Prolonged delivery of compstatin analogues from gels

FIELD: medicine.

SUBSTANCE: group concerns a prolonged delivery of a compstatin analogue, as well as an optional additional active agent when released from a microscopic gel-like inclusion formed if a liquid composition containing the compstatin analogue is introduced into an extravascular space, such as a vitreous chamber of eye in a mammalian body. The invention also refers to a method of treating an individual suffering age-related macular degeneration (ARMD).

EFFECT: improving the compstatin delivery system for the complement system inhibition required for treating ARMD.

58 cl, 4 ex, 6 tbl, 5 dwg

 

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority based on provisional applications U.S. patent No. USSN 60/976,919 dated October 2, 2007, and USSN 61/026,460, 5 February 2008. The content of these applications is incorporated into the present application by reference.

PREREQUISITES TO the CREATION of INVENTIONS

[0002] the complement System consists of more than 30 serum and cellular proteins involved in three major signaling pathways, known as the classical, alternative and pectin way. A classical way, as a rule, starts with the binding of a complex of antigen and antibody IgM or IgG with C1 (though other certain promoters can initiate this cascade). Activated C1 cleaves C4 and C2 with education Sa and C4b, and Sa and C2b. C4b and Sa combined with the formation of the C3-convertase, which cleaves C3 to education Sa and C3b. Linking 3b with C3-convertase leads to the formation of C5-convertase, which cleaves C5 to Sa and C5b. Sa, Sa and Sa are anaphylatoxins and mediate different responses in acute inflammatory response. Sa and Sa are also chemotactic factors that attract cells of the immune system such as neutrophils.

[0003] the Activation of alternative pathways initiate the surface of bacterial cells and a variety of complex polysaccharides. In this cascade C3b, is razuysya the cleavage of C3, which occurs spontaneously at a low level, binds to the target, for example, with the surfaces of cells and forms a complex with factor B, which is later cleaved by factor D to form C3-Mac. The cleavage of C3 and bind other molecules with C3b C3-convertase leads to the formation of C5-convertase. The activity of the C3 - and C5-Mac this pathway is regulated proteins CR1, DAF, MCP and fH. The action of these proteins is related to either their ability to enhance the activity of decay (i.e., the ability to break down convertase), or with the ability to serve as a cofactor of factor I in the cleavage of C3b or C4b, or so, and the others.

[0004] C5-convertase, which are formed during both paths, split C5 with education Sa and C5b. C5b then contacts C6, C7 and C8 with the formation of the C5b-8, which is a catalyst for the polymerization of C9, leading to the formation of C5b-9 membrane attack complex (MAC). The MAC itself is embedded in the membrane of target cells and induces lysis of the cells. The presence of small amounts MAC on the cell membrane can result in a number of events other than death of a cell.

[0005] Leckenby way of complement is initiated by the binding of mannose-binding lectin (MBL) and MBL-associated serine protease (MASP) with carbohydrates. Gene MBL-1 (the man known as LMAN-1) encodes an integral membrane protein type I localized the intermediate region between the endoplasmic reticulum and Golgi apparatus. Gene MBL-2 encodes a soluble mannose-binding protein found in serum. The person in pectin cascade MASP-1 and MASP-2 are involved in the proteolysis of C4 and C2, leading to the formation of C3-convertase described above.

[0006] the Activity of the complement system in mammals is regulated by various proteins, which are proteins control complement (SSR) or proteins-regulators of activation of complement (RCA) (U.S. Patent No. 6,897,290). These proteins differ in their specificity for the ligand and the mechanisms of suppression of complement. They can accelerate the disintegration Mac and/or act as cofactors for factor I in enzymatic cleavage of C3b and/or C4b into smaller fragments. SSR are characterized by the presence of several (usually 4-56) homologous motifs, which are called short consensus repeats (SCR)modules squirrel control of complement (SSR) or SUSHI domains. For these domains, consisting of approximately 50-70 amino acids, usually about 60 amino acids, characterized by the conserved motif, which includes four cysteine connected by disulfide bonds (two disulfide bonds), Proline, tryptophan and many hydrophobic residues. A family of proteins SSR includes the complement receptor type 1 (CR1; C3b:C4b receptor), complement receptor type 2 (CR2), membrane cfactory protein (MCP; CD46), f is a torus, accelerating decomposition (DAF-factor), complement factor H (fH) and C4b-binding protein (C4bp). CD59 is associated with membrane regulator of complement, not structurally related to SSR.

[0007] More information about the complement system and its activation can be found in the following references: Makrides SC, Pharm Rev., 50(1): 59-87, 1998;

Lisczewski, MK and Atkinson, JP, The Human Complement System in Health and Disease, Volanakis, JE and Frank, MM, eds., Dekker, New York, pp.149-66, 1998; Kuby Immunology, 2000; Paul, W.E., Fundamental Immunology, Lippincott Williams & Wilkins; 5thed., 2003; and Walport MJ., Complement. First of two parts. N Engi J Med., 344(14): 1058-66, 2001.

[0008] Along with the fact that activation of the complement system plays an important role in the reactions of innate and acquired immunity, increasing evidence indicates that the complement system is involved in the processes of tissue damage in various ischemic, inflammatory and autoimmune diseases (Makrides, SC, Pharm Rev., 50(1): 59-87, 1998; Lisczewski, MK and Atkinson, JP, in The Human Complement System in Health and Disease, Volanakis, JE and Frank, MM, eds., Dekker, New York, pp.149-66, 1998). Inhibition of the complement system has been proposed as a therapeutic strategy in many of these diseases. Konstatin and its analogues are cyclic peptides that are associated with Sz and inhibit its activation. In this area there is a need for new compositions and methods introduction analogues constatine. There is also a need in the POPs is assured of an improved system of drug delivery.

A BRIEF DESCRIPTION of the INVENTION

[0009] the Present invention provides new compositions and methods of administration of analogues of constatine slow release in the mammalian organism. In one aspect of the invention features a liquid composition containing an analogue of constatine sufficient for the formation of a macroscopic gel-like structure at its introduction into the extravascular space in the body of a mammal. In some embodiments of the invention extravascular space is a vitreous chamber of the eyeball. In other embodiments of the extravascular space is a subconjunctival space. In some embodiments of the extravascular space is a retro-bulbar, subconjunctival, subtenon or subretinal space. In some embodiments of the invention, the analog constatine has an activity of at least 100 times greater activity constatine. In some embodiments of the invention similar constatine has an activity of at least 150 times greater activity constatine. In some embodiments of the invention, the analog constatine has an activity of at least 200 times greater activity constatine. In some embodiments, R is the realization of the invention, the analog constatine has an activity of at least 250 times greater activity constatine. In some embodiments of any of the methods according to the invention, comprising the step of introducing the composition to the subject, the subject is a monkey. In some embodiments of any of the methods according to the invention, comprising the step of introducing the composition to the subject, the subject is the man.

[0010] In some embodiments of the invention the liquid composition comprises an analog of constatine and the second active agent in addition to the analog constatine. The second active agent can predstavlaete a polypeptide, peptide, low-molecular ones substance, nucleic acid, etc. In some embodiments, the second active agent is an inhibitor of the complement. In some embodiments, the second active agent is an inhibitor of angiogenesis.

[0011] the Invention additionally provides a method of treating a complement-mediated disease in a mammal, comprising the administration to a subject any of the above liquid compositions. In one aspect of the invention features a method of treating a complement-mediated disease comprising the step of introducing liquid composition that contains an effective amount of an analogue of constatine, in the extravascular space in the body of a subject, where the specified effective amount is sufficient for the formation of macroscopically gel-like structure, containing analogue of constatine in the extravascular space. In some embodiments of the invention specified effective amount is sufficient for formation of a macroscopic gel-like structure that diminishes in size and becomes virtually not detectable at least for 2 weeks. In some embodiments of macroscopic gel-like structure decreases over time in size and delivers the analog constatine in the active form with the achievement of therapeutic concentration in the extravascular space of at least 2 weeks, at least 4 weeks, at least 2 months, at least 3 months, at least 6 months, 9 months or 12 months. In some embodiments, a macroscopic gel-like structure remains easily detectable, for example, at least 2 weeks, at least 4 weeks, at least 2 months, at least 3 months, at least 6 months, 9 months or 12 months, up to 18 or 24 months. Similar constatine in the "active form" retains the ability to bind to C3 and to inhibit its cleavage.

[0012] In some embodiments of the invention, the composition is injected into the floor is here vitreous chamber of the eyeball of the subject, are at risk or suffering from age-related macular degeneration (AMD). In some embodiments of the subject is at risk or suffering from atrophic AMD. In some embodiments of the subject is at risk or suffering from diabetic retinopathy, uveitis, glaucoma, or pigmentary degeneration of the retina.

[0013] In some embodiments of the invention, the composition is injected into the intrathecal cavity in the body of the subject. The subject may suffer from a spinal cord injury or chronic pain.

[0014] In some embodiments of the composition is introduced into the cranial cavity of a subject, for example, in the ventricle of the brain. The subject may suffer from various sclerosis, Parkinson's disease, Alzheimer's disease, or stroke.

[0015] In some embodiments of the composition is injected into the synovial cavity or synovial bag of the subject. The subject may suffer from arthritis, for example rheumatoid arthritis, psoriatic arthritis, Reiter syndrome, juvenile arthritis or gout. [0016] In one aspect of the invention also provides methods of making compositions according to the invention.

[0017] Some aspects of the invention offer a method of treating a complement-mediated disease comprising the step of introducing a liquid composition containing effective to icesto analog constatine, in the extravascular space of the subject, where an effective amount is sufficient for the formation of discrete macroscopic gel-like structure containing analogue of constatine specified in the extravascular space. In some embodiments of the invention the effective amount is sufficient for formation of a macroscopic gel-like structure that diminishes in size and remains easily detectable at least for 2 weeks. In some embodiments of the invention the effective amount is sufficient for formation of a macroscopic gel-like structure that diminishes in size and delivers the analog constatine in the active form at least 2 weeks. In some embodiments of the invention the effective amount is sufficient for formation of a macroscopic gel-like structure that diminishes in size and remains easily detectable at least for 3 months. In some embodiments of the invention the effective amount is sufficient for formation of a macroscopic gel-like structure that diminishes in size and delivers the analog constatine in the active form at least use the e 3 months. In some embodiments of the invention the effective amount is sufficient for formation of a macroscopic gel-like structure that diminishes in size and delivers the analog constatine in the active form with the achievement of therapeutic concentrations of the specified analog constatine in the extravascular space or adjacent tissue at least 2 weeks. In some embodiments of the invention the effective amount is sufficient for formation of a macroscopic gel-like structure that diminishes in size and delivers the analog constatine in the active form with the achievement of therapeutic concentrations of the specified analog constatine in the extravascular space or adjacent tissue for at least 3 months. In some embodiments of the analog constatine includes a peptide sequence which comprises a sequence selected from the sequences SEQ ID NO: 3, 4, 5, 6 and 7. In some embodiments of the analog constatine has at least 100 times greater activity than the activity of sequence SEQ ID NO: 8. In some embodiments of the analog constatine has at least 200 times more activity than the activity of sequence SEQ ID NO: 8. In some var is the preferable implementation of the analog constatine has the sequence selected from SEQ ID NO: 14, 21, 28, 29, 30, 31, 32, 33, 34 and 36. In some embodiments of the analog constatine has a sequence selected from SEQ ID NO: 28, 32 and 34. In some embodiments of the analog constatine represents the peptide sequence SEQ ID NO: 14. In some embodiments of the analog constatine represents the peptide sequence SEQ ID NO: 28. In some embodiments of the analog constatine represents the peptide sequence SEQ ID NO: 32. In some embodiments of the analog constatine is a peptide of posledovatelnosti SEQ ID NO: 33. In some embodiments of the analog constatine represents the peptide sequence SEQ ID NO: 34. In some embodiments of the concentration of the specified analog constatine in the liquid composition is in the range from 1 mg/ml to 50 mg/ml In some embodiments of the concentration of the specified analog constatine in the liquid composition is in the range from 2 mg/ml to 25 mg/ml In some embodiments of vitreous chamber glaznov Apple injected from 50 μg to 5000 μg analog constatine. In some implementations into the vitreous chamber of the eyeball injected from 400 mcg to 1500 mcg analog constatine. In some implementations into the vitreous chamber of the eyeball is administered to about 450 mcg similar to what Platina. In some implementations into the vitreous chamber of the eyeball is injected approximately 1050 µg analog constatine. In some implementations into the vitreous chamber of the eyeball introduce the analogue of constatine in a volume of 25 ál 125 ál. In some embodiments of the analog constatine injected into the vitreous chamber of the eyeball in the volume of about 50 μl. In some embodiments of the analog constatine injected into the vitreous chamber of the eyeball in the volume of approximately 75 ml. In some embodiments of the subject suffering from age-related macular degeneration, and the liquid composition is injected into the vitreous chamber of the eyeball. In some embodiments of the liquid composition contains an effective amount of a second therapeutic agent. In some embodiments, the second therapeutic agent is an inhibitor of complement, angiogenesis inhibitor, a steroid, an anti-inflammatory agent, anti-infective agent or analgesic. In some embodiments of the composition is administered by injection into the vitreous body. In some embodiments of the composition contains a lot of microparticles and nanoparticles. Microparticles and nanoparticles can contain a therapeutic agent that may be, but not necessarily, equivalent to constatine; and if terapevticheskii is analogous to constatine, he can represent the same, but not necessarily, similar constatine, which forms a gel. In some embodiments of at least some of the microparticles or nanoparticles are incorporated into a gel after administration of the composition.

[0018] the Invention provides gel-like structure containing analogue of constatine and at least one endogenous polypeptide, normally present in the extravascular space of the subject. In some embodiments of the invention, the polypeptide is a polypeptide that is present in the extravascular space, selected from the following group: the vitreous chamber of the eyeball, the subconjunctival space, the tenon's space, subretinal space, synovial cavity and the spinal cavity.

[0019] the Invention provides a liquid composition containing an analogue of constatine, wherein the composition is characterized by the ability to form a macroscopic gel-like structure at its introduction into the vitreous chamber of the eyeball of the subject is a mammal. In some embodiments of the analog constatine is present in a quantity sufficient for the formation of discrete macroscopic gel-like structure at its introduction into the vitreous chamber of the eyeball of the subject by injection, for example the EP, in the vitreous body, in the amount of from about 50 μl to 100 μl. In some embodiments of the analog constatine is present in a quantity sufficient for the formation of a macroscopic gel-like structure that diminishes in size and remains easily detectable at least for 2 weeks. In some embodiments of the analog constatine is present in a quantity sufficient for the formation of a macroscopic gel-like structure that diminishes in size and delivers the analog constatine in the active form at least 2 weeks. In some embodiments of the analog constatine is present in a quantity sufficient for the formation of a macroscopic gel-like structure that diminishes in size and remains easily detectable at least for 3 months. In some embodiments of the analog constatine is present in a quantity sufficient for the formation of a macroscopic gel-like structure that diminishes in size and delivers the analog constatine in the active form at least 3 months. In some embodiments of the analog constatine is present in a quantity sufficient for the formation of a macroscopic gel-like structure, kotoraya time is reduced in size and remains easily detectable at least for 6 months. In some embodiments of the analog constatine is present in a quantity sufficient for the formation of a macroscopic gel-like structure that diminishes in size and delivers the analog constatine in the active form at least within 6 months. In some embodiments of the analog constatine is present in a quantity sufficient for the formation of a macroscopic gel-like structure that diminishes in size and delivers the analog constatine in the active form with the achievement of therapeutic concentrations of the specified analog constatine in the vitreous chamber of the eyeball or the adjacent tissue at least 2 weeks. In some embodiments of the analog constatine is present in a quantity sufficient for the formation of a macroscopic gel-like structure that diminishes in size and delivers the analog constatine in the active form with the achievement of therapeutic concentrations of the specified analog constatine in the vitreous chamber of the eyeball or the adjacent tissue for at least 3 months. In some embodiments of the analog constatine contains a peptide sequence which comprises a sequence selected from the following group of sequences: SEQ ID NO: 3, 4, 5, 6 and 7. In some of the which embodiments of activity similar constatine exceeds the activity sequence SEQ ID NO: 8 at least 100 times. In some embodiments of activity similar constatine exceeds the activity sequence SEQ ID NO: 8 with at least 200 times. In some embodiments of the sequence of analog constatine includes a sequence selected from SEQ ID NO: 14, 21, 28, 29, 30, 31, 32, 33, 34 and 36. In some embodiments of the sequence of analog constatine includes a sequence selected from SEQ ID NO: 28, 32 and 34. In some embodiments of the sequence of analog constatine includes SEQ ID NO: 28. In some embodiments of the sequence of analog constatine includes SEQ ID NO: 32. In some embodiments of the sequence of analog constatine includes SEQ ID N0: 34. In some embodiments of the specified number of analog constatine in the liquid composition is in the range from 1 mg/ml to 50 mg/ml In some embodiments of the specified number of analog constatine in the liquid composition is in the range from 3 mg/ml to 25 mg/ml In some embodiments of the composition contains from 150 μg to 5000 μg analog constatine. In some embodiments of the composition contains from 250 μg to 2000 μg analog constatine. In some embodiments of the composition contains from 400 mcg to 1500 mcg analog constatine. In some embodiments of the composition has a volume of 25 ál 125 ál. In some is that embodiments of the composition contains from 150 μg to 2000 μg analog constatine in a volume of 50 μl to 100 μl. In some embodiments of the composition essentially consists of analog constatine in the water. In some embodiments of the composition essentially contains no excipients. In some embodiments of the composition comprises a component selected from the following groups: sugar alcohols and amino acids. In some embodiments of the presence of the specified component modulates the rate of disappearance enable in vivo. In some embodiments of the composition contains histidine. In some embodiments of the composition contains a buffer. In some embodiments of the composition contains sodium acetate. In some embodiments of the composition contains mannitol. In some embodiments of the liquid composition further comprises an effective amount of a second therapeutic agent. In some embodiments, the second therapeutic agent is an inhibitor of complement, angiogenesis inhibitor, a steroid, an anti-inflammatory agent, protivojinfectionnaya agent or analgesic.

[0020] the Invention also provides a method of making a composition for prolonged delivery of a therapeutic agent, comprising preparing a liquid composition containing therapeutic agent and the analog constatine, where the analogue of constatine is present in sufficient amount to HUF is investing macroscopic gel-like structure with the introduction of the composition into the extravascular space of the subject is a mammal. In some embodiments of the method further includes the introduction of liquid composition in the extravascular space in the body of a mammal. In some embodiments of the extravascular space is a vitreous chamber. In some embodiments of the extravascular space is a vitreous chamber, in this case the subject suffering from AMD.

[0021] Some aspects of the invention offer a method of treating subjects at risk of or suffering from AMD, including the introduction of a liquid composition containing an analogue of constatine directly into the vitreous chamber of the eyeball of the subject, where the liquid composition contains a sufficient number of analog constatine for the formation of a macroscopic gel-like structure after injection. In some embodiments of the analog constatine contains a peptide sequence which includes the sequence from the following group: SEQ ID NO: 3, 4, 5, 6 and 7. In some embodiments of activity similar constatine exceeds the activity sequence SEQ ID NO: 8 at least 100 times. In some embodiments of the analog constatine has an activity of at least 200 times higher than the activity of SEQ ID NO: 8. In some embodiments of the sequence of analog constatine PEFC includes the sequence, selected from SEQ ID NO: 14, 21, 28, 29, 30, 31, 32, 33, 34 and 36. In some embodiments of the sequence of analog constatine includes a sequence selected from SEQ ID NO: 28, 32 and 34. In some implementations, the number of analog constatine is in the range from 2 mg/ml to 20 mg/ml In some embodiments of from 100 μg to 2000 μg analog constatine is injected into the eye. In some embodiments of from 250 μg to 1500 μg analog constatine is injected into the eye. In some embodiments of from 400 to 1200 µg µg analog constatine is injected into the eye. In some embodiments of the liquid composition further comprises an inhibitor of angiogenesis.

[0022] the Invention provides a liquid composition containing an analogue of constatine and water, where the concentration of analog constatine is in the range from 3 to 50 mg/ml In some embodiments of concentration equivalent to constatine is in the range from 5 to 30 mg/ml In some embodiments of concentration equivalent to constatine is in the range from 8 to 25 mg/ml In some embodiments of any of these compositions similar constatine contains a peptide selected from the group comprising the sequences: SEQ ID NO: 3, 4, 5, 6 and 7. In some embodiments of any of these compositions, the active analog constatine exceeds the activity of SEQ ID NO: 8 at least 100 times. In some embodiments of any of these compositions, the active analog constatine exceeds the activity of SEQ ID NO: 8 with at least 200 times. In some embodiments of any of these compounds sequence of analog constatine includes a sequence selected from the sequences SEQ ID NO: 14, 21, 28, 29, 30, 31, 32, 33, 34 and 36. In some embodiments of any of these compounds sequence of analog constatine includes a sequence selected from the sequences SEQ ID NO: 28, 32 and 34. In some embodiments of any of these compositions, the composition essentially consists of analog constatine and water. In some embodiments of any of these composition the composition further comprises excipients selected from amino acids and sugar alcohols. The invention provides a liquid composition containing an analogue of constatine and water, where the concentration of analog constatine is in the range from 100 to 2000 mg/ml, for example from 100 mg/ml to 1000 mg/ml, or from 100 to 500 mg/ml, where the composition contains a component that changes the properties of the composition so that the gel formed upon introduction of the composition into the extravascular space (e.g., the vitreous chamber of the eyeball), decomposes or dissolves faster than in the absence of the component. In some embodiments of the components is NT represents an excipient, selected from a sugar alcohol, and amino acids. In some embodiments of the amino acid is a basic amino acid, such as histidine. In some embodiments of the sugar alcohol is a mannitol. In some embodiments of the component is a buffer, for example, sodium acetate.

[0023] Unless otherwise specified, the present invention uses standard methods of molecular biology, chemistry, cell culture, animal husbandry, medical and veterinary tests, etc. and are received in the corresponding field value terms. This application contains links to various patents and publications. A list of all scientific articles, books, patents, patent applications and other publications cited in this application are included in this application by reference: Current Protocols in Molecular Biology, Current Protocols in Immunology, Current Protocols in Protein Science, and Current Protocols in Cell Biology, John Wiley & Sons, N.Y., edition as of July 2002; Sambrook, Russell, and Sambrook, Molecular Cloning: A Laboratory Manual, 3rded., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, 2001; Kuby Immunology, 4thed., Goldsby, R.A., Kindt, T.J., and Osbome, B. (eds.), W.H. Freeman, 2000, Goodman and Oilman''s The Pharmacological Basis of Therapeutics, 10thEd., McGraw Hill, 2001, Katzung, B. (ed.) Basic and Clinical Pharmacology, McGraw-Hill/Appleton & Lange; 9th edition (December 2003); Goldman & Ausiello, Cecil Textbook of Medicine, 22nded., W.B. Saunders, 2003. In the event of conflict or inconsistency, m is waiting for any included links and the present invention, the description of the present invention (including any amendments) shall prevail. In this application we use common abbreviations for amino acids, unless otherwise stated.

BRIEF DESCRIPTION of FIGURES

[0024] Figure 1 shows a macroscopic gel-like structure, isolated from the vitreous of the rabbit, which was introduced active analog constatine by injection into the vitreous body.

[0025] Figure 2 shows the results of ultrasound scanning of animals 12 days (left) and 8 weeks (right) after injection of the active analog constatine. 12 days after the injection there is a major turn on, and much smaller, but still detectable incorporation observed after 8 weeks.

[0026] Figure 3 shows the proteins included in the composition include, along with active analog constatine identified by the color of the gel after SDS-electrophoresis. Also shown are proteins found in the vitreous body after the introduction of the analog constatine or without its introduction.

[0027] Figure 4 presents a graph showing that the analogue of constatine present in inclusion, remains stable and retains complement inhibitory activity over time. The blue area is the analogue of constatine (standard); the green area is the analogue of constatine obtained from the gel izvlechenij is from the vitreous of the rabbit 6 weeks after injection; brown spots - analog constatine obtained from the gel, extracted from the vitreous of the rabbit after 8 months after injection; red ovals: inactive control peptide G8A.

