the extracellular matrix. Perhaps the most notable is the commonly observed phenomenon that the microorganisms in the biofilm environment do not exhibit the same sensitivity to antimicrobial agents such as antibiotics, antifungal and antibacterial means, and immune defenses of the host or clearance mechanisms. It is believed that this stability is due to the barrier effect of the extracellular matrix and/or phenotypic changes in the microorganisms. For example, when the formation of biofilms, more antibodies are not associated with microorganisms (e.g. bacteria) in the biofilm. Experiments have shown that antibodies tightly cover the biofilm on the outside, but they are not present in the biofilm. Studies of leukocyte activity against biofilms showed similar results. Toxicobrain may also differ in planktonic organisms and its equivalent in biofilm colonies, suggesting about phenotypic changes in microorganisms. In addition, suppose that the microorganisms in biofilms can grow slower and the slower pohlad the TB antimicrobial agents.

Biofilms are easily formed on the water surrounding surfaces and formed of microbial colony on any surface exposed to water (any "wet" the surface), will be almost sure to exist in the form of biofilm structure. Besides, now it becomes obvious and increasingly documented that biofilms can also form in the case of microbial infections, i.e. inside an infected host or on it. Thus the formation of biofilms can also be "physiological" or "biological" surface, i.e. on the living or biotic surfaces, or surfaces on the infected organism is the owner or in it (for example, human or animal subject, other than man), for example on the inner or outer surface of the body or tissue. Increasingly, it is believed that this biofilm (or infection) in the tissues of the body participates in various infectious diseases, including, for example, congenital endocarditis heart valve (mitral, aortic, right atrioventricularis, pulmonary heart valves), acute otitis media (middle ear), chronic bacterial prostatitis (prostate), cystic fibrosis (lung), pneumonia (respiratory tract), periodontitis (tissue that supports the teeth, such as gums, periodontal ligament, alveolar bone). Of course, both of these biofilm niches are present at the implantation of medical devices, and the formation of biofilms on implanted such ("embedded") devices may cause clinical problems with infection in these areas, such as the artificial valve endocarditis and infection associated with a device, such as intrauterine devices, contact lenses, prostheses (e.g., prosthetic joints), and in areas catheterization, for example in the presence of a Central venous or urinary catheters.

A significant problem and the risk, the presence of these biofilm infections is that the microorganisms (or, more specifically micro-colonies) can break or detach from the biofilm and to invade other tissues, including much of the circulation. Such circulating microorganisms from biofilms can cause additional infections and cause significant clinical problems, especially in view of the fact that separated circulating microorganisms can have all the stability characteristics of the parent community.

Biofilm infection usually develops gradually, and the obvious symptoms may develop slowly. However, after the formation of biofilms, as noted above, difficult to purify, and biofilm infection is usually the C is stable and rarely decomposed by the host immune system or immune mechanisms even in individuals with a healthy innate and adaptive immune responses. Active master answers can actually be detrimental, such as cell-mediated immunity (e.g., invasive neutrophils) can cause collateral damage to adjacent healthy tissue of the host. Biofilm infections respond only briefly to treatment with antibiotics. Thus, while planktonic microbial cells can be eliminated by using antibodies or phagocytes and they are more sensitive to antimicrobial agents, microorganisms in biofilms are resistant to antibodies, phagocytes and antimicrobial agents. Phagocytes attach to the biofilm, but phagocytosis is not completed. Phagocytic enzymes, however, are released and can damage the tissue around the biofilm. Planktonic bacteria can be released from the biofilm, and this release may cause them to spread and acute infection in the surrounding tissues.

The surface of the body or tissues that are lost or damaged (for example, necrotic or inflamed), are particularly susceptible to biofilm infection. Wounds are susceptible to infection, and wounds, which, for a short time not healed, may be the formation of biofilms. Wounds are an ideal environment for the formation of biofilms due to their susceptibility to bacterial colonization, DOS is upnote substrate and a surface for attachment of the biofilm. Problematically, wound infection often additionally delays the healing process and thus makes the wound more susceptible to the formation of biofilms and the resulting infection. Wound healing is delayed (so-called chronic wounds), are objects of special attention in relation to the formation of biofilms. A chronic wound is in an inflammatory state, with elevated levels of proinflammatory cytokines. The effect of these cytokines is the creation of an area of accumulation of immune cells (neutrophils and macrophages). If such a system because something is delayed (as in chronic wounds), bacteria or other microorganisms have time to attach to the surface and enter into the biofilm stage of growth. Growing evidence that both chronic and short-term wounds may be areas of biofilm infection, with evidence of the presence of diverse microbial communities or populations in wounds, especially chronic wounds, including anaerobic bacteria in chronic wounds. Chronic wound infections have two important common characteristic with other biofilm infections: a sustainable infection, which is not eliminated by the immune system of the host, even in persons with healthy innate and adaptive immune responses, and increased stable is epostl to systemic and local antimicrobial agents. Thus, biofilm infections very difficult to treat and biofilm infection is very difficult to eliminate. Frequent readjustment is one of the most clinically effective treatments for the relief of chronic wounds. It is an effective treatment partly in view of the fact that it is physically removes biofilm from the wound. This is similar in principle to the destruction of infections from colonized by biofilms medical devices introduced (e.g., catheters), where antibiotic therapy is ineffective, the most effective approach is the removal or replacement of an infected biofilm device.

Chronic wounds represent a major health problem worldwide and require significant clinical resources. Three main types of chronic wounds are diabetic foot ulcers, venous ulcers of the leg and decubitus ulcers, although other wounds, including surgical wounds may become chronic. Care for these wounds requires great expense and patience, and as a consequence effective antiviolence treatment or virtually any treatment that promotes or facilitates the treatment of biofilms and thus accelerates or facilitates wound healing, will be of great value.

In a broader sense, this Shiro is the first spread of biofilms and medical, environmental, industrial, or other commercial issues that cause them any ways to improve or endowment struggle with the biofilms will be very important both from a clinical and from a commercial point of view.

Therefore, there is a need for new ways of dealing with biofilms in clinical and industrial or commercial situations, and the present invention is directed to satisfying this need.

In particular, and as noted above, it was found that a specific class of alginates, namely some alginate oligomers, is effective as antiviolence agents. Alginate oligomers can interact with extracellular polymers in biofilms and thereby weaken it, making possible or facilitating its removal or destruction (or destruction), and/or facilitating access to the biofilm antimicrobial agents, thereby enhancing their effectiveness against biofilms. Thus, according to the present invention proposes a new method or means of combat biofilm, including the use of alginate oligomers.

Alginates are linear polymers (1-4)-linked β-D-mannurone acid (M) and/or C5-epimer, α-L-guluronic acid (G). Primary structure of alginates may vary considerably. the action M and G can be arranged in the form of homopolymer blocks neighboring M or G residue, in blocks alternating M and G residues and in the intervals of such block structures can discover a single M - or G - residues. The alginate molecule may contain some or all of such structures, and such structures can be unevenly distributed throughout the polymer. In the limiting case, there is a homopolymer guluronic acid (polyglutamate) or a homopolymer mannurone acid (polymannuronate).

Alginates have been isolated from marine brown algae (for example some species of Durvillea, Lessonia and Laminaria) and bacteria such as Pseudomonas aeruginosa and Azotobacter vinelandii. Others found (for example, Pseudomonas fluorescens, Pseudomonas putida and Pseudomonas mendocina) retain the genetic ability to produce alginates, but in nature they do not produce detectable levels of alginate. Through mutations such neproducyruth found to encourage to develop a consistently large number of alginate.

Alginate is synthesized in the form of polymannuronate, and G-residues are formed under the action of epimers (especially C5-epimerase) on M-residues in the polymer. In the case of alginates extracted from algae, G-residues mainly organized in the form of G-blocks, as the enzymes involved in the biosynthesis of alginate in seaweed, mainly enter G in adjacent position with another G, thus turning plots M-residues in G-blocks. Pony is the use of such biosynthetic systems allows us to produce alginates with specific primary structures (WO 94/09124, Gimmestad, M. et al, Journal of Bacteriology, 2003, Vo1 185(12) 3515-3523 and WO 2004/011628).

Alginates are usually isolated from natural sources in the form of large high molecular weight polymers (for example, an average molecular weight in the range of from 300,000 to 500,000 Da). It is known, however, that such large alginate polymers may be destroyed or split, for example, by chemical or enzymatic hydrolysis to obtain alginate structures with lower molecular weight. Alginates are used in industrial scale, typically have an average molecular weight in the range from 100,000 to 300,000 Yes (i.e. such alginates are still considered large polymers), although alginates with an average molecular weight of approximately 35,000 used in pharmaceuticals.

It was found that alginate oligomers may affect the extracellular matrix of biofilms. Without wanting to be bound to any specific theory it is believed that this effect causes the destruction of the extracellular matrix of the biofilm, and it is, therefore, leads to physical destruction of biofilms. In addition, the destruction intensifies the impact on the microorganisms in the biofilm (or their immunogenic components such as LPS and peptidoglycans patterns) immune system of the infected host and/or any antimicrobial agents who used the ü or will be applied. In addition, the destruction reduces the close link between the extracellular matrix and microorganisms, and this increases the sensitivity of microorganisms to antimicrobial agents on the phenotypic level.

Thus, in the invention, a method of combating biofilm, including interaction specified biofilms with alginate oligomer.

As noted above, alginates are usually in the form of polymers with an average molecular weight of at least 35000 Yes, that is approximately lasts 175-190 Monomeric residues, although usually much higher, and alginate oligomer according to the present invention can be defined as a substance obtained by fractionation (i.e. smaller) alginate polymer, usually natural alginate. Alginate oligomer can be considered as alginate with an average molecular weight less than 35000 Yes (i.e. less than approximately 190 or less than 175 Monomeric residues), in particular alginate with an average molecular weight of less than 30,000 Da (i.e. less than approximately 175 or less than 150 Monomeric residues), more specifically with an average molecular weight of less than 25,000 or 20,000 Da (i.e. less than approximately 135 or 125 Monomeric residues, or less than about 110 or 100 Monomeric residues).

Alternatively, the oligomer contains mostly 2 or Bo is her unit or residue, and alginate oligomer for use in the invention typically contains from 2 to 100 monomer residues, preferably from 2 to 75, preferably from 2 to 50, more preferably from 2 to 40, 2 to 35, or from 2 to 30, i.e. alginate oligomer for use in the invention usually has an average molecular weight of from 350 to 20,000 Da, preferably from 350 to 15,000 Da, preferably from 350 to 10,000 and more preferably from 350 to 8000 Yes, from 350 to 7000 Da, or from 350 to 6000 Da.

Alternatively, the alginate oligomer may have a degree of polymerization (DP), or srednecenovogo degree of polymerization (DPn) from 2 to 100, preferably from 2 to 75, preferably from 2 to 50, more preferably from 2 to 40, 2 to 35, or from 2 to 30.

As stated above, usually formed biofilms on surfaces or interfaces, and biofilm, which is treated according to the present invention can be placed on any surface or surfaces of the partition. Thus, in the method according to the invention, the biofilm can be on any living or non-living (or biotic or abiotic) surface, i.e. any living surfaces or surfaces derived from living matter (such as dead or damaged tissue, such as necrotic tissue), (the term "live" is used in this description of the invention as including any living surface is or any surface, originating from living matter, in particular the living surface, which died), or any inert or inanimate surface (the surface which in the past was not alive or lively).

The term "bringing into contact" covers any means of delivery alginate oligomer to the biofilm, both directly and indirectly, and thus, any method of applying the alginate oligomer on the biofilm or the exposure of the biofilm to the impact of the alginate oligomer, for example the application of the alginate oligomer directly to the biofilm, or the introduction of alginate oligomer to a subject with a biofilm infection. Therefore, you should understand what is included both in vitro and in vivo methods.

More specifically, the biofilm will be in contact with an effective amount of alginate oligomer, more specifically with the amount of alginate oligomer effective to combat biofilm.

Alginate oligomer, as noted above, includes (or contains) residues or units of guluronate or guluronic acid (G), and/or mannuronate or mannurone acid (M). Alginate oligomer according to the invention preferably consists of the same or essentially the same (i.e. essentially consists of them) residues uronate/Bronevoy acid, more specifically the same or essentially the same G and/or M-residues. In the alternative is actively, in the alginate oligomer for use in the present invention at least 80%, more specifically at least 85, 90, 95 or 99% of the Monomeric residues may represent the remains of uronate/Bronevoy acid, or, more specifically, G - and/or M-residues. In other words, preferably the alginate oligomer does not contain other residues or units (e.g., other sacharine residues, or, more specifically, other remnants of Bronevoy acid/uronate).

Alginate oligomer preferably is a linear oligomer.

More specifically, in the preferred embodiment at least 30% of the Monomeric residues of the alginate oligomer are G-residues (i.e. guluronate or guluronate acid). In other words, alginate oligomer contains at least 30% residue of guluronate (or guluronic acid). Specific embodiments thus include alginate oligomers (for example, containing from 30 to 70% G-(guluronate) residues or from 70 to 100% G (guluronate) residues. Thus, a typical alginate oligomer for use in the present invention may contain at least 70% G-residues (i.e. at least 70% of the Monomeric residues of the alginate oligomer are G-residues).

Preferably at least 60%, more specifically at least 70% or 75%, even more is specifically at least 80, 85, 95 or 99% of the Monomeric residues are guluronate. In one embodiment of the alginate oligomer may be oligohaline (i.e. homologoues G or 100% G)

In another preferred embodiment described above alginates according to the invention have a primary structure, where most of G-residues are located in the so-called G-blocks, preferably at least 50%, more preferably at least 70 or 75% and most preferably at least 80, 85, 90 or 95% of the single G-residues are G-blocks. G-unit is a continuous sequence of at least two G residues, preferably at least 3 adjacent G residues, more preferably at least 4 or 5 adjacent G residues, most preferably at least 7 adjacent G residues.

In particular at least 90% of G-residues are connected by a link 1-4 with the other G-residue. More specifically at least 95%, more preferably at least 98% and most preferably at least 99% G-residues of alginate are connected by a link 1-4 with the other G-balance.

Alginate oligomer for use in the invention is preferably 3-35-dimensional, more preferably 3-28-dimensional, in particular 4-25-dimensional, especially 6-22-dimensional, in particular 8-20-dimensional, especially 10-15-dimensional, for example having the molecular is th weight in the range from 350 to 6400 Yes, or from 350 to 6000 Da, preferably from 550 to 5500 Yes, preferably from 750 to 5000 and particularly from 750 to 4500 Da.

It can be a single compound or may be a mixture of compounds, for example with an interval of degrees of polymerization. As noted above, the Monomeric residues in the alginate oligomer may be the same or different, and not everyone should carry electrically charged groups, although it is preferable that the majority (e.g. at least 60%, preferably at least 80%, more preferably at least 90%) were carrying them. Preferably, a substantial majority, for example at least 80%, more preferably at least 90% charged groups have the same polarity. In the alginate oligomer ratio of hydroxyl groups to the charged groups is preferably at least 2:1, more specifically at least 3:1.

Alginate oligomer for izobreteniyam to have a degree of polymerization (DP) or srednecenovogo degree of polymerization (DPn) 3-28, 4-25, 6-22, 8-20, or 10-15, or from 5 to 18, or from 7 to 15, or from 8 to 12, especially 10.

The distribution of molecular weight is preferably one in which not more than 5 mol.% have DP two times higher than the corresponding upper limit for DPn. Similarly, preferably, not more than 5 mol.% had a DP lower than the number in twice the smaller is its corresponding lower limit for DP n. Suitable alginate oligomers described in WO 2007/039754, WO 2007/039760 and WO 2008/125828, descriptions of which are expressly incorporated by reference into this specification in their entirety.

Typical suitable alginate oligomers have DPnin the range of 5-30, the fraction of guluronate/galacturonase (FGat least 0,80, the fraction of mannuronate (Fm) : not more than 0.20 and at least 95 mol.% with DP not more than 25.