[0028] Figure 5 presents a graph showing the measurement result of the concentration of analog constatine in serum and vitreous bodies cynomolgus macaques 14 days after injection into the vitreous body 0, 450, 1050 or 2100 mg of composition. All estimates are presented as mean ± standard error; n=4 for each of the vitreous body, and n=8 for serum. It should be noted that the actual values for serum concentration 100 times lower than shown in the graph, i.e. each value of the concentration in serum is about 100 times lower than the corresponding concentration in the vitreous body at the same dose injection.

DEFINITION

[0029] Before proceeding to a detailed description of the invention, it is necessary to clarify that the present invention is not limited to the specific examples of systems or parameters, which, of course, may vary. You must also understand that the use in this application terminology is intended only to describe the specific modes of the invention and is in no way intended to limit the scope of the invention. The following definitions are presented for the convenience of the reader, and them, and the use is not contrary to the use of such terms in the relevant field, unless otherwise specifically stated.

[0030] the Terms "inhibitor of angiogenesis and antiangiogenic agent" are used interchangeably and are used in this application to describe agents that can inhibit or slow down one or more associated with the formation, growth and/or development of new blood vessels processes that include proliferation of endothelial cells, migration of endothelial cells, capillary formation, but are not limited to. In addition, these agents can inhibit the exudation of fluid from blood vessels.

[0031] the Term "antagonist" refers to a compound that inhibits (prevents, reduces, decreases, blocks or cancels) the effect of a given molecule. The antagonist is able to act a certain way on a certain activity of the molecule so that the biological activity of the molecule is reduced or blocked by the principle of antagonism (e.g., anti-competitive or reverse action) in relation to one or more natural action of the molecule. Non-limiting examples of antagonists include antibodies or their antigen-binding fragments, proteins, peptides, nucleic acids (such as mediators of Rnci, ribozymes, antisense sequence), or small molecules.

[0032] the Term "antibody" refers immun who globulin or its derivative, containing immunoglobulin domain are able to bind to the antigen. Antibodies can be of any origin, for example, human, murine, rabbit, goat, chicken, etc. Antibody may be representative of any class of immunoglobulins, including any of the classes of human immunoglobulins: IgG, IgM, IgA, IgD and IgE, or their subclasses, such as IgGI, IgG2, etc. In various embodiments of the invention the antibody is a fragment such as Fab', F(ab')2, scFv (single-chain variable) or another piece that retains the binding site of the antigen, or scFv fragment obtained by recombination, as an example obtained by recombination of fragments. See, for example, Alien, T., Nature Reviews Cancer, Vol.2, 750-765, 2002, and references are included. The antibody may be monovalent, bivalent or multivalent. The antibody may be a chimeric or "humanized" antibody, in which, for example, the variable region of the rodent to be grafted to the constant region of human origin, while retaining the specificity of the antibodies of the rodent. The area of human origin does not necessarily have to be obtained from a person, i.e. to be first synthesized in the human body. Instead, "human" area can be synthesized in the body of a mammal whose genome includes genes human immunoglobu the ins. See, for example, Vaughan, et al., (1998), Nature Biotechnology, 16: 535-539. The antibody may be partially or fully humanitarian. The antibody may be polyclonal or monoclonal, although for the purposes of the present invention, monoclonal antibodies are generally preferred. Methods for producing antibodies that specifically associated with virtually any molecule known in the field. For example, monoclonal or polyclonal antibodies can be isolated from blood or ascitic fluid of an animal that produces an antibody (for example, after natural exposure or immunization molecule or antigenic fragment)can be obtained using recombinant technologies in cell culture or by using transgenic organisms, or can be created at least partially by chemical synthesis.

[0033] the Terms "approximately" or "about" in relation to numeric values, as a rule, involve numbers that are within ±10%, in some embodiments ±5%, in some embodiments ±1%, in some embodiments ±0.5% of the number unless otherwise stated or the context (except where such number exceed 100% of a possible value).

[0034] the Term "biocompatible" is used in accordance with its use in this field, and refers to the ve is estu, which is essentially non-toxic to cells in vitro; for example, in some implementations, if its addition to cells in culture in quantities close to concentrations suitable for use in vivo, leads to the destruction of not more than 20% of the cells. The substance is considered biocompatible for the recipient if it is essentially non-toxic to cells and tissues of the recipient used in quantities and applications, and also not causes or does not cause a significant adverse or unfavorable effect on the body of the recipient, for example, immune or inflammatory reaction, the formation of scar tissue, etc.

[0035] the Term "biodegradable" means that the substance is able to break down physically and/or chemically inside the cells or extracellular component in the body of the subject, for example, by hydrolysis under physiological conditions and, as a result of natural biological process, such as the action of enzymes present within cells or within the body, and so on, with the formation of more low molecular weight chemical compounds that can be metabolized to probable reuse in the body, and/or displayed or otherwise disposed of. Substances that dissolve, split, or break up into small fragments, for example, soluble molecules is or supramolecular complexes, under physiological conditions, refer to "biodegradiruemym" substances. Preferred biodegradative substance is biocompatible.

[0036] a "Biological macromolecule" means a large molecule composed of smaller subunits that occur in biological systems. Examples of biological macromolecules include polypeptides, nucleic acids and polysaccharides. Typically, a biological macromolecule contains at least 3 subunits (e.g., amino acids, nucleosides, sugars etc). Biological macromolecules can be, but not necessarily, natural polypeptides, nucleic acids, or polysaccharides. Biological macromolecule can be modified, for example, it may be associated with non-biological molecule such as a synthetic polymer, etc.

[0037] Component of complement" or "protein complement" means a molecule that is involved in activation of the complement system or participates in one or more complement-mediated processes. Components of the classical pathway of complement include, for example, C1q, C1r, C1s, C2, C3, C4, C5, C6, C7, C8, C9 and C5b-9 complex, also known as the membrane attack complex (MAC) and active fragments or products of enzymatic degradation of any predecessors (for example, Sa, C3b, Sa, C4b, Sa, etc). Components al is ernatives path includes, for example, factors b, D, I and properdin. Components of pectin pathways include, for example, MBL2, MASP-1 and MASP-2. Complement components also include cellular receptors soluble components of complement. Such receptors include, for example, the receptor CA (C5aR), receptor CA (C3aR), complement receptor 1(CR1), complement receptor 2 (CR2), complement receptor 3 (CR3), etc. Need to understand that the term "component of complement" does not refer to those molecules and molecular structures that are "initiators" to activate complement, for example, complexes of antigen-antibody, foreign patterns on the surfaces of bacterial cells or on artificial surfaces, etc.

[0038] "complement Regulatory protein" is a protein involved in the regulation of the activity of complement, such as complement factor H (CFH) in mammals.

[0039] a "Protein control of complement" is a protein that regulates the complement system, containing many SCR sections.

[0040] Complement-like protein" is a protein sequence which is at least 20% of its overall length is identical to the sequence of the protein or protein complement control of complement, and/or a protein that special way competes with proteins of the complement or proteins control complement for binding to their target, for example, has at least 10% greater AF is innosti. Genes encoding such proteins can be detected in the immediate vicinity of genes encoding complement proteins or proteins control the complement, having the identical sequence. For example, the CFH gene cluster contains numerous CFH - like genes (for example, CFHR1, CFHR1, CFHR3, CFHR4 and CFHR5).

[0041] the Term "associated with complement protein is common to components of the complement proteins that regulate the complement, and the complement-like proteins; however, it should be understood that the invention encompasses embodiments, which relate specifically to the components of the complement proteins that regulate complement, complement-like proteins and their combinations.

[0042] the Term "effective amount" of an active agent, such as an inhibitor of complement, refers to the amount of active agent sufficient to induce a desired biological response (or to suppress undesirable biological response). Specialists in this field it is clear that the absolute amount of a particular agent, which is effective may vary depending on such factors as the desired biological response, the applicable agent, the target tissue, etc. Specialists in this field it is also clear that "effective amount" can be achieved by a single administration, or by the introduction neskolkih. For example, the effective amount may be an amount sufficient to alleviate at least one symptom of the disease. The effective amount may be an amount sufficient to slow the development of chronic or progressive disease, for example, to increase the period of time prior to the manifestation of one or more symptoms or signs of disease or to increase the period of time before the disease will lead to a certain degree of destruction of the body. The effective amount may be an amount sufficient to provide a faster and more effective recovery from the disease than in the absence of the agent.

[0043] the Term "identical" refers to the area in which the sequence of two or more amino acids or polypeptides are the same. The percentage identity between the target sequence and the second sequence according to certain criteria, for example, the length of the desired sequence can be calculated by the sequence alignment, which determines the number of residues (nucleotides or amino acids) within the analyzed area in front of identical residue, and the use of spaces can improve the identity, divided by the total number of residues of interest, the overall sequence or the second sequence (the which is more)that fall in the analyzed area, and multiplied by 100. Under the space is meant a sequence region that is not occupied by the remainder. When calculating the number of identical residues to determine the percentage identity results are rounded to the nearest whole number. Percent identity can be calculated using known in the field of various computer programs. For example, such computer programs as BLASTN, BLASTP, Gapped BLAST and so on, align the desired sequence by any of a variety of sequences from a shared database and calculate the percentage identity between them. The algorithm of Karlin-Altshul (Karlin and Altschul, Proc. Natl. Acad. Sci. USA 87:22264-2268, 1990), modified by Karlin and Altschul, Proc. Natl. Acad. Sci. USA 90:5873-5877, 1993, included in the program NBLAST and XBLAST of Altshuler and others (Altschul, et al. Nucleic Acids Res. 25: 3389-3402, 1997). When using the programs BLAST and Gapped BLAST can be used the parameters set by default. These programs can be found on the Internet website URL www.ncbi.nlm.nih.gov. In some embodiments, the percent identity between the target sequence and the second sequence is calculated using BLAST2 using the default settings.

[0044] the Term "isolated" means (1) separated from at least some of the components, to the which is normally associated in the natural environment; 2) obtained or purified by a process that involves human intervention and/or 3) not found in nature. For example, the molecule remote from the cell, which it produces, is "isolated". Chemically synthesized molecule is "isolated".

[0045] the Term "related", when used in respect of two or more compounds, means that these compounds are physically combined or linked together with the formation of the molecular structure, which is stable enough to make the connection remained United in the formation of the connection and, preferably, in the conditions of the new molecular structure, for example, physiological conditions. In some preferred embodiments of the invention, the bond is covalent bond. In other embodiments of communication is non-covalent. Connections can be connected directly or not directly. If two connections are connected directly, then they are either covalently linked to each other, or are near enough to each other to intermolecular forces between them could maintain their relationship. When two nets are not connected directly, each of them is linked either covalently or ecovalence with the third connection, which supports communication between TLD is I these elements. In General, when you specify that two connections are connected through a "linker" or "linking group" or "the connecting section, the relationship between these two compounds is indirect, and, as a rule, each of the related compounds covalently linked to the linker. The linker can be any suitable group that reacts with two link connections within a sufficient time and under conditions that are compatible with the stability of compounds (which may optionally be protected, depending on conditions), and in a quantity sufficient to obtain an acceptable result.

[0046] a "Liquid composition" is a composition containing at least one chemical substance which is liquid at room temperature. The liquid composition may contain a therapeutic agent. therapeutic agent can be, for example, dissolved, suspended or dispersed in it. therapeutic agent may be present as individual molecules mixed with the molecules of the liquid or in the form of microscopic or macroscopic aggregates of particles and so on, provided that such aggregates or particles do not obstruct the free flow of the composition in accordance with its liquid state.

[0047] the Term "topical application or local injection" in relation to the delivery of the composition or AG the NTA refers to the delivery, which is not mediated transport of the composition or agent to its intended target tissue or target area through a system of vessels. The composition or the agent may be delivered directly to the intended target tissue or site, or to an adjacent area, for example, in the immediate vicinity of the proposed site or target tissue. For example, the composition can be delivered by injection or infusion of the composition. As a result, local injection in the immediate vicinity of the target tissue or area of the target composition, or one or more of its components can diffuse to the intended target tissue or area. It should be understood that in one local introduction of a fraction of a therapeutic agent (typically, only a small fraction of injected dose) can get into the bloodstream and transported to another area, including return transport to the intended target tissue or area.

[0048] "Local activation" refers to activation of complement, which occurs outside the vascular bed.

[0049] "Many" means more than one.

[0050] the Term "polypeptide" when used in this application refers to an amino acid polymer, typically including one or two similar amino acids. Protein is a molecule consisting of one or more polypeptides. The peptide is relatively short polypeptide, the AK rule, from 2 to 60 amino acids in length, for example, from 8 to 40 amino acids in length. The terms "protein", "polypeptide" and "peptide" may be used interchangeably. Polypeptides used according to this application may contain amino acids that occur in proteins in nature, amino acids that are not found in proteins in nature, and/or analogs of amino acids, which are not amino acids. In this context, "similar" amino acids may be a different amino acid, which structurally resembles the amino acid, or a substance which is not an amino acid, which structurally resembles the amino acid. There are a large number of generally accepted analogues of the 20 amino acids commonly found in proteins ("standard" amino acids). One or more amino acids in the polypeptide can be modified, for example, by attaching to them the chemical structure, such as a carbohydrate group, a phosphate group, Farnesina group, separately group, a group of fatty acids, a linker for conjugation, functionalization, or other modification, etc. of a Specific non-limiting examples of suitable analogues and modifications described in international PCT publications WO 2004026328 and WO 2007062249. The polypeptide can be acetiminophen for example, N-end, and/or minioven, for example, on the C-end. Mo is eficacia can occur in any part of the polypeptide, including the peptide bond, the side chain of amino acid and amine end or carboxyl end. Specific polypeptide may contain many types of modifications. The polypeptides may be branched and/or cyclic, branched or unbranched. Polypeptides can be isolated from natural sources, obtained in vitro or in vivo in suitable expression systems using recombinant DNA technology (e.g., recombinant host cells or in transgenic animals or plants), synthesized by chemical methods, such as traditional solid-phase peptide synthesis, and/or methods, including chemical ligation of synthesized peptides (see, for example, Kent, S., J Pept Sci., 9(9):574-93, 2003), or obtained through combinations of the above methods. These methods are well known, and the person skilled in the art can select and apply the appropriate method for the synthesis of peptides and polypeptides described in this application. The polypeptide can include one or more sites of subsidies, as described, for example in the publication U.S. No. 20040115774. In some embodiments of the invention, the polypeptide is modified with a polymer using one or more of the methods described in this application or incorporated into it by reference. The terms "polypeptide the I sequence" or "amino acid sequence" when used in this application may refer to a polypeptide material per se and not to carry information about the sequence, characterizing the polypeptide biochemically (i.e., a sequence of letters or three-letter codes, the selected letters or codes that represent abbreviations of the names of the amino acids). Polypeptide sequence used in this application are provided in the direction from N-Terminus to the C-end, unless otherwise specified.

[0051] the Term "purified" when used in this application means that the compound or substance is separated from one or more other compounds or substances with which it was before it was cleaned. Compound or substance may be partially cleared, almost cleared or clean. A substance or compound, such as a nucleic acid or polypeptide, is clean, if it is isolated from almost all other compounds and substances which are not solvents or any ions contained in the solution; i.e. when it is at least about 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more than 99%, of the dry weight of the mixture. Partially or substantially purified compound or substance, such as a nucleic acid or polypeptide can be isolated from at least 50% by dry weight, at least 60%, at least 70%or at least 80% of the material with which they occur in nature, for example, cleocin the first material, such as cellular proteins and/or nucleic acids. In some embodiments of the invention, the amount of purified nucleic acid or polypeptide is at least 10%, 20%, 30%, 50%, 60%, 70%, 80%, 90%, 95%, 99%, or even more, from the dry weight of the total amino acid or polypeptide in the mixture, respectively. Methods of estimation of purity known in this field and include chromatography methods, immunological methods, electrophoretic methods, etc. Any of the methods in this application of polynucleotides or polypeptides can be purified.

[0052] the Term "reaction-sposobna functional group" as used in this application refers to groups, including but not limited to, olefins, acetylene, alcohols, phenols, ethers, oxides, halide, aldehydes, ketones, carboxylic acids, esters, amides, cyanate, isocyanates, thiocyanates, isothiocyanates, amines, diamides, hydrazines, hydrazides, diazo group, diezani, nitrogen, NITRILES, mercaptans, sulfides, disulfides, sulfoxidov, sulfones, sulfonic acid, sulfinate acids, acetals, ketals, anhydrides, sulfates, sulfenamide acid, isonitrile, amidine, imides, imidate, Netanya hydroxylamine, oximes, hydroxamic acids, digitoxigenin acid, alley, orthoepy, sulfites, enamines, inamine, urea, pseudoceramides, semicarbazides, carbodiimide carbamates, imine, azides, uzasadnienie, isoxazolidinone and nitroso compounds. Reaction-sposobnye functional groups also include groups that are often used to obtain bioconjugate, for example, N-hydroxysuccinimide, esters, maleimide, sulfhydryl and so on (see, for example, Hermanson, G., Bioconjugate Techniques, Academic press, San Diego, 1996). Methods of obtaining each of these functional groups are well known in this field, and their application or modification for a particular purpose are within the experience of specialists (see, for example, Sandier and Karo, eds. ORGANIC FUNCTIONAL GROUP PREPARATIONS, Academic Press, San Diego, 1989).

[0053] the Term "RNA interference" or "MKI" used in this application in accordance with their application in this field, for example, these terms refer to any method, which may be a reduced expression of the gene or the amount of gene product with the inside, or when the expression inside the target cell or organism a double-stranded RNAS (dsrnas), called "mediators of Rnci", the sequence of which corresponds to the sequence from the gene of interest, especially short double-stranded RNA (miRNA)containing a sequence that is complementary to the messenger RNA of the gene of interest.

[0054] the Term "specific binding" generally refers to the physical connection between the polypeptide-target (or, more obobs the NGOs, molecule-target) and a binding molecule such as an antibody or ligand. Linking, as a rule, depend on the structural features of the target, such as antigenic determinant, epitope binding pocket or gap recognized by the binding molecule. For example, if the antibody specific to the epitope And the presence of a polypeptide containing the epitope, or the presence of free unlabeled epitope And in reactions containing free labeled epitope As, and binding his molecule, will reduce the number associated with the binding molecule labeled epitope A. you Must understand that the specificity is not absolute, and usually refers to certain conditions, which are binding. For example, as is well known in this area, many of antibodies cross-react with other epitopes in addition to those already present epitopes of the molecule target. Such cross reaktsionnosposobnykh may be acceptable depending on the application of this antibody. Average person is able to pick up antibodies or ligands that have a sufficient degree of specificity in order to operate properly when a particular application (for example, to determine the target molecules for therapeutic purposes etc). You must also understand that specificity can activates into account additional factors, such as the affinity binding molecules against targets and the affinity binding molecules against other targets, for example, competitive. If the binding molecule exhibits a high affinity to the molecule of interest target and low affinity to other molecules, such antibody will likely be suitable reagent. If the specificity of the binding molecules is set in a certain condition or conditions, such a molecule may be used in other, preferably similar conditions without the need for revaluation of its specificity. Linking two or more molecules can be considered specific if the equilibrium dissociation constant (Kd) is 10-3M or less, preferably 10-4M or less, more preferably 10-5M, or less, for example, 10-6M or less, 10-7M or less, 10-8M or less, or 10-9M, or less, in experimental conditions, such as physiological conditions.

[0055] a "Substantial sequence identity" in relation to the amino acid sequence means that the sequence of at least about 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 95% identical to the control sequence. In particular embodiments of the term in relation to amino acid serial is a major means, that sequence at least about 70%, 80%, 85%, 90%, 95%, 98% or 99% identical to the control sequence. In particular embodiments, at least 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 95% identical amino acids are conservative substitutions of amino acid control sequence. Conservative substitutions may be determined in accordance with Stryer, L., Biochemistry, 3rd ed., 1988, on which the following groups of amino acids have similar properties of the side chains, such as charge, hydrophobicity, degree of aromatization, etc. (1) Aliphatic side chains: G, A, V, L, I; (2) Aromatic side chains: F, Y, W; (3) Gray-containing side chains: s, M;

(4) Aliphatic hydroxyl side chains: S, T; (5) Basic side chains: K, R, N;

(6) Acidic amino acids: D, E, N, Q; (7) Cyclic aliphatic side chain: P, which can also be considered as belonging to the group (1). In accordance with another adopted classification conservative substitutions occur within the following groups of amino acids: (1) non-Polar: A, L, I, V, G, P, F, W, M; (2) Polar: S, T, S, Y, N, q (3) Key: K, R, N; (4) Acid: D, that is, Amino acids with small side chain (G, A, S, T, M) also form a group, within which can happen to a conservative replacement. Can be used other ways of classification, known in this area. In addition, in accordance with this, the options can be classified analogues of amino acids and not natural amino acids.

[0056] the Term "subject" as used in this application refers to the individual who introduces the agent, for example, for experimental, diagnostic and/or therapeutic purposes. Unless otherwise specified, the subjects are mammals, for example, Pets allowed, such as dogs, cats, rabbits, etc., monkeys or humans.

[0057] the Term "system" when used in this application with respect to components of the complement refers to the complement proteins, which are synthesized in the liver by hepatocytes and enter the bloodstream, or synthesized by circulating macrophages or monocytes and secreted into the bloodstream.

[0058] the Term "therapeutic agent" when used in this application refers to any pharmacologically active agent used for the treatment of the disease. The term includes any pharmaceutically acceptable salts, prodrugs, salts of prodrugs and derivatives thereof, known in the field, or could be easily obtained using standard methods known in this field. The term "prodrug" refers to a predecessor agent, this in itself is a prodrug is a pharmacologically active (or has a weakened or different action than the desired action of the medicinal agent), but after the introduction into the body transformed (for example, metabolites the m path) in a pharmaceutically active drug agent. therapeutic agent can be, but not necessarily, a small molecule or a biological macromolecule, such as a polypeptide, antibody, nucleic acid, for example, an aptamer, an agent of Rnci, such as small interfering RNA (siRNA), etc. In this application therapeutic agent is sometimes called "active agent" or "drug". A small molecule, usually an organic compound having a molecular weight of 1,500 daltons (Da)or less, for example, 1,000 Da, or less, for example, 500 Da or less, and having multiple carbon-carbon connection.

[0059] the Term "treatment" when used in this application refers to any medical or surgical procedure on the subject. The treatment may be carried out in order to reverse, alleviate, inhibit or prevent the progression to eliminate or reduce the probability of occurrence of one or more symptoms or manifestations of disease, disorders or pathological conditions. "Prevent" means to cause such action to the disease, disorder, pathological condition or symptoms, or symptoms thereof, did not appear for at least some time at least in some subjects. Treatment may include the step of introducing the subject agent after developing one or more SIM the volume or manifestations, indicative of the complement-mediated pathological conditions; for example, to reverse, alleviate, reduce the severity of the pathological condition, and/or to inhibit or prevent the progression of pathological conditions, and/or to reverse, alleviate, reduce the severity and/or inhibit one or more symptoms or manifestations of pathological conditions. The composition according to the invention can be administered to a subject suffering from a complement-mediated disorder, or those at high risk of developing this disorder compared to the average person. The composition according to the invention can be introduced with the aim of prevention, i.e. until the development of any symptoms or manifestations of pathological conditions (e.g., a subject with at least one risk factor for pathological conditions).

[0060] the Term "alkyl" when used in this application refers to a saturated linear, branched or cyclic hydrocarbon which contains from about 1 to 22 carbon atoms (including all combinations and podnominatsii sequences and specific numbers of carbon atoms), preferably from about 1 to about 12 or from about 1 to about 7 carbon atoms in some embodiments of the invention. Alkyl the s groups include, but not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, t-butyl, n-pentyl, cyclopentyl, isopentyl, neopentyl, n-hexyl, isohexyl, cyclohexyl, cyclooctyl, substituted, 3-methylpentyl, 2,2-dimethylbutyl and 2,3-dimethylbutyl.

[0061] the Term "halogen" as used in this application refers to F, Cl, Br or I.

[0062] the Term "alkanoyl" when used in this application refers to a possibly substituted linear or branched acyclic aliphatic residue having from about 1 to 10 carbon atoms (including all combinations and podnominatsii sequences and specific numbers of carbon atoms), for example, from about 1 to about 7 carbon atoms. Alcoholnye groups include, but are not limited to, formyl, acetyl, propionyl, butyryl, isobutyryl, pentanoyl, isopentanol, 2-methyl-butyryl, 2,2-dimethoxypropane, hexanoyl, heptanoyl, octanoyl etc. the Term "lower alkanoyl" refers to substituted linear or branched acyclic aliphatic residue having from about 1 to 5 carbon atoms (including all combinations and podnominatsii sequences and specific numbers of carbon atoms). Such groups include, but are not limited to, formyl, acetyl, propionyl, butyryl, isobutyryl, pentanoyl, isopentanol etc.

[0063] the Term "aryl" when is ispolzovanie in this application refers to a possibly substituted mono - or bicyclic system of aromatic rings, having from about 5 to 14 carbon atoms (including all combinations and podnominatsii sequences and specific numbers of carbon atoms), preferably from about 6 to about 10 carbon atoms. For non-limiting examples include phenyl and naphthyl.

[0064] the Term "aralkyl" when used in this application refers to alkyl radicals having aryl substituents and having from about 6 to about 22 carbon atoms (including all combinations and podnominatsii sequences and specific numbers of carbon atoms), and in some embodiments of the invention are preferred radicals with from about 6 to about 12 carbon atoms. Kalkilya groups can be substituted. For non-limiting examples include benzyl, naphthylmethyl, diphenylmethyl, triphenylmethyl, phenylethyl and diphenylether.

[0065] the Terms "alkoxy" and "alkoxyl" when used in this application may refer to a substituted group alkyl-O-, where alkyl is defined above. Typical alkoxy and CNS groups include methoxy, ethoxy, n-propoxy, and-propoxy, n-butoxy, epoxy group.

[0066] the Term "carboxy" as used in this application refers to the group-C(=O)HE.

[0067] the Term "alkoxycarbonyl" when used in this application refers to the group-C(=O)O-alkyl, where alkyl is of marginal previously.

[0068] the Term "aroyl" when used in this application refers to the group-C(=O)-aryl, where aryl is defined above. Typical aroline groups include benzoyl and naphtol.

[0069] typically, the substituted chemical groups include one or more substituents which are bonded to a hydrogen atom. Typical substituents include, for example, halogen, alkyl, cycloalkyl, aralkyl, aryl, sulphydryl, hydroxyl (-OH), alkoxyl, cyano (-CN), carboxyl (-COOH), -C(=O)O-alkyl, aminocarbonyl (-C(=O)NH2), -N-substituted aminocarbonyl (-C(=O)other”), CF3, CF2CF3etc. Each particle R” have the above-mentioned substituents independently can represent, for example, H, alkyl, cycloalkyl, aryl or aralkyl.

[0070] the Term "L-amino acid" when used in this application refers to any naturally occurring levogyrate alpha-amino acid present in the natural environment in proteins, or to alkilany esters of these alpha-amino acids. The term "D-amino acid" refers to Pervouralsk alpha-amino acids. Unless otherwise indicated, all amino acids specified in this application are L-amino acids.

[0071] When used in this application, the term "aromatic amino acid" refers to amino acid, which comprises at least one aromatic ring, for example, the aryl group.

[0072] When using the years in this application, the term "similar aromatic amino acid" refers to similar amino acids, which includes at least one aromatic ring, for example, the aryl group.

A DETAILED DESCRIPTION of several VARIANTS of REALIZATION of the INVENTION

[0073] the Prolonged release of analog constatine of gel-like structures

[0074] the Present invention relates to prolonged delivery of analog constatine with the release of analogue constatine of the gel-like structure which is formed by introducing a liquid composition containing an analogue of constatine, in the extravascular space in the body of a mammal, such as the vitreous chamber of the eyeball. The invention also relates to prolonged delivery of analog constatine with the release of analogue constatine of the gel-like structure that is formed by contact of the liquid composition containing the equivalent of constatine, with the substance of the body of the subject. The substance of the body is usually liquid or liquid-containing substance, which is not a blood (or its derivatives, such as plasma or serum) or urine. For example, the substance may be vitreous body.

[0075] the Substance of the body usually contain protein (protein), inorganic or organic ion (ions) and/or glycosaminoglycan. One or more of these components can initiate the formation of a gel-like structure, or to participate in teaching the enterprise. In some embodiments of the invention gellike structure contains one or more endogenous proteins or glycosaminoglycan present in the substance of the body where these endogenous proteins or glycosaminoglycan included in the gel-like structure at its formation in vivo.

[0076] the Invention also relates to prolonged delivery of therapeutic agent upon introduction of a liquid composition containing therapeutic agent and the analog constatine, in the extravascular space in the body of the mammal with the formation of a gel-like structure. The invention also relates to prolonged delivery of therapeutic agent in the release of therapeutic agent from the gel-like structure that is formed by contact of the liquid composition containing the equivalent of constatine and therapeutic agent, with the substance of the body, for example, protein-containing substance of the body. The substance of the body, usually a liquid or liquid-containing substance, non-blood (or liquid derivatives, such as plasma or serum) or urine. For example, the substance may be liquid part of the vitreous body.

[0077] the Invention also relates to a gel-like structure formed ex vivo upon contact of the liquid composition containing the equivalent of constatine with one or Bluebelle, ions or glycosaminoglycans, or combinations thereof, sufficient to initiate the formation of a gel. Proteins, ions or glycosaminoglycan may be identical to those present in vivo in the spaces of the body, such as the vitreous chamber of the eyeball, synovial cavity, etc. or may substantially coincide with them on the structure or sequence. Gel-like structure may also contain additional therapeutic agent. The invention also relates to prolonged delivery of analog constatine and possibly the second therapeutic agent with the introduction of gel-like structures needy subject, for example, by injection, implantation or deposition on the body area where you want to inhibit complement.

[0078] In some embodiments of the invention, the liquid composition is characterized by the fact that it contains no other gel-forming substance, in addition to analog constatine. When removing analog constatine of such composition the gel structure is not formed after the introduction of the liquid composition to a subject under conditions in which the gel-like structure would be formed easily in the presence of analog constatine in sufficient quantity, for example, after injection inside the vitreous body. In some embodiments of the invention, the liquid composition of ha is acterized fact, that contains the gel-forming substance, and those substances are present in quantities insufficient for the formation of gel in the absence of analog constatine under conditions in which the gel-like structure would be formed easily in the presence of analog constatine in sufficient quantity, for example, after injection into the vitreous body. In some embodiments of the invention the liquid composition, thus, differs from the compositions which contain gel-forming substances such as collagen, synthetic polymers, etc. that can be incorporated into other recipes analog constatine to form a gel in sufficient quantities in the absence of analog constatine.

[0079] In some embodiments of the invention a liquid composition essentially consists of analog constatine and one or more chemical substances that are liquid at room temperature (e.g., water). In some embodiments of at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, at least 99% of the dry weight of the composition is similar constatine. In some embodiments of the liquid composition contains water and an organic solvent. In some embodiments of the liquid the composition in addition to the analog constatine contains inorganic ion, buffer, dissolving agent, or combination thereof, where the composition forms a gel-like structure at its introduction in the absence of analog constatine. Additionally, the liquid composition may include analog constatine and one or more therapeutic agents.

[0080] the Invention also relates to methods of modulating the formation of a gel from a liquid composition containing an analogue of constatine, and/or methods of modulating properties of the gel formed from a liquid composition containing an analogue of constatine, and to methods of investigation agents to determine their ability to modulate the desired one or more properties of the gel, and/or one or more parameters of the formation of the gel (for example, the rate at which the gel is formed or disappears, density, consistency, content analog constatine, the rate of release of analog constatine etc). The methods include preparing a liquid composition containing an analogue of constatine, a liquid substance (e.g. water) and the investigational agent; introducing the liquid composition in the extravascular space of the subject is a mammal (e.g., the vitreous chamber of the eyeball) and the study of one or more properties of the composition in vivo after its introduction. The method is used to identify agents that desired affect one or more properties of the gel and/or the formation of the tion. Investigational agents can contain standard excipients, viscosity modulators, inorganic and organic ions, buffers, etc.

[0081] One aspect of the invention is a liquid composition containing an analogue of constatine, a liquid substance and a modulator of one or more properties of the gel or gel formation.

[0082] Constatin is a cyclic peptide that binds to a component of the complement NW and inhibits the activation of complement. In U.S. patent No. 6,319,897 described a peptide which has the sequence Ile-[Cys-Val-Val-Gln-Asp-Trp-Gly-His-His-Arg-Cys]-Thr (SEQ ID NO: 44), by a disulfide bond between the two cysteine indicated by brackets. It is necessary to clarify that the name "constatin" was not used in U.S. patent No. 6,319,897, but was later accepted in the scientific and patent literature (see, for example, Morikis, et al. Protein Sci., 7(3):619-27, 1998) to determine the peptide having the same sequence as SEQ ID NO: 2 described in U.S. patent No. 6,319,897, but amidinophenoxy to-end, as shown in Table 1 (SEQ ID NO: 8). The term "constatin" is used in this application in accordance with its use (i.e., in relation to SEQ ID NO: 8). Were opened analogues of constatine that have a higher complement inhibitory activity than constatin. See, for example, international publication WO 2004/026328 (PCT/US 2003/029653), Morikis, D., et al., Biochem Soc Trans. 32 (Pt 1):28-32, 004, Mallik, B., et al., J. Med. Chem., 274-286, 2005; Katragadda, M., et al. J. Med. Chem., 49: 4616-4622, 2006; international publication WO 2007062249 (PCT/US 2006/045539); WO 2007044668 (PCT/US 2006/039397), USSN 11/544,389 (Compstatin and Analogs Thereof for Eye Disorders) and description below.

[0083] the Present invention has been at least partly as a result of the unexpected discovery that the introduction of liquid composition containing the active analog constatine and water, in the vitreous body leads to the formation of a macroscopic gel-like structure, also referred to as "inclusion" or just "gel" (see Figure 1). The term "gel" as used in this application refers to a gel or any structure of a material with a characteristic gel physical properties (e.g., gelatin). Thus, the term "gel" is used to describe the physical nature gelatinous or gelatinases patterns, and not to describe a particular physical structure. Specific physical properties are viscosity, elasticity, mechanical strength and compressibility. The term "gel" refers to topics, but not limited to, substances which have a homogeneous structure, which is usually attributed to the gels experts in the field of colloidal chemistry, where the gel can be represented as a colloidal system in which a porous network of interconnected particles (typically of nanometer-sized) Zap the play volume of a liquid medium. Extravascular space may contain the substance of the body, which can have varying degrees of viscosity. The substance may be a liquid body. The substance itself can at least partly be gel-like in consistency. In some embodiments of the present invention gellike structure formed after the introduction of the analog constatine, is more viscous and more dense texture than the contents of the extravascular space in which it is formed.

[0084] the Inventors hypothesized that the inclusion could serve as a unique system of prolonged delivery of analog constatine. It was found that the inclusion contains a significant fraction of the injected analog constatine, and that the substance remains active and is gradually released. Inclusion is a discrete structure, which can be easily separated from the material of the vitreous body in the preparation process shortly after it was formed, and by using a relatively large concentrations. Over time, and/or when using lower concentrations, the gels are soft and not very heavy and easy to break during handling, along with their destruction over time. In some embodiments of the invention gels are almost speeches what they and essentially transparent. In some embodiments of the invention macroscopic gel-like structure over time is reduced in size and/or becomes less dense (based on ultrasound) and releases the analog constatine in the active form with the achievement of therapeutic concentrations of at least 2 weeks, at least 4 weeks, at least 2 months, at least 3 months, at least 6 months, at least 9 months, at least 12 months (for example, up to about 3, 6, 9, 12, 18 or 24 months in different implementations). Extravascular space may be a separate chamber, compartment or cavity, which may contain liquid or semi-liquid substance in the body. Space may be the vitreous chamber of the eyeball, synovial cavity, intrathecal space, etc. Chamber, cavity or compartment may contain fabric, or may be at least partially covered with a cloth, on which the complement has an unwanted effect in the complement-mediated disorder. In some embodiments of the fabric does not contact directly with the contents of the space, but is near enough to the inhibitor of complement could diffuse in the extracellular fluid surrounding tissue. For example,the fabric may be contiguous, for example, to be within 10-20 mm, in some embodiments in the range of 5-10 mm, in the range of 1-5 mm, in the range of 0.5-1 mm, in the range of 0.1-0.5 mm, or within 0.1 mm, and less from the lining compartment; and not to be separated from compartment barrier that would prevent diffusion of the inhibitor of complement in the fabric. Therapeutic concentrations can be measured in substance inside the extravascular space (e.g., vitreous body), or in the lining of his cloth, or in surrounding tissues.

[0085] In some embodiments of macroscopic gel-like structure remains easily detectable for at least 2 weeks, at least 4 weeks, at least 2 months, at least 3 months, at least 6 months, at least 9 months or at least 12 months (for example, up to about 3, 6, 9, 12, 18 or 24 months in various implementations). You must understand that there may be differences between subjects, and the above estimates may reflect average values for a group of study subjects. The term "easily detectable" means that the inclusion can be detected (i) with standard Ophthalmoscope examination; (ii) during preparation without increasing and/or (iii) on images obtained PR is ultrasound. In some embodiments of the inclusion has an initial diameter (or longest axis dimension, or the greatest distance between two points on the surface) of about 5 mm, but may be less or more, depending, for example, from the volume of the liquid composition and concentration of the analogue constatine. In some embodiments of macroscopic gel-like structure has a diameter of at least 1 mm (or the long axis dimension, or the greatest distance between two points on the surface) for at least 2 weeks, at 4 weeks, at least 2 months, at least 3 months, at least 6 months, at least 9 months or at least 12 months (for example, up to about 3, 6, 9, 12, 18, or 24 months in various implementations).

[0086] typically, the inside of the turn is captured only part of the input analog constatine. With increasing concentration, and the total number of input connections increase in the fraction of injected dose found later in inclusion. In some embodiments of the invention at least 25% of the introduced initially, the dose remains within the inclusion. For example, in some embodiments of from 25% to 50%, or from 50% to 75%, of the input analog constatine remains inside the inclusion. In some embodiments of the invention from 7% to 90% of the originally introduced version of constatine remains inside the inclusion. You must understand that there may be differences between subjects, and the above estimates may be average values for the group of study subjects.

[0087] the Inclusion, of course, is not toxic at the doses employed, does not alter the normal physiology of the eye and does not affect vision. In addition to analog constatine it contains proteins normally present in the vitreous body. Inclusion is a long period of time, during which it releases the analog constatine. The term "release" when used in this application generally means doing an active agent (e.g., analog constatine) available outside the inclusion (for example, inside the body), but does not imply any particular mechanism by which this process occurs. Release may, for example, occur in the degradation or destruction of incorporation, by diffusion of the agent from the incorporation, etc. of Redundant analog constatine retains significant activity. In some embodiments of redundant analog constatine retains at least 50% activity (per mol) input connection (for example, from 50% to 75%, from 75% to 95%, from 95% to 100% of the initial activity).

[0088] Thus, the inclusion ensures deposition of analog constatine and offers new ways of prolonged pulling the I. It is assumed that the gel-like inclusions similar inclusions formed after introduction of the composition into the vitreous body, can be formed in vivo, when a liquid composition containing an analogue of constatine is entered into the extravascular space containing body fluids such as synovial fluid cavity, synovial bags, the cavity of the skull, ventricles in the skull cavity, intrathecal space, etc. Such inclusions may contain similar constatine and one or more substances, such as ions, proteins, or glycosaminoglycan present in the liquid of the corresponding space. It is also assumed that the gel-like inclusions can be generated ex vivo from a liquid composition containing an analogue of constatine and one or more proteins, ions or glycosaminoglycan that is similar or identical to those present in vivo in the extravascular space.

[0089] the Invention also provides a method of treating a complement-mediated disorders, comprising the step of introducing a liquid composition containing an effective amount of an analogue of constatine, in the extravascular space of the subject, where the specified effective amount is sufficient for the formation of discrete macroscopic gel-like structure containing analogue of constatine within the specified Vneshaudit the space. In some embodiments of the invention, the composition is injected into the vitreous chamber of the eyeball, for example, by injection into the vitreous body. In some embodiments of a needle is used 27-30 G.

[0090] the Invention also offers gel-like material containing analogue of constatine and one or more proteins, ions, and/or glycosaminoglycan (GAG)present in the natural conditions in the extracellular space of mammals. In one embodiment, the implementation of proteins, ions and/or glycosaminoglycan is naturally present in the vitreous body. In another embodiment, the implementation of proteins, ions and/or glycosaminoglycan is naturally present in synovial fluid. In another embodiment, the implementation of proteins, ions and/or glycosaminoglycan is naturally present in the cerebrospinal fluid (CSF). In some embodiments of the invention, the gel-like material contains at least 2, 3, 4, 5, 6, 7, 8, 9, 10, or more individual proteins and/or GAG. In some embodiments of the invention the protein or GAG is not a substance that is commonly used in this area for the preparation of compounds with slow release. In some embodiments of the invention the protein or GAG is not therapeutic agent known in this field.

[0091] In some embodiments of the invention helipad BNY material is either present in the body of the subject, or extracted from it. In such embodiments, the gel material may contain proteins, ions and/or glycosaminoglycan that are endogenous to this particular subject (i.e., they are normal found in a specific subject). In some embodiments of gel-like material is formed ex vivo. In the latter case, the inclusion may contain proteins, ions or glycosaminoglycan, which are similar or identical proteins, ions or glycosaminoglycans subject, which entered the gel, but not checked out this subject. For example, they can be isolated from natural sources, synthesized using chemical technologies or technologies of recombinant synthesis, etc.

[0092] In some embodiments of the invention, the analog constatine is introduced under conditions suitable for the formation of gel-like inclusions in the vitreous chamber of the eyeball for the treatment of a subject at risk of or suffering from eye disease. In some embodiments of the invention, the disease is a disease in which there is increased activation of complement (for example, high concentrations of products of activation of complement) in the eyes of subjects suffering from the disease, compared with activation of complement in the eyes of not suffering from the disease of individuals. In some embodiments of the invention, the disease is one of the diseases, in which there is increased formation of MAC in the eyes of suffering from disease subjects compared with the number of MAC in the eyes of not suffering from the disease of individuals.

[0093] In some embodiments of the disease is a macular degeneration, for example age-related macular degeneration (AMD). In some embodiments of the disease is an exudative AMD. In some embodiments of the disease is a atrophic AMD. In some embodiments of the subject suffers from geographic atrophy (GA). In some embodiments of the subject suffers from age maculopathy, which belongs to the early or intermediate stages of atrophic AMD, in which HA is not developing. In some embodiments of the disease is a diabetic retinopathy. In some embodiments of the disease is a glaucoma. In some embodiments of the disease is a posterior uveitis or keratitis. In some embodiments of the disease is a retinal pigments. For more information about these and other eye diseases can be found in concurrently filed patent applications USSN 11/247,886, USSN 11/544,389 and USSN 11/612,751, and Kanski J. Clinical Ophthalmology: A Systematic Approach, Butterworth-Heinemann; 6 editio (May 4, 2007). In some embodiments of the composition is introduced into the vitreous body, for example, by injection into the vitreous body. In some embodiments of the composition is introduced into the anterior chamber of the eye.

[0094] the Extravascular space can be chosen according to the specific disease. For example, if the abnormal condition is arthritis, inhibitor of complement may be injected directly into the synovial cavity. Examples of the joints, which can be liquid compositions according to the invention, include hip, knee, elbow joint, wrist joint, sternoclavicular, fist, predpljusnevye, ankle or any other joint subject to arthritic diseases. The compositions are also suitable for the introduction of joint bag. Examples of joint capsules, which can be entered compositions according to the invention include acromial, douglasa beam, tubetorial Delta, infrapatellar, sciatic and other joint bags, well-known experts in this field. If the pathological condition is damage to the spine or chronic pain in the spine, the composition can be entered intrathecal. If the pathological condition is Alzheimer's disease, Parkinson's disease or stroke, the composition can be entered into the system of the ventricles (e.g., third, fourth, or side gelado the EC), which includes a number of structures in the brain, which is a continuation of the Central spinal canal. Can be used known in the field methods of introducing therapeutic agents into such spaces of the body (see below).

[0095] Suitable number of analog constatine can be selected at least partially based on the level of proteins of complement and/or activation of complement within interest extravascular space or adjacent tissues. To determine the level based on measurements conducted on a group of subjects, or individual in need of treatment of the subject. In some embodiments of the invention, the approximate dosage analog constatine is selected at least partly on the basis of this information and it is possible by estimating the total number of proteins of the complement taking into account the approximate amount of space. For example, can be selected approximate dose equivalent of constatine to achieve average or constant concentration in space, sufficient to bind at least 50%, 60%, 70%, 80%, 90%, 95%, or more C3 present. In some embodiments of the dose is selected to achieve an average or constant concentration, greater in 1, 2, 5, 10, 20, 50 time, or any number of times from 1 to 50, greater concentration C3. In some Islands Ianto implementation of the invention similar constatine released from enable to maintain an effective average or constant concentration over a prolonged period of time, for example, within 2-4 weeks, 4-6 weeks, 1-3 months, 3-6 months, 6-12 months, 12-24 months. In some embodiments, the implementation of the selected dose appropriate to achieve the speed of release in the vitreous chamber of the eyeball about 0.01 μg/day to about 3-5 mcg/day, for example, from about 0.05 μg/day to about 3 mg/day, from about 0.1 μg/day to about 3 mg/day, from about 0.5 μg/day to about 3 mg/day, from about 0.5 μg/day to about 2 mg/day, etc. for at least 2 weeks, for example, 2-4 weeks, 4-6 weeks, 1-3 months, 3-6 months, 6-12 months, 12-24 months. In some embodiments, the implementation of the selected dose appropriate to achieve the speed of release in the vitreous chamber of the eyeball from about 0.5 μg/day to 1 μg/day, or from about 1 μg/day to about 3 mg/day, at least 2 weeks, for example, 2-4 weeks, 4-6 weeks, 1-3 months, 3-6 months, 6-12 months, 12-24 months.

[0096] the Invention provides unit dosage compositions described in this application, containing, as a rule in the vessel, a sufficient amount of liquid composition (or solids which can be mixed with liquid pharmaceutical carrier to obtain a liquid composition to provide the desired therapeutic effect on the subject, i.e. enough for a single injection nordausen is camping in the treatment of the subject. In one embodiment, the implementation of the dosing unit is sterile and freeze-dried. In another embodiment, the implementation of the dosing unit is sterile and cooked in liquid form suitable for administration to a subject, for example, by injection or infiltration. In another embodiment, the implementation of the dosing unit is represented by a suspension or dispersion in a liquid suitable for administration to a subject, for example, by injection, infiltration, etc. In some embodiments of the liquid composition contains at least 1 mg analogue of constatine 1 ml of liquid, for example, from about 1 mg to about 300 mg analogue of constatine 1 ml of liquid. Similar constatine do not have to be completely dissolved in the liquid, for example, at least part of the analog constatine may be present in the form of particles. In some embodiments of concentration equivalent to constatine in the composition is in the range from 2 mg/ml to 20 mg/ml In some embodiments of concentration equivalent to constatine in the composition is in the range from 2 mg/ml to 10 mg/ml, for example, from 2 mg/ml to 5 mg/ml In some embodiments of concentration equivalent to constatine in the composition is in the range from 8 mg/ml to 25 mg/ml In some embodiments of the concentration of dissolved analog constatine is in the range from 2 mg/ml to 20 mg/ml In some embodiments of the concentration of dissolved analog constatine is in the range from 2 mg/ml to 10 mg/ml, for example, from 2 mg/ml to 5 mg/ml In some embodiments of the concentration of dissolved analog constatine is in the range from 8 mg/ml to 25 mg/ml

[0097] In some embodiments of the analog constatine is injected into the vitreous chamber of the eyeball (for example, by injection into the vitreous body) in the concentration component of at least 1 mg/ml, for example, from 1 mg/ml to 10 mg/ml, for example, from 2 mg/ml to 5 mg/ml, in a volume of at least 25 µl. In some embodiments of the volume is 25 ál, 150 ál, for example, from 40 ál 125 ál, for example, from 50 μl to 100 μl. In some embodiments of the volume of approximately 50 μl, for example, from 45 to 55 ál ál. In some embodiments of volume approximately 100 ál, for example, from 90 to 110 ál ál. In some embodiments of injected from 50 μg to 2 mg analogue of constatine, for example, from 100 μg to 500 μg. In some embodiments of injected from 500 μg to 1 μg analog constatine. In some embodiments of introduced from 1 mg to 1.5 mg analogue of constatine. In some embodiments of injected from 1.5 mg to 2.0 mg analogue of constatine.