In addition, appropriate alginate oligomers have srednecenovogo degree of polymerization in the range of 7-15 (preferably 8-12), the fraction of guluronate/galacturonase (FGat least 0,85 (preferably at least 0,90), the fraction of mannuronate (Fm) not more than 0.15, preferably not more than 0.10) and have at least 95 mol.% with a degree of polymerization less than 17, and preferably less than 14).

In addition, appropriate alginate oligomers have srednecenovogo degree of polymerization in the range of 5-18 (especially 7-15), the fraction of guluronate/galacturonase (FGat least 0,80 (preferably at least 0,85, especially at least 0,92), the fraction of mannuronate (Fm) not more than 0.20, preferably not more than 0.15, especially not more than 0.08) and have at least 95 mol.% with a degree of polymerization less than 20 (preferably less than 17).

In addition, appropriate alginate oligomers have srednecenovogo frame the polymerization in the range of 5-18, the fraction of guluronate/galacturonase (FGat least 0,92, the fraction of mannuronate (Fm) not more than 0.08 and have at least 95 mol.% with a degree of polymerization less than 20. In addition, appropriate alginate oligomers have srednecenovogo degree of polymerization in the range of 5-18 (preferably 7-15, more preferably 8 to 12, especially about 10), the fraction of guluronate/galacturonase (FGat least 0,80 (preferably at least to 0.85, more preferably at least about 0.90, in particular at least 0,92, especially at least 0,95), the fraction of mannuronate (Fm) : not more than 0.20, preferably not more than 0.15, more preferably not more than 0.10, in particular not more than 0.08, particularly not more than 0.05) and have at least 95 mol.% with a degree of polymerization less than 20 (preferably less than 17, more preferably less than 14).

In addition, appropriate alginate oligomers have srednecenovogo degree of polymerization in the range of 7-15 (preferably 8-12), the fraction of guluronate/galacturonase (FGat least 0,92 (preferably at least 0.95)is the fraction of mannuronate (Fm) not more than 0.08, preferably not more than 0.05) and have at least 95 mol.% with a degree of polymerization less than 17, and preferably less than 14).

In addition? suitable alginate oligomers have srednecenovogo degree of polymerization in which intervale 5-18, the fraction of guluronate/galacturonase (FGat least 0,80, the fraction of mannuronate (Fm) : not more than 0.20, and have at least 95 mol.% with a degree of polymerization less than 20.

In addition,appropriate alginate oligomers have srednecenovogo degree of polymerization in the range of 7-15, the fraction of guluronate/galacturonase (FGat least 0,85, the fraction of mannuronate (Fm) not more than 0.15, and have at least 95 mol.% with a degree of polymerization less than 17.

In addition, appropriate alginate oligomers have srednecenovogo degree of polymerization in the range of 7-15, the fraction of guluronate/galacturonase (FGat least 0,92, the fraction of mannuronate (Fm) not more than 0.08 and have at least 95 mol.% with a degree of polymerization less than 17.

Alginate oligomer usually incurs a charge and, therefore, the counterions for the alginate oligomer may be any physiologically acceptable ion, especially those which are usually used for charged medicinal substances, for example sodium, potassium, ammonium, chloride, mesilate, meglumin and so on. You can also use ions that contribute to the gelation of alginate, for example metal ions 2 groups.

Although alginate oligomer may be a synthetic material formed by polymerization of the appropriate number of residues gulur the Nata and mannuronate, alginate oligomers for use according to the invention can be conveniently acquire, produce or manufacture from natural sources, such as sources mentioned above, namely matter - the natural sources of alginate.

Cleavage of the polysaccharide to oligosaccharide to obtain alginate oligomer used in accordance with the present invention, can be performed using conventional methods of lysis polysaccharides, such as enzymatic cleavage and acid hydrolysis. Then the oligomers can be chromatographically separated from the products of destruction of polysaccharides using ion-exchange resin or by fractional precipitation or solubilization, or filtering. In the US 6121441 and WO 2008/125828, which is expressly incorporated by reference in this description of the invention in its entirety, describes a method suitable for obtaining alginate oligomers for use according to the invention. For more information and discussion can be found, for example, in "Handbooks of Hydrocolloids", Ed. Phillips and Williams, CRC, Boca Raton, Florida, USA, 2000, which is expressly incorporated into this description by reference in its entirety.

Alginate oligomers may also be chemically modified, including, but not limited to, modification to add charged groups (such as karboksilirovanie or karboksimetilirovaniya Glik is s), and alginate oligomers modified with the aim of changing the plasticity (for example, through oxidation periodata).

Alginate oligomers (for example, oligomenorrhoea acid), which are suitable for use according to the invention can conveniently be obtained by acid hydrolysis of alginic acid from Laminaria hyperbora and Lesson/a nigrescens, but not limited to, by dissolution at neutral pH, adding an inorganic acid to reduce the pH to 3.4 for precipitation of the alginate oligomer (olgagolovaneva acid), washing with a weak acid, resuspendable at neutral pH and freeze-drying.

Alginates to obtain alginate oligomers of the invention can also be obtained directly from the appropriate bacterial sources, such as Pseudomonas aeruginosa or Azotobacter vinelandii, although it is expected that algal sources are most suitable due to the fact that the alginates produced by these organisms usually have a primary structure in which the majority of G-residues organized in G-blocks, and are not in the form of individual residues.

Was cloned and described the molecular apparatus that participates in the biosynthesis of alginate in Pseudomonas fluorescens and Azotobacter vinelandii (WO 94/09124; Ertesvag, H., et al., Metabolic Engineering, 1999, Vol. 1, 262-269; WO 2004/011628; Gimmestad, M., et al (see above); Remminghorst and Rehm, Biotechnology Letters, 2006, Vol. 28, 1701-1712; Gimmestad, M. et al., Jurnal of Bacteriology, 2006, Vol. 188(15), 5551-5560), and alginates with specified primary structures can be easily obtained by controlling such systems.

Contents G alginates (for example, the substance is the source of alginate) can be increased by epimerization, for example, using C5-epimerase Manorama from A.vinelandii or other epimerase enzymes. Thus, for example, the epimerization of in vitro can be performed using epimerase isolated from Pseudomonas or Azotobacter, for example AlgG from Pseudomonas fluorescens or Azotobacter vinelandii or enzymes AlgE (AlgE1 to AlgE7) from Azotobacter vinelandii. Use epimerase from other organisms, which have the ability to produce alginate, in particular algae, also specifically considered. In vitro, the epimerization of alginates with low content of G using AlgE epimerase Azotobacter vinelandii is described in detail in Ertesvag et al. (see above) and Strugala et al. (Gums and Stabilisers for the Food Industry, 2004, 12, The Royal Society of Chemistry, 84-94). Preferred is the epimerization with one or more epimerase AlgE Azotobacter vinelandii, other than AlgE4, as these enzymes are capable of producing G-block structure. Mutant variants or homologues from other organisms also specifically considered as useful for the application. In WO 94/09124 described recombinant or modified enzymes C5-epimerase of mannuronate (enzymes AlgE), for example encoded epimerase on what sledovatelnot, in which the DNA sequence encoding different domains or modules epimerase been moved or deleted and recombined. Alternatively, you can use the natural mutants epimerase enzymes (AlgG or AlgE), obtained for example by site-directed or random mutagenesis of genes AlgG or AlgE.

Another approach is the creation of organisms Pseudomonas and Azotobacter, which are subjected to mutations in some or all of their epimerase genes in such a way that these mutants produce alginates with the required product structure of alginate oligomer, or even alginate oligomers with the required structure and size (or molecular weight). The creation of a number of organisms Pseudomonas fluorescens mutant genes AlgG is described in detail in WO 2004/011628 and Gimmestad, M., et al., 2003 (see above). The creation of a number of organisms Azotobacter vinelandii mutant genes AlgE disclosed in Gimmestad, M., et al., 2006 (see above). The specialist can use this guide to create new mutants, which could produce alginate oligomers of the invention without great costs.

Another approach is to remove or inactivate endogenous epimerase genes from one organism Azotobacter or Pseudomonas, and then introducing one or more exogenous epimerase genes, which may or may not be mutated (i.e. can be wild type or modifitsirovannymi) and the expression of which can be controlled, for example, through the use of inducible or other "managed promoters". Choosing the right combination of genes, it is possible to produce alginates with a given primary structure.

Another approach could be the introduction of some or all of the mechanisms of biosynthesis of alginate in Pseudomonas and/or Azotobacter in neproducyruth alginate body (for example, E Li) and induction of the production of alginate these genetically modified organisms.

When using such systems on the basis of culture on the primary structure of the alginate or alginate oligomer can be influenced by culture conditions. In the framework of the training specialist can adjust the cultivation parameters such as temperature, osmolarity, levels/sources of nutrients and atmospheric parameters to manipulate the primary structure of alginates produced by a specific organism.

Links on "remnants" and "M-remnants/M", or guluronate acid, or mannurone acid, or guluronate, or mannuronate should be understood interchangeably as referring to guluronate acid/guluronic and mannurone acid/mannuronate (specifically α-L-guluronate acid/guluronic and P-D - mannurone acid/mannuronate) and also include their derivatives in which one or more of the available side chains of the Il the groups was modified, not causing antibiotica activity becomes essentially lower than the unmodified polymer. The normal group, modifying the saccharide may include acetyl group, sulfate, amino, deoxy, alcohol, aldehyde, ketone, ether, anhydrous. Alginate oligomers can also be chemically modified to include charged groups (such as karboksilirovanie or karboksimetilirovaniya glikana) and to modify plasticity (for example, through oxidation periodata). Specialist known additional chemical modifications that can be made on the monosaccharide subunits of oligosaccharides and which can be applied to the alginates of the invention.

Under the "biofilm" refers to a community of microorganisms, characterized by the prevalence of sessile cells that are attached to the base or to the surface of the section, or to each other (there may be some motile cells) and which are embedded in a matrix of extracellular polymers (more specifically, extracellular polymers, which they produce), characterized in that the microorganisms of this colony show a modified phenotype in relation to growth rate and gene transcription (for example, compared with their "semiplenary" or free-floating or planktonic counterparts).

the Term "combat biofilm" is widely used in this description of the invention and includes any effect of destruction, reduction or destruction of the biofilm (i.e., "attack" on an existing biofilm) or bring it to a state more sensitive to the action of an antimicrobial agent or of the host immune response, and suppressing, reducing, delaying or preventing the formation of biofilms. Thus, the "struggle" includes any treatment of a biofilm, which has a negative effect on the biofilm.

"Combating biofilm", therefore, includes both the preventive and counter measures or treatment. Combating biofilm, therefore, includes the prevention of the formation of biofilms, the elimination of the biofilm, reducing the size of the biofilm, reducing the number of microorganisms in biofilm colonies, reducing or stopping the growth rate of the biofilm, reducing or stopping the rate of increase in the number of microorganisms in biofilm colonies, reducing the physical integrity of the biofilm, increasing the sensitivity of the microorganisms in the biofilm colony to antimicrobial agent or mechanism of immune defense of the host and increase the permeability of the biofilm to antimicrobial agent or mechanism of immune defense of the host.

Thus, the method according to the invention can be used clinically, for example in the treatment of biofilm infections, or it can use the AMB in cleaning or disinfecting any surface for example, commercial or industrial surfaces.

The size, structure, integrity, and number of microorganisms in the biofilm can be explored in any convenient way. For example, to estimate the size, integrity and structure of biofilms is often used scanning and transmission electron microscopy. Histochemical staining of microorganisms and/or components of the extracellular matrix is also routine (for example, dye 630/650-X, SE, BODIPY™ for the components of the matrix of biofilms of Pseudomonas and dye FM™ 1-43 for cell membranes Pseudomonas) and can be used to estimate the number of microorganisms and the structure and integrity of the biofilm visually or by means of devices for sorting cells, confocal microscopes or EPI-fluorescence microscopes. Analysis MWES (minimum concentration eradication of biofilms), Moskowitz SM, et a) (2004) J Clin Environ, 42: 1915-1922, and described in more detail in the Examples, can be used to assess the sensitivity of the microorganisms in the biofilm to antimicrobial agent. Donlan and Costerton, 2002, Clin. The Mic. Rev, Vol. 15(2), 167-193 suggested additional examples.

Biofilms, which can be treated according to the invention, is not limited from the point of view of the microorganisms in biofilms, because alginate oligomer according to the invention, inter alia, focused on the extracellular matrix. Sootvetstvenno is, biofilm may contain any class, genus or species of microorganism, namely any microorganism, which can form a biofilm. These microorganisms usually include bacteria, covering any genera or species of bacteria. Thus, bacteria may be gram-positive or gram-negative or non-staining Gram. They can be aerobic or anaerobic. Bacteria can be pathogenic or non-pathogenic, or cause rotting or indicator bacteria.

Examples of genera or species of bacteria include Abiotrophia, Achromobacter, Acidaminococcus, Acidovorax, Acinetobacter, Actinobacillus, Actinobaculum, Actinomadura, Actinomyces, Aerococcus, Aeromonas, Afipia, Agrobacterium, Alcaligenes, Alloiococcus, Alteromonas, Amycolata, Amycolatopsis, Anaerobospirillum, Anaerorhabdus, Arachnia, Arcanobacterium, Arcobacter, Arthrobacter, Atopobium, Aureobacterium, Bacteroides, Balneathx, Bartonella, Bergeyella, Bifidobactehum, Bilophila Branhamella, Borrelia, Bordetella, Brachyspira, Brevibacillus, Brevibacterium, Brevundimonas, Brucella, Burkholderia, Buttiauxella, Butyrivibrio, Calymmatobacterium, Campylobacter, Capnocytophaga, Cardiobacterium, Catonella, Cedecea, Ceilulomonas, Centipeda, Chlamydia, Chlamydophila, Chromobacterium, Chyseobacterium, Chryseomonas, Citrobacter, Clostridium, Collinsella, Comamonas, Corynebacterium, Coxielta, Cryptobacterium, Delftia, Dermabacter, Dermatophilus, Desulfomonas, Desulfovibrio, Dialister, Dichelobacter, Dolosicoccus, Dolosigranulum, Edwardsiella, Eggerthella, Ehrlichia, Eikenella, Empedobacter, Enterobacter, Enterococcus, Erwinia, Erysipelothrix, Escherichia, Eubacterium, Ewingella, Exiguobacterium, Facklamia, Fili factor, Flavimonas, Flavobacterium, Francisella, Fusobacterium, Gardnerella, Gtobicatella, Gemella, Gordona, Haemophilus, Hafnia, Helicobacter, Helococcus, Holdemania, Ignavigranum, Johnsonella, Kingella, Klebsiella, Kocuria, Koserella, Krthia, Kytococcus, Lactobacillus, Lactococcus, Lautropia, Leclercia, Legionella, Leminorella, Leptospira, Leptotrichia, Leuconostoc, Listeria Listonella Megasphaera, Methylobacterium, Microbactehum, Micrococcus, Mitsuokella, Mobiluncus, Moellerella, Moraxella, Morganella, Mycobacterium, Mycoplasma, Myroides, Neisseria, Nocardia, Nocardiopsis, Ochrobactrum, Oeskovia, Oligella, Orientia, Paenibacillus, Pantoea, Parachlamydia, Pasteurelia, Pediococcus, Peptococcus, Peptostreptococcus, Photobacterium, Photorhabdus, Plesiomonas, Porphyrimonas, Prevotella, Propionibacterium, Proteus, Providencia, Pseudomonas, Pseudonocardia, Pseudoramibacter, Psychrobacter, Rahnella, Ralstonia, Rhodococcus, Rickettsia Rochaiimaea Roseomonas, Rothia, Ruminococcus, Salmonella, Selenomonas, Serpulina, Serratia, Shewenella, Shigella, Simkania, Slackia, Sphingobacterium, Sphingomonas, Spirillum, Staphylococcus, Stenotrophomonas, Stomatococcus, Streptobacillus, Streptococcus, Streptomyces, Succinivibrio, Sutterella, Suttonella, Tatumella, Tissierella, Trabutsiella, Treponema, Tropheryma, Tsakamurella, Turicella, Ureaplasma, Vagococcus, Veillonella, Vibrio, Weeksella, Wolinelia, Xanthomonas, Xenorhabdus, Yersinia, and Yokenella; for example gram-positive bacteria, such as M. tuberculosis, M. bovis, M. typhimurium, M. bovis strain BCG, substanti BCG, M. avium, M. intracellulare, M. afhcanum, M. kansasii, M. marinum, M. ulcerans, M. avium subspecies paratuberculosis, Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcus equi, Streptococcus pyogenes, Streptococcus agalactiae, Listeria monocytogenes, Listeria ivanovii, Bacillus anthracis, B. subtilis, Nocardia asteroides, Actinomyces israelii, Propionibacterium acnes, and Enterococcus species, and gram-negative bacteria, such as Closthdium tetani, Closthdium perfringens, Clostridium botulinum, Pseudomonas aeruginosa, vibrio cholerae, Actinobacillus pleuropneumoniae, Pasteurelia haemolytica, Pasteurelia multocida, Legionella pneumophila, Salmonella typhi, Brucella abortus, Chlamydi trachomatis, Chlamydia psittaci, Coxiella burnetti, E. Li, Neiserria meningitidis, Neiserria gonorrhea, Haemophilus influenzae, Haemophilus ducreyi, Yersinia pestis, Yersinia enterolitica, Escherichia coli, E. hirae, Burkholderia cepacia, Burkholeria pseudomallei, Francisella tularensis, Bacteroides fragilis, Fusobascterium nucleatum, Cowdria ruminantium, but are not limited to.