[0098] In some embodiments of from about 150 μg to PR is approximately 250 µg analog constatine is injected into the vitreous chamber of the eyeball. In some embodiments of from about 250 μg to about 350 μg analog constatine is injected into the vitreous omerprazole Apple. In some embodiments of from about 350 μg to about 450 mcg of analog constatine is injected into the vitreous chamber of the eyeball. In some embodiments of from about 450 μg to about 550 μg analog constatine is injected into the vitreous chamber of the eyeball. In some embodiments of from about 550 μg to about 650 μg analog constatine is injected into the vitreous chamber of the eyeball. In some embodiments of from about 650 μg to about 750 μg analog constatine is injected into the vitreous chamber of the eyeball. In some embodiments of from about 750 μg to about 850 μg analog constatine is injected into the vitreous chamber of the eyeball. In some embodiments of from about 850 μg to about 950 μg analog constatine is injected into the vitreous chamber of the eyeball. In some embodiments of from about 950 μg to about 1050 µg analog constatine is injected into the vitreous chamber of the eyeball. In some embodiments of from about 1050 to 1150 µg µg analog constatine is injected into the vitreous chamber of the eyeball. In some embodiments of from about 1150 μg to about 120 μg analog constatine is injected into the vitreous chamber of the eyeball. In some embodiments of from about 1250 μg to about 1350 μg analog constatine is injected into the vitreous chamber of the eyeball. In some embodiments of from about 1350 μg to about 1450 μg analog constatine is injected into the vitreous chamber of the eyeball. In some embodiments of from about 1450 μg to about 1550 ág analog constatine is injected into the vitreous omerprazole Apple. In some embodiments of from about 1550 μg to about 1650 µg analog constatine is injected into the vitreous chamber of the eyeball. In some embodiments of from about 1650 μg to about 1750 µg analog constatine is injected into the vitreous chamber of the eyeball. In some embodiments of from about 1850 μg to about 1950 ág analog constatine is injected into the vitreous chamber of the eyeball. In some embodiments of from about 1950 μg to about 2050 µg analog constatine is injected into the vitreous chamber of the eyeball.

[0099] As described in Example 3 and presented in Figure 5, experiments have shown that it is possible to measure the concentration of analog constatine in serum after his introduction in StegoVideo body. It was found that the serum concentration correlates well with the concentration in the vitreous body. The invention proposes a method of Enki local concentration of analog constatine after local injection in the extravascular space, including the measurement of the concentration of analog constatine in the blood after injection of analogue constatine in the extravascular space and define its local concentration in the specified space on the basis of the known correlation between the concentration in the blood and the concentration of ukcasinos space. The invention provides a method of determining the concentration of analog constatine in the vitreous body, including the measurement of the concentration of analog constatine in the blood after administration into the eye (for example, in the vitreous body) analog constatine and determining the concentration in the vitreous body based on a known correlation between blood concentration and the concentration in the vitreous body. Such methods offer a convenient way to monitor the local concentration (for example, the concentration in the vitreous body) analog constatine and information necessary for the purpose of re-treatment. For example, when the concentration in serum (and, therefore, the local concentration of, for example, in the vitreous body) becomes lower than the desired concentration, the subject of prescribed re treatment. Thus, this method allows the professional practitioner to adapt the time interval between the insertion doses of the composition for treatment of a specific subject.

[00100] the Invention thus comprises the em method monitoring entity after local injection (e.g., injection into the eye) analog constatine. It should be understood that the analysis can be performed on plasma or serum using various methods, for example, using the method of high performance liquid chromatography (HPLC)with detection using a detection or UV or fluorescence detection, jointly or separately, from mass spectrometric analysis (LC-MS).

[00101] in Addition, it was found that the gel-like structure is detected with ultrasound (Figure 2). You can observe the size and density of inclusions over time, thereby providing a second method of monitoring the subject for possible re-appointment of treatment. For example, re-treatment can be carried out, when the diameter or density of the inclusions becomes less than a predetermined limit. To monitor the size of the inclusion and information relevant to the decision to re-treat, you can use the serum in conjunction with ultrasound, with or without ultrasound.

[00102] you Must understand that the analogue of constatine can form gel-like turning on some subjects, but not necessarily in all subjects, at a certain dose. You must also understand that the analogue of constatine can form is its gel-like inclusion in the introduction to some extravascular space, but not everything. Dose and specificity of analogue constatine are selected according to the circumstances. For example, can be selected such similar constatine, and in a dose, at which, as shown, inclusions form at least 50%, at least 60%, at least 70%, at least 80%, at least 90% or at least 95% of the subjects (for example, monkeys or humans) with the introduction of specific extravascular space.

[00103] Konstatin and its analogs

[00104] As described above, the invention appeared at least partly the result of a random and unexpected discovery that a liquid composition containing a sufficient amount and/or concentration of the active analog constatine, form a macroscopic gel-like structure at its introduction into the vitreous chamber of the eyeball mammals. Not wanting to be limited by any theory, the inventors suggest that this property may be of various analogues of constatine. The person skilled in the art can easily verify manifest whether certain analogues of constatine similar properties when introducing them into the vitreous chamber of the eyeball or other extravascular space, based on the discovery described in this application. Aspects of the invention are liquid compositions comprising such analogs of constatine, and applications of the texts of such compositions for the introduction of analogues of constatine and possibly additional active agent. In some embodiments of the liquid composition contains the first and second counterparts of constatine, where at least one of the analogues of constatine forms a gel-like inclusion at its introduction in the extravascular space. In some embodiments of the liquid composition comprises an analog of constatine and a second active agent, which is an analogue of constatine.

[00105] the Term "analogue of constatine" when used in this application refers to constatino and any similar, the inhibitory complement. You must understand that when used in this application, the term "analog constatine" to describe the aspect of the invention, or when referring to the aspect of the invention, the invention includes a variant in which the analog constatine is the compound described in the Examples (SEQ ID NO: 32), unless otherwise specified. The term "analog constatine" includes Konstatin and other compounds that are created or identified on the basis of constatine, and inhibiting complement activity which is at least 50% activity constatine, according to the measurement results, for example, using any method of estimating the activation of complement, adopted in this area, or similar or essentially equivalent analyses. The method can be, for example, based on the assessment of red blood cell lysis mediated classic or and Iternational way. In some embodiments of used ELISA analysis. Ways to evaluate the ability of the compound to inhibit activation of the complement, among other references, are described in international publication WO 2004/026328, Morikis, above, Mallik, above, Katragadda 2006, above. Concatemer and multimer of constatine or inhibiting the complement analogues (with suitable modification of the N - and/or C-end) are also used in the present invention.

[00106] the Activity of the analog constatine can be expressed through the IC50(the concentration of compound at which activation of complement is inhibited by 50%), for example, through a specific concentration in plasma, where the low IC50meets the highest activity, as shown in this area. Activity preferred for use in the present invention analog constatine is at least the same as the activity of constatine. Some modifications are known to reduce or block the complement inhibitory activity and can definitely be omitted from any implementations of the invention. You must understand that the exact Malinina IC50measured for a specific analog constatine will vary depending on experimental conditions. Can be used a reference, for example, experimentally obtained, in which the IC50determined is undertaken for many different compounds under essentially the same conditions. In one implementation, the IC50analog constatine not more than IC50constatine. In some embodiments of the invention, the activity of the analog constatine in 2-99 times more activity constatine (i.e. has similar IC50less than IC50constatine, 2-99 times). For example, the activity may be 10-50 times more activity constatine, or 50-99 times greater than the activity of constatine. In some embodiments of the invention, the activity of the analog constatine in 99-264 times more activity constatine. For example, the activity may be in 100, 110, 120, 130, 140, 150, 160, 170,180, 190, 200, 210, 220, 230, 240, 250, 260 or 264 times more than the activity of constatine. In some embodiments of the invention, the analog constatine is the active counterpart to constatine, such as an analogue with activity at least 100 times greater activity constatine. In some embodiments of the invention, the activity of the analog constatine at least 150, at least 200 or at least 250 times greater than the activity of constatine. Table 1 shows the sequence of some of these analogues. In some embodiments of the invention involves the use of analogues, which activity in 264-300, 300-350, 350-400, or 400-500 times greater than the activity of constatine. The invention also involves primenenie analogues is of constatine, with activity, 500 and 1000 times greater activity constatine.

[00107] For various analogues of constatine was measured Todbinding to C3 by using the method of isothermal titration calorimetry (Katragadda et al., J. Biol. Chem., 279(53), 54987-54995, 2004). Binding ability of various analogues of constatine with C3 was correlated with their activity considering the fact that the lower Kdas is known, indicates a higher binding capacity. Linear correlation between the binding ability and activity was shown for some of the studied analogues of constatine (Katragadda, 2004, above; Katragadda 2006, above). In some embodiments of the invention KD of binding to C3 analog constatine takes values from 0.1 μm to 1.0 μm, from 0.05 μm to 0.1 μm, from 0.025 μm to 0.05 μm, from 0.015 μm to 0.025 μm, 0.01 μm to 0.015 μm, or from 0.001 μm to 0.01 μm. In some embodiments of the IC50analog constatine is from approximately 0.1 μm to 0.2 μm. In some embodiments of the IC50analog constatine is from about 0.05 microns to about 0.1 microns. In some embodiments of the IC50analog constatine is from about 0.001 microns to about 0.05 microns.

[00108] Connection created or identified on the basis of constatine" include, but are not limited to, compounds that contain aminoxy the pilot circuit, the sequence which is obtained by (i) modifying the sequence of constatine (for example, substitution of one or more amino acid sequence constatine another amino acid or an analogue of the amino acids, the insertion of one or more amino acids or analogs of amino acids in the sequence of constatine or deletions of one or more amino acids or analogs of amino acids in the sequence of constatine); (ii) selection of phage display collections of protein sequences, in which one or more amino acids of constatine are randomly distributed and further modified in accordance with the method (i); or (iii) the detection of the sequence using the screening of compounds that compete for binding to C3 or its fragment with konstatinos or any of their analogues, obtained by the methods (i) or (ii). Many used analogues of constatine include hydrophobic cluster, β-turn and a disulfide bridge.

[00109] In some embodiments of the invention, the sequence of analog constatine includes the sequence, or essentially consists of a sequence, which is obtained by using 1, 2, 3 or 4 substitutions in the sequence of constatine, i.e. when 1, 2, 3 or 4 amino acids in the sequence of constatine replaced by another standard amino acid or Nestan artney amino acid. In some embodiments of the invention, the modified amino acid at position 4. In some embodiments of the invention, the modified amino acid at position 9. In some embodiments of the invention modified amino acids at positions 4 and 9. In some embodiments of the invention only change the amino acids in positions 4 and 9. In some embodiments of the invention, the modified amino acid at position 4 or 9, or, in some embodiments, changed both amino acids at positions 4 and 9, and up to 2 amino acids located in positions selected from 1, 7, 10, 11, or 13. In some embodiments of the invention modified amino acids at positions 4, 7 and 9. In some embodiments of the invention, the modified amino acid at position 2 or 12, or changed both amino acids, provided that the compound retains the ability to cyclization. This change (change) at position 2 and/or 12 can be additional to a change (changes) in the position 1, 4, 7, 9, 10, 11 and/or 13. The sequence of any of the analogues of constatine obtained by substitution of one or more amino acids in the sequence of constatine may also include up to 1, 2 or 3 additional amino acids at the C-end. In one embodiment, the implementation of additional amino acid is a Gly. The sequence of any similar compstat is a, obtained by substituting one or more amino acids in the sequence of constatine may also include up to 5 or up to 10 additional amino acids at the C-end. You must understand that the analogues of constatine can have any of the characteristics or properties inherent in the various options outlined in this application and the characteristics or qualities inherent in any of the options, can also refer to any other option, described in this application, unless otherwise specified or the context. In some embodiments of the invention, the sequence of analog constatine comprises or essentially consists of a sequence identical to the sequence constatine, except for the positions corresponding to positions 4 and 9 in sequence constatine.

[00110] Konstatin and some analogues of constatine with a little more activity than constatin contain only standard amino acids ("standard amino acids" include glycine, leucine, isoleucine, valine, alanine, phenylalanine, tyrosine, tryptophan, aspartic acid, asparagine, glutamic acid, glutamine, cysteine, methionine, arginine, lysine, Proline, series, Tramin and histidine). Some analogues of constatine with increased activity, include one or more non-standard amino acids. Used n the standard amino acids include once and multiple halogenated (for example, f) amino acids, D-amino acids, Homo-amino acids, N-alkylated amino acids, dehydrolinalool, aromatic amino acids (not related to phenylalanine, tyrosine and tryptophan), ortho-, meta - or para-aminobenzene acid, phospholinoleate, methoxylamine amino acids and α,α-disubstituted amino acids. In some embodiments of the invention, the analog constatine obtained by substitution of one or more L-amino acids in the analog constatine described earlier in this application, the corresponding D-amino acid. Such compounds and methods of their use are aspect of the invention. Typical used nonstandard amino acids include 2-nafcillin(2-NaI), 1-nafcillin (1-NaI), 2-intergrity carboxylic acid (2Ig1), dihydrostreptomycin (Dht), 4-benzoyl-L-phenylalanine (BPA), 2-α-aminobutyric acid (2-Abu), 3-α-aminobutyric acid (3-Abu), 4-α-aminobutyric acid (4-Abu), cyclohexylamine (Cha), homecollection (hCha), 4-fluoro-L-tryptophan (4fW), 5-fluoro-L-tryptophan (5fW), 6-fluoro-L-tryptophan (6fW), 4-hydroxy-L-tryptophan (40H-W), 5-hydroxy-L-tryptophan (50H-W), 6-hydroxy-L-tryptophan (60H-W), 1-methyl-L-tryptophan (IMeW), 4-methyl-L-tryptophan (4MeW), 5-methyl-L-tryptophan (5MeW), 7-Aza-L-tryptophan (7aW), α-methyl-L-tryptophan (αMeW), β-methyl-L-tryptophan (βMeW), N-methyl-L-tryptophan (NMeW), 5-methoxy-tryptophan, ornithine (from), citrulline, norleucine, γ-glut minbuy acid, etc.

[00111] In some embodiments of the invention, the analog constatine includes one or more analogs of Trp (for example, at position 4 and/or 7 relative to the sequence constatine). Typical analogs of Trp listed above. Cm. also Beene, et. al. Biochemistry 41: 10262-10269, 2002 (described, including, single and multiple halogenated analogs of Trp); Babitzke & Yanofsky, J. Biol. Chem. 270: 12452-12456, 1995 (described, including methylated and halogenated Trp, and other analogs of Trp and indole); and U.S. patent№№6,214,790, 6,169,057, 5,776,970, 4,870,097, 4,576,750 and 4,299,838. Other analogues include Trp variants, substituted (for example, methyl group) and or P carbon, and possibly also one or more of the position of the indole ring. As analogs of Trp can be used amino acids comprising two or more aromatic rings, including substituted, unsubstituted and substituted alternative variants of the rings. In some embodiments of the invention similar to Trp, for example, in position 4 is 5-methoxy, 5-methyl-, 1-methyl - or 1-formyl-tryptophan. In some embodiments of the invention is similar Trp (for example, position 4), including 1-alkyl substitution, for example, replacement of lower alkyl (for example, C1-C5). In some embodiments, the implementation uses N(α)-methyl-tryptophan or 5-methyl-tryptophan. In some embodiments of use is facilitated by the similar, including Deputy 1-alkanols, for example, the lower alkanoyl (for example, C1-C5). Examples include 1-acetyl-L-tryptophan and L-β-tryptophan.

[00112] In some embodiments of the analog of Trp has increased hydrophobicity compared to Trp. For example, the indole ring may be substitution of one or more alkyl (e.g., metal) groups. In some embodiments of the analog of Trp participates in hydrophobic interactions with C3. This analog of Trp may be located, for example, at the position 4 in the sequence of constatine. In some embodiments of the analog of Trp component includes substituted or unsubstituted bicyclic aromatic ring or two or more components substituted or unsubstituted monocyclic aromatic ring.

[00113] In some embodiments of the analog of Trp has an increased ability to form hydrogen bonds with C3 compared to the Trp, but has not increased hydrophobicity compared to Trp. The analog of Trp can have a greater polarity compared to the Trp and/or increased ability to participate in electrostatic interactions with the donor hydrogen bond at C3. Specific typical Trp analogues with increased ability to form hydrogen bonds include electrometrically Deputy in indole coleta analog of Trp may be located, for example, at the position 7 in the sequence of constatine.

[00114] In some embodiments of the invention, the analog constatine includes one or more Ala analogues (for example, in polozhenii on sequence constatine), for example, analogues of Ala, which is identical to Ala, except that they include one or more CH2group in the side chain. In some embodiments of the analogue Ala is an unbranched once methylated amino acid, such as 2-Abu. In some embodiments of the invention, the analog constatine includes one or more analogs of Trp (for example, in position 4 and/or 7 in the sequence of constatine) and the analog of Ala (for example, at the position 9 in the sequence of constatine).

[00115] In some embodiments of the invention, the analog constatine is a compound that includes a peptide having the sequence (X (aa)n-Gin-Asp-XAA-Gly-(X aa)m, (SEQ ID NO: 2), where each X aa and each X”AA represents an independently selected amino acids or analogs of amino acids, where XAA represents Trp or an analog of Trp, and where n>1 and m>1 and n+m takes values between 5 and 21. The peptide has the crustal sequence Gln-Asp-XAA-Gly, where XAA represents Trp or an analog of Trp, for example, the analog of Trp, which has improved the ability to form hydrogen tie the donor hydrogen bonds compared to Trp, but in some implementations not having increased hydrophobicity compared to Trp. For example, the analog can be similar, with the substitution in the indole ring of Trp electronegative group, for example halogen, such as fluorine. In one embodiment, the implementation XAA represents 5-fluoro-tryptophan. The person skilled in the art it is obvious that as the contrary is not proved, then any non-naturally occurring peptide sequence which includes this crustal sequence, and which suppresses the activation of complement and/or binds C3, must be received on the basis of sequence constatine. In alternative implementations Xaa represents an amino acid or similar amino acids other than analog of Trp, which allows the peptide Gin-Asp-XAA-Gly to form a β-turn.

[00116] In some embodiments of the invention, the peptide has the crustal sequence X aa-GIn-Asp-XAA-Gly (SEQ ID NO: 3), where X aa and XAA is selected from Trp and analogs of Trp. In some embodiments of the invention, the peptide has the crustal sequence X aa-GIn-Asp-XAA-Gly (SEQ ID NO: 3), where X aa and XAA is selected from Trp and analogs of Trp and other amino acids or amino acid analogues comprising at least one aromatic ring. In some embodiments of the invention crustal sequence forms a β-turn in point is adalah peptide. β-turn can be flexible, allowing the peptide to adopt two or more conformations, as shown, for example, using the method of nuclear magnetic resonance (NMR spectroscopy). In some embodiments of X'aa is an analog of Trp, which includes substituted or unsubstituted bicyclic component aromatic ring or two or more substituted or unsubstituted monocyclic components of the aromatic ring. In some embodiments of the invention X aa selected from the group comprising 2-nafcillin, 1-nafcillin, 2-intergrity, carboxylic acid, dihydrolipoate and benzylpenicillin. In some embodiments of the invention X aa represents the analog of Trp, which has a higher hydrophobicity compared to Trp. For example, X'aa can be a 1-methyltryptophan. In some embodiments of the invention, XAA is an analog of Trp, which has a high ability to form hydrogen bonds compared to the Trp, but in some embodiments of the invention do not have an increased hydrophobicity compared to Trp. In some embodiments of the invention, the analog of Trp, which has a high ability to form hydrogen bonds compared to the Trp includes the modification of the indole ring of Trp, for example in position 5, Taku is how the substitution of a hydrogen atom by a halogen atom in position 5. For example, XAA may be a 5-fluoro-tryptophan.

[00117] in some embodiments of the invention, the peptide has the crustal sequence X aa-Gln-Asp-XAA-Gly-X”aa (SEQ ID NO: 4), where X aa and XAA are each independently selected from Trp and analogs of Trp, and X”AA is selected from His, Ala, Ala analogues, Phe and Trp. In some embodiments of the invention X aa represents the analog of Trp, which has a higher hydrophobicity compared with Trp, such as 1-methyltryptophan or other similar Trp having an alkyl substituent in the indole ring (for example, in position 1, 4, 5 or 6). In some embodiments of X'aa is an analog of Trp, which includes substituted or unsubstituted bicyclic component aromatic ring or two or more substituted or unsubstituted monocyclic components of the aromatic ring. In some embodiments of the invention X aa selected from the group comprising 2-nafcillin, 1-nafcillin, 2-intergrity carboxylic acid, dihydrolipoate and benzylpenicillin. In some embodiments of the invention, XAA is an analog of Trp, which has a high ability to form hydrogen bonds with C3 compared to the Trp, but in some embodiments, the implementation has not increased hydrophobicity compared to Trp. In some embodiments of the invention, the analog of Trp, to the which has a high ability to form hydrogen bonds compared to Trp, includes modification of the indole ring of Trp, for example, in position 5, such as the replacement of the halogen atom of hydrogen in position 5. For example, XAA may be a 5-fluoro-tryptophan. In some embodiments of X"AA represents Ala or an analog of Ala, such as Abu, or different unbranched once methylated amino acid. In some embodiments of the invention, the peptide has the crustal sequence X aa-Gln-Asp-XAA-Gly-X”aa (SEQ ID NO: 4), where X aa and XAA are each independently selected from Trp, Trp and analogs of amino acids or amino acid analogues comprising at least one aromatic side chain, and X”AA is selected from His, Ala, analogues of Ala, Phe, and Trp. In some embodiments, X”AA selected from analogs of Trp, aromatic amino acids, and analogs of aromatic amino acids.

[00118] in some preferred embodiments of the invention, the analog constatine is cyclic. The peptide can be cycletour using the relationship between any two amino acids, one of which is (H AA)nand the other is located inside (X AA)m. In some embodiments of the cyclic portion of the peptide has a length of 9-15 amino acids, for example, a length of 10-12 amino acids. In some embodiments of the cyclic portion of the peptide has a length of 11 amino acids with communication (for example, by a disulfide bond between amino is a slot in positions 2 and 12. For example, the peptide can have a length of 13 amino acids with communication between amino acids at positions 2 and 12, the circular section has a length of 11 amino acids.

[00119] In some embodiments of the peptide comprises or consists of the sequence X aa1's Aa2's Aa3's AA-Gln-Asp-Xaa-Gly-X”aa1-X”Aa2-X”Aa3-X”a-X”AA (SEQ ID NO: 5). In some embodiments of H a and Xaa is selected from Trp and analogs of Trp, and X aa1, 's Aa2, 's Aa3, X”Aa1, X”Aa2, X”Aa3, X”a, and X”AA independently selected from amino acids and amino acid analogues. In some embodiments of H a and XAA is selected from aromatic amino acids and aromatic amino acid analogues. Any or all of X aa1, 's Aa2, 's Aa3, X”Aa1, X”Aa2, X”Aa3, X”a and X”AA may be identical amino acids in the corresponding positions of the sequence of constatine. In one embodiment, X”Aa1 represents Ala or once methylated non-branched amino acid. The peptide can be cycletour using the covalent bond between (i) X aa1, 's Aa2, 's or Aa3 and (ii) X”Aa2, X”Aa3, X”a or X”AA. In one embodiment, the implementation of the peptide cycletour using covalent bonds between H Aa2 and X”AA. In one embodiment, the implementation of covalently linked amino acids each are Cys, and a covalent bond is a disulfide (S-S) bond. In one embodiment, the implementation of the covalent bond is predstavlyaet a, C-O, C-S or C-N bond. In some embodiments of one of the covalently linked residues represents an amino acid or similar amino acids having a side chain that includes a primary or secondary amine; another covalently linked residue is an amino acid or similar amino acids having a side chain that includes a group of the carboxylic acid, and the covalent bond is an amide bond. Amino acids or analogs of amino acids having a side chain that contains a primary or secondary amine include lysine and diaminobenzidine acids with the General structure of NH2(CH2)nCH(NH2)COOH, such as 2,3-diaminopropionic acid (dapa), 2,4-diaminobutyric acid (daba) and ornithine (from), where n=1 (dapa), 2 (daba), and 3 (h), respectively. Examples of amino acids having a side chain containing the group of carboxylic acids, dicarboxylic acids include amino acids such as glutamic acid and aspartic acid. Can also be used such analogues as beta-hydroxy-L-glutamic acid.

[00120] in some embodiments of the analog constatine is a compound that includes a peptide having the sequence:

[00121] Xaa1-Cys-Va1-Xaa2-Gin-Asp-Xaa2*-Gly-Xaa3-His-Arg-Cys-Xaa4 (SEQ ID NO: 6), where:

Xaa1 is Not, Val, Leu, B1-Ile, B11-Leu or a dipeptide comprising Gly-Ile or B1-Gly-Ile, and1represents the first blocking group;

Xaa2 and Xaa2* are independently selected from Trp and analogs of Trp; Xaa3 represents His, Ala or an analog of Ala, Phe. similar Phe, Trp or an analog of Trp;

Xaa4 is a, L-Thr, D-Thr, He, Val, Gly, a dipeptide selected from Thr-Ala and Thr-Asn, or a Tripeptide comprising Thr-Ala-Asn, where the carboxyl end IT at any of the L-Thr, D-Thr, lie, Val, Gly, Ala, or Asn possibly replaced by a second blocking group2; and the two Cys residue linked by a disulfide bond.

[00122] In other embodiments of Xaa1 is absent or any amino acid or analogous to amino acid and Xaa2, Xaa2*, Xaa3 and Xaa4 are defined above. If Xaa1 is absent, to the N-terminal Cys residue can be attached to the blocking group1.

[00123] in another embodiment, the implementation Haa represents any amino acid or similar amino acids, and Xaa1, Xaa2, Haha* and Hua defined above. In another embodiment, the implementation Haa is a dipeptide selected from the group comprising Thr-Ala and Thr-Asn, where the carboxyl end HE Ala or Asn possibly replaced by a second blocking group2.

[00124] In any embodiments of the analog constatine of SEQ ID NO: 6 Xaa2 can be a Trp.

[00125] In any embodiments of the analog constatine of SEQ ID NO: 6, Xaa2 can not only what to be an analogue of Trp, includes substituted or unsubstituted component bicyclic aromatic ring or two or more substituted or unsubstituted components monocyclic aromatic ring. For example, the analog of Trp can be selected from 2-naphthylamine (2-Nal), 1-naphthylamine (1-Nal), 2-intergrity carboxylic acid (Ig1), dihydrothiophene (Dht) and 4-benzoyl-L-phenylalanine.

[00126] In any embodiments of the analog constatine of SEQ ID NO: 6 Xaa2 can be a to be the analogue of Trp with increased hydrophobicity compared to Trp. For example, the analog of Trp can be selected from 1-methyltryptophan, 4-methyltryptophan, 5-methyltryptophan and 6-methyltryptophan. In one embodiment, the implementation of similar Trp is a 1-methyltryptophan. In one embodiment, the implementation Xaa2 represents a 1-methyltryptophan, Xaa2* represents Trp, Haa represents Ala, and other amino acids identical to the corresponding amino acid sequence constatine.

[00127] In any of the variants of realization analogues constatine of SEQ ID NO: 6 Xaa2* may represent an analogue of Trp, such as an analog of Trp, which has improved the ability to form hydrogen bonds with Sz compared to the Trp, but which in some implementations is not increased hydrophobicity compared to Trp. In some embodiments of the analog of Trp includes power is otricatelniy Deputy in the indole ring. For example, the analog of Trp can be selected from

5-percription and 6-perception.

[00128] In some embodiments of the invention Xaa2 represents Trp, Xaa2* is the analog of Trp, which has improved the ability to form hydrogen bonds with C3 compared Trp, but which in some implementations is not increased hydrophobicity compared to Trp. In some embodiments of the analog constatine of SEQ ID NO: 6 Haa is an analog of Trp, with increased hydrophobicity compared to Trp, such as an analog of Trp, selected from 1-methyltryptophan, 4-methyltryptophan, 5-methyltryptophan, and 6-methyltryptophan; and Haa* is the analog of Trp, which has improved the ability to form hydrogen bonds with C3 compared to the Trp, but which in some implementations is not increased hydrophobicity compared to Trp. For example, in one implementation Haa is methyltryptophan, and Haa* represents 5-perception.

[00129] In some of the above embodiments of Haa represents Ala. In some of the above embodiments of Haa is a once methylated non-branched amino acid, for example, Abu.

[00130] in some embodiments of the invention Haa represents Phe.

[00131] in n is which embodiments of the invention use the analog constatine of SEQ ID NO: 6, described above, where Ha and Ha* independently selected from Trp, Trp analogues, and other amino acids or amino acid analogues, which include at least one aromatic ring; and Haa represents His, Ala or an analog of Ala, Phe, Trp, similar to Trp or another aromatic amino acid or similar aromatic amino acids.

[00132] In some embodiments of the invention a blocking group at the N - or C-end of any analogue of constatine described in this application, is any compound that protects the peptide from degradation that would occur in the absence of such groups, in the blood or vitreous body of the mammal (e.g. human or monkey). For example, a blocking group1can be any group that changes the structure of the N-Terminus of the peptide to inhibit the cleavage of the peptide bond between the N-end amino acids of the peptide and the neighboring amino acid. The blocking group2can be any group that changes the structure of the C-end of the peptide to inhibit the cleavage of the peptide bond between the C-terminal amino acid of the peptide and the neighboring amino acid. Can be used with any suitable blocking group, known in this area. In some embodiments of the invention the blocking group' includes acyl group (i.e., part of carbon the strong acid, which remains after removal of the-Oh groups), which in this application is also called "alkanoyl". Acyl group generally contains from 1 to 12 carbon atoms, for example, from 1 to 6 carbon atoms.

For example, in some embodiments of the invention the blocking group' is selected from the following group: formyl, acetyl, propionyl, butyryl, isobutyryl, valeryl, isovaleryl, etc. In one embodiment, to implement a blocking group1represents an acetyl group, i.e. Xaal is an Ac-He, Ac-Val, Ac-Leu or Ac-Gly-Ile.

[00133] In some embodiments of the invention the blocking group2is a primary or secondary amine (-NH2or other1where R is an organic group such as an alkyl group).

[00134] In some embodiments of the invention the blocking group1is any group that neutralizes or reduces the negative charge, which in the absence of such groups may be present on the N-end at physiological pH values. In some embodiments of the invention the blocking group2is any group that neutralizes or reduces the negative charge, which in the absence of such groups may be present on the s-end at physiological pH values.

[00135] In n is which embodiments of the invention, the analog constatine acetiminophen or amitirova on the N-end and/or end, respectively. Similar constatine can be acetiminophen at N-end, amitirova to-end, or acetiminophen on the N-end and amitirova-end. In some embodiments of the invention, the analog constatine does not include acetyl group, and an alkyl or aryl group at N-end.

[00136] In some embodiments of the analog constatine is a compound that includes a peptide having the sequence:

[00137] Xaa1-Cys-Va1-Haa-Gin-Asp-Xaa2*-Gly-Xaa3-His-Arg-Cys-Xaa4 (SEQ ID NO: 7); where:

Xaa1 is a lie, Va1, Leu, Ac-He, Ac-Va1, Ac-Leu or a dipeptide comprising Gly-Ile or Ac-Gly-Ile;

Xaa2 and Xaa2* are independently selected from Trp and analogs of Trp;

Xaa3 represents His, Ala or an analog of Ala, Phe, or an analogue of Phe, Trp, or an analog of Trp;

Xaa4 is a, L-Thr, D-Thr, He, Val, Gly, a dipeptide selected from Thr-Ala and Thr-Asn, or a Tripeptide comprising Thr-Ala-Asn, where the carboxyl end IT any of the L-Thr, D-Thr, lie, Val, Gly, Ala or Asn possibly substituted-NH2 group, and two Cys residue linked by a disulfide bond.

[00138] Xaa1, Xaa2, Xaa2*, Xaa3, and Xaa4 are defined above for the different variants of realization of SEQ ID NO: 6. For example, in some embodiments of Xaa2* represents Thru some embodiments of Haa is an analog of Trp, which has a higher hydrophobicity compared to the Trp, for example, 1-methyltryptophan. In some embodiments of Haa is the Wallpaper Ala. In some embodiments of Haa is a once methylated non-branched amino acid.

[00139] In some embodiments of the invention Xaa1 represents Ile, and Ha is an L-Thr.

[00140] In some embodiments of the invention Xaa1 represents Ile, Haa* represents Trp, and Ha is an L-Thr.

[00141] in some embodiments of the invention use the analog constatine of SEQ ID NO: 7, as described above, where Ha and Ha* independently selected from Trp, Trp analogues, other amino acids or analogues of aromatic amino acids; and Haa represents His, Ala or an analog of Ala, Phe, or an analogue of Phe, Trp, similar to Trp or another aromatic amino acid or similar aromatic amino acids.

[00142] in some embodiments of any of the analog constatine described in this application, Xaa3 is analogous to His. In some embodiments of any of the analog constatine described in this application, Xaa3 represents Phe.

[00143] In the Table 1 presents a non-limiting list of analogues of constatine used in various embodiments of the present invention. Analogues written as abbreviations in the left column indicating the specific modifications in the indicated positions (1-13) compared to the original peptide, konstatinos. is consistent with use in the field, the term "constatin" when used in this application, and the activity of constatine, which compares the activity of analogues of constatine described in this application relate to the peptide constatine, aminirovanie-end. Unless otherwise indicated, peptides lidirovali-end. Bold font is used to highlight some modifications. Assess the relative activity of the peptides compared to the activity of constatine based on published data and analyses (e.g., WO 2004/026326, Mallik, 2005; Katragadda, 2006; WO 2007/062249). In case, if there are links to several publications on the evaluation of activity of the peptides, presents the most recent published estimate, and it is necessary to understand that the estimates can be averaged in case of discrepancies between analyses. It should also be understood that the peptides listed in Table 1, when used in the compositions and methods according to the invention, generally cyklinowanie using a disulfide bond between two Cys residues. However, there may be other ways of cyclization of peptides.

ND = no data

[00145] In some embodiments of the compositions and methods according to the invention similar constatine has a sequence selected from the sequences 9-36. In some embodiments, the implementation status is Bob and methods according to the invention similar constatine has the sequence selected from the sequences SEQ ID NO: 14, 21, 28, 29, 32, 33, 34 and 36. In some embodiments of the compositions and methods according to the invention similar constatine has the sequence of SEQ ID NO: 28. In some embodiments of the compositions and methods according to the invention similar constatine has the sequence of SEQ ID NO: 29. In some embodiments of the compositions and methods according to the invention similar constatine has a sequence selected from SEQ ID nos: 30 and 31. In some embodiments of the compositions and methods according to the invention similar constatine has the sequence of SEQ ID NO: 32. In some embodiments of the compositions and methods according to the invention similar constatine has the sequence of SEQ ID NO: 33. In some embodiments of the compositions and methods according to the invention similar constatine has the sequence of SEQ ID NO: 34. In some embodiments of the compositions and methods according to the invention similar constatine has the sequence of SEQ ID NO: 36.

[00146] In other embodiments of used analog constatine having the sequence shown in Table 1, but where the AC group substituted by a blocking group1as explained above. In other embodiments of used analog constatine having the sequence shown in Table 1, n is where the group - NH2replaced by a blocking group2as explained above.

[00147] in one embodiment, the implementation of the analog constatine binds essentially the same plot β-chain of human C3, and Konstatin. In one implementation, the analog constatine is a compound that binds to a fragment of the C-terminal site of the β-chain of human C3 with a molecular weight of approximately 40 kDa, which binds Konstatin (Soulika, A.M., et al., Mol. Immunol., 35:160, 1998; Soulika, A.M., et al., Mol. Immunol. 43(12):2023-9, 2006). In some embodiments of the analog constatine is a compound that binds to a binding site of constatine identified in the structure of constatin-C3, for example, the crystal structure obtained by the method of three-dimensional NMR spectroscopy. In some embodiments of the analog constatine represents a connection that can displace constatin in the structure of constatin-C3 with the formation of essentially the same intermolecular bonds with C3, which is typical for constatine. In some embodiments of the analog constatine is a compound that binds to a binding site peptide having the sequence shown in Table 1, for example, SEQ ID NO: 14, 21, 28, 29 or 32, in the structure of the peptide-C3, for example, crystalline structure. In some embodiments, re is the implementation of the analog constatine represents a connection, which binds to the binding site of the peptide having SEQ ID NO: 30 or 31 in the structure of the peptide-C3, for example, crystalline structure. In some embodiments of the analog constatine is a connection that could substitute the peptide sequence of SEQ ID NO: 9-36, for example, SEQ ID NO: 14, 21, 28, 29, 30,32, 34 or 36, in the structure of the peptide-C3 with the formation of essentially the same intermolecular bonds with C3, which is characteristic of the peptide.

[00148] the Average specialist when using routine experimental methods can easily determine whether the analog constatine to contact the fragment C-terminal site of the β-chain of C3. For example, the person skilled in the art can synthesize capable potassiun option analog constatine by including in the connection sequence of amino acids capable of potassiun, such as β-benzoyl-L-phenylalanine (BPA), for example, in the C-terminal site of the sequence (Soulika, A. M., et al, above). Optionally, you can include additional amino acids, for example, a plot of epitope such as FLAG or PA, for more rapid detection of the connection, for example, by the method of Western blotting. At incubation analog constatine with a fragment of activated stitching. Joint localization analog constatine and C3 fragment indicates their binding. The methods of the surface plasma resonance can also be used to determine the ability of linking analogue constatine with the binding site of constatine NW or its fragment. To predict whether the connection to form essentially the same intermolecular connection with C3, which forms constatin or peptide having a sequence of one or more peptides from Table 1, for example, SEQ ID NO: 14, 21, 28, 29, or 32, or, in other implementations, SEQ W N0: 30 or 31, the person skilled in the art can use the program for molecular modeling.

[00149] the Analogues of constatine can be obtained by different synthetic methods to create peptides known in this field, by condensation of amino acid residues, for example, in accordance with common methods of peptide synthesis, can be obtained from the appropriate coding their sequences of nucleic acids by expression in vitro or in living cells using methods known in this field. For example, peptides can be synthesized using standard solid-phase methods described in Malik, above, Katragadda, above, and/or WO 2004026328. Potentially reactive group such as amino group and a carboxyl group, a reactive functional group, etc. can be protected with a subsequent removal of their protection by means of various protective groups and methods known in this field. See, for example, "Protective Groups in Organic Synthesis", 3rded. Greene, T. W. and Wuts, P.G., Eds., John iley & Sons, New York: 1999. Peptides may be purified using standard methods such as reverse-phase HPLC. If desired, can be carried out by separation of the diastereomeric peptides using known techniques such as reverse-phase HPLC. The compositions can be optionally dried and subsequently dissolved in a suitable solution, for example, in the water. the pH of the final solution can be increased, for example, to physiological pH values using a base, such as NaOH. Sample peptides may optionally be analyzed using the method of mass spectrometry, for example, to confirm their mass and/or the presence of disulfide bonds. See, for example, Mallik, 2005, and Katragadda, 2006.

[00150] the Structure of constatine known in this field, and also known patterns of a number of analogues of constatine having activity greater than the activity of constatine that were detected by NMR spectroscopy (Malik, above). Information about the structure can be used to generate mimetics of constatine. In one embodiment, the implementation of a mimetic of constatine is any compound that competes with konstatinos or any similar constatine (for example, analog constatine, the sequence of which is presented in Table 1) for binding to C3 or a fragment (such sacrament β-chain weight of 40 kDa, associated with constatin), and which has activity equal to or greater activity constatine. A mimetic of constatine can be a peptide, nucleic acid, or a molecule with a low molecular weight. In some embodiments of the mimetic of constatine is a compound which binds to the binding site of constatine identified in the structure of constatin-C3, for example, the crystal structure or the three-dimensional structure obtained by NMR spectroscopy. In some embodiments of the mimetic of constatine is the connection, the ability to replace constatin in the structure of constatin-C3 with the formation of essentially the same intermolecular bonds with C3, which forms constatin. In some embodiments of the mimetic of constatine is a compound which binds to the binding site of the peptide having the sequence shown in Table 1, in the structure of the peptide-C3. In some embodiments of mimetic of constatine is ones relationship, but has a side chain that represents the sequence is designed based on the sequence constatine.

[00151] For specialists in this area it is obvious that if one was established some desired conformation of a short peptide, it is easy to find ways to create the project for a peptide or peptidomimetic with the same conformation. See, for example, G.R.Marshall (1993), Tetrahedron, 49: 3547-3558; Hruby and Nikiforovich (1991), Molecular Conformation and Biological Interactions, P.Balaram & S.Ramasehan, eds., Indian Acad. of Sci., Bangalore, PP. 429-455), Eguchi M, Kahn m, Mini Rev Med Chem., 2(5):447-62, 2002. Design of peptide analogues can also be optimized in consideration of the contribution of the side chains of amino acid residues, for example, the effects of functional groups or steric factors, which are described in this field for constatine and its analogues, along with other peptides.

[00152] the Experts obviously, the mimetic peptide may serve as well as the peptide, if we are talking about a specific frame in the conformation and functionality of the side chains required for binding to C3 and suppressing activation of complement. Thus it is understood that in the framework of the present invention addresses the receipt and application of the C3-binding inhibiting the complement of connections when using natural amino acids, derivatives of amino acids and amino acid analogues, or molecules that are not amino acids, is able to unite with the formation of a suitable frame conformation. Ones analogues or analogues, including peptide and ones of the components in this application are sometimes referred to as "peptidomimetics" or "esoteric mimic" to describe substituents or derived peptide, on adusa largely the same major conformational characteristics and/or other functional properties, to be quite similar to typical peptides that inhibit the activation of complement. More generally, mimetic of constatine is any compound that has pharmacophore like pharmacophore of constatine, even if it has a different skeleton.

[00153] the Use of peptidomimetics to create high-affinity peptides is well known in this field. Given the limitations of rotation, similar to those for amino acid residues in the peptide, analogs, including diaminotoluene compounds can be analyzed, and their conformational motifs can be installed, using schemes of Ramachandra (Hruby & Nikiforovich 1991), along with other known methods. To identify mimetics of constatine that are associated with C3, can be applied methods virtual screening. In such methods can be used matching algorithms to combine, compare and, if necessary, to classify many of the structures of the candidate with the aid of a computer. To conduct virtual screening can be used any available program from their huge diversity. Typical programs used for flexible connection of molecules, include DOCK 4.0, FlexX 1.8, AutoDock 3.0, GOLD 1.2, ICM 2.8 and later versions. [00154] the person skilled in the art will easily be able to find a suitable screening the final analysis to determine additional mimetics of constatine and to select those that have the desired inhibitory activity. For example, canstatin or its analog may be labeled (e.g., using radioactive or fluorescent label) and preincubation with C3 at different concentrations of the investigated compounds. Evaluates the ability of the compounds to reduce the binding of the analogue constatine with C3. The investigational compound that reduces the binding of the analogue constatine with C3, is a candidate for the role of mimetica of constatine. For example, an investigational compound that reduces the equilibrium concentration of the complex analogue of constatine-C3 at least 25%or at least 50%, is a candidate for the role of mimetica of constatine. Experts obviously, can be used a number of variants of this screening study. Compounds selected for screening analysis may include natural products library of aptamers, libraries, phage display libraries of compounds synthesized using combinatorial chemistry, etc. of the Invention includes the creation of combinatorial libraries of compounds on the basis of cow sequence described above, and screening the library to identify mimetics of constatine. Any of these methods can also be used to identify new analogues of constatine with higher inhibitor Akti is ness, than previously studied analogues of constatine.

[00155] Combination therapy and compositions

[00156] the Invention provides liquid compositions comprising: (a) analog constatine sufficient for the formation of a macroscopic gel-like structure after his introduction in the extravascular space in the body of the mammal; and (b) an additional therapeutic agent, where the specified therapeutic agent is an analogue of constatine. The present invention involves the use of analogues of constatine together with one or more other secondary agents effective for the treatment of disorders. Agents can act on the same target (s) or path (the path), or to other targets or path. Similar constatine and the secondary agent can act additively or synergistically (when combined activity of agents that is greater than the sum of their activities in separate introduction). In some embodiments of the secondary agent is introduced to ensure violations are not mediated through the activation of complement, i.e. a gel-like structure is used essentially as a delivery system for the secondary agent with its slow release. In some embodiments, the implementation for the formation of a gel-like structure is used, the peptide comprising a sequence selected from SEQ ID NO: 3,4, 5, 6 or 7, where the peptide has a lower complement inhibitory activity than constatin. In some embodiments of the activity of the peptide is 50%or less, from the activity of constatine.

[00157] the Secondary agent (agents) are selected in accordance with the specific disorder that should be treated. Suitable agents include anti-inflammatory agents such as corticosteroids, nonsteroidal anti-inflammatory agents, leukotriene or antagonists of leukotriene receptor, cytokine or receptor antagonists of cytokine (e.g., anti - α agents, such as antibodies or soluble receptors TNFα, or its fragments, which bind α), anti - IgE agents (e.g. antibodies or antibody fragments that bind to IgE or IgE receptor), angiogenesis inhibitors, analgesics and anti-infective agents.

[00158] In some embodiments of the secondary agent is a neuroprotective agent, or an antioxidant, or a compound that inhibits or slows down the cycle of transformations of rhodopsin.

[00159] In some embodiments of the invention, the additional active agent is an inhibitor of angiogenesis. Uses a variety of inhibitors of angiogenesis. In some embodiments of the invention, the inhibitor of angiogenesis is specifically associated with the Noi or more isoforms or receptors of growth factor vascular endothelial (VEGF). Angiogenesis inhibitor can be an antibody, antibody fragment, polypeptide, peptide, nucleic acid, aptamer or siRNA. In some embodiments of the invention, the angiogenesis inhibitor specifically binds to one or several or all isoforms or receptors of growth factor vascular endothelial (VEGF). In some embodiments of the secondary therapeutic agent is humanitariannet monoclonal antibody, which binds to the growth factor vascular endothelial (VEGF), such as an antibody described in Presta LG, et al., Cancer Res., 57, 4593-4599 (1997), or antigen-binding fragment, or antibody or antibody fragment that binds to the same epitope. In some embodiments of the invention, the inhibitor of angiogenesis comprises, or is a peptide or polypeptide mammals, such as the factor derived from the pigment epithelium (FPPA), angiostatin, endostatin etc., or a fragment that retains angiogenic activity. Angiogenesis inhibitor can be an inhibitor that is commonly used for the treatment of AMD and/or neovascularization of choroidea or neovascularization of the retina, such as Lucentis® (ranibizumab), Avastin® (bevacizumab) or Macugen® (sodium pegaptanib). In some embodiments of ranibizumab, bevacizumab or p is captaib used in the concentration (or amount), approximately 0.5-5 times greater than the concentration, some for its clinical use as a single agent in the treatment of exudative AMD. In typical implementations, the number is approximately 0.3 mg, 0.5 mg, 1.0 mg or 1.5 mg In some embodiments of the liquid composition containing the equivalent of constatine and inhibitor of angiogenesis, is introduced into the eye of a subject suffering from the neovascularization choroidea and/or neovascularization of the retina, for example, a subject suffering from exudative AMD. The inhibitor can be created, for example, using recombinant technology, hybridoma technology, chemical synthesis or isolated from natural sources.