Thus, as a typical example, the biofilm may contain bacteria of the genus Staphylococcus, Pseudomonas, Legionella, Mycobacterium, Proteus, Klebsiella, Fusobacterium, or other intestinal or coliform bacteria.

Biofilms can also contain fungi, including, for example, fungi of the genus Candida, Aspergillus, Pneumocystis, Penicillium and Fusanum. Typical fungal species include Candida albicans, Candida dubliniensis, Cryptococcus neoformans, Histoplama capsulatum, Aspergillus fumigatus, Coccidiodes immitis, Paracoccidiodes brasiliensis, Blastomyces dermitidis, Pneomocystis camii, Penicillium marneffi, Altemaria alternate, but are not limited to.

In addition, the biofilm may contain algae, and typical species of algae include Chaetophora, Chlorella protothecoides, Coleochaete scutata, Coleochaete soluta, Cyanidioschyzon merolae Aphanochaete, Gloeotaenium, Oedogonium, Oocystis, Oscillatoria, Paradoxia multisitia, Phormidium, Chroococcus, Aphanothece, Fragillaria, Cocconis, Navicula, Cymbelia, Phaeodactylum, and cyanobacteria (blue-green algae and diatoms such as Nitzschia palea.

Biofilms can also contain other organisms, such as parasites, such as protozoa such as Toxoplasma, such as Toxoplasma gondii, Plasmodium species such as Plasmodium falciparum, Plasmodium vivax, Plasmodium malariae. Trypanosoma brucei, Trypanosoma cruzi, Leishmania species, such as Leishmania major, Schistosoma, such as Schistosoma mansoni and Entamoeba histolytica.

Usually biofilm contains a mixed population of micro-organisms, and thus biofilm, podvergshayasya alginate oligomers according to the invention, can contain any number of the above mentioned species. Preferably at least two, more preferably at least 5 and most preferably at least 10.

Preferably biofilm colony contains microorganisms at least one of the following genera: Citrobacter, Enterobacter, Escherichia, Hafnia, Serratia, Yersinia, Peptostreptococcus, Bactehodes, Pseudomonas, Legionella, Staphylococcus, Enterococcus, Streptococcus, Klebsiella, Candida, Proteus, Burkholderia, Fusobacterium and Mycobacterium, such as Staphylococcus aureus, Staphylococcus epidermidis, Legionella pneumophila, Candida albicans, Pseudomonas aeruginosa, Burkholderia cepacia and Streptococcus Pyogenes.

As noted above, the biofilm may be present on the surface. The surface is not limited and includes any surface that can meet the microorganism, in particular, as noted above, the surface exposed to water or humidity. The surface may be biotic or abiotic, non-living (or abiotic) surfaces include any surface that may be exposed to microbial contact or contamination. Thus, specifically the surface of the equipment, including industrial equipment, or any surface exposed to the aquatic environment (for example, marine equipment, or ships or boats, or their parts or components), or any surface exposed to any frequent the environment, for example, the pipe or casing.

Such inanimate surfaces exposed to microbial contact or contamination include, in particular, any part of machinery or equipment for the processing, preparation, storage or dispensing food or beverages, installation of air conditioning, industrial machinery, for example in the chemical or biotechnological processing plants, storage tanks, and medical or surgical equipment. Any installation or equipment to move, or transport, or delivery of substances, which can be exposed to water or humidity, prone to the formation of biofilms. Such surfaces include, in particular, pipes (this term is widely used in this description of the invention and includes any pipe or line). Typical non-living or abiotic surfaces include, without limitation, equipment or surface for processing, storage, dispensing or food preparation, storage tanks, conveyors, floors, drains, coolers, freezers, equipment surfaces, walls, valves, belts, pipes, pipes for air conditioning, refrigeration, line dispensing food or beverage, heat exchangers, ship hulls, or any h is the motion of the ship structure, which is exposed to water, dental technical water supply, pipelines for oil drilling, contact lenses and containers for storage.

As noted above, medical or surgical equipment or devices are a special class of surfaces, which can be formed biofilm. It can include any kind of lines, including catheters (e.g., Central venous and urinary catheters), prosthetic devices such as heart valves, artificial joints, artificial teeth, dental crowns, dental dams and implants for soft tissues (for example, implants for breast, buttocks and lips). Included any type of implantable (or "put") medical devices (e.g. stents, intrauterine devices, pacemakers, tube for intubation, prostheses or prosthetic devices, lines or catheters). "Entered" medical device may include a device in which any part of it is inside a body, the device may be fully or partially "introduced."

The surface may be made of any material. For example, it may be a metal, such as aluminum, steel, stainless steel, chromium, titanium, iron, and their alloys, and the like. Also, the surface may be a plastic, naprotechnology (for example, polyethylene, ultrahigh-molecular weight polyethylene, polypropylene, polystyrene, poly(meth)acrylate, Acrylonitrile, butadiene, ABS (acrylonitrilebutadienestyrene), Acrylonitrile-butadiene, and so forth.), polyester (e.g. polyethylene terephthalate) and polyamide (e.g. nylon), combinations thereof and the like. Other examples include acutally copolymer, polyphenylsulfone, polysulfone, polythermal, polycarbonate, polyetheretherketone, polyvinylidene fluoride, poly(methyl methacrylate) and poly(tetrafluoroethylene). In addition, the surface can be represented as bricks, tiles, ceramics, porcelain, wood, vinyl, linoleum or carpeted floor, combinations thereof and the like. In addition, the surface can be a food, such as beef, poultry, pork, vegetables, fruits, fish, shellfish, combinations thereof and the like.

Biotic or living surface may include any surface or surface section in the body or on the body. Thus, as noted above, it can be seen as a "physiological" or "biological" surface. It can be any internal or external surface of the body, including any tissue which may include hematologic or hematopoietic tissue (e.g. blood). As noted above, dead, or dying (for example, necrotic the definition), or damaged (for example, diseased or destroyed or torn fabric especially vulnerable to the growth of biofilms, and the fabric covered by the term "living" or "biotic". The surface may be a mucous or nessisity surface.

Typical biotic surfaces include, without limitation, by them, any surface in the oral cavity, such as teeth, gums, gingival sulcus, periodontal pocket, reproductive path (e.g., cervix, uterus, fallopian tubes, peritoneum, middle ear, prostate gland, urinary tract, the inner lining of blood vessels, mucous membranes of the eye, the cornea tissue, respiratory tract, lung tissue (for example, bronchial and alveolar), heart valves, gastrointestinal tract, skin, scalp, nails and the interior of wounds, especially chronic wounds, which can be local or internal wounds.

In one aspect, the surface is not mucous or, more specifically, has no mucus coating with high viscosity. The expert is able to determine when the mucous on the surface has a high viscosity. In one embodiment the surface is not a surface, secreting mucus tissue. More specifically, in this embodiment, the surface is not a surface of the fabric, covered with mucus. The region is tis common knowledge known tissue, which secrete mucus, and tissue that are covered with slime.

Accordingly, it can be seen that the invention offers medical use of alginate oligomers, which are defined in this description of the invention, for treating or preventing biofilm infection in a subject (e.g., biofilm infection of any microorganism, including bacteria, viruses, fungi or parasites, such as protozoa). The infection may represent a pathogenic infection. Typical examples of microorganisms that can cause infection described above. It should be noted infections caused by Citrobacter, Enterobacter, Escherichia, Hafnia, Serratia, Yersinia, Peptostreptococcus, Bacteriodes, Pseudomonas, Legionella, Sfaphylococcus, Enterococcus, Streptococcus, Kiebsiella, Candida, Proteus, Burkholderia, Fusobacterium and Mycobacterium, such as Staphylococcus aureus, Staphylococcus epidermidis, Legionella pneumophila, Candida albicans, Pseudomonas aeruginosa, Burkholdeha cepacia and Streptococcus pyogenes. It should be noted infections caused by Pseudomonas, such as Pseudomonas aeruginosa.

The term "subject" is widely used in this description of the invention and includes a biofilm infection that takes place inside of the subject or on the subject, for example on the outer surface of the body. Biofilm infection can be chronic (i.e. can be a chronic biofilm infection, such as infection that persists for at least 5 or at least 10 days, the particular at least 20 days, more specifically, at least 30 days, and most specifically at least 40 days. Chronic infections often manifest in the form of biofilm infections, but biofilm infection may not be a chronic infection, as defined in this specification.

This aspect of the invention biofilm infection can occur on the inside surface of the subject or on the subject (i.e. biotic surfaces, as noted above) and/or on the surface of medical devices, particularly implantable or "put" medical device.

Thus, this aspect of the invention proposed alginate oligomer (which may be any alginate oligomer as defined in this description of the invention) for use in the treatment or prevention of biofilm infection in a subject.

Alternatively, in this aspect of the invention features the use of alginate oligomer for the manufacture of a medicinal product for use in the treatment or prevention of biofilm infection in a subject.

This aspect of the invention is also a method of treating or preventing biofilm infection in a subject, comprising the introduction of a pharmaceutically effective amount of alginate oligomer to a subject in need of it.

In addition, PR is alagaesia the use of alginate oligomer in the treatment or prevention of biofilm infection in a subject.

The subject may be any person or animal subject, not a person, but, more specifically, may be a vertebrate, such as mlekopitayushchie the subject, bird, fish or reptile. Subjects-people are preferred, but the subject can represent, for example, any livestock or a domestic animal or an animal in the zoo. Thus, typical animals include dogs, cats, rabbits, mice, Guinea pigs, hamsters, horses, pigs, sheep, goats, cows, birds and fish. Thus, covered by the application of the invention in veterinary medicine. The subject can be regarded as a patient.

Biofilm infection can occur in any individual, but some subjects are more susceptible to infection than others. Subjects who are susceptible to biofilm infections include, without limitation, entities, whose epithelial and/or endothelial barrier is weakened or broken, the subjects, based on whose secretion mechanisms of protection against infection by microorganisms dismantled, broken, weakened or undermined, and the entities that have immune disorders, immunodeficiency or depressed immunity (i.e. entities in which any part of the immune system is not working properly or not work properly, other the words, in which any part of the immune response or immune activity is reduced or weakened or as a result of disease or clinical impact or other treatment, or otherwise).

Typical examples of subjects exposed to biofilm infections include, but without limitation, subjects with previously developed infection (for example, by bacteria, viruses, fungi or parasites such as protozoa), especially in subjects with HIV subjects with sepsis and subjects with septic shock; subjects with immune deficiency, such as subjects, preparing for, undergoing, or recovering from chemotherapy and/or radiotherapy, subjects (including patients with autograft, allograft and xenograft) with organ transplantation (e.g., bone marrow, liver, lung, heart, heart valve, kidneys and so on), subjects with AIDS; subjects in a health facility such as hospital, mainly of the subjects in the intensive care unit or in a critical state (i.e. in units relevant to the life-support systems, or maintain body of the patient); subjects suffering from trauma; subjects with burns, subjects with acute and/or chronic wounds; newborn subjects; elderly subjects; subjects with cancer ill the of (broadly defined in this description of the invention and including any neoplastic condition as malignant, or non-cancerous), especially patients with cancer of the immune system (eg leukaemia, lymphoma and other hematological cancers); subjects suffering from autoimmune conditions such as rheumatoid arthritis, diabetes type 1 diabetes, Crohn's disease, especially in subjects undergoing immunosuppressive treatment of these diseases; subjects with reduced or eliminated epithelial or endothelial secretion (e.g., mucus, tears, saliva) and/or clearance of secrets (e.g., subjects with subfunctions cilia on the mucous tissue), and/or patients with increased mucus viscosity (for example, smokers) and subjects with COPD (chronic obstructive pulmonary disease), bronchitis, cystic fibrosis, emphysema, lung cancer, asthma, pneumonia or sinusitis) and subjects with established medical device.

Thus, the entities, which, in particular, with biofilm infection can be treated according to the present invention, include patients who remain weakened or due to poor perfusion, repeated trauma, malnutrition, inadequate oxygenation or dysfunction of cells.

Especially noteworthy subjects who have suffered physical injury. The injury itself may cause weakening or breaking rules is a high functioning epithelial and/or endothelial barrier of the subject or the immune system of a subject may become weakened in response to trauma (shock response). The term "injury" in the broad sense refers to damage to the cells as foreign bodies and/or physical damage to the cells. In foreign bodies included microorganisms, particles, chemical agents, and the like. In physical damage included mechanical damage, thermal damage, such as obtained from overheating or overcooling; electrical damage, such as caused by contact with sources of electrical voltage; and radiation damage, caused, for example, long-term extensive exposure to infrared, ultrafioletowego or ionizing radiation.

It should also be noted subjects with burn. Any burn, especially severe burns, has a significant impact on the integrity of epithelial and/or endothelial barrier of the subject, and the subject often becomes immunocompromising in response to the burn (shock response).

Typical cause burn agents represent the extremes in temperature (for example, fire and fluids and gases, extreme temperatures), electricity, corrosive chemicals, friction and radiation. The degree and duration of exposure, together with the intensity/power of the agent lead to burns of varying severity. Obvalivanie (i.e. trauma associated with high temperature liquids and/or gases) believed the W burn.

The severity of epidermal burns are usually classified in two ways. The most famous is the classification according to the degree. Burns first degree is usually limited to erythema (redness) in the core area of injury and a white spot of hemolysis at the site of injury. Cellular injury such burns applies only to the depth of the epidermis. Second degree burns also manifest as erythema in the core area of injury, but with surface blisters on the epidermis. Cellular injury, second-degree burns covers the superficial (papillary) dermis and may also include the deep (reticular) dermis layer. Third degree burns are burns, in which the epidermis is lost with damage to the hypodermis. Damage is usually marginal, including charring. Sometimes there is a burn scab (dry, black necrotic tissue). Third degree burns may require a graft. In burns of the fourth degree has disastrous damage to the hypodermis, for example, completely lost the hypodermis, and the damage extends to the underlying muscle, tendon, and ligament tissue. There are charring and burn scab. Transplantation of tissue is required if the burn was not fatal.

Another well-known system of classification is the classification in thickness. Burns "surface the thickness" corresponds to burns of the first degree. Range of second-degree burns covered two classes of burns "partial thickness". "Partial thickness-superficial" are burns that affect the epidermis only on the depth of the papillary dermis. "Partial thickness-deep" are burns that affect the dermis to the deep reticular dermis. "Full thickness" burns correspond to the burns of the third and fourth degree.

Some physical damage, for example, some burns, and cellular damage by foreign objects lead to the formation of the wound. More specifically, the wound can be considered as a break in the tissue or exposed tissue. Wounds can also be caused spontaneously formed by the lesion, such as a skin ulcer (for example, venous, diabetic ulcer or bedsore), anal another or ulcers of the mouth.

Wounds usually defined either as acute or chronic. Acute wounds are wounds that are properly developed through three recognizable stages of the healing process (i.e. inflammatory phase, proliferative phase, and the remodeling phase) without prolonged dynamics. Chronic wounds, however, are those wounds that do not perform an ordered sequence of biochemical events of the healing process, as the wound stops at one of the stages of healing. Usually chronic wounds stop what I'm in the inflammatory phase. According to a particular aspect of the present invention is a chronic wound is a wound that does not shivsena for at least 40 days, especially at least 50 days, more specifically at least 60 days, and most specifically at least 70 days.

As noted above, the wounds are an ideal environment for infection, including biofilm infection, especially chronic biofilm infection, due to a defective epithelial barrier, and the availability of substrate, and surface colonization and biofilm attachment. Problematic that wound infection is often additionally delays the healing process and thus leads to the fact that the wound be more susceptible to the formation of biofilms and the resulting infection. Alginates according to the invention, thus, are effective in the treatment and prevention of biofilm infection of the wounds, and chronic wounds represent one preferred aspect of the present invention.

Thus, in one embodiment of the invention, a method for treating or preventing biofilm infections, including chronic biofilm infection of the above subjects, in particular in subjects with respiratory diseases or disorders, such as cystic fibrosis, wounds, burns and/or trauma, VK is uchumi introduction to the subject of the pharmaceutically effective amount of alginate oligomer, as defined in this specification.

In one aspect of particular importance alginate oligomers can be used for treating or preventing biofilm infection in wounds, such as burns, for example in the treatment of infected wounds, such as burns.

Due to the ability to treat and prevent biofilm infection of wounds alginate oligomers, as defined in the description of the invention, can eliminate one of the obstacles to wound healing, and thus alginate oligomers, as defined above, are also effective in promoting healing of acute and chronic wounds.

Under the stimulation of healing understand that treatment accelerates the healing process of the wound in question (i.e. the passage wound through three recognizable stages of the healing process). Acceleration of the healing process can manifest itself in the form of increased rate of passage through one, two or all stages of healing (i.e. inflammatory phase, proliferative phase and/or phase of remodeling). If the wound is a chronic wound, which stopped at one of the stages of healing, acceleration can be manifested in the form of a resume after a stop of the linear sequential process of healing. In other words, treatment shifts the wound from nasatelevision state, where p is on begins to develop through the stages of healing. This development after the resumption can occur with normal speed or even slower speed compared with the speed of normal healing of acute wounds.

The alginate oligomers may be used to treat biofilm infections everywhere, wherever they were in the body or on the body. Thus, in one embodiment of the biofilm infection can be a infection of medical devices, in particular "entered" medical device.

As noted above, biofilms are found on the teeth, for example in the form of plaque. Alginate oligomers can be used according to the present invention as an oral therapeutic agents, for example in the fight against plaque, for example in removing it or reducing it or to prevent, reduce or delay its development. They can also be used in the treatment and prevention of infections or infectious diseases that may occur in the oral cavity, such as gingivitis and periodontitis.

As noted above, while the treatment of biofilm infections of the lungs and respiratory tract and all areas of the body mostly covered by the present invention, in one embodiment of the medical use according to the invention is not directed to the treatment of (1) biofilms in the Airways of patients, the article is adusa from COPD (chronic obstructive pulmonary disease), in particular, nasal sinuses, and lungs, in particular in the treatment of cystic fibrosis, chronic obstructive pulmonary disease, emphysema, bronchitis and sinusitis; (2) the middle ear of patients suffering from exudative otitis media; or (3) reproductive tract female patients with impaired ability to conceive; or (4) digestive path patients with dysfunction of the digestive tract (e.g., constipation).

In specific embodiments of the invention the alginate oligomers may be used in the treatment of congenital valve endocarditis, acute inflammation of the middle ear, chronic bacterial prostatitis, pneumonia, dental plaque, periodontitis, biofilm infections in respiratory diseases, which may include cystic fibrosis and asthma, and infections associated with the device associated with the implantable or prosthetic medical devices such as artificial valve endocarditis, or infection of the lines or catheters, or artificial joints, or substitutions tissue.

"Pharmaceutically effective amount of alginate is the number of alginate, which provides a measurable impact on the target biofilm (as defined above) and/or measurable impact on the target state. This quantity can be defined according to the traditional methods of determining dozer the wok, and the expert is able to detect evidence of successful treatment on the basis of their experience and using standard tests available that are designed to monitor the size, structure, integrity of the biofilm and the number of colonies (e.g., described above), and tests designed to monitor the target state.

The appropriate dose of alginate will vary from subject to subject and can be determined by a physician or a practicing veterinarian in accordance with the weight, age and gender of the subject, the severity of the condition, by way of introduction, as well as selected specific alginate oligomer. Usually alginate oligomers according to the invention is applied to the biofilm in the local concentrations of up to 10%, preferably up to 6%, more preferably up to 4% and most preferably up to 2%.

"Treatment", when used in relation to biofilm infection (i.e. in relation to the treatment of the medical condition/disease in the subject as opposed to that applied to the biofilm), is widely used in this description of the invention and includes any therapeutic effect, i.e. any positive impact on the state or against biofilm infections. Thus, included not only the elimination or destruction of the infection, or CoE is of the subject or infection, but also improvement in the infection or condition of the subject. Thus, included, for example, the improvement of any symptom or sign of infection, or any clinically acceptable indicator of infection/condition (for example, reducing the size of the wound and accelerate healing time). Treatment, therefore, includes both curative and palliative therapy, such as previously developed or diagnosed infection/condition, that is, the counter treatment.

"Warning", when used in this description, refers to any preventive effect. Thus, it includes delaying, limiting, reducing or preventing a condition or start condition, or one or more symptoms, for example in terms of condition or symptom before preventive treatment. Prevention, therefore, expressly includes as an absolute prevention of the emergence and development of the state, or its symptoms, and any delay or developmental condition or symptom, or reduce or limit the development or progression of the condition or symptom.

Specifically, alginates according to the invention can be selected as a preventive treatment, for example to prevent or at least information, minimize the risk of biofilm infections (for example, underthe influence of the pathogen). This aspect of the invention is particularly useful in the care of hospitalized patients, because the risk of contact with nosocomial infection (commonly known as hospital/acquired infection or infection associated with a medical institution), for example Staphylococcus aureus, methicillin-resistant Staphylococcus aureus (MRSA), Pseudomonas aeruginosa, Acinetobacter baumannii, Stenotrophomonas maltophilia, Clostridium difficile, Mycobacterium tuberculosis and vancomycin-resistant Enterococcus, can be minimized through preventive mode alginate oligomers described in this description of the invention. This aspect of the invention is also particularly useful in the care of the subjects suffering from trauma, subjects with burns and subjects with wounds, all of which, as noted above, are more susceptible to pathogen infection than the subject who is not affected in this way.

Basically, subjects in need of treatment or prevention according to the invention, diagnosed as suffering from or at risk target status or identify as having a biofilm infection or a risk of development of biofilm infections.

Specifically, the alginate oligomers of the invention can be selected as a prophylactic treatment to prevent or at least minimize the risk of biofilm infections, including, for example, in zirovnice RAS, congenital valve endocarditis, acute inflammation of the middle ear, chronic bacterial prostatitis, periodontitis, infection of the respiratory tract and lungs (for example, cystic fibrosis or other respiratory disease, plaque, pneumonia or infection of medical (e.g., "put") medical devices.

In one preferred embodiment of the invention the alginate oligomers may be used together or in combination with antimicrobial agent. In the context of medical applications such agent may be any clinically useful antimicrobial agent and, in particular, an antibiotic. In the context of non-clinical applications of antimicrobial agent can also be any antimicrobial agent used for such purposes, for example, any disinfectant or antiseptic, or cleaning or sterilizing agent. Agents can be used individually or together in a single composition, simultaneously or sequentially, or separately with any desired time interval.

Thus, as a typical example, the antimicrobial agent can be used after the alginate oligomer, but in some circumstances it may be useful prior to or simultaneous use.

You can use any antimicrobial agent, which is the first aim of at least one of the microorganisms in the biofilm target. The choice of antimicrobial agent of course should be suitable for the surface undergoing treatment, but, for example, can be used antimicrobial agents such as antibiotics, antifungal agents, antiseptics and/or sterilizing conditions, such as irradiation (e.g. UV, x-ray, gamma), extreme temperatures and extreme pH values.

Typical antibiotics include, without limitation, aminoglycosides (e.g., amikacin, gentamicin, kanamycin, neomycin, netilmicin, streptomycin, tobramycin); carbacephem (for example, loracarbef); cephalosporins 1st generation (for example, cephalo-Smoking, Cefazolin, cephalexin); cephalosporins 2nd generation (e.g., cefaclor, cefamandole, cephalexin, cefoxitin, cefprozil, cefuroxime); cephalosporins 3rd generation (e.g., cefixime, cefdinir, cefditoren, cefoperazone, Cefotaxime, cefpodoxime, ceftazidime, ceftibuten, ceftizoxime, Ceftriaxone); cephalosporins 4th generation (such as cefepime); macrolides (eg, azithromycin, clarithromycin, dirithromycin, erythromycin, troleandomycin); carbapenems (e.g., aztreonam); penicillins (eg, amoxicillin, ampicillin, carbenicillin, cloxacillin, dicloxacillin, nafcillin, oxacillin, penicillin G, penicillin V, piperacillin, tikarcillin); the polypeptide antibiotic and (for example, bacitracin, colistin, polymyxin B); quinolones (eg, ciprofloxacin, enoxacin, Gatifloxacin, levofloxacin, lomefloxacin, moxifloxacin, norfloxacin, ofloxacin, trovafloxacin); sulfonamides (for example, mafenide, sulfacetamide, sulfamethizole, sulfasalazin, sulfisoxazole, trimethoprim-sulfamethoxazole), tetracyclines (e.g., demeclocycline, doxycycline, minocycline, oxytetracycline, tetracycline); carbapenems (e.g. imipenem, Meropenem, ertapenem, doripenem, panipenem/betamipron, biapenem, PZ-601); chloramphenicol; clindamycin, ethambutol; fosfomicin; isoniazid; linezolid; metronidazole; nitrofurantoin; pyrazinamide; inupristin/dalfopristin; rifampin; spectinomycin; and vancomycin. The antibiotics vancomycin, tobramycin, Meropenem, ciprofloxacin, piperacillin, colistin, aztreonam ciprofloxacin and azithromycin are preferred.

Typical preservatives include, but without limiting them, chlorine bleach (sodium hypochlorite), Quaternary ammonium compounds (e.g. benzalkonium chloride, cetyltrimethylammonium bromide, cetylpyridinium chloride, hydrogen peroxide, phenolic compounds (e.g., TCP (trichlorophenyl)), alcohols (for example ethanol), Virkon™, iodine compounds (e.g., povidone-iodine), silver compounds (e.g., nano/micro-particles of elemental silver).

Tipichnytakzhe substances include, but without their limitations, the polyene (e.g., natamycin, rimoldi, filipin, nystatin, amphotericin b, condicin); imidazoles (eg, miconazole, ketoconazole, clotrimazole, econazole, bifonazol, butoconazole, fenticonazole, isoconazole, oxiconazole, sertaconazole, sulfonate, tioconazole); triazoles (e.g., fluconazole, Itraconazole, isavuconazole, ravuconazole, Posaconazole, voriconazole, terconazole); allylamine (e.g., terbinafine, amorolfine, naftifine, butenafine); and echinocandin (for example, anidulafungin, caspofungin, micafungin).

The antimicrobial agent can be conveniently applied before, simultaneously with the alginate, or after it. It is convenient to apply the antimicrobial agent is essentially at the same time as alginate, or immediately after it. For example, the antimicrobial agent is applied at least 1 hour, preferably at least 3 hours, more preferably at least 5 and most preferably at least 6 hours after administration of alginate oligomer. To optimize antimicrobial effect of the antimicrobial agent, it is possible to give (for example, to enter or to deliver again in times that are appropriate for your agent. The expert is able to develop a suitable dosage or regimen. When long-term treatment of alginate can also be reused. His mo is but to use as often as an antimicrobial agent, but usually less. Desired frequency will depend on the localization of biofilm infection, the composition of the colony and used antimicrobial agent, and the expert is able to optimize the dosage or use for best results.

In the preferred embodiment the antimicrobial agent can be used or applied after physical removal or reduction (e.g., processing) of the biofilm from the surface.

After removing or attempting to remove biofilm surface can be brought into contact with the alginate oligomers within 0-24 hours, in particular 2 to 12 hours, more specifically 4-8 hours, most specifically 5-7 hours, for example 6 hours. Then, optionally, you can use an antimicrobial agent. Such a scenario may be desirable or particularly applicable in the clinical setting. In the case of wounds infected with biofilm, the duration of incubation may be conveniently designed to fit the replacement schedule of the wound dressing.

Physically removing biofilms can be performed using any suitable surgical, mechanical or chemical means. Easy to use liquid, gel, helium-solavie, semi-solid compositions or gas applied under pressure to the biofilm, sonication, l is serum or using abrasive device. The composition used to remove, by itself or in the form of a washing solution before, during or after conveniently may contain alginate oligomer.

Thus, in one particular embodiment features a cleansing or washing compositions, for example the solution for wounds containing alginate oligomer, in particular any alginate oligomer as defined in this description of the invention. This cleansing composition is usually a sterile solution, particularly aqueous sterile solution or sterile solution or oil-based, and can optionally contain proteolytic enzymes (e.g. collagenase, trypsin, pepsin, elastase), abrasive solid phase (for example, colloidal silica, ground pumice, ground vegetable or animal shell).

It may be useful to use in combination or together with other agents that destroy the biofilm. Destructors biofilms include, but without limiting them, proteases, such as serine protease, metalloprotease and cysteine protease (examples of these types of proteases are listed in ER, the full contents of which are incorporated in this description by reference); nucleases, such as Tnkase I and II, Mcasa a, H, I, II, III, P, PhyM, R; lipase enzymes that can destroy polysaccharides, gelsolin, Tilney restorer, acetylcystein is, uncharged low molecular weight polysaccharide (e.g. dextran), or the anionic polyaminoamide (for example, poly-ASP or poly-GLU).

I can mention alginate-liasu, and combined its use with alginate oligomer as defined in this description of the invention, represents one possible specific embodiment of this aspect of the invention.

Use in combination or together with immunostimulating agents may also be useful in the treatment of biofilms in clinical conditions. Such immunostimulatory agents can be conveniently used in the moments corresponding to that described above in relation to antimicrobial agents, and possibly can be used in combination with alginate oligomer and an antimicrobial agent. Suitable immunostimulatory agents include, but are not limited to, cytokines, such as TNF, IL-1, IL-6, IL-8, and immunostimulatory alginates, such as alginates with a high content of M, as described, for example, in U.S. patent 5169840, WO 91/11205 and WO 03/045402, which is expressly incorporated by reference into this specification in their entirety, but including any alginate with immunostimulating properties.