[00160] the Invention also provides a method including: (a) introduction to the subject is a mammal of similar constatine sufficient for the formation of a macroscopic gel-like structure at its introduction in the extravascular space of the subject; and (b) the introduction of an inhibitor of angiogenesis in the body of the subject. In various embodiments of the invention, the inhibitor of angiogenesis can be entered or not entered s in the composition of the liquid composition. If the inhibitor of angiogenesis is introduced as part of another structure, it can be entered either in the same extravascular space, or in another area. The separate introduction of liquid composition may shall be entered in advance essentially at the same time, or after administration of an inhibitor of angiogenesis. In some embodiments of the inhibitor of angiogenesis is introduced first, and the liquid composition according to the invention is inserted through a certain time interval. The time interval may be, for example, to 1, 2 or 4 weeks after administration of the inhibitor of angiogenesis, or up to 2 or 3 months after administration of the inhibitor of angiogenesis. In some embodiments of the liquid composition according to the invention is introduced into the eye of a subject suffering from AMD, only after the subject has a better view, and/or a decrease in the thickening of the retina, and/or reducing the exudation inside the eye (for example, when measured using optical coherence tomography and fluorescence angiography). Angiogenesis inhibitor may be used in accordance with standard dosages and routes of administration for such an agent (for example, by way of introduction into the vitreous body).

[00161] the Invention provides liquid compositions comprising: (a) analog constatine sufficient for the formation of a macroscopic gel-like structure when introduced into the extravascular space of the subject is a mammal; and (b) an additional inhibitor of complement, where a specific inhibitor of the complement is not an analogue of constatine. In various who's implementations secondary inhibitor of complement is a peptide, the polypeptide ones molecule with a low molecular weight, aptamer, antibody or nucleic acid. In some embodiments of the invention, the secondary agent is a cyclic peptide. In some implementations, the agent is an antagonist of the receptor CA (C5aR). Typical receptor antagonists Sa include a variety of small cyclic peptides, such as peptides described in U.S. patent No 6,821,950; USSN 11/375,587 and/or PCT/US 06/08960 (WO 2006/099330). In some embodiments of the invention uses a cyclic peptide comprising the sequence [OPdChaWR] (SEQ ID NO: 33). In some embodiments of the invention uses a cyclic peptide comprising the sequence [KPdChaWR] (SEQ ID NO: 37). In some embodiments of the invention is a peptide comprising the sequence (XAA)n[OPdChaWR] (SEQ ID NO: 38), where XAA represents an amino acid residue, and n takes values from 1 to 5. In some embodiments of the invention is a peptide comprising (Xaa)n[KPdChaWR] (SEQ ID NO: 39), where XAA represents an amino acid residue, and n takes values from up to 5. In some embodiments of the invention n is equal to 1. In some embodiments of the invention n is equal to 1, and XAA is a standard or non-standard aromatic s is nomikoto. For example, can be used peptides F-[OPdChaWR] (SEQ ID NO: 40), F-[KPdChaWR] (SEQ ID NO: 41), Cin-[OPdChpWR] (SEQ ID NO: 42) and HCin-[OPdChaWR] (SEQ ID NO: 43). Perhaps the free end contains a blocking group, for example, terminal amino acid may be azetilirovanna. (Abbreviations: O - ornithine; Cha - cyclohexylamin; Cin - cynnamoyl; Hcin - hydrocinnamic; square brackets denote an internal peptide bond).

[00162] the Composition containing particles

[00163] In some embodiments of the liquid composition in addition to sufficient for formation of a gel-like structure to the number of analog constatine contains many particles that include a therapeutic agent, the composition is capable of releasing therapeutic agent. Such compositions and gels containing such compounds, are aspect of the invention. Particles can be, for example, microparticles or nanoparticles. Microparticles or nanoparticles may represent a particle-based polymer (for example, containing synthetic organic polymers, polypeptides, etc.), particles created on the basis of the lipids or lipid-containing particles, such as liposomes, Nozomi, micelles, etc. In some embodiments of the invention therapeutic agent is an inhibitor of the complement. Particles are retained in the gel-like structure and are released with vremenem the process of destruction or dissolution of the gel-like structure. Particles included in the gel, can at least partially break down or dissolve. The inhibitor of the complement can be an analog constatine, which may be either the same analog constatine, which forms a gel-like structure, or other similar constatine. Other commonly used therapeutic agents discussed above.

[00164] the Nanoparticles or microparticles can be created using any method known in this field, including, but not limited to, methods, spray drying, phase separation, single or double emulsion solvent evaporation, solvent extraction solvent and a simple or complex koatservatsii. The dispersed polymer compositions can also be obtained by granulation, extrusion and/or spheronization. See, for example, the publication of the application for U.S. patent No. 20040092470. How to create liposomes and the other based on the lipid particles known in the field. In some embodiments of the particles consist essentially of one or more analogues of constatine. In some embodiments of the particles comprise at least 50% of the analogue (analog) constatine on a dry weight basis. It is possible that such particles may contain one or more auxiliary substances.

[00165] the Composition may contain nanoparticles or microcast is s, having different compositions and/or properties. The conditions used to obtain particles can be selected to obtain particles with the desired size or characteristics (e.g., hydrophobicity, hydrophilicity, external morphology, density, hardness, "stickiness", shape etc). Used a method of obtaining particles and used conditions (e.g. solvent, temperature, concentration, flow rate of air, etc. may also depend on therapeutic agent and/or composition of the polymer matrix. Generally, it is desirable to avoid extreme temperatures or pH values, which can lead to significant degradation of the inhibitor of the complement. Obviously, the degree of degradation may depend on certain conditions,' and the time during which the inhibitor of the complement being exposed to these conditions, as well as the structure and properties of the agent. For example, a stable peptide, such as an analogue of constatine, can have significant benefits. Compounds can be tested on the subject of retaining sufficient efficiency in obtaining them using the selected method. In some embodiments of the use of the technology selected according to the method yields a composition which, following technology, saves at least 10%, preferably 20%, 50%, or more, from the activity and the output connection.

[00166] Used a method of obtaining particles and conditions (e.g. solvent, temperature, concentration, flow rate of air, etc. may also depend on some active agents, and other components included in the composition. If the particles obtained one of the above methods, have a size beyond the desired size, the particles can be calibrated in size, for example, by sieving, grinding, etc. Can be used a combination of methods.

[00167] the Microparticles and nanoparticles used in the invention can have a range of values. In General, microparticles have a diameter of 500 microns or less, for example, from 1 μm to 500 μm, from 50 μm to 500 μm, from 100 μm to 250 μm, from 20 μm to 50 μm, from 1 μm to 20 μm, from 1 μm to 10 μm, and so on; and the nanoparticles have a diameter less than 1 micron, for example, from 10 nm to 100 nm, from 100 nm to 250 nm, from 100 nm to 500 nm, 250 nm, 500 nm, 250 nm to 750 nm, from 500 nm to 750 nm. In some embodiments of the microparticles have a diameter in the range from 5 to 750 micrometers. In some embodiments of the microparticles have a diameter in the range from 10 to 500 micrometers. In some embodiments of the microparticles have a diameter in the range from 20 to 200 micrometers. In some embodiments of the nanoparticles have a diameter in the range from 5 to 750 nanometers. In some embodiments, implementing the tion of the nanoparticles have a diameter in the range from 10 to 500 nanometers. In some embodiments of the nanoparticles have a diameter of from 20 to 200 nanometers. In some embodiments of the particle size selected to reduce or prevent their transport across the capillary walls, thus minimizing the likelihood of their release into the vascular system.

[00168] In some embodiments of the microparticles or nanoparticles are formed from a polymer selected from the group consisting of hyaluronic acid, chitosan, collagen, gelatin, alginate, poly(lactides), poly(glycolide), poly(lactide-co-glycolide), poly(milk acid), poly(glycol acid), poly(lactic acid-co-glycol acid), polycaprolactone, polycarbonates, polyetherimides, polyanhydrides, poly(amino acids), poly(arteparon), Polyacetals, polycyanoacrylates, poliferation, poly(dioxanone), poly(alkilenkarbonatov), copolymers of polyethylene glycol and polyarthra, biodegradable polyurethanes, blends and copolymers. For example, the polymer may be a polylactic acid (PLA)polyglucosan acid (PGA) or poly(lactic acid-co-Gileva acid). Particles may be essentially the same size (e.g. diameter) or shape, or may be heterogeneous in size and/or shape. They may be essentially spherical, or may have other forms, in lednum if the particle size is determined not by the diameter, as the largest direct distance between two points on the particle surface. Many particles can be represented approximately 20% to 100% of the particles falling within the scope of any of the following ranges of values, for example, about 40%, 50%, 60%, 70%, 80%, 90% etc.

[00169] the Liquid compositions and methods for their preparation

[00170] In General, inhibitors of complement and other therapeutic agents obtained using standard methods known in this field and suitable for compounds of this class. Peptides, such as analogs of constatine, and other peptides described in this application can be obtained using standard technologies solid-phase synthesis of peptides. For example, the analogue of constatine can be created using the method of solid-phase synthesis of protected peptide using Fmoc chemistry with the release of the peptide from the resin and removal of the protective groups of the side chains, along with the formation of disulfide bonds between Cys2 and Cysl2, with subsequent purification and transformation of oxidized peptides in the acetate form. If necessary, the main product liabilitiesa. Manufacturers peptides include such companies as Advanced Chemtech, Ambiopharm, American Peptide, Dalton Pharma Services, GenScript, Integrated Biomolecule, Lonza, New England Peptide, Peptide 2.0, Synthetech, etc. Recombinant polypeptides can be obtained using standard recomb nantah technologies nucleic acids, as described, for example in USSN 11/247,886 and PCT/US 2005/36547 (WO 2006042252). For more detailed information regarding the preparation of recombinant polypeptides and purification of polypeptides, see, for example, Hardin, S., et al., (Eds.), "Cloning, Gene Expression and Protein Purification: Experimental Procedures and Process Rationale", Oxford University Press, Oxford, 2001. Antibodies, for example, monoclonal antibodies can be obtained from hybridomas or created by recombinant methods known in this field. Nucleic acids such as siRNA, aptamers etc, are synthesized using standard methods. Chemical modification, such as conjugation with PEG, can be obtained using standard methods.

[00171] the Liquid composition according to the present invention contains an analog of constatine and a pharmaceutically acceptable carrier. The term "pharmaceutically acceptable carrier" refers to non-toxic carrier, which does not inhibit the pharmacological activity of the compounds, the bearer of which he is. Pharmaceutically acceptable carriers or diluents which can be used in the compositions according to the invention include liquids such as water, saline, etc. but are not limited to them. In some embodiments of the invention, the pharmaceutically acceptable carrier is a water. In some embodiments of the pH of the liquid is about composition takes values from 3.5 to 6.5. In some embodiments of the pH of the liquid postava takes values from 4.0 to 4.5. In some embodiments of the pH of the liquid composition takes values from 4.5 to 5.0. In some embodiments of the pH of the liquid composition takes values from 5.0 to 5.5. In some embodiments of the pH of the liquid composition takes values from 5.5 to 6.5.

[00172] In some embodiments of the invention the liquid composition may contain various additional components. For example, can include buffers, pH modifiers, solvents, agents regulating osmolarity (e.g., sugar), etc. Can be used with standard excipients known in this field. In some embodiments of the analog constatine dissolved in a liquid, e.g. water, to which have been added one or more buffers or excipients. For example, this may be a solution containing such excipients as amino acid, or polyhydric alcohol (for example, sugar alcohol), and/or buffer. Similar constatine is added in powder form and dissolved. If necessary, the solution is filtered.

[00173] Can be used pharmaceutically acceptable salt analogue, constatine, such as derivatives, pharmaceutically acceptable inorganic or organic acids and bases. In addition to the, you must understand that the invention includes a solvate, hydrates, enantiomers, conformation, tautomers, polymorphic forms, etc. of the active agents described in this application. In some embodiments of the analog constatine as counterion comprises acetate.

[00174] the Number and concentration of analogue (analog) constatine in the composition may vary depending on a number of factors, including but not limited to, the degree of identity analogue (analog) constatine, specific pathological condition and its severity and so on, provided that such amount and concentration are sufficient for formation of a macroscopic gel-like structure with the introduction of the composition. The duration of release can be controlled by selecting the appropriate number and/or concentration of the input analog constatine. You must also understand that the minimum quantity and/or concentration of analog constatine required to form a macroscopic gel-like inclusions may vary depending on such factors as the type of the subject, age of the subject, etc. the person skilled in the art will easily be able to find the appropriate values. In addition, the composition does not necessarily have to form a gel-like inclusion in the body of each entity subject to its de the Union.

[00175] In some embodiments of the invention provides a composition containing an analogue of constatine, where the composition is characterized by the fact that after his introduction into the vitreous body of the primacy it forms a gel, which remains detectable by ultrasound and/or ophthalmic research at least 3 months and at least 75% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, or more) eyes, which was introduced in the composition. In some embodiments of the invention provides a composition containing an analogue of constatine, where the composition is characterized by the fact that after his introduction into the vitreous body of the primacy of the composition forms a gel, which remains detectable by ultrasound and/or ophthalmological research for at least 6 months at least 75% of the eyes, which was introduced in the composition. In some embodiments of the invention provides a composition containing an analogue of constatine, where the composition is characterized by the fact that after his introduction into the vitreous body of the primacy of the composition forms a gel, which remains detectable. using ultrasonic and/or ophthalmological research for at least 9 months, at least 75% of the eyes, which was introduced in the composition. In some embodiments, re is the implementation of the APE is an APE, for example, cynomolgus macaque. In some embodiments of the Primate is a human. In some embodiments of the invention, the gel becomes undetectable by ultrasound after 12, 15, 18, or 24 months after the introduction of at least 75% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, or more) eyes. In some embodiments of the gel releases from 1 μg to 5 μg analog constatine a day, for example, about 3 μg per day for at least 3 months, for example, within 3-6, 6-9 or 9-12 months.

[00176] the Invention provides liquid formulations containing analogue of constatine and one or more excipients or buffers. In some embodiments of the excipient is an amino acid. In some embodiments of the amino acid is an arginine, histidine or series. In some embodiments of the auxiliary substance is a "sugar alcohol" (also called "polyhydric alcohol"). In some embodiments of the auxiliary substance selected from the group comprising ethylene glycol, glycerol, aritra, Arabic, xylitol, ribitol, mannitol, sorbitol, isomalt, ▫ maltitol and lactic. In some embodiments of the auxiliary substance is present in a concentration from 1 mm to 250 mm, n is the sample, from 10 mm to 100 mm, or from 10 mm to 50 mm, for example, approximately 20-45 mm. In some embodiments of the composition contains two or more excipients. In some embodiments of the buffer is selected from acetate buffer and phosphate buffer. In some embodiments of the buffer is a sodium acetate. In some embodiments of the concentration of the buffer takes values from 10 mm to 1M, for example, from 20 mm to 500 mm, for example, from 50 mm to 200 mm.

[00177] As described in Example 4, it was found that the presence of certain excipients and/or buffers in the liquid composition leads to the formation of gels, which have different and more delicate texture than gels formed from the composition containing the same number of analog constatine and water only. In addition, these gels disappear in less time after the introduction of the composition into the vitreous body than gels formed from the composition containing the same number of analog constatine and water only. The inventors have shown that the use of such auxiliary substances and buffers allows you to modulate various physical properties of the gel and the rate at which the analog constatine released from the gel in vivo. The invention provides a method of modulating the rate of release of analog constatine of Geel is, containing the specified analog constatine, the method includes the preparation of the specified analog constatine in the composition of the liquid composition containing excipient or buffer, where the presence of an excipient or buffer modulates (e.g., increases or decreases) the speed with which formed after administration of the composition in vivo, the gel dissolves or degrades.

[00178] One aspect of the invention is the discovery that the use of auxiliary substances that increase the rate of dissolution/destruction of the gel in vivo, allows the introduction of increased amounts and/or higher concentrations of analog constatine without generating too stable to release the desired quantity of analog constatine over time gel.

[00179] the Assessment of inhibiting the complement capabilities

[00180] For analysis of the ability of the agent to inhibit the activation of the complement system (or any of the other important properties) may use any suitable method. A number of tests can be conducted in vitro. For example, the ability of the agent to inhibit the classical or alternative pathway of complement can be estimated by measuring the indirect complement hemolysis red blood cells (e.g. erythrocytes of rabbit or sheep sensitized Ileana sensitized to the antibody) adding human serum or set of components of complement in the presence or absence of the agent. The agent capable of inhibiting complement, if this analysis it reduces hemolysis to a statistically significant level (p<0.05). The ability of the agent to communicate with one or more components of complement, such as C3, C5, factor B, factor D, can be analyzed using isothermal titration calorimetry or other techniques suitable for implementation in a liquid medium. In another embodiment, the implementation of the ability of the agent to communicate with a component of the complement is measured using ELISA analysis. For example, wells tiralongo microplate covered by the agent. Inhibitor of complement can be functionalitywith in order to speed up linking it with the tablet. For example, the agent can be biotinylated in the case of streptavidin coated tablet. Component (s) of complement is added to the wells. After the incubation period, the wells are washed, and bound complement components are detected using antibodies to the interest component of complement. Other methods include surface plasma resonance, equilibrium dialysis, etc.

[00181] the Methods for measuring systemic or local activation of complement in vitro or in vivo, and methods for evaluating the ability of an inhibitor of complement suppressing such activation is known in this field. For example, measuring the number of products is tov activation of complement, such as C3a, Sa, C3bBb, C5b-9, the covalent bond between the recognizable molecule of the classical pathway (C1q) and activated C4, and so on, allows you to assess the degree of activation of complement. Reducing the number of such products indicates the suppression of the activation of complement in some embodiments of the invention. In some embodiments of the measured ratio between the active cleavage product and its inactive desArg form (e.g., C3a/C3adesArg). The expert can distinguish the activation of the classical, alternative and pectinophora ways when measuring correctly selected product (s) activation of complement and/or selection of appropriate activators of complement, such as zymosan, lipopolysaccharide, immune complexes, etc. Other methods include measurement mediated by complement hemolysis of red blood cells due to the formation of the terminal complex.

[00182] the Activation of complement in vivo and/or its inhibition by inhibitors of complement can be measured in a corresponding biological sample. For example, systemic activation of complement and/or its suppression inhibitor of complement can be measured in the blood sample or blood plasma. Local activation and/or suppression in the vitreous body can be measured in a sample of the vitreous body. Local activation and/or suppression in the stomach is positive paths can be measured in the specimen. Local activation and/or suppression in the CNS can be measured in the CSF sample. Serial measurements started prior to the introduction of the inhibitor of complement, allow us to estimate the degree of suppression of the activation of complement inhibitor, as well as the dynamics and duration of inhibition. It is obvious that the reduction product activation is possible to speak only if the observed reduction in the number of products activation detectable before the introduction of the inhibitor of complement, or their disappearance.

[00183] Suitable methods are described in several references listed in this application (U.S. patent No 5,157,110; 6,551,595; U.S. patent. No 6,319,897; WO 2004/026328 (PCT/US 2003/029653), USSN 10/937,912; Morikis, 2004; Mallik, 2005; Katragadda, M., 2006).

[00184] the Monitoring of the degradation of the gel-like inclusions containing analogue of constatine

[00185] As noted above, certain gel-like inclusions according to the present invention are identified in vivo using ultrasound. If necessary, the degradation of inclusion can also be measured with the introduction of a contrast agent in a liquid composition prepared for injection. When used in this application, the term "contrast agent" refers to a substance, for example, the molecule or molecular complex that can be detected if present in vivo using a particular method or methods. Generally, the method of detection is the NR is SNIM and non-invasive, i.e., the method does not include damage to the skin or other accessible body surface or inside the body. In some embodiments of the detection of a contrast agent can be used to estimate the mass or volume of the composition for prolonged delivery device or prolonged delivery, which is stored within "X" time after injection; and/or to estimate the mass or volume of the composition for prolonged delivery device or prolonged delivery, which degrades within "X" time after injection. If it is detected that a previously defined mass (or volume) of the composition is decreased or remained in the same state, the subject can repeat the treatment within a reasonable period of time. For example, repeated treatments may be scheduled after 1, 2, 3, or 4 weeks from the moment when detected, or it is expected that the composition for prolonged delivery degraded by at least 70%, 80%, 90%, 95%, 99% or 100%. A method for the detection of a contrast agent can alternatively or additionally be used to estimate the amount of therapeutic agent, which is stored in the composition for prolonged delivery device or prolonged delivery, i.e. not yet freed.

[00186] the Methods of treatment and the choice of the subject

[00187] the Invention before aget treatment of the subject, involving the introduction of a liquid composition containing an analogue of constatine, in the extravascular space of the subject in an amount sufficient for the formation of a macroscopic gel-like inclusions. The methods according to the invention can include the choice of the subject, which will be introduced in the composition according to the invention. The subject, as a rule, is at risk or suffering from macular degeneration, for example age-related macular degeneration. In some embodiments of the subject suffers from at least one complement-mediated disease, which is characterized by inflammation of the eye. In some embodiments, the subject is at risk or suffering from at least one complement-mediated disease, in addition to diseases of the eye characterized by macular degeneration or inflammation of the eye.

[00188] the Composition typically administered to a subject to treat disorders or preventing the development of such disorders. Thus, the subject usually is at risk or suffering from such a condition. To identify a subject at risk of or suffering from disorders of interest according to this application can be used any suitable research and criteria. Methods of diagnosis of diseases, and representing the interest according to this application, and the ways to assess response to therapy known in this field.

[00189] In some embodiments of the invention, the method of treatment includes identifying the subject of a genetic polymorphism associated with the progression of the disease Or an increased risk of developing the disease. The term "identification" in this case refers to the finding that the subject has a polymorphism that increases the risk of developing the disease, either by holding or processing of a suitable research, or using results of research conducted or processed other, where research shows whether the subject of the polymorphism. It should be understood that a successful genetic research is not necessarily 100%accurate. Polymorphism can be within a gene that encodes a component of complement.

[00190] the Results of genetic studies have shown a link between certain alleles of the genes encoding various associated with complement proteins, and predisposition to the development of AMD and/or increased likelihood of developing complications from AMD. The allele that is associated with an increased likelihood of developing disease or pathological condition, and/or an increased likelihood of developing complications of the disease or pathological condition, or value the probability of adverse outcome of the disease or pathological condition, called in this application the "risk allele" of the disease or pathological condition; and a gene called "modifier risk of the disease or pathological condition. Some of the risk alleles are alleles of the genes that encode the complement factor H (CFH), where alleles have polymorphism, leading to the appearance isoforms CFH, which contains His instead of Tyr at position 402 (Tyr 402 His polymorphism). Not wanting to be limited by any theory, the inventors suggest that the Tyr402His variant of CFH may be less effective in controlling the activation of complement and/or may be modified tissue localization, adversely affecting the ability of the tissue to control the complement. In subsequent studies it was found that other isoforms CFH closely associated with the risk of developing AMD (Klein, R.J. et al. Complement Factor H Polymorphism in Age-Related Macular Degeneratipn. Science (2005); Edwards, A.O. et al. Complement Factor H Polymorphism and Age-Related Macular Degeneration. Science (2005); Haines, J. L. et al. Complement Factor H Variant Increases the Risk of Age-Related Macular Degeneration. Science (2005). In addition, it was shown that variants of genes encoding complement proteins C2, C3, factor B, C7 and MBL-2, are also associated with the risk of developing AMD (Gold, B. et al. Variation in factor B (BF) and complement component 2 (C2) genes is associated with age-related macular degeneration. Nat. Genet. 38, 458-462 (2006); Dinu, V. et al. Evidence for Association between Multiple Complement Pathway Genes and AMD. Genet. Epidem. 31, 224-237 (2007) Yates, J.R.W., Complement NW Variat and the Risk of Age-Related Macular Degeneration, N. Engl. J. Med., 357: 19-27, 2007). In addition, it was shown that some variants of genes encoding CPHR1, CFHR3 and CFI associated with the risk of developing AMD. Can be used in the methods, at least in part based on the identification of such options.