The use of alginate oligomers or in combination with growth factors such as PDGF (platelet-derived growth factor), FGF (fibroblast growth factor), EF (epidermal growth factor), TGF (transforming growth factor), hGF and enzymes may also be useful in medical applications according to the invention. Typical examples of suitable enzymes include, but without limiting them, proteases, such as serine protease, metalloprotease and cysteine protease (examples of these types of proteases are listed in EP 0590746, the full contents of which are included in this description by reference); nucleases, such as Gnkazy I and II, RNase A, N, I1II, III, P, PhyM, R; lipase enzymes that can destroy polysaccharides.

The use of alginate oligomers in combination or together with a physiologically acceptable agent in order to decrease mucosal viscosity, can also be useful, for example an enzyme that breaks down nucleic acid (e.g., Tnkase, such as Tnkase I), gelsolin, Tilney restorer, acetylcysteine, sodium chloride, uncharged low molecular weight polysaccharide (e.g. dextran), arginine (or other precursors or stimulators of the synthesis of nitric oxide) or anionic polyaminoamide (for example, poly-ASP or poly-GLU), Ambroxol, bromhexin, carbocisteine, domodal, eprazinone, erdosteine, latesteijn, mesna, altenessen, Cabrera, stepronin, tiopronin are known specific mukolitikami. Use Gnkazy is especially preferred.

As noted above, alginate oligomers may can be used with any other therapeutically active agent, which it is desirable to use, for example anti-inflammatory agent. The combined use of alginate oligomer with an additional therapeutically active agent (e.g., antimicrobial or anti-inflammatory agent) preferably can provide a dose reduction (e.g., normal or normal dose) additional therapeutically active agent, for example it can be used in its normal or usual dose or a lower dose, for example up to 50% (or 50%) of its normal dose.

This invention includes the use of one alginate oligomer or a mixture of (a variety of/many different alginate oligomers. Thus, for example, you can use a combination of different alginate oligomers (for example, two or more).

In the case of medical applications alginates according to the invention it is possible to introduce the subject in any suitable form or by any suitable means, such as local, oral, parenteral, enteral, parenteral routes, or by inhalation. Preferably the alginate enter local, oral or parenteral way or by inhalation.

The specialist is able to produce alginates according to the invention in the form of pharmaceutical compositions that are adapted for these routes of administration according to L. the favorite color of conventional ways, known in the art and widely described in the literature. Solely as a guide in Examples 11 and 12 describe two possible compositions (local composition and the cleaning liquid).

The present invention thus also features a pharmaceutical composition for use in the treatment or prevention of biofilm infections containing alginate oligomer as defined in this description of the invention, together with at least one pharmaceutically acceptable carrier, diluent or excipient.

The active ingredient may include, possibly in conjunction with other active agents with one or more conventional carriers, diluents and/or excipients, obtaining conventional galenic preparations such as tablets, pills, powders (e.g., inhalation powders, lozenges, sachets, starch pills, elixirs, suspensions, emulsions, solutions, syrups, aerosols (as a solid or in a liquid medium), sprays (e.g., nasal sprays), compositions for use in nebulizers, ointments, soft and hard gelatin capsules, suppositories, sterile injectable solutions, sterile packaged powders, and the like.

Examples of suitable carriers, excipients and diluents are lactose, dextrose, sucrose, sorbitol, mannitol, to omaly, Arabian gum, calcium phosphate, inert alginates, tragakant, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water syrup, water, mixture of water/ethanol, water/glycol, water/polyethylene, hypertonic salt water, glycol, propylene glycol, methylcellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, liquid paraffin or fatty substances such as hard fat or suitable mixtures. Preferred excipients and diluents are mannitol and hypertonic saline water (saline).

The composition can additionally include lubricating agents, moistening agents, emulsifying agents, suspendresume agents, preservatives, sweeteners, flavoring agents and the like.

As noted above, the alginate oligomers, proposed for use according to the invention can be used in combination with other therapeutic agents, for example, to enter together in one pharmaceutical preparation or composition, or separately (that is, for separate, sequential or simultaneous administration). Thus, alginates according to the invention can be combined with a second (or additional) therapeutically active agent, for example in the pharmaceutical kit or as a combined product ("combination").

Thus, in an additional aspect, the present invention features a product containing an alginate oligomer as defined in this description of the invention and a second active agent, as a combined preparation for separate, simultaneous or sequential application to the biofilm and/or administration to a subject for use in combating biofilm, and/or of treating or preventing biofilm infection in a subject or of any state specified in the description,

Additional therapeutically active agents can be included in pharmaceutical compositions, as noted in relation to combined therapies above.

Parenteral input forms, such as intravenous solutions, should be sterile and free from fisiologica unacceptable agents, and should have a low osmolarity to minimize irritation or other side effects when introduced, and, therefore, the solutions should preferably be isotonic or slightly hypertonic, such as hypertonic salt water (saline). Suitable carriers include aqueous media, typically used to administer parenteral solutions, such as sodium chloride injection, ringer's solution for injection, dextrose injection, dextrose and sodium chloride injection, rest the R ringer's lactate for injection and other solutions such as described in Remington''s Pharmaceutical Sciences, 15th ed., Easton: Mack Publishing Co., 1405-1412 and 1461-1487 (1975) and The National Formulary XIV, 14th ed., Washington: American Pharmaceutical Association (1975). The solutions can contain preservatives, antimicrobial agents, buffers and antioxidants commonly used for parenteral solutions, excipients and other excipients that are compatible with biopolymers and which will not interfere with the manufacture, storage or use of the products.

For the local introduction of the alginate oligomer can be creams, ointments, gels, transdermal patches and the like. Alginate oligomers can also be entered in the medical dressings, such as wound dressings, such as woven (e.g., tissue) or non-woven headbands headbands (for example, gels or dressings with gel component). Known application of alginate polymers in bandages, and dressings, or in fact any bandages may additionally include alginate oligomers according to the invention.

Thus, in an additional specific embodiment of the invention further provides a wound dressing containing alginate oligomer (which may be any alginate oligomer as defined in this description of the invention).

In addition, the local system, which, as expected, are appropriate, represent sistemistica medicines in situ, for example gels, where the solid, semi-solid, amorphous or liquid crystalline gel matrix formed in situ, and which may contain alginate oligomer. Such matrices can be conveniently designed to regulate the release of alginate oligomer from the matrix, for example, the release may be detained and/or continuously during the selected time period. Such systems can form gels only when in contact with biological tissues or fluids. Typically, the gels are bioadhesive. Delivery in any area of the body, which can save or be adapted to save predelivery composition, may be targeted by such method of delivery. Such systems are described in WO 2005/023176.

For application to oral, buccal and dental surface specifically mentioned toothpastes and solutions for rinsing the mouth. Thus, in one particular aspect include compositions for the care of the oral cavity or oral hygiene containing alginate oligomer (which may be any alginate oligomer as defined in this description of the invention), in particular liquid mouth rinse or toothpaste.

As noted above, a preferred composition according to the invention is a composition for cleansing to the Yu use in the purification process for the removal of biofilms, for example, from tissue. Typically, such a composition is a liquid, but you can use gel, helium-solavie or semi-solid composition. The composition can be used for cleansing wounds from biofilms (for example, printability under pressure) and/or can be used for washing fabric before, during and/or after cleansing in other ways, such as surgical, mechanical or chemical methods. The specialist can easily prepare a composition for purification in accordance with the invention.

In the case of biofilms on the still surface of the alginate oligomer may be deposited on the surface to be processed in any suitable composition or drug, or any suitable way. Thus, the alginate oligomer may be in liquid, gel, helium-Polevoy, semi-solid or solid form (e.g., solutions, suspensions, homogenates, emulsions, pastes, powders, aerosols, vapor). Typically, the compositions for treatment of such biofilms on inanimate surfaces will be pharmaceutically unacceptable composition. The choice of the form of the composition will be dictated by the structure of the biofilm, composition and localization of the colony. For example, if the biofilm is localized in the pipeline for liquids, it may be convenient to apply a liquid composition. Can also be preferably used in order to use the composition, which remains on the surface to be processed, but which will not leach into the liquid under normal use, for example adhesive gel. The specialist can easily prepare suitable compositions based on General knowledge. For example, alginate oligomer can be added to the composition of the paint and apply to the surface to be processed, such as a boat hull or other parts of the vessel, which is subject to the action of water, or structure, or any part of, the tank (for example, a reservoir for storage or processing) or on almost any part of any industrial mechanism. Such compositions also can contain an antimicrobial agent as described above, for example chlorine bleach, TCP, ethanol, Virkon™, povidone-iodine, silver compounds, and so forth. Because of the composition may not be pharmaceutically acceptable, more stringent anti-microbial agents can be considered from the point of view of surface damage, pollution, security, user and contamination of the treated surface, and interaction with other components of the composition.

The composition of the invention can be prepared in the form of the drug to provide fast, slow or delayed release of the active ingredient after administration to the subject of/who and the surface using, well known in the art. The adhesive compositions are also preferred. Adhesive composition, the composition for slow and/or delayed release may be particularly suitable.

In an additional aspect of the invention are exposed to biofilm colonization, whose susceptible to colonization surfaces were pre-treated alginate oligomer as defined in this description of the invention. Non-limiting examples of products and surfaces exposed to biofilm colonization described above. Particularly noteworthy is the equipment for processing, storage or dispensing of food and beverages and medical devices. Pre-processing can be performed by any appropriate means, for example by any application of the alginate oligomer on the surface, especially the application of the surface coating, for example by spray drying, the polymer coating polymer comprising alginate oligomer, and staining, polishing or varnishing using coloring, polishing agent or lakiruyuschii compositions containing an alginate oligomer. This "covering" composition (e.g., paint, means for glossing or varnish)containing alginate oligomer, represents more the second aspect of the present invention. Alternatively, alginate oligomer, you can enter into the substance in the manufacture of the surface. This approach is suitable for surfaces made of polymers such as plastics and silicones, such as medical devices described above.

The invention is additionally described with reference to the following non-limiting Examples in which:

Figure 1 shows the bacterial growth in biofilms of Pseudomonas formed overnight and then treated with mucin (2.5 g/l) and G-fragments (0, 1%, 2% or 6%) during the night, at 0 h, 6 h and 24 h after treatment with amikacin overnight (4096-0 µg/ml).

Figure 2 shows the bacterial growth in biofilms of Pseudomonas formed overnight and then treated with mucin (2.5 g/l) and G-fragments (0, 1%, 2% or 6%) during the night, at 0 h, 6 h and 24 h after treatment with oxytetracycline during the night (4096-0 µg/ml).

Figure 3 shows the bacterial growth in Pseudomonas biofilms formed in the presence of mucin (2.5 g/l) and G-fragments (0, 1%, 2% or 6%) during the night, at 0 h, 6 h and 24 h after treatment with oxytetracycline during the night (4096-0 µg/ml).

Figure 4 shows the bacterial growth in Pseudomonas biofilms formed in the presence of mucin (2.5 g/l) for 6 h and then treated with mucin (2.5 g/l) and G - fragments (0 or 6%) during the night, at 0, 6 and 24 h after treatment and during the night with amikacin, tobramycin, oxytetracycline and the and amikacin+oxytetracycline" (4096-0 µg/ml).

Figure 5 shows the bacterial growth in biofilms of Pseudomonas RAO formed within 6 h without mucin and then processed G-fragments (or About 6%) without mucin, 0 h, 6 h and 24 h after treatment and during the night by amikacin (4096-0 μg/ml) or tobramycin (1024-0 µg/ml).

Figure 6 shows the bacterial growth in biofilms of Pseudomonas RAO formed in the presence of mucin (2.5 g/l) for 6 h and then treated with mucin (2.5 g/l) and "G-unit #0802" (0 or 6%) during the night, at 0, 6 and 24 h after treatment and during the night by amikacin (4096-0 μg/ml) or tobramycin (1024-0 µg/ml).

7 shows the bacterial growth in biofilms of Staphylococcus aureus ATCC 6538, formed in the presence of mucin (2.5 g/l) for 6 h and then treated with mucin (2.5 g/l) and G-fragments (0 or 6%) during the night, at 0, 6 and 24 h after an overnight treatment with oxytetracycline (4096-0 µg/ml).

On Fig shows the bacterial growth in biofilms wound isolate "1103" MRSA formed in the presence of mucin (2.5 g/l) for 6 h and then treated with mucin (2.5 g/l) and G-fragments (0 or 6%) adding during the night, at 0 h, 6 h and 24 h after treatment and during the night tobramycin (1024-0 µg/ml).

Figure 9 shows the effect of G-fragments and mucin for the attachment of Candida albicans ATCC 90028 and Candida dubliniensis SW1in biofilms formed with mucin (2.5 g/l) and G-fragments at a concentration of About 2% during the night.

figure 10 presents e of microsemi biofilms of Pseudomonas, formed in the presence of mucin (2.5 g/l) for 6 h and then treated with mucin (2.5 g/l) and G-fragments with 0 or 2% within 24 hours

EXAMPLES

Example 1 - Matter and standard methods

Bacterial strains.

Two strains from culture collections, Pseudomonas aeruginosa RAO (ATCC 15682, Raney isolate) and Staphylococcus aureus (ATCC 6538)was used for analysis MWES (analysis of the minimum concentration eradication of biofilms) in conjunction with the clinical isolate of chronic venous ulcers of the leg, S. aureus (MRSA) "1103". Two types of strain Candida, .albicans ATCC 90028 and .dubliniensis D36tused for analysis of the attachment.

Chemical and bacterial protection.

Bacterial colonies were grown on the basis of blood agar No. 2 (VA; Lab15, LabM, Bury, UK) with the addition of 5% sheep blood and used for insulinopenia tripton-soy broth (TSB, CM0129, Oxoid, Basingstoke, UK) for growth during the night. Biofilms were formed in the b + on the cation composition of the broth Miller-Hinton (SUNW; Lab114, LabM). All used antibiotics were pure pharmaceutical grade (Sigma-Aldrich, Gillingham, UK) and included amikacin, oxytetracycline and tobramycin. Glycoprotein mucin gastric mucus pigs (cleaned by using Jeff Pearson, Newcastle University) and alginate oligomers CF-5/20 ("G-fragments"; 2600 Yes, %90-95 G) and G-unit #0802 (6400 Yes, %G 91) were provided Algipharma AS, Sandvika, Norway.

Analysis of the minimum to which centralimediato biofilms (MWES).

Method used MWES was the adaptation Moskowitz SM, et al. (2004) J Clin Environ 42: 1915-1922. After treatment with storage at -80°C. bacterial isolates were grown on BA and then grown overnight in TSB. After dilution of the bacterial cultures to 0.5 on the McFarland in SMNW in the presence or in the absence of mucin (2.5 g/l) 100 ál was transferred into the wells of flat-bottomed 96-well microtiter tablet. In Example 3, the bacterial culture was diluted to 0.5 on the McFarland in SAMNU with mucin (2.5 g/l) and alginate and 100 µl was transferred into the wells of flat-bottomed 96-well microtiter tablet.

The tablets were then wrapped in parafilm to prevent loss of moisture and incubated at 37°C, allowing to form the biofilm. Incubation time and conditions were varied as described below.

After the formation of the biofilm planktonic cells and the supernatant was removed and each well was then washed with sterile phosphate-saline buffer solution (PBS). After washing, the cells were treated with combinations of alginates and/or antibiotics in the presence or in the absence of mucin (2.5 g/l) in 100 μl of SMNW.

The tablets were then wrapped in parafilm and incubated at 37°C With careful shaking. Incubation time and conditions were varied as described below. Used antibiotics and the concentration ranges shown below.

Moons and were washed in PBS and 100 μl of each concentration serial dilution of the antibiotic in SAMNU then added in two Parallels. The tablets again wrapped up in parafilm and incubated at 37°C With careful swirling in the night.

In all analyses MWES a finite number of cells was determined as follows. The wells were washed in PBS, and biofilm resuspendable in 100 μl of SMNW through intensive pipetting. The optical density was measured at 620 nm (OD620) with a microplate reader (FLUOstar OPTIMA, BMG LABTECH) immediately (0 h) and after incubation at 37°C in 6 h and 24 h

Is MWES represent such a concentration of antibiotic that inhibits any growth of bacteria in the sample. Bacterial growth was measured as the increase in light absorption by the sample. Therefore, the decrease in the value MWES is an indication that the sensitivity of the sample to the antibiotic increased (i.e., fewer antibiotic to prevent bacterial growth).

Used antibiotics and spacing concentration

td align="center"> The tobramycin
AntibioticInterval concentration (μg/ml)
Amikacin4-4096
Amikacin + Oxytetracycline4-4096
Oxytetracycline4-4096
4-4096

Analysis of the minimum concentration eradication of biofilms (MWES) without mucin.

Pseudomonas aeruginosa RAO ADS 15682) was used to determine the values MWES without the addition of mucin. Was guided by the Protocol MWES, which is described above, but without addition of mucin to the growth environment. Experienced two antibiotics: amikacin, and tobramycin.

Analysis of the insertion of the yeast.

Used analysis attachment was an adapted version of Djordjevic et al., (2002) Appl Environ Environ 68: 2950-2958. .albicans ADS 90028 and .dubliniensis CD36trepresented the Candida strains used for the analysis of attachment. Candida was grown on agar Saburo with dextrose (Lab33, LabM) and night broth cultures were grown in liquid nutrient medium Saburo (Lab9, LabM). After adding 5 ál of the overnight culture, 95 μl of SMNW with the addition of mucin (2.5 g/l) and G-fragments (with concentrations of 0, 2%, 6% or 10%) was added to the wells. Tablets wrapped up in parafilm and incubated at 37°C during the night, allowing you to form biofilm.

Planktonic cells and the supernatant liquid was removed from the wells before washing the obtained biofilms (3) sterile dH2O. Then the tablets were dried at 56°C for 45 minutes and Then each well was stained with 150 ál of 1% (vol./about.) crystal violet (in water) for 45 minutes Tablets e is e times washed (3x) dh20, then add 200 ál of 95% ethanol. After 5 min, 100 μl from each well was transferred to a new microtiter plate. OD was then measured on a plate reader at 540 nm.

The growth of biofilms for rendering.

After removal from storage at -80°C. bacterial isolates were grown on BA and then grown overnight in TSB. After dilution of the bacterial cultures to 0.5 for McFariand in SAMNU with mucin (2.5 g/l) 100 ál was transferred into the wells of flat-bottomed 96-well microtiter tablet. The tablets were then wrapped in parafilm to prevent loss of moisture and incubated at 37°C for 6 h to ensure the formation of biofilms. After the formation of the biofilm planktonic cells and the supernatant was removed and each well was then washed with sterile phosphate-saline buffer solution (PBS). After washing the cells were treated with G-fragments and mucin (2.5 g/l) in 100 μl of SMNW. Then the tablets were wrapped in parafilm and incubated at 37°C for 24 h with careful shaking.

Scanning electron microscopy (SEM) of Pseudomonas biofilms.

The glutaraldehyde (2%) was added to the biofilms treated with G-fragments, and were fixed at room temperature for 24 hours. The samples were obezvozhivani in the sequence graded concentrations of ethanol, dried in a device for drying in a critical point (Balzers PD 030, Germany), mounted on aluminum ribs, covered with gold in the device for spraying in vacuum (EMscope model AE 1231, UK) and then examined by scanning electron microscope (FEI-Philips XL-20, The Netherlands).

Confocal microscopy of intact biofilms using BODIPY®630/650-X SE

Biofilms treated with G-fragments, washed with sterile distilled water and stained with BODIPY dye®630/650-X, SE (BODIPY®630/650-X, SE, Invitrogen Ltd), which selectively stains the matrix components (EPS) in Pseudomonas biofilms.

BODIPY®630/650-X, SE was added (100 µl (10 µg/ml)) to each sample biofilms. The preparation is incubated in the dark for 1 hour and then analyzed using CLSM.

Example 2 Measurement values IVJBEC for night biofilms of Pseudomonas aerug'snosa, pre-treated G-fragments

Was guided by the analysis MWES described above. Biofilms were generated in tablets during the night without mucin. After washing PBS biofilms were incubated with 0, 1, 2 or 6% G-fragments and mucin during the night. After PBS washing, the cells were incubated overnight with antibiotics (amikacin or oxytetracycline) and without mucin. The results are graphically presented in figures 1 and 2 and are shown in Tables 2 and 3 below. As you can see, pre -, during the night, the processing biofilms G-fragments caused the Nigeria at 6 h and 24 h values MWES for amikacin or oxytetracycline. 6 h values MWES for amikacin and oxytetracycline was reduced by half using 1% G-fragments and accounted for one-fourth with 2 and 6% G-fragments. 24 h values MWES for oxytetracycline was halved by all concentrations of G-fragments. 24 h values MWES for amikacin was decreased, although it was not possible to quantify this reduction. This means that pre-treatment during the night G-fragments increases the sensitivity of Pseudomonas aeruginosa in biofilms to these antibiotics.

Table 2 - Summary of values MWES 6 hours after exposure to antibiotics during the night. Biofilms of Pseudomonas generated during the night. The mucin (2.5 g/l) and G-fragments at concentrations of 0, 1, 2 or 6% was added to the formed biofilms. Values were expressed as μg/ml of antibiotic.

Table 3 - Summary of values MWES in time 24 hours after exposure to antibiotics during the night. Biofilms of Pseudomonas generated during the night. The mucin (2.5 g/l) and G-fragments at concentrations of 0, 1%, 2% or 6% was added to the formed biofilms. Values were expressed as μg/ml of antibiotic.

The key

Example 3 - Dimension values MWES for Pseudomonas aeruginosa biofilms formed in the presence of G-fragments

Rukovodstvo the analysis were MWES, described above. Biofilms were generated in tablets over night in the presence of mucin and 0, 1, 2 or 6% G-fragments. After washing biofilms were exposed to the effect of oxytetracycline (without mucin) during the night. The results are presented graphically in figure 3 and listed in Tables 4 and 5 below. As you can see, at all concentrations tested G-fragments, the generation of biofilms in the presence of G-fragments of half-reduced 24-hourly values MWES. 6-hourly values MWES was halved when used 2% and 6% G-fragments. 1% G-fragments did not cause a decrease. These data show that Pseudomonas aeruginosa in biofilms generated in the presence of G-fragments are more sensitive to oxytetracycline than Pseudomonas aeruginosa in biofilms generated in the absence of G-fragments.

Table 4 - summary of values MWES at 6 h after exposure to antibiotics during the night. Biofilms of Pseudomonas generated with mucin (2.5 g/l) and G-fragments at concentrations of 0, 1, 2 or 6%. Values are expressed as μg/ml of antibiotic

Table 5 - summary of values MWES 24 h after exposure to antibiotics during the night. Biofilms of Pseudomonas generated with mucin (2.5 g/l) and G-fragments at concentrations of 0, 1, 2 or 6%. Values are expressed as μg/ml of antibiotic

The key

Example 4 Measurement values MWES for Pseudomonas aeruginosa biofilms generated for 6 h and pretreated G-fragments

Was guided by the analysis MWES described above, with the mucin present throughout the analysis. Biofilms were generated in the presence of mucin within the 6-hour incubation, washed, and incubated with G-fragments and mucin during the night. After washing with PBS, the cultures were exposed to the effect of antibiotics (amikacin, tobramycin, oxytetracycline, or a combination of amikacin and oxytetracycline) without mucin. The results are graphically presented in figure 4 and in tables 6 and 7. As you can see, pre-treatment 6 h biofilms 6% G-fragments leads to 6 h values PSES for all the tested antibiotics was equal to at least one quarter, i.e. the sensitivity of Pseudomonas aeuroginosa in a biofilm to these antibiotics was increased at least four times. In fact, 6% G-fragments caused the recession to 1/8 of the control values 6 h values MWES for oxytetracycline. 24-hour values MWES for amikacin and tobramycin were halved. 24-hour value MWES for oxytetracycline and mixtures amikacin/oxytetracycline showed no change of values MWES.

Table 6 - summary of values MWe is at 6 h after exposure to antibiotics during the night. Biofilms of Pseudomonas generated in the environment with the added mucin within 6 h, were subjected to 0 or 6% G-fragments during the night and then were exposed to the effect of antibiotics. Values were expressed as μg/ml of antibiotic

Table 7 - Summary of values MWES 24 h after exposure to antibiotics during the night. Biofilms of Pseudomonas generated in the environment with the added mucin within 6 h, were subjected to 0 or 6% G-fragments during the night and then were exposed to the effect of antibiotics. Values are expressed as μg/ml of antibiotic

The key

Example 5 Measurement values MWES for Pseudomonas aeruginosa biofilms formed within 6 h without mucin and pre-treated G-fragments without mucin

The Protocol of Example 4 was repeated, using tobramycin and amikacin, but without the addition of mucin. The results are shown in Figure 5. As you can see, in the absence of mucin G-fragments were still able to halve all values MWES except 24 h values MWES for amikacin. This is an indication that the mucin does not play a major role in the effects observed in the Examples above.

Example 6 Measurement values MWES for Pseudomonas aeruginosa biofilms formed within 6 h and pretreated different and chinatime the oligomers

Analysis MWES described in Example 4 was repeated with the alternative alginate oligomer, G-unit (#0802) (molecular weight of 6400 compared to G-fragments CF-5/20, molecular weight 2600) and using tobramycin and amikacin. Is MWES in 24 h for amikacin decreased four times in the pre-treatment of biofilms 6% G-unit (#0802). The same treatment resulted in a decrease in half-24 h values MWES for tobramycin. These data show that another alginate oligomer may cause increased sensitivity of Pseudomonas aeruginosa RA in biofilms to tobramycin and amikacin.

Example 7 - Measurement values MWES for 6 h biofilms containing other bacteria pre-treated G-fragments

The impact of G-fragments on biofilms of Staphylococcus aureus was studied using analysis MWES described in Example 4, and oxytetracycline. As can be seen in Fig.7, the pre-treatment of biofilms containing S. aureus was ATSS 6538, 6% G-fragments, twice reduced values MWES 6 and 24 h for oxytetracycline. As you can see in Fig, pre-treatment of biofilms containing Raney isolate "1103" MRSA, 6% G fragments, twice reduced the value MWES in 24 h for tobramycin. These data show that other bacteria, usually found in biofilms, can be made more sensitive to oxytetracycline and tobramycin is here pre-treatment of such biofilms G-fragments.

Example 8 the Effect of G-fragments for the attachment of yeast in biofilm

The influence of G-fragments for the attachment of Candida albicans and Candida dubliniensis in the biofilm were studied using analysis of attachment described above. Reducing the attachment of both Candida species was observed when biofilms containing these yeast were formed in the presence of 2% G-fragments and mucin compared with one mucin as a control (Figure 9). These data indicate that G-fragments can affect the attachment of yeast cells in the developing biofilms.

Example 9 - Microscopic analysis of the structure of biofilms of Pseudomonas and influence of G-fragments

The General structure of Pseudomonas biofilms was monitored using scanning electron microscopy (SEM). Figure 10 shows the effect of 2% G-fragments on biofilm structure. Extracellular polysaccharide (EPS), covering the cell surface is impaired in the presence of 2% G-fragments.

Example 10 - Microscopic analysis of the structure of biofilms of Pseudomonas and influence of G-fragments

The influence of G-fragments on the structure of the biofilm matrix Pseudomonas studied using confocal microscopy of intact biofilms labeled with the fluorescent dye BODIPY® 630/650-X SE. This dye selectively stains the components of the matrix (EPS) in Pseudomonas biofilms. Subtle fragmentation biofilm Matri is s became visible with increasing concentrations of G-fragments compared with control only mucin".

Example 11 - Local composition comprising alginate oligomer

An example of a local composition (moisture lotion for skin care body)containing alginate oligomer, get with the following ingredients.

Oil phase:
Vaseline oil3%
Cyclomethicone4%
Isopropylmyristate3%
Stearic acid1,8%
Cetyl alcohol1,0%
Literallayout1,5%
Aqueous phase:
Carbomer 9840,10%
Glycerin3%
Triethanolamine0,90%
Alginate oligomer0,1%
Waterthe point 81.60%

Example 12 a Composition for PTS is recorded, containing alginate oligomer

An example of a liquid composition for cleansing containing alginate oligomer, get with the following ingredients, %:

Castor oil77,8
Peruvian balsam, refined10
Collagenase0,2
ZnCl0,5
Water5
Polyoxyethylene(10)alerby ether4
Colloidal silicon2
Alginate oligomer0,5

Example 13 Composition in the form of a sterile aqueous solution containing, per cent:

Alginate oligomer according to the invention 10

Saline9
Sterile purified water100

This solution is prepared by mixing the components in accordance with known methods manufactured by Garrett, Borg is tion.

Examples 14-15 - Sets containing alginate oligomer

Example 14

1 vial containing a sterile aqueous solution of alginate oligomer from Example 13; and

1 vial containing a sterile aqueous solution of azithromycin.

These solutions were obtained by mixing the components in accordance with known manufacturing methods.

Example 15

1 vial containing a sterile solution of alginate oligomer-based oils in Example 12; and

1 vial containing a sterile aqueous solution of benzalkonium chloride.

These solutions were obtained by mixing the components in accordance with known manufacturing methods.

1. In vitro method of combating biofilm containing gram-negative bacteria, gram-positive bacteria or yeast, including the conversion of the specified biofilms in contact with the alginate oligomer having an average molecular weight less than 20,000 daltons, and at least 80% of the residues of G (α-L-guluronate acid).

2. The In vitro method according to claim 1, where the biofilm is on the surface, selected from surfaces of machinery or equipment for the processing, preparation, storage or dispensing food or beverages, the surfaces of air-conditioning units, the surfaces of industrial machinery, surfaces of storage tanks, the surface effect is skogo or surgical equipment, surface water/marine equipment or surfaces of buildings and other structures.

3. The In vitro method according to claim 2, where the surface is selected from surfaces of equipment for processing, preparation, storage or dispensing, food, tanks, conveyors, floors, drains, coolers, freezers, equipment surfaces, walls, valves, belts, pipes, air conditioning systems, refrigeration units, lines, dispensing food or beverage, heat exchangers, boat hulls, dental technical water supply, pipelines for oil drilling, contact lenses, containers for storing contact lenses, catheters, prosthetic device or implanted medical devices.

4. The In vitro method according to claim 1, where the specified alginate oligomer used in combination c:
(1) an antimicrobial agent; or
(2) another destroying biofilm agent and/or decreasing mucosal viscosity agent.

5. The In vitro method according to claim 4, where the antimicrobial agent is an antibiotic, an antifungal agent, an antiseptic, disinfecting, sterilizing or cleaning agent.

6. The In vitro method according to claim 4, where the specified one destroying the biofilm agent and/or reducing mucosal viscosity agent is selected from proteases, nucleases, lipases, the farm is tov, able to break down polysaccharides, gelsolin, reductants of thiol, utilzation, sodium chloride, uncharged low molecular weight polysaccharide or anionic polyaminoamide and predecessor or stimulator of the synthesis of nitric oxide.

7. The In vitro method according to claim 4, where the specified one destroying the biofilm agent and/or reducing mucosal viscosity agent is an enzyme alginate-liasu and/or Tnkase.

8. The In vitro method according to any one of claims 1 to 7, where alginate oligomer has an average molecular weight of less than 15000, 10000, 8000 or 7000 Yes.

9. The In vitro method according to any one of claims 1 to 7, where alginate oligomer has srednecenovogo degree of polymerization 2-100, preferably 2-75, 2-50, 2-35 or 2-30.

10. The In vitro method according to any one of claims 1 to 7, where alginate oligomer has up to 100 Monomeric residues and is preferably 2-35-dimensional, 2-30-dimensional, 3-35-dimensional, 3-28-dimensional, 4-25-dimensional, 6-22-dimensional, 8-20-dimensional or 10-15-dimensional.