[001911 the Present invention includes the analysis of the genotype of the subject on the subject of any gene and/or polymorphisms described in the above references, or variants of any other gene (genes)encoding associated with complement proteins (see above); and identification of a suitable subject for the introduction of a liquid composition containing an analogue of constatine, at least in part on the basis of the results of these analyses (for example, the composition will be administered to the subject if the results of the analysis indicate that the subject is at increased risk for the development or manifestation of complement-mediated diseases such as AMD). It should be understood that the polymorphisms, such as SNPS can be in nonequilibrium coupling (LD) with other SNPs, for example, other SNP located on the same chromosome. Such SNP may be present in haplotypes. For example, some SNPS may be linked for up to 100 TPN or even 150 TPN, and more (Reich, D.E., et al., Nature, 411, 199-204, 2001). Thus, in some embodiments, the methods according to the invention include determining whether the haplotype individuo least one polymorphic variant, associated with an increased risk of developing a complement-mediated disease, where the specified polymorphism is located in a gene associated with complement. In some embodiments, the haplotype comprises encoding Tyr 402 His variant CFH gene. In some embodiments, the haplotype does not include encoding Tyr402His variant of CFH gene (see, e.g., Li, et al., above).

[00192] the Genotype of an individual can be determined using various methods. By applying a certain technique is not critical to the present invention, and there is no necessity to describe in this application; such methods are well known in this field. The methods typically use a biological sample obtained from the individual, where the sample contains nucleic acids and/or proteins. When used in this application, the term "biological sample" refers to any of the following: a cell or cells, tissue, liquid body, such as blood, urine, saliva, cerebrospinal fluid, etc. the Term "biological sample" also refers to any material obtained in the processing of biological material corresponding to the previous definition, for example, for isolating or purifying DNA, RNA and/or protein from a sample, amplification of the sample or part thereof, during the digestion of restriction enzymes, etc. usually used clicks the set of technical documents for blood or tissue sample. Methods may include analysis of the DNA of the individual on the content of the desired allele or polymorphism. Can also be used RNA, if interested polymorphism is located in the transcribed portion of a gene. In some embodiments, the methods include the identification of specific nucleotides, where polymorphism (SNPs) in the provision of such nucleotides associated with an increased risk of adverse outcome of the disease and/or susceptibility to AMD or other complement-mediated disease.

[00193] the Methods of conducting such tests are well known in the field and include, for example, the isolation of DNA or RNA, their possible amplification (for example, using polymerase chain reaction (NDP) or PCR (reverse transcription) and a number of methods such as allele-specific elongation of the primers, allele-specific hybridization, digestion with restriction enzymes, sequencing, etc. In some embodiments of genotyping is carried out using microchip or "chip". In some embodiments of genotyping is performed using analyses that use pellets, for example, using the Luminex platform. Other methods include ligation of oligonucleotide probes (U.S. patent No. 5,185,243, 5,679,524, 5,573,907), degradation analysis, analysis of labeled heteroduplexes (the MG), etc. Examples include the analysis of the Taqman® assay. Applied Biosystems (U.S. patent No. 5,723,591). Technology caliraya hybridization sample (TCGP), which is a system for determining nucleic acid-based amplification of the signal or probe, and no amplification of the target (U.S. patent No. 5,011,769, 5,403,711, 5,660,988, and 4,876,187), can also be used. Can also be used invasive methods of cleavage, for example, Invader.RTM. (Third generation technology)described in the Eis, P.S. et al., Nat. Biotechnol. 19:673, 2001. Can be studies based on molecular beacons (U.S. patent No. 6,277,607; 6,150,097; 6,037,130) or energy transfer fluorescent signal (PEPS). Publication of an application for U.S. patent No. 20050069908 and references in this application to give an idea of the variety of other methods that can be used to identify nucleic acids. U.S. patent No. 5,854,033, 6,143,495 and 6,239,150 describe compositions and methods for amplification and multiple detection of molecules of interest, including replication type "rolling ring". The method is used for the simultaneous detection of multiple nucleic acids in the sample. If desired, nucleic acid sequeiros. Publication of an application for U.S. patent No. 20050026180 describes methods for multiplex reactions of nucleic acids, including amplification, detection and Genot the bright feast, which can be adapted to determine the sequence in specific interest provisions to determine whether the individual's genotype associated with increased risk of adverse outcome of the disease.

[00194] To a General non-limiting examples of suitable methods include methods based on hybridization, the use of molecular beacons, SNP microarray, enzymatic methods, such as methods based on length polymorphism restriction fragments; methods based on PCR methods using flap-endonuclease, elongation of the primer, 5'-nuclease, analysis, ligation of oligonucleotides; other post-amplification methods based on physical properties of DNA polymorphism single-stranded conformation, methods gel electrophoresis using a temperature gradient, denaturing high-performance liquid chromatography and sequencing. Highly efficient sequencing is convenient from the point of view of speed, and sequencing is likely to become a routine method for use in genotyping purposes. Can be used in methods based on prosecution, in situ sequencing, sequencing of the granules (US 2007087362), etc. Cm. also PCT/US 2006/029449 (WO 2007014338) for detailed information and related op what edelenyi.

[00195] In some embodiments of the solution is injected directly into the eye, for example, by intraocular injection. Can be used standard methods of introduction into the eye, such as injection into the vitreous body, the injection in the anterior chamber of the eye, etc. In some embodiments of the invention, the composition is injected into subtenons space, injection retrobulbar space or subconjuctival space.

[00196] In some embodiments of the subject suffering from ocular revascularization (e.g., exudative AMD or proliferative diabetic retinopathy), previously to the introduction of the composition according to the invention the treatment with the inhibitor of angiogenesis to prevent bleeding and/or exudate. In some embodiments of the subject for treatment with an inhibitor of angiogenesis, and the composition according to the invention is inserted through 1-6 weeks after administration of the inhibitor of angiogenesis. [00197] In some embodiments of the invention, the solution is injected directly into the joint or periarticular space.

[00198] the delivery can be made by injection (e.g., using a needle 25, 27 or 30 G, and others), through a catheter, etc.

[00199] In some embodiments of the invention, the composition containing similar is onstation, delivered in one or more cavities or spaces of the Central nervous system containing the CSF, for example, in the ventricles or mostomozzhechkovogo tank. Delivery agent in the ventricle or mostomozzhechkovogo the tank can be performed using an infusion pump at such implantation of the catheter, when it opens in the ventricle or mostomozzhechkovogo tank releasing the composition of the end, which leads to the formation of a gel-like inclusions. The agent, which is an analog of constatine, diffuses out of the ventricles and mostomozzhechkovogo tank. Therefore, the introduction of these cavities can deliver the agent in a relatively wide area of the brain, not ogranichilasj specific area. In some embodiments of the invention, the delivery composition in the cavity containing the CSF, produced during implantation of the catheter surgically inside the skull so that one end of the catheter had access to the cavity. The other end of the catheter in this case is connected with a reservoir (e.g., vessel, Ommaya), which is located under the scalp (subcutaneously).

[00200] the Methods for intrathecal injection is well known in this field. If the subject suffers from a spinal injury, the catheter is implanted so that the releasing part of the end lies in the intrathecal space, while the other end is connected with the reservoir is the Varos pump. Such methods are mainly used for the treatment of chronic pain and are routine ways of prolonged delivery of analgesic agents for months. Such methods are used in the present invention for delivery of a liquid composition containing an analogue of constatine in a quantity which is effective for the formation of a macroscopic gel-like inclusion in the intrathecal space.

[00201] In one implementation of the present invention to a subject suffering from traumatic brain injury, stroke or spinal cord injury, treatment is carried out by introducing an analogue of constatine under conditions suitable for formation of a macroscopic gel-like inclusions containing analogue of constatine. In some embodiments of neuroprotective or neurotropic agent is also administered systemically and/or locally in various embodiments of the invention. In some embodiments of neuroprotective or neurotropic agent is introduced together with an inhibitor of complement in the same composition.

[00202] the dosing Interval (i.e. the time between the introduction to the subject the composition according to the invention) and the dose equivalent of constatine delivered at each introduction, may vary. In some embodiments of the delivery composition is carried out with an interval of more than 6 weeks, for example, 2, 3, 4, 5, 6 or 7 months, or in some intermediate number of weeks, for example, 8, 10, 12, 14, 16 weeks, etc. In some embodiments of the composition is delivered with even greater time intervals, for example intervals of 7, 8, 9, 10, 11, or 12 months. In other embodiments of the time interval is 6 weeks or less, for example, 1, 3, 4, 5, or 6 weeks. For example, the composition may be injected on average every 2 weeks, every 4 weeks, every 30 days, etc. of Course, the time interval may vary. For example, the interval between dose can vary from 6 weeks or less, and up to 6 weeks, and more. In some embodiments of the average time interval between the introduction of the composition according to the invention is at least 6 weeks, for example, 2, 3, 4, 5 or 6 months, or any intermediate number of weeks, for example, 8, 10, 12, 14, 16 weeks, etc. In some embodiments of the invention, the composition is injected several times with a mean time interval of at least 6, 8, 10 or 12 weeks, or 3, 4, 6, 8, 12, 15, 18 or 24 months difference, etc. Composition may be at least 1, 2, 5, 10, 15, 20, or more times. The composition may be an unlimited number of times with different time intervals to a subject at risk of or suffering from a complement-mediated disorder, e.g the measures of AMD. EXAMPLES

[00203] Example 1: Formation of gel-like inclusions after the introduction, active analog constatine in the vitreous body of the apes [00204] the Synthesis of an analogue of constatine

[00205] the Synthesis of the active analog constatine (Examples 1-3 also called "connection") was conducted in accordance with the solid-phase method described by Merrifield (J. Amer. Chem. Soc. 85, 2149 (1963)). the α-Amino group of each amino acid was protected with Fmoc-group. Functional groups of the side chains were protected by various suitable protective groups. The peptide chain is formed by embedding the C-terminal amino acids (i.e. Thr) amide in the polymer Market with further successive joining of amino acids, removing the protective groups of the side chains and the selection of the polymer. When the sequence of the peptide was completely formed, the N-terminal peptide polymer has azetilirovanie (using kairouseki solution consisting of Ac2O/CH2Cl2/DIEA in a volume ratio of 6:50:3), then the polymer was sequentially rinsed with solutions Meon and DMF. After the separation of the peptide from the polymer using standard methods, between residues of the peptide Cys2and Cys12formed disulfide bridge by I2the oxidation. First linear peptide Restoral a 20% aqueous solution of acetonitrile to obtain a solution of PEP the IDA with a concentration of 1 mg/ml Next, to a solution of the peptide when it is mixing one drop of solution was added to I2/Nal (Nal used to increase the solubility of I2in N2About). First adding I2its yellow color disappeared upon contact with the solution. Preservation yellow I2testified to that was added a sufficient amount of I2the solution in the flask was stirred for another 30 minutes to complete the oxidation. To neutralize the excess of I2and quenching the reaction solution was added ascorbic acid (0.1M). The reaction was monitored by high performance liquid chromatography (HPLC).

[00206] After completion of the oxidation of the prepared reaction mixture to purification by the method of preparative HPLC by filtering through a glass fiber filter with pores of 1 μm. The filtered solution of the peptide was loaded into a column for preparative HPLC, filled with C18polymer reverse phase, which was ruled by a system of preparative HPLC (Varian HPLC). The column was Polyanovo Buffer "And" [0.1% TFU in H2O - 1:1000 (V/V)] and a linear gradient of Buffer "B" [0.1% TFU in acetonitrile -1:1000 (V/V)].

[00207] the Fractions collected from the preparative column, were analyzed using an analytical HPLC system (Varian HPLC)equipped with an analytical column HPLC (Kromasil C18 5 μm). Fractions that satisfy orally requirements of purity, were then collected for the next stage of the process. Fractions that did not meet the requirements of cleanliness, were re-purified to achieve the necessary degree of purity, and then collected. The process was repeated to achieve at least 95% purity. The collected purified fractions were then re-loaded into the preparative HPLC column, washed ammonium acetate buffer and suirvey appropriate buffer system for the exchange of salt of the peptide on the necessary salt, in this case, the acetate salt. Fractions were again analyzed using analytical HPLC, and the fractions that corresponded to the final criterion of purity, were collected and prepared for lyophilization. Used lyophilizator-type collector, and each container lyophilizate was placed not more than 350 ml of purified peptide after freezing. Freeze-drying was carried out for more than 3 days. The final volume of the peptide was analyzed for purity by reverse-phase HPLC.

[00208] the Preparation of liquid formulations

[00209] Were prepared with two different formulations of the composition when dissolved in water for injection at different concentrations: (i) 0.299 mg/ml, denoted by LD, and (ii) 4.51 mg/ml, denoted by HD; and sterilization through the filter Millipore Durapore 47 mm, 0.22 μm. Liquid compositions were poured into separate vials of 250 μl each). The pH value of the composition of the LD was 5.48, while the pH value of the composition of the HD was 6.0.

[00210] the Introduction inside the vitreous body

[00211] In experiments involving the introduction of analog constatine, the synthesis of which is described above (SEQ ID NO: 32), in water for injection into the vitreous body of the apes, it was found that the connection after its injection into the vitreous body is able to form a gel-like almost spherical inclusions within the vitreous body of a Primate (cynomolgus macaques). Inclusions were formed in the injection area and only when the larger of the two studied concentrations of the compounds. High dose consisted of injection of intravitreal (IVT) connection in an approximate dose of 150 µg compounds in a volume of 50 μl of water for injection, whereas low dose contained an approximate dose of 3 μg of the compound in a volume of 50 μl of water for injection. Upon examination of the anterior eye using a slit lamp 2 and 15 days after administration of the compound was not detected anomalies. When reverse binocular ophthalmoscopy on day 2 revealed diffuse vitreous opacity in the right eye, one animal received a dose of IVT in a concentration of 3 μg per eye, and the four animals that received a dose of IVT in a concentration of 150 μg per eye. By day 15 the vitreous opacity was still observed in four of these animals, and others who Vuh animal was first seen vitreous opacity. There was no reason to believe that the connection caused any obvious adverse reactions within the vitreous chamber of the eyeball. Indicators ERG were within normal limits, and tonometry did not reveal changes in intraocular pressure caused by the introduction of composition.

[00212] At necropsy, most male animals (10 of 12)who received the first dose of IVT connection in 150 mcg on the eyes, during the preparation of the eye were found gellike spherical inclusions within the vitreous body. These inclusions were isolated and examined for the content of the connection. The results showed that the content of the compounds ranged from 7 to 72 µg µg for inclusion. One of the two animals euthanized 2 weeks after injection of a single dose of IVT, was not detected spherical inclusions in the vitreous body, and therefore, was not identified compounds. Similarly, one animal received a dose of IVT in 150 mcg on the eyes, sleeping 4 hours after injection, there was a spherical inclusions in the vitreous body. Thus, inclusions were first detected in the first control point time - four hours after administration of the compounds and were still detected in one of the two animals euthanized two weeks. When histopathological studies have found links the data with the introduction of composition anomalies in the eyes of animals, receiving doses connection with the subsequent formation or without the formation of inclusions in the vitreous body.

[00213] Inclusion were examined for the content of the composition, and the number of detected composition was measured using HPLC. It was found that the inclusions contain a large number of connections. In all animals, in the vitreous body which administered the compound in the amount of 150 μg per eye in any checkpoint time (from 4 hours to 2 weeks after injection) in the vitreous body was discovered the connection. These results gave reason to believe that inclusion in the formation of the released connection over time.

[00214] Example 2: Characterization of gel-like inclusions formed in the vitreous body of the rabbit with the introduction of active analog constatine

[00215] Research on new Zealand white rabbits (NZW) were conducted to provide more detailed definitions of the parameters of the formation of inclusions in the formation of inclusion after injection into the vitreous body, and also to evaluate the acute toxic properties of a compound after its injection into the vitreous body.

[00216] Methods

[00217] the Preparation of a solution of connections

[00218] the composition of the HD was prepared as described in Example 1 and diluted with water for injection (WFI) in sterile conditions for which Holocene different doses of the compounds. 50 μl of the composition was administered injectable into the vitreous body of the rabbit.

[00219] in Experimental animals

[00220] the Study was performed to assess the ability of the compound to form inclusion within the eye after its injection into the vitreous body in concentrations from 0 to 200 μg per eye(0, 25, 50, 75, 100, 125, 150,175, 200 μg) (table 2). Injection of the compound in each of the concentrations was performed in three eyes. To determine the effect of dose were used thirteen animals. Five additional animals were used for histological analysis.

[00221] the Methods for conducting experiments on animals

[00222] Anesthesia: Animals were anestesiologi using intramuscular injection of a solution of ketamine (50 mg/kg) + xylazine (10 mg/kg).

td align="center"> 1312 OE, DIS
Table 2
A study to determine the effect of dose on the formation of inclusions in the vitreous body of the eye of new Zealand white rabbits
Connection IVT (μg per eye)ProcedureAndID animal
Left eyeRight eye
2525OE, DIS
2550OE, DIS1334
5050OE, DIS1335
7575OE, DIS1344
75100OE, DIS1345
100100OE, DIS1346
125125OE, DIS1313
125150OE, DISA
150150OE, DIS1338
175175OE, DIS1339
175200OE, DIS1340
2002001341
00OE, DIS1342
00OE, HISTO1390
00OE, HISTO1391
2525OE, HISTO1337
5050OE, HISTO1343
100100OE, HISTO1336
AThese procedures were carried out in 27 hours after administration of the compound
OE = Ophthalmic research
DIS = Dissection
HISTO = Histological analysis

[00223] Primyenyeniye procedures: Directly in the eye, put 2 drops of local anesthetic (0.5% procainamide), and then 2 drops of 5% ophthalmic solution povidone-iodine. Special attention was paid to a solution of povidone-iodine was covered with the location of the planned injection. The excess fluid out OK logosnitrog area cleaned dry gauze cloth. Local anesthetic and a solution of povidone was left to act for at least 5 minutes before injection into the vitreous body.

[00224] injections: Injections into the vitreous body was performed using a syringe for injection of tuberculin 0.5 ml (with an integral needle 29G long1/2inches); 0.2 ml of a solution of compounds selected from the ampoule. The excess liquid was forced out so that the syringe contained only the required amount of liquid for a single injection (50 µl). Eyes were kept open using the expander eye of the century. The needle was introduced in verkhneisetsky quadrant at a distance of 3.5-4.0 mm from the limbus, avoiding the horizontal Meridian and directing the needle to the center of the eye. Only half (1/4inches) from the total length of the needle (1/2inch) was injected into the eye. The injection was performed slowly to avoid a sharp increase in intraocular pressure. In order to be sure that the entire volume of the solution is inside the eye, and avoid reverse flow through the injection, the needle was removed slowly.

[00225] Injection procedure: Immediately after injection, the eyes were washed with saline. The excess fluid in the peri-orbital area cleaned dry using disposable napkins 4×4. Then put two drops of antibacterial ophthalmic solution (Vigamo®, moxifloxacin, 0.5% ophthalmic solution).

[00226] the Ophthalmic examination: All animals until their slumber passed ophthalmologist-tested (using indirect ophthalmoscopy) the practitioner clinical ophthalmologist. Assessment criteria of the analysis are shown in Table 3.

[00227] Euthanasia: Euthanasia was performed on the shot rabbits by injection into the ear vein approximately 2-3 ml of Beuthanasia-D (Schering Plough) using needle 22 G.

Table 3
Assessment criteria indirect ophthalmologic analysis
ScoreAnalysis of the vitreous body
0Transparent, there are no inclusions, knots, haze
1Light haze, no cloudy inclusions or opacities of the vitreous body
2Moderate turbidity, or muddy the knot, or the inclusion of
3Significant vitreous opacity
4Absolute POM is teenie
ScoreAnalysis of the retina
0Smooth, no damage, protrusions or defects
1Nonsmooth; has folds, projections or roughness
2Weak inflammation or bleeding
3Retinal detachment, moderate bleeding
4The destruction of the retinal tissue (HA, neovascularization of choroidea, retinal rupture)

[00228] eye Enucleation for preparation of the eyeballs: One hand held wide open eyelids, conjunctiva was separarely on the edge of the cornea using a blade of a scalpel. External eye muscles cut clockwise, also cut and removed a nictitating membrane. Optic nerve was cut with scissors for enucleation with the introduction of their eye camera with nasal side. The eyeball was removed "monolith" when trimming structures of the retro-bulbar Department of the eyes. Each enableireland.ie the eyeball was purified from related eye membranes, eye, colon muscles and extraocular muscles with small curved scissors.

[00229] the EOS of the liquid vitreous of the rabbit, spherical inclusions from the vitreous body and the layers of the retina from the isolated eyeballs: Cleaned eyeball rabbit was placed in a sterile Petri dish. When capturing the eye with thin 1x2-tooth clip was made a neat incision through the sclera at a distance of 5 mm from the limbus by the scalpel blade No. 11. Then when trimming the sclera and the retina around the perimeter of the eye separates the upper part. The inclusion of carefully collected with tweezers, then transferred in Eppendorf tubes and kept on ice. If the inclusion was too fragile, it had been scraped with a scalpel No. 15. The vitreous body is grasped with toothed forceps, and then transferred into the Eppendorf tube and kept on ice. The retinal layer from each eyeball has been scraped with the back surface of the eye with a scalpel No. 15 and immediately transferred into the Eppendorf tube with a solution of RNA Later (Ambion, Austin, TX) and kept on ice. All collected materials (liquid vitreous, inclusion and retinal layer) were frozen and stored at -20°C.

[00230] the evaluation Criteria inclusions: If in the process it was discovered inclusion, it was recovered and carried out its evaluation. Evaluation criteria for inclusions shown in Table 4.

4
Table 4
Evaluation criteria include
ScoreSizeTransparencyDensity
0No activationNo activationNo activation
1diameter ≤ 1 mmTransparentCrumble at a touch; or in the preparation process of the eye
Apple reveals many fragile pieces
2the 1-2 mm in diameterHas blue/white hue; transparentEasily deformed tweezers; the weak force causes permanent deformation of the inclusions (inclusions)
3the diameter of 2-4 mmHas a pronounced blue/white shade; not quite transparentEasily deformed with tweezers, but elastic; enable (enable) may return to the initial form
Enable (enable) too
diameter ≥ 4 mmOpaquedense for its deformation tweezers

[00231] Histopathological analysis: After a survey conducted euthanasia of animals, the eyes were nuclearman, fixed in a solution of Davidson, were obtained from histological sections and painted Mr. E. Slices from both eyes of animals treated with 25, 50 or 100 μg of the compound eye (table 2, last three animals)were analyzed recognized by the American Board of pathology a pathologist and re-examined by a veterinary surgeon-pathologist with professional certification and is a recognized expert. [00232] the Study by HPLC:

[00233] the Preparation of the sample inclusion

Before analysis, the inclusion was freeze-dried

- Activate suspended in 50 μl of a solution of 80% acetic acid/20% water

- Took 8 ál of solution gel

- Added 20 μl of stock solution of internal standard (dilution in the mobile phase And 1:16)

- Added 52 ál of mobile phase And

Preparation of sample of the vitreous body

Sample preparation vitreous was conducted in accordance with SOP-D 0002v1 with modifications. In this case, until the sample to 50 μl of vitreous was added 20 μl of internal standard is a (significantly similar sequence with connection) (dilution in water 1:8).

[00234] the Analysis sample

[00235] the Samples as inclusions, and vitreous body were analyzed by HPLC using reversed-phase column with phase Cig Hypersil Gold AQ, 2.1 x 150 mm, 5 μm, Thermo Electron), with UV detection at a wavelength of 214 nm. Were built separate calibration curves for analysis of inclusion and vitreous body.