11. The In vitro method according to any one of claims 1 to 7, where alginate oligomer has at least 85%, 90%, 95% or 99% of the residues G.

12. The In vitro method according to any one of claims 1 to 7, where alginate oligomer has a primary structure in which at least 90% of the residues of G are connected by a link 1-4 with another residue G.

13. The use of alginate oligomer in the manufacture of a medicinal product for use in the treatment or prevention of biofilm is Peccei the subject, where biofilm infection comprises gram-negative bacteria, gram-positive bacteria or yeast and where alginate oligomer has an average molecular weight of less than 20,000 and at least 80% of residues G.

14. Use item 13, where the biofilm is inside or on the inner or on the outer surface of the body.

15. The application 14, where the internal or external body surface selected from a surface of the oral cavity, genital tract, urinary tract, respiratory tract, gastrointestinal tract, peritoneum, middle ear, prostate gland, the inner lining of blood vessels, mucous membranes of the eyes, corneal tissue, lung tissue, heart valves, skin, scalp, nails or the inner part of the Russian Academy of Sciences.

16. Use item 13, where the subject is selected from a subject with a previously developed infection of the subject with a weakened immune system, receiving intensive or intensive care treatment of the subject, the subject is suffering from injury, the subject of a burn, a subject with acute and/or chronic wound, a newborn of a subject, the subject is elderly subject with cancer, a subject suffering from an autoimmune condition, a subject with reduced or eliminated epithelial or endothelial secretion and/or clearance of secret or entity entered medical device.

<> 17. The application of article 16, where the subject is selected from a subject with a condition selected from HIV (human immunodeficiency virus), sepsis, septic shock, AIDS (acquired immunodeficiency syndrome), cancer of the immune system, rheumatoid arthritis, diabetes type 1 diabetes, Crohn's disease, COPD (chronic obstructive pulmonary disease), bronchitis, cystic fibrosis, emphysema, lung cancer, asthma, pneumonia and sinusitis, subject, prepared to undergoing, or recovering from chemotherapy and/or radiotherapy, subjects with organ transplantation, the subject in the health care institution, or the smoker.

18. Use item 13, where the specified drug is used to treat and/or prevent biofilm infections of wounds and/or burns or "put" medical device.

19. Use item 13, where biofilm infection is a Pseudomonas, such as Pseudomonas aeruginosa infection.

20. Use item 13, where the drug used in the treatment and/or prevention of dental plaque, gingivitis, periodontitis, congenital valve endocarditis, acute inflammation of the middle ear, chronic bacterial prostatitis, pneumonia, asthma or related device infections associated with implantable and/or prosthetic medical devices or tissue transfers.

21. Use item 13, where the specified drug is used in combination c:
(1) an antimicrobial agent;
(2) another destroying biofilm agent and/or decreasing mucosal viscosity agent; or
(3) additional therapeutically active agent.

22. Use item 21, where the specified antimicrobial agent is an antibiotic, an antifungal agent, an antiseptic, disinfecting, sterilizing or cleaning agent.

23. Use item 21, where the specified one destroying the biofilm agent and/or reducing mucosal viscosity agent is selected from proteases, nucleases, lipases, enzymes that can destroy polysaccharides, gelsolin, reductants of thiol, utilzation, sodium chloride, uncharged low molecular weight polysaccharide, anionic polyaminoamide and predecessor or stimulator of the synthesis of nitric oxide.

24. Use item 13, where the specified drug is used in combination with enzyme alginate-liati and/or Dnazol.

25. Use item 21, where the specified additional therapeutically active agent is an immune-stimulating agent, a growth factor or an anti-inflammatory agent.

26. The use according to any one of p-25, where the alginate oligomer has an average molecular weight of less than 15000, 10000, 8000 or 7000 Yes.

27. The application of l is the Boma from PP-25, where alginate oligomer has srednecenovogo degree of polymerization 2-100, preferably 2-75, 2-50, 2-35 or 2-30.

28. The use according to any one of p-25, where the alginate oligomer has up to 100 Monomeric residues and is preferably 2-35-dimensional, 2-30-dimensional, 3-35-dimensional, 3-28-dimensional, 4-25-dimensional, 6-22-dimensional, 8-20-dimensional or 10-15-dimensional.

29. The use according to any one of p-25, where the alginate oligomer has at least 85%, 90%, 95% or 99% of the residues G.

30. The use according to any one of p-25, where the alginate oligomer has a primary structure in which at least 90% of the residues of G are connected by a link 1-4 with another residue G.

31. A set containing the alginate oligomer and the second active agent for use in combating biofilm or in the treatment or prevention of biofilm infection in a subject, where biofilm infection comprises gram-negative bacteria, gram-positive bacteria or yeast, where alginate oligomer has an average molecular weight of less than 20,000 and at least 80% of residues G and where specified the second active agent is a:
(1) an antimicrobial agent;
(2) another destroying biofilm agent and/or reducing mucosal viscosity agent;
(3) an immunostimulating agent;
(4) a growth factor; or
(5) anti-inflammatory agent.

32. Set p where specified antimicrobial agent p is ecstasy an antibiotic, an antifungal agent, an antiseptic, disinfecting, sterilizing or cleaning agent.

33. Set p where the specified one destroying the biofilm agent and/or reducing mucosal viscosity agent is selected from proteases, nucleases, lipases, enzymes that can destroy polysaccharides, gelsolin, reductants of thiol, utilzation, sodium chloride, uncharged low molecular weight polysaccharide, anionic polyaminoamide and predecessor or stimulator of the synthesis of nitric oxide.

34. Set p where the specified one destroying the biofilm agent and/or reducing mucosal viscosity agent is an enzyme alginate-liasu and/or Tnkase.

35. Set p in the form of hypertonic saline solution inhalation solution, inhalation powder, nasal spray, cleansing compositions, local songs, toothpaste or liquid for rinsing the mouth.

36. Set according to any one of p-35, where the alginate oligomer has an average molecular weight of less than 15000, 10000, 8000 or 7000 Yes.

37. Set according to any one of p-35, where the alginate oligomer has srednecenovogo degree of polymerization 2-100, preferably 2-75, 2-50, 2-35 or 2-30.

38. Set according to any one of p-35, where the alginate oligomer has up to 100 Monomeric residues and is preferably 2-35-dimensional, 2-30-dimensional, 3-35-dimensional, 3-28-dimensional, 4-25-dimensional, 6-2-dimensional, 8-20-dimensional or 10-15-dimensional.

39. Set according to any one of p-35, where the alginate oligomer has at least 85%, 90%, 95% or 99% of the residues G.

40. Set according to any one of p-35, where the alginate oligomer has a primary structure in which at least 90% of the residues of G are connected by a link 1-4 with another residue G.

41. Machinery or equipment for processing, storage or dispensing food or beverages, covered with alginate oligomer for combating biofilm with an average molecular weight of less than 20,000 and at least 80% of residues G.

42. Machinery or equipment under paragraph 41, where the alginate oligomer has an average molecular weight of less than 15000, 10000, 8000 or 7000 Yes.

43. Machinery or equipment under paragraph 41, where the alginate oligomer has srednecenovogo degree of polymerization 2-100, preferably 2-75, 2-50, 2-35 or 2-30.

44. Machinery or equipment under paragraph 41, where the alginate oligomer has up to 100 Monomeric residues and is preferably 2-35-dimensional, 2-30-dimensional, 3-35-dimensional, 3-28-dimensional, 4-25-dimensional, 6-22-dimensional, 8-20-dimensional or 10-15-dimensional.

45. Machinery or equipment under paragraph 41, where the alginate oligomer has at least 85%, 90%, 95% or 99% of the residues G.

46. Machinery or equipment under paragraph 41, where the alginate oligomer has a primary structure in which at least 90% of the residues of G are connected by a link 1-4 with another residue G.

47. Installation of air conditioning, covered with alginate oligomer for combating biofilm with an average molecular weight of less than 20,000 and at least 80% of residues G.

48. Installation p, where alginate oligomer has an average molecular weight of less than 15000, 10000, 8000 or 7000 Yes.

49. Installation p, where alginate oligomer has srednecenovogo degree of polymerization 2-100, preferably 2-75, 2-50, 2-35 or 2-30.

50. Installation p, where alginate oligomer has up to 100 Monomeric residues and is preferably 2-35-dimensional, 2-30-dimensional, 3-35-dimensional, 3-28-dimensional, 4-25-dimensional, 6-22-dimensional, 8-20-dimensional or 10-15-dimensional.

51. Installation p, where alginate oligomer has at least 85%, 90%, 95% or 99% of the residues G.

52. Installation p, where alginate oligomer has a primary structure in which at least 90% of the residues of G are connected by a link 1-4 with another residue G.

53. Industrial machinery or equipment covered alginate oligomer for combating biofilm with an average molecular weight of less than 20,000 and at least 80% of residues G.

54. Machinery or equipment item 53, where the alginate oligomer has an average molecular weight of less than 15000, 10000, 8000 or 7000 Yes.

55. Machinery or equipment item 53, where the alginate oligomer has srednecenovogo degree of polymerization 2-100, preferably -75, 2-50, 2-35 or 2-30.

56. Machinery or equipment item 53, where the alginate oligomer has up to 100 Monomeric residues and is preferably 2-35-dimensional, 2-30-dimensional, 3-35-dimensional, 3-28-dimensional, 4-25-dimensional, 6-22-dimensional, 8-20-dimensional or 10-15-dimensional.

57. Machinery or equipment item 53, where the alginate oligomer has at least 85%, 90%, 95% or 99% of the residues G.

58. Machinery or equipment item 53, where the alginate oligomer has a primary structure in which at least 90% of the residues of G are connected by a link 1-4 with another residue G.

59. Tank or storage vessel, covered with alginate oligomer for combating biofilm with an average molecular weight of less than 20,000 and at least 80% of residues G.

60. The tank or vessel by p, where alginate oligomer has an average molecular weight of less than 15000, 10000, 8000 or 7000 Yes.

61. The tank or vessel by p, where alginate oligomer has srednecenovogo degree of polymerization 2-100, preferably 2-75, 2-50, 2-35 or 2-30.

62. The tank or vessel by p, where alginate oligomer has up to 100 Monomeric residues and is preferably 2-35-dimensional, 2-30-dimensional, 3-35-dimensional, 3-28-dimensional, 4-25-dimensional, 6-22-dimensional, 8-20-dimensional or 10-15-dimensional.

63. The tank or vessel by p, where alginate oligomer has at least 85%, 90%, 95% or 99% of the residues G.

64. The tank or vessel by p, where the algae is more oligomer has a primary structure, in which at least 90% of the residues of G are connected by a link 1-4 with another residue G.

65. Medical or surgical equipment or device is covered with alginate oligomer for combating biofilm with an average molecular weight of less than 20,000 and at least 80% of residues G.

66. Equipment or device on p, where alginate oligomer has an average molecular weight of less than 15000, 10000, 8000 or 7000 Yes.

67. Equipment or device on p, where alginate oligomer has srednecenovogo degree of polymerization 2-100, preferably 2-75, 2-50, 2-35 or 2-30.

68. Equipment or device on p, where alginate oligomer has up to 100 Monomeric residues and is preferably 2-35-dimensional, 2-30-dimensional, 3-35-dimensional, 3-28-dimensional, 4-25-dimensional, 6-22-dimensional, 8-20-dimensional or 10-15-dimensional.

69. Equipment or device on p, where alginate oligomer has at least 85%, 90%, 95% or 99% of the residues G.

70. Equipment or device on p, where alginate oligomer has a primary structure in which at least 90% of the residues of G are connected by a link 1-4 with another residue G.

71. Water or marine equipment, coated alginate oligomer for combating biofilm with an average molecular weight of less than 20,000 and at least 80% of residues G.

72. Equipment p, where alginate oligomer has an average m is molecular weight of less than 15,000, 10000, 8000 or 7000 Yes.

73. Equipment p, where alginate oligomer has srednecenovogo degree of polymerization 2-100, preferably 2-75, 2-50, 2-35 or 2-30.

74. Equipment p, where alginate oligomer has up to 100 Monomeric residues and is preferably 2-35-dimensional, 2-30-dimensional, 3-35-dimensional, 3-28-dimensional, 4-25-dimensional, 6-22-dimensional, 8-20-dimensional or 10-15-dimensional.

75. Equipment p, where alginate oligomer has at least 85%, 90%, 95% or 99% of the residues G.

76. Equipment p, where alginate oligomer has a primary structure in which at least 90% of the residues of G are connected by a link 1-4 with another residue G.

77. Refrigeration equipment covered alginate oligomer for combating biofilm with an average molecular weight of less than 20,000 and at least 80% of residues G.

78. Equipment p, where alginate oligomer has an average molecular weight of less than 15000, 10000, 8000 or 7000 Yes.

79. Equipment p, where alginate oligomer has srednecenovogo degree of polymerization 2-100, preferably 2-75, 2-50, 2-35 or 2-30.

80. Equipment p, where alginate oligomer has up to 100 Monomeric residues and is preferably 2-35-dimensional, 2-30-dimensional, 3-35-dimensional, 3-28-dimensional, 4-25-dimensional, 6-22-dimensional, 8-20-dimensional or 10-15-dimensional.

81. Equipment p, where alginate oligomer has at least 85, 90%, 95% or 99% of the residues G.

82. Equipment p, where alginate oligomer has a primary structure in which at least 90% of the residues of G are connected by a link 1-4 with another residue G.

83. A cleansing composition for combating biofilm containing
(1) alginate oligomer having an average molecular weight of less than 20,000 and at least 80% of the residues of G; and
(2) at least one proteolytic enzyme and/or at least one abrasive solid phase,
where this composition is a sterile aqueous solution or a sterile solution or oil-based.

84. Cleansing composition according p, where alginate oligomer has an average molecular weight of less than 15000, 10000, 8000 or 7000 Yes.

85. Cleansing composition according p, where alginate oligomer has srednecenovogo degree of polymerization 2-100, preferably 2-75, 2-50, 2-35 or 2-30.

86. Cleansing composition according p, where alginate oligomer has up to 100 Monomeric residues and is preferably 2-35-dimensional, 2-30-dimensional, 3-35-dimensional, 3-28-dimensional, 4-25-dimensional, 6-22-dimensional, 8-20-dimensional or 10-15-dimensional.

87. Cleansing composition according p, where alginate oligomer has at least 85%, 90%, 95% or 99% of the residues G.

88. Cleansing composition according p, where alginate oligomer has a primary structure in which at least 90% of the residues of G are connected by a link 1-4 with another G.



 

Same patents:

Organic compounds // 2489439

FIELD: biotechnologies.

SUBSTANCE: invention is related to a compound of a formula

, where R' is the below formula, and R" is hydrogen, in free form or in the form of a pharmaceutically acceptable acid-additive salt. Proposed compounds of the formula (IA) inhibit activity of DPP-IV (dipeptidylpeptidase-IV).

EFFECT: compounds are justified for application as medicinal agents designed for inhibition of DPP-IV and for treatment of conditions mediated by DPP-IV, such as achrestic diabetes.

11 cl, 2 ex, 2 dwg, 7 tbl

FIELD: organic chemistry, medicine, pharmacy.

SUBSTANCE: invention relates to geranyl compounds represented by the following formulas (I-1) , (I-2) or (I-3) wherein R1 means compounds of the following formulas: or R2 means a group remaining after removing all carboxyl groups presenting in carboxylic acid chosen from group consisting of malic acid, citric acid, succinic acid, fumaric acid and others; m = 1, 2 or 3; n = 0, 1 or 2, and m + n represent a number of carboxylic groups presenting in indicated carboxylic acid; R3 means p-hydroxyphenyl or mercapto-group. Also, invention relates to derivatives of mevalonic acid represented by the following formula (I-4): wherein R means -CH2OH or CH3. Also, invention to an antitumor agent comprising as an active component geranyl compound of formulas (I-1), (I-2) or (I-3) or derivative of mevalonic acid of the formula (I-4), and optionally a pharmaceutically acceptable carrier or solvent. Also, invention relates to a method for treatment of liver cancer based on using geranyl compound of formulas (I-1), (I-2) or (I-3), or derivative of mevalonic acid of the formula (I-4) and using proposed compounds in manufacturing an antitumor agent. Invention provides using geranyl compounds or derivatives of mevalonic acid as antitumor agents.