[00236] the Analysis of samples of inclusions in the vitreous body of the rabbit by electrophoresis in polyacrylamide gel in the presence of sodium dodecyl sulfate:

After the dissolution of the frozen-dried incorporation in vitro of Eppendorf in solution: FSB: Acetic acid (1:1), sample (samples) include vitreous of the rabbit was Liquefiable (samples inclusions of animals No. 1341, A and 1339).

- Added the paint to load samples (4) and beta mercaptoethanol.

The samples were boiled 5 minutes on thermopride.

- Cooled samples on ice for several minutes.

- Centrifuged samples at 10,000 rpm for 2 minutes

- In holes for polyacrylamide gel with sodium dodecyl sulfate were loaded separately in two volumes (5 μl and 20 μl) of each sample, leaving between one empty hole.

- Downloaded the marker SeeBlue to control.

The samples were driven at a constant voltage of 200V for 1 hour.

The gel was removed from the device and okriashvili in Kumasi blue for 30 minutes

- ViDi the s protein bands were cut from the gel, were placed in separate Eppendorf tubes and sent for mass spectrometry analysis.

[00237] the Results

[00238] In Table 5 presents an overview of the research results.

Table 5
Results studies
ID animal and doseIndirect analysisEvaluation of the gel (after preparation)
AnimalEyesDoseProcedureArticle bodyRetin-aSizeTransparencyRaft-
ness
The overall score
342Left0Dissection100.00.00.00.0
342Right 0Dissection100.00.00.00.0
312Left25Dissection100.00.00.00.0
312Right25Dissection100.00.00.00.0
334Left25Dissection100.00.00.00.0
334*Right50Dissection 201.01.00.02.0
1335*Left50Dissection100.50.50.01.0

Dissection
1335Right50Dissection200.00.00.00.0
1344Left75Dissection100.00.00.00.0
1344Right75Dissection2 00.50.50.51.5
1345Left75Dissection100.00.00.00.0
1345Right100Dissection200.50.00.00.5
1346Left100Dissection300.51.00.01.5
1346Right100Dissection200.00.0 0.00.0
1310Left125Dissection100.50.00.51.0
1313Left125Dissection102.01.01.04.0
1313Right125Dissection102.02.02.06.0
1310Right150Dissection202.02.02.06.0
1338 Left150Dissection103.03.03.09.0
1338Right150Dissection102.03.03.08.0
1339Left175Dissection221.01.01.03.0
1339Right175Dissection102.00.02.04.0
1340Left175401.01.01.03.0
1340Right200Dissection102.02.02.06.0
1341Left200Dissection103.03.02.08.0
1341Right200Dissection103.03.03.09.0
* The inclusion of too small to get its sample

[00239] the results of the indirect analysis and evaluation of inclusions are shown in the Table is e 5, along with the numbers of animals and doses. Inclusions were found in the eyes of all animals that received a dose equal to or greater than 125 mcg connections on the eyes. Inclusions were not detected in the eyes of animals afflicted received 25 μg of the compound eye. At other concentrations, inclusion was formed with a different frequency. In this experiment inclusion could not be precisely identified by the indirect analysis. It should be noted that the ophthalmologist could see a slight darkening even used at lower concentrations than the concentrations at which is formed inclusion.

[00240] Indirect ophthalmologic examination did not reveal any pathological changes associated with injection of compounds into the vitreous body.

Characteristics of the ocular fundus were normal, except for one animal, No. 1339, which was observed retinal hemorrhage, in the opinion of the ophthalmologist caused by damage to the needle during injection. Vitreous opacity was detected in 9 out of 32 analyzed eye. The presence of turbidity did not correlate with the dose of a compound.

[00241] Histopathological evaluation

[00242] The five animals in this experiment was also performed histopathological study. Animals and treatment they conducted are presented in Table 6. The eyes of the animal which were examined by two pathologists. Both the pathologist concluded that all the tested eye is normal. The cornea and iris, and ciliary body, and lens, and sclera, and retina, and vitreous body were normal.

[00243] the Analysis of inclusions: Inclusions were analyzed by HPLC to confirm the presence of compounds and to assess its quantity. Also conducted the analysis using polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate on the subject of the possible presence of other proteins that could not be identified using the HPLC Protocol (identifies mainly peptides with low Mr). When UV detection was not identified other components, in addition to connections. The data confirmed the hypothesis that the content of the connection inside of the turn correlates with the amount of the compounds injected into the vitreous body. Was measured concentration in the vitreous body of the rabbit after injection into the vitreous body of each dose of the compound. After 27 hours after injection was not observed correlative between the present compound and injected the connection.

[00244] In the Figure 3 presents the result of analysis using polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate inclusions in the eyes of rabbits: 1341, right eye (200 µg connection), 1310, right eye (150 µg connection) and 139, right eye (175 µg connection). Protein bands from lane 2 were cut and sent for identification using mass spectrometry. All visible protein bands from samples include vitreous of the rabbit were successfully identified using the method MS/MALDI. The results are shown in the table below.

[00245] In conclusion, the formation of inclusion appeared to be dose-dependent. Turn on never formed at the dose of injection of 25 μg per eye, and reliability was formed in the eyes of the animals receiving a dose of 125 μg per eye or more (all in a volume of 50 ál). Analysis of spherical inclusions in the vitreous bodies showed that the active compound is the main component of inclusion, and that the protein content of the inclusions is limited to proteins that are present in the vitreous body of the eye of rabbits. 24 hours after injection was carried out histopathological study of the eyes of rabbits treated with a single injection into the vitreous body dose of compound 25, 50 and 100 μg per eye. Both the pathologist came to the conclusion that all the studied sections of the eyes were normal.

[00246] In a subsequent study, new Zealand white rabbits were kept for more than 8 months after the introduction of the compounds into the vitreous body. The compound extracted from the gels, izvlechennykh from the vitreous of a rabbit in the moments of time in the range from 6 weeks to 8 months after the introduction, remained stable and maintained a significant complement inhibitory activity in standard assays (Figure 4).

[00247] In another paper rabbits was conducted repeated treatment 3 months after primary treatment (after witnessing the disappearance of the gel-like inclusions formed during the first treatment). Re-treatment led to the formation of a new start, which confirmed the feasibility of re-treatment using such composition sustained release.

[00248] Example 3: a Further study of inclusions from analog constatine in monkeys

[00249] Were carried out further research to further study the behavior of inclusions in the eyes of monkeys over time. It was found that inclusion can be detected using the Ophthalmoscope research, and to monitor them over time by means of ultrasound, as in rabbits.

[00250] In one study, Javanese makaka were administered 0, 150, 450, 1050 or 2100 µg connection in 50 μl of water for injection by injection into the vitreous body. It was found that 2 weeks after injection inclusions were detected in the eyes of all animals that were administered a dose of 150 μg, and in the eyes of the animals that were administered higher doses.

[00251] Some of the animals that were administered 0, 450, 1050 or 2100 µg connect the tion, were euthanized 14 days after injection and measured the concentration in serum and vitreous body. The measurement was carried out using HPLC. As shown in Figure 5, the compound is slowly released from the inclusions of the vitreous body of the apes at least two weeks. If the introduction is too low for the formation of inclusion concentrations connection disappears from the vitreous body cynomolgus macaques with T½ approximately 22 hours. Consequently, it is obvious that measured in this experiment the concentration of compound 2 weeks after its introduction should reflect the amount released from the incorporation of the connection instead of the residual connection, never was part of the inclusion.

[00252] Further research was aimed at studying the behavior of inclusion over time. Inclusions formed during dose injection of 150 µg remained detectable for more than 2 months, but decreased in size. Some include completely disappeared within 3 months, while some remained for 6 months. Inclusions formed at the dose of injection 450, 1050 or 2100 µg remained detectable for more than 6 months. It was obvious that they were reduced in size and/or density to the reference point of time in 6 months. Inclusions formed at the dose entered what I 450, 1050 and 2100 µg, in most cases remained visible in the reference point of time at 37 weeks and before the checkpoint time in 1 year, in which the study was over. Inclusion was observed both when Ophthalmoscope and an ultrasound. As it turned out, over time they continued to decrease in size and/or broken up into a number of smaller inclusions. In addition, during this time there was a prolonged release of analog constatine, which was determined based on the measurement of the concentration of analog constatine in human blood serum samples obtained from these animals.

[00253] In an independent study after the introduction of the compounds in water by injection into the vitreous body in such a low concentration as 1 mg/ml (50 μg in 50 μl), gel-like inclusions were observed and Ophthalmoscope, and ultrasound. Enable ceased to be detected using these methods, approximately 1 week after dosing.

[00254] it Was observed that the results on the formation and disappearance rates inclusions were more reproducible in monkeys than in rabbits. In General, it was found that the inclusions formed in rabbits, denser and longer retained than inclusions formed at the same doses in monkeys. Neither result is stomatologicheskogo research neither during physical studies on monkeys and rabbits did not reveal adverse effects of inclusions.

[00255] Example 4: Modulation properties including

[00256] Were tested with various excipients, buffers and pH to assess their ability to modify properties that affect the formation of the gel analog constatine used in Examples 1-3, and to assess their influence on the properties of gels in vitro and in vivo. Were investigated amino acids, including arginine, series and histidine. It was found that formulations containing histidine, possess the best properties from the viewpoint of stability of the resulting gel compared with formulations containing arginine or series.

[00257] In some experiments investigated the effects of different concentrations of sodium acetate (NaCH3COO), histidine and mannitol in vitro and/or in vivo, either individually or in various combinations. The concentration of histidine and mannitol ranged from 10 to 50 mm. In General, it was observed that the use of compositions containing any of these substances led to the formation of gels, which were more fragile and quickly reduced in size in vivo than gels formed using compositions containing only analog constatine and water. It was concluded that the addition of these excipients to pozvoljaetrealizovat speed reduce the size of the inclusions and to modulate the rate of release of analog constatine of inclusion. Such modulation could potentially allow you to enter a higher total dose equivalent of constatine.

[00258] Typical compounds included the following:

[00259] A. 100 mm sodium acetate solution (pH=5.10) in water for injection.

[00260] B. 100 mm sodium acetate + 25 mm histidine (rn=5.2)

[00261] C. 100 mm sodium acetate + 45 mm mannitol (pH=5.05)

[00262] the pH Range ~ (5-5 .50)

[00263] Substance:

1. Sodium acetate:

Stock solution: 3 M

Manufacturer: Ambion

pH=5.5

The final concentration in the composition: 100 mm

The final pH value of the composition: 5.1

2. Histidine:

DL-Histidine hydrochloride, 98% min Molecular weight: 209.64 Manufacturer: Acros Organics

3. D-Mannitol

The powder according to the USP monograph

manufacturer: Fisher Scientific

[00264] the Protocol of preparation of the composition:

To prepare 10 ml of a solution:

1. Dilute sodium acetate in distilled water from 3 M to 100 mm.

2. To weigh a certain number of histidine/mannitol to achieve the required concentration.

3. To determine the final value of the pH and osmolarity of the solution. Prepare a composition similar constatine:

1. Weigh the empty tubes Eppendorf

2. Add the required number of analog constatine in these tubes

3. Add the solution (water/sodium acetate/sodium acetate + mannitol/sodium acetate+histidine) to achieve neobhodimo the concentration of analog constatine

4. Filter the solution through a sterile filter (0.22 μm)

5. Measure the optical density of the final solution, to check the final concentration equivalent to constatine

6. Measure the pH of the final solution.

[00265] In some experiments similar constatine was added to 100 mm sodium acetate solution (pH=5.10) in water for injection. Composition (1050 µg analog constatine in 50 μl of fluid) was administered by injection into the vitreous body 3 monkeys. All three animals from 2 to 6 week were included. On the 9th week of explicit inclusions were not found. One of 3 animals was observed in the remainder of the turn. In contrast, in animals that received the same number of formulas analogous to constatine comprising dissolved in water for injection analog constatine were observed inclusion on the 9th week. The presence of sodium acetate, thus, increased the rate of dissolution of inclusions.

******

[00266] the Professionals in this field is obvious, or they may verify using nothing more than standard experiments that there are many equivalents of the specific variants of the invention described in this application. The amount of the claims of the present invention is not limited to the above description, but rather is defined by the attached claims. It should be understood that the Fig is giving is not limited to specific results, obtained in any particular example or any particular implementation of the invention. In the claims the nouns used in the singular can mean one subject or more, unless specified otherwise in contradiction to the context or otherwise evident from the context. The claims or the description, which include the conjunction "or" between one or more members of the group, satisfy options where one, more than one, or all group members are present, used, or otherwise relate to a given product or method, unless otherwise specified in contradiction to the context or does not follow from it. The invention includes embodiments, in which one particular member of the group is used or otherwise related to that product or process. For example, and without limitation, it should be understood that where the claims or descriptions indicate that the residue at a certain position can be selected from a particular group of amino acids or amino acid analogues, the invention includes specific embodiments, in which the residue at this position is any of the following amino acids or amino acid analogues. The invention also includes embodiments, in which more than one or all group members are present, used or otherwise relate to Yes the resultant product or method. In addition, you must understand that the invention encompasses all variations, combinations and transformations, in which one or more limitations, elements, conditions, descriptive terms, etc., from one or more of the listed items are included in another item. In particular, any claim that is dependent on another claim, may be amended by the inclusion of one or more elements or limitations contained in any other clause, dependent on the same basic point. In addition, you must understand that if the item relates to the composition, it includes the introduction of the composition according to any method described in this application and obtaining the composition according to any one of the ways to get outlined in this application, unless otherwise specified, or if the specialist is not obvious contradiction or inconsistency.

[00267] you Must understand that where elements are represented as lists, for example, in the form of Markush structures, each sub-group elements are also included, and any element (s) may be excluded from the group. For brevity, only some of these implementation options have been specifically described individually and specifically in this application, but the invention includes all such variations. Also it should be understood that, in General, where the invention or aspects of the invention relate to including OS is especially items signs, etc., some implementations of the invention or aspects of the invention comprise or essentially consist of such elements, attributes, etc.

[00268] the Inclusion of phase "security" in some ways the implementation of the invention is directed to an indication that the composition is introduced for the treatment of disorders described in the method. Thus, the subject will have or be at risk of violations, and the composition is introduced for the treatment of disorders, usually on the advice of a doctor or surgeon practice, which may or may not be the same person who enters the composition. The invention includes embodiments, in which the step of providing not openly included and embodiments, in which phase of the software included. The invention also includes embodiments, in which the step of identifying the subject as being at risk or suffering from a complement-mediated disorders is included.

[00269] where constraints are specified, the invention includes embodiments, in which the endpoint is enabled, the embodiments in which both endpoints are excluded, and the embodiments, in which one extreme point is included and the other excluded. It should be assumed that unless otherwise specified, both endpoints are included. In addition, you must understand that unless otherwise specified or the e follows from the context and understanding of the average person skilled in the art, estimates are expressed as ranges, can be any specific value or subrange within established ranges in different embodiments of the invention, to one decimal place number of the bottom of the range, unless the context indicates otherwise. The time period of 1 month is understood as 30 days. A time period of 1 year as understood as 365 days. For any embodiments of the invention, in which a numeric evaluation is preceded by "about" or "approximately", the invention includes an embodiment in which specific assessment is described. For any embodiments of the invention, in which a numeric evaluation is not preceded by "about" or "approximately", the invention includes an embodiment in which the assessment is preceded by "about" or "approximately".

[00270] you Must understand that any some variant of implementation, the characteristic or aspect of the present invention can be explicitly excluded from any one or more claims. For example, any particular composition, compound or class of compounds, place, way, or route of administration, dose, formulation, or a complement-mediated disorder may be excluded from any one or more claims.

1. A method of treating a complement-mediated disease comprising the step of introducing liquid composition, sod is rasego analog constatine, in the extravascular space of the subject, wherein the number of the specified analog constatine is sufficient for the formation of discrete macroscopic gel-like structure in the extravascular space with the introduction in the absence of other gel-forming substances.

2. The method according to claim 1, characterized in that the said effective amount is sufficient for formation of a macroscopic gel-like structure that diminishes in size and remains easily detectable for at least 2 weeks, preferably for at least 3 months.

3. The method according to claim 1, characterized in that the said effective amount is sufficient for formation of a macroscopic gel-like structure that diminishes in size and delivers the analog constatine for at least 2 weeks, preferably for at least 3 months.

4. The method according to claim 1, characterized in that the said effective amount is sufficient for formation of a macroscopic gel-like structure that diminishes in size and delivers the analog constatine in the active form with the achievement of therapeutic concentrations of the specified analog constatine in the extravascular space or priloga the tissue at least 2 weeks, preferably at least 3 months.

5. The method according to claim 1, characterized in that the analogue of constatine includes a peptide sequence which contains a sequence selected from the following group: SEQ ID NO: 3, 4, 5, 6 and 7.

6. The method according to claim 1, characterized in that the activity of the analog constatine exceeds activity sequence SEQ ID NO: 8 at least 100 times, preferably at least 200 times.

7. The method according to claim 1, characterized in that the analogue of constatine has a sequence selected from SEQ ID NO: 14, 21, 28, 29, 30, 31, 32, 33, 34 and 36, preferably a sequence selected from SEQ ID NO: 28, 32, and 34.

8. The method according to claim 1, characterized in that the number of the specified analog constatine in the liquid composition is in the range from 1 mg/ml to 50 mg/ml, preferably in the range from 2 mg/ml to 25 mg/ml

9. The method according to claim 1, characterized in that the vitreous chamber of the eyeball injected from 50 μg to 5000 μg analog constatine, preferably from 150 μg to 2000 μg analog constatine, more preferably from 400 mcg to 1500 mcg analog constatine.

10. The method according to claim 1, characterized in that the analogue of constatine injected into the vitreous chamber of the eyeball in a volume of 25 μl 125 μl, preferably in a volume of about 50 μl.

11. The method according to claim 1, characterized in that the entity with whom radet age-related macular degeneration, and this liquid composition is administered into the vitreous chamber of the eyeball.

12. The method according to claim 1, characterized in that the liquid composition further comprises an effective amount of a second therapeutic agent.

13. The method according to item 12, wherein the second therapeutic agent is an inhibitor of complement, angiogenesis inhibitor, a steroid, an anti-inflammatory agent, anti-infective agent or analgesic.

14. The method according to claim 1, characterized in that the said composition is administered by injection into the vitreous body of the eyeball.

15. The method according to claim 1, characterized in that the composition includes a lot of microparticles or nanoparticles.

16. Gellike structure for slow release of the analog constatine, characterized in that the gel-like structure contains similar constatine sufficient for the formation of gel-like structures in the absence of other gel-forming substances, and at least one endogenous polypeptide, normally present in the extravascular space of the subject.

17. Gellike structure according to item 16, wherein the extravascular space selected from the following group: the vitreous chamber of the eyeball, the subconjunctival space, subtenons space, subretinal space, sinovi is supplemented flax cavity and the spinal cavity.

18. A liquid composition for the formation of inclusion analog constatine containing the specified analog constatine and liquid pharmaceutically acceptable carrier, wherein the composition is characterized by the ability to form a macroscopic gel-like structure at its introduction into the vitreous chamber of the eyeball of the mammal in the absence of other gel-forming substances.

19. A liquid composition for p, characterized in that the analogue of constatine is present in a quantity sufficient for the formation of discrete macroscopic gel-like structure at its introduction into the vitreous chamber of the eyeball of the subject by injection into the vitreous body.

20. A liquid composition for p, characterized in that the analogue of constatine is present in a quantity sufficient for the formation of a macroscopic gel-like structure that diminishes in size and remains easily detectable for at least 2 weeks, preferably for at least 3 months, more preferably for at least 6 months.

21. A liquid composition for p, characterized in that the analogue of constatine is present in a quantity sufficient for the formation of a macroscopic gel-like structure that diminishes in size which frees up the equivalent of constatine in the active form for at least 2 weeks, preferably for at least 3 months, more preferably for at least 6 months.

22. A liquid composition for p, characterized in that the analogue of constatine is present in a quantity sufficient for the formation of a macroscopic gel-like structure that diminishes in size and delivers the analog constatine in the active form with the achievement of therapeutic concentrations of the specified analog constatine in the vitreous chamber of the eyeball or the adjacent tissue for at least 2 weeks, preferably within 3 months.

23. A liquid composition for p, characterized in that the analogue of constatine includes a peptide sequence which comprises a sequence selected from the following group: SEQ ID NO: 3, 4, 5, 6 and 7.

24. A liquid composition for p, characterized in that the activity of the analog constatine exceeds activity sequence SEQ ID NO: 8 at least 100 times, preferably 200 times.

25. A liquid composition for p, characterized in that the sequence of analog constatine includes a sequence selected from SEQ ID NO: 14, 21, 28, 29, 30, 31, 32, 33, 34 and 36, preferably a sequence selected from SEQ ID NO: 28, 32 and 34.

26. A liquid composition for p, characterized in that the number of the specified analog constatine in the liquid composition is e is in the range from 1 mg/ml to 50 mg/ml, preferably from 3 mg/ml to 25 mg/ml

27. A liquid composition for p, characterized in that the composition contains from 150 μg to 5000 μg analog constatine, preferably from 250 μg to 2000 μg analog constatine, more preferably t 400 mcg to 1500 mcg analog constatine.

28. A liquid composition for p, characterized in that the composition has a volume of 25 ál 125 ál.

29. A liquid composition for p, characterized in that the composition contains from 150 μg to 2000 μg analog constatine in a volume of 50 μl to 100 μl.

30. A liquid composition for p, characterized in that the composition consists essentially of analog constatine in the water.

31. A liquid composition for p, characterized in that the composition essentially contains no excipients.

32. A liquid composition for p, characterized in that the composition contains a component selected from the group consisting of polyhydric alcohols and amino acids.

33. A liquid composition for p, characterized in that the presence of the component modulates the rate at which the inclusion disappears in vivo.

34. A liquid composition for p, characterized in that the composition contains histidine.

35. A liquid composition for p, characterized in that the composition contains a buffer.

36. A liquid composition for p, characterized in that the composition contains sodium acetate.

37. Liquid is a composition p, characterized in that the composition contains mannitol.

38. A liquid composition for p, characterized in that the composition additionally contains an effective amount of a second therapeutic agent.

39. The liquid composition according to § 38, characterized in that the second therapeutic agent is an inhibitor of complement, angiogenesis inhibitor, a steroid, an anti-inflammatory agent, anti-infective agent or analgesic.

40. The liquid composition according to § 38, characterized in that the second therapeutic agent is an antibody or antibody fragment.

41. The method of preparation of a composition for delivery of a therapeutic agent over a prolonged period of time, comprising: preparing a liquid composition containing therapeutic agent and the analog constatine, characterized in that the analogue of constatine is present in a quantity sufficient for the formation of a macroscopic gel-like structure in the absence of other gel-forming substances, with the introduction of the composition into the extravascular space in the body of a mammal subject.

42. The method according to paragraph 41, further comprising introducing the liquid composition in the extravascular space in the body of a mammal.

43. The method according to paragraph 41, wherein the specified extravascular space is the FDS is th vitreous chamber of the eyeball.

44. The method according to paragraph 41, wherein the specified extravascular space is a vitreous chamber of the eyeball, and the specified subject suffering from age-related macular degeneration (AMD).

45. The method of treatment of a subject suffering from AMD or at risk, including the introduction of a liquid composition containing an analogue of constatine directly into the vitreous chamber of the eyeball of the subject, characterized in that the liquid composition contains the number of analog constatine, enough to form a macroscopic gel-like structure in the absence of other gel-forming substances after its introduction.

46. The method according to item 45, wherein the specified analog constatine contains a peptide sequence which comprises a sequence selected from the group of sequences SEQ ID NO: 3, 4, 5, 6 and 7.

47. The method according to item 45, wherein the specified analog constatine has at least 100 times greater activity than the activity of SEQ ID NO: 8, preferably specified analog constatine has at least 200 times more activity than the activity of SEQ ID NO: 8.

48. The method according to item 45, wherein the specified analog constatine has a sequence selected from SEQ ID NO: 14, 21, 28, 29, 30, 31, 32, 33, 34 and 36, or preferably