EFFECT: valuable medicinal properties of compounds and pharmaceutical composition.

7 cl, 3 tbl, 17 ex

The invention relates to a method for producing D-glucuronic acid by heating salts 1,2-D-isopropylidene - D-glucuronic acid in aqueous solution in the presence of acid agents

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to pharmaceutical industry and represents a method for increasing biocidal and therapeutic action of a suspension-cream with metronidazole containing metronidazole 10 mg and chlorhexidine 0.5 mg in gel 1 g, consisting in detoxification and polymerisation of the suspension-cream with 0.1% glutaric aldehyde with 0.1% alkyldimethylbenzylammonium chloride, 2% aethonium, 0.2% metronidazole and 0.1% dimethylsulphoxide.

EFFECT: invention provides biocidal action on viruses, aerobic and anaerobic microorganisms, spores, mould fungi, faster regeneration and biodegradation for 8-12 hours.

5 ex, 1 tbl

FIELD: medicine.

SUBSTANCE: group of inventions refers to veterinary science and aims at normalising metabolic processes, stimulating immune system and blocking mechanisms of infectious process with a risk of endogenous infection activation. A method for producing a combination immunometabolic preparation with anti-infectious activity involves dissolving succinic acid and levamisole in demineralised water with formalin added. According to the other aspect of the invention, the immunometabolic preparation additionally contains polyethylene glycol. The ingredients are used in the declared ratio.

EFFECT: using this group of inventions provides producing an injection form of the immunometabolic preparation with anti-infectious activity.

2 cl, 3 tbl

FIELD: medicine.

SUBSTANCE: treating a wound surface with dioxidine is followed by an infrared laser light with a permanent magnetic field not earlier than 5 days after the operation. Magnetic induction intensity is within the range of 20-50 mT; a laser pulse repetition frequency is within the range of 80 Hz, and a power is 0.25-0.5 W. The whole postoperative area is subject to the daily distant labile exposure to a defocused beam at 0.5 cm for 30-60 seconds. That is followed by applying tissues with hypertonic solution 3 to 5 times a day; the therapeutic course is 10-15 procedures.

EFFECT: method enables providing higher effectiveness and reducing a length of treatment ensured by the integrated exposure to the antibacterial agents and magnetic laserophoresis in the presented regimen, preventing developing postoperative complications.

2 ex

FIELD: medicine.

SUBSTANCE: group of inventions refers to medicine, namely to medical microbiology, and can be used for treating intestinal yersiniosis, or pseudotuberculosis, or salmonellosis experimentally. That is ensured by simulating one of the above pathologies in experimental animals followed by oral administration of therapeutic preparations in therapeutically effective doses with controlling the content of pathological microflora for a period of time providing elimination of the disease. Stimbifid or a culture fluid of lactic acid bacilli is administered orally as the therapeutic preparation into each animal for the first 2 hours after infection once a day as a monotherapeutic preparation promoting the colonisation resistance of the intestinal mucosa to an agent. Stimbifid is administered into each animal in a dose of 13 mg for 5 days from the moment of the infection, while the culture fluid is administered in an amount of 0.2 ml throughout 7 days from the moment of the injection. What is also presented is a method for preparing an agent for treating intestinal yersiniosis, or pseudotuberculosis, or salmonellosis experimentally.

EFFECT: group of inventions provides a high percentage of pathology elimination by using the prebiotic preparation and the liquid probiotic complex with no accompanying negative consequences.

4 cl, 3 tbl, 3 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention relates to methods of treating or relieving severity of disease in patient, where disease is selected from mucoviscidosis, hereditary emphysema, chronic obstructive pulmonary disease (COPD), "dry eye" disease. Methods include introduction of effective amount of N-(5-hydroxy-2,4-di-tert-butylphenyl)-4-oxo-1H-quinoline-3-carboxamide or pharmaceutical composition, containing said compound, to patient.

EFFECT: treatment of relief of disease severity in patient, where disease is selected from mucoviscidosis, hereditary emphysema, chronic obstructive pulmonary disease (COPD), "dry eye" disease.

16 cl, 15 tbl

FIELD: medicine, pharmaceutics.

SUBSTANCE: claimed invention provides pharmaceutical compositions, which include an effective quantity of ceftaroline fosamile or its pharmaceutically acceptable salt and an antibacterial agent or its pharmaceutically acceptable salt. The antibacterial agent is selected from a group, consisting of meropenem, piperacillin plus tazobactam, amikacin and astreonam.

EFFECT: compositions by the invention produce a synergistic effect on bacteria strains, are used in treatment of bacterial infections.

7 cl, 2 dwg, 17 tbl, 3 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: in formula R1 is H or (1-6C alkyl); R2 represents NRbRc, (1-4C)alkyl, (1-4C)fluoroalkyl, CF3, (1-4C)hydroxyalkyl, -(1-4Calkyl)hetAr1, -(1-4Calkyl)NH2, -(1-4C alkyl)NH(1-4Calkyl), -(1-4Calkyl)N(1-4Calkyl)2, hetAr2, hetCyc1, hetCyc2, phenyl substituted where applicable by NHSO2(1-4Calkyl) or (3-6C)cycloalkyl, substituted where applicable by (1-4C alkyl), CN, OH, OMe, NH2, NHMe, N(CH3)2, F, CF3, CO2(1-4C alkyl), CO2H; C(=O)NReRf or C(=O)ORg; Rb is H or (1-6C alkyl); Rc represents H, (1-4C)alkyl, (1-4C)hydroxyalkyl, hetAr3 or phenyl, wherein the above phenyl is substituted where applicable by one or more substitutes independently from halogen, CN, CF3 and -O(1-4C alkyl); Re represents H or (1-4C)alkyl; Rf represents H, (1-4C)alkyl or (3-6C)cycloalkyl; Rg represents H or (1-6C)alkyl; X is absent or represents -CH2-, -CH2CH2-, -CH2O- or -CH2NRd; Rd represents H or (1-4C alkyl); R3 represents H or (1-4C alkyl); and n is equal to 0-6. The radical values NRbRc, Y, hetAr1, hetAr2, hetAr3, hetCyc1, hetCyc2, NReRf, R4 are specified in the patent claim. The invention also refers to a pharmaceutical composition containing the above compounds, to a method of treating Trk kinase mediated diseases and conditions, such as pain, cancer, inflammation, neurodegenerative disease, Typanosoma cruzi infection, osteolytic disease, and to a method of preparing the above compounds.

EFFECT: invention refers to new derivatives of pyrazolo[1,5-a]pyrimidines possessing an inhibitory activity on tropomyosin-related kinases (Trk).

42 cl, 1 tbl, 105 ex

FIELD: veterinary medicine.

SUBSTANCE: method of emergency prevention and treatment of brucellosis, comprising simultaneous intramuscular administration of antibiotic resistance version of the vaccine strain of Brucella and antibrucellar preparation, characterised in that as the particular preparation the cyprolet- resistant version of strain of Brucella B.abortus 82 CR is used at a dose of 1.109 m.c. , and as the antibacterial preparation the ciprofloxacin (cyprolet) is used which is administered at a dose of 1 and 3 mL/kg, 2 times per day for 5-7 days. The cyprolet- resistant version of Brucella B.abortus 82 CR is obtained by growing a stock culture of Brucella B.abortus 82 on MPGGA, containing 5, 10, 20 mg per 1 ml of medium. Prevention and treatment of brucellosis is carried out 1-10 days prior to the period of greatest risk of infection and in the period of greatest risk of infection.

EFFECT: method increases effectiveness of treatment and prevention of brucellosis due to interference of avirulent strain of Brucella in relation to brucellosis pathogen, induction by cyprolet- resistant version of vaccine strain of mediators of immunogenesis - activators of phagocytosis-cytokines, powerful antibacterial agent cyprolet, which together provide the suppression of reproduction of microbial DNA and death of parasite.

3 cl, 8 ex

FIELD: veterinary medicine.

SUBSTANCE: method comprises hyperimmunisation of bulls-producers with inactivated antigens C1. perfringens, comprising toxoids and somatic antigens of bacteria of serotypes A, C and D in the culture medium with the following ratio of components per 1 litre of antigen: toxoid and somatic antigen of strain No 28 (type A) in the culture medium with a concentration of 1012 · (9-10) m.c. in 1 cm3, cm3 - 300.0-350.0; toxoid and somatic antigen of strain No 392 (type C) in the culture medium at a concentration of 1012 · (9-10) m.c. in 1 cm3, cm3 - 300.0-350.0; toxoid and somatic antigen of strain No 213 (type D) in the culture medium at a concentration of 1012 · (9-10) m.c. in 1 cm3, cm3 - 300.0-350.0; formalin, cm3 - 4.0-5.0. The additional hyperimmunisation of bulls-producers is carried out with inactivated antigens of E.coli, comprising somatic and adhesive antigens K99, A20 in saline, TL-, TS-toxoids of E.coli in the culture medium in the following ratio of components per 1 litre of antigen: somatic and adhesive antigen of strain KV-1 (K99) in saline with a concentration of 1012 · (14-15) bln.c. in 1 cm3, cm3 - 300.0-350.0; somatic and adhesive antigen of strain PZ-3 (A20) in saline with a concentration of 1012 · (14-15) m.c. in 1 cm3, cm3 - 300.0-350.0; formalin, cm3 - 4.0-5.0; thermostable and thermolabile toxoids of E. coli strains KV-1 and PZ-3 in the culture medium (DPR) 1:8-1:16, l - to 1. Hyperimmunisation of bulls-producers is carried out four times by subcutaneous injection in the neck region on one side of the antigen dose C1. perfringens and in the neck region on the other side - E.coli antigen dose, according to the following scheme: on the first day - 8 cm3, after 14 days - 10 cm3, after 28 days - 15 cm3, and after 42 days - 20 cm3 from blood, then serum is separated and preserved.

EFFECT: invention with the further intake enables to obtain the hyperimmune serum, which provides treatment and prevention of mixed infection - anaerobic enterotoxaemia and colibacillosis diarrhoea of calves.

5 tbl, 7 ex

FIELD: medicine.

SUBSTANCE: invention refers to veterinary science and aims at preventing and treating the porcine respiratory disease complex PRDC. What is declared is using a recombinant protein ORF2 PCV2 or an immunogenic composition containing the recombinant protein ORF2 PCV2, for preparing a therapeutic agent for preventing and treating the porcine respiratory disease complex (PRDC) caused by Mycoplasma hyorhinis and/or a PRDC-associated clinical symptom of a notable increase of a death rate in the middle to late feeding phase caused by PCV2, and at least one another pathogen causing PRDC in animals, wherein one pathogen causing PRDC is specified in a group containing PRRSV, Mycoplasma hyopneumoniae, Bordetella bronchiseptica, swine influenza virus, Actinobacillus pleuropneumoniae, Mycoplasma hyorhinis. Streptococcus suis and/or Pasteurella multocida.

EFFECT: invention is high-effective for treating and preventing the porcine respiratory disease complex PRDC.

2 dwg, 3 tbl, 3 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention relates to the pharmaceutical industry, in particular to a peroral pharmaceutical composition, which contains polysaccharides from platyclades of Opuntia Ficus Indica, an extract of Olea Europeae leaves, alginate and sodium bicarbonate in a specified ratio. Components of the composition described above act with respect to reduction of gastroesophageal reflux.

EFFECT: peroral pharmaceutical composition is intended for the prevention and treatment of gastroesophageal reflux and GERD.

4 cl, 9 tbl, 6 dwg

FIELD: chemistry.

SUBSTANCE: group of inventions relates to biotechnology and medicine. Disclosed is a polysaccharide which is isolated from the Bifidobacterium infantis NCIMB 41003 strain and has the structure [-β(1,3)-D-GalpNAc-β(1,4)-D-Glcp-]n, where said disaccharide unit repeats n times, which yields a polysaccharide with molecular weight greater than 100000 Da. The polysaccharide exhibits immunomodulating activity and is used in preparing medicinal agents for treating or preventing undesirable inflammatory activity, undesirable gastrointestinal inflammatory activity, rheumatoid arthritis and autoimmune disorders.

EFFECT: pharmaceutical composition for treating and preventing inflammatory disorders and a food product containing the isolated polysaccharide are disclosed.

9 cl, 6 dwg, 3 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to pharmaceutical industry, namely to a method for preparing water-soluble fractions of mannoproteins and β-glucan. A method for preparing the water-soluble fractions of mannoproteins and β-glucan consisting in the fact that yeast biomass is prepared by mechanical activation in activators and mills; the prepared mechanical complex is added with a solution of enzymic complex showing β-glucanase or protease activity; that is followed by hydrolysis; the prepared hydrolysate is divided into mannoprotein and β-glucan fractions to be subject to purification under certain conditions.

EFFECT: method provides the more effective hydrolysis and higher yield of the end product.

2 cl, 4 dwg, 10 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to pharmaceutical industry and represents pharmaceutical composition for treating gastroesophageal reflux disease, containing at least one proton pump inhibitor and at least one probiotic, wherein the proton pump inhibitor is taken in the amount of 0.05-25 wt % in the composition; and the probiotic is taken in the amount of 10-95 wt %; additive agents up to 100 wt %.

EFFECT: provided preventing Hpylori translocation, avoiding the necessity of Hpylori detection and antibacterial course of eradication, higher safety of the prolonged therapy with the proton pump inhibitors and avoided gastric mucosa atrophy, and a risk of gastric cancer.

5 cl, 10 tbl

FIELD: medicine.

SUBSTANCE: invention refers to medicine. What is described is using the material for the purpose of neural dysfunction recovery with the above material containing a polysaccharide derivative hydrogel wherein 0.5 wt % of the aqueous solution contains a complex module in the amount of 1 to 1000 N/m2, while a loss factor makes 0.01 to 2.0 that is measured at angular velocity 10 rad/sec with using a dynamic viscoelasticity meter. The above material for neural dysfunction recovery may represent hydrogel injected with using a syringe and has an excellent body residence, and has a restorative effect on the damaged or degenerated nerve function.

EFFECT: preparing the material for neural dysfunction recovery.

19 cl, 6 dwg, 6 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: group of inventions relates to medicine, particularly to ophthalmology. An ophthalmic composition for treating keratoconjunctival damages and inflammations consists of an aqueous solution containing arabinogalactan 1 to 10 wt %, one or more preserving agents specified in a group consisting of sodium merthiolate, thimerosal, phenylmercuric nitrate or phenylmercuric acetate, phenylethyl alcohol, methyl-, ethyl-, propyl parabene, chlorhexidine acetate or gluconate or chlorobutanol, and containing no benzalkonium chloride. The ophthalmic composition is applied as a lacrimal substitute recommended for people wearing contact lenses.

EFFECT: group of inventions provides treating corneal erosions caused by wearing contact lenses.

16 cl, 5 tbl, 7 dwg

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to chemical-pharmaceutical industry and represents a composite enterosorbent of a silicone polymer specified in a group containing methyl monosilane acid xerogel or methyl monosilane acid hydrogel, differing by the fact that contains at least one ingredient specified in a group: lactulose, inulin, lignin, fructooligosaccharide, alginic acid in the form of pharmaceutically acceptable salts, chitosan, pectin, gum resin, beta-glucan in the amount of 0.1 to 10 portions per 1 weight portion of monosilane acid hydrogel or xerogel.

EFFECT: invention provides creating an agent to provide normalising the intestinal microflora and relieving the manifesting intoxication.

3 cl, 6 ex

Up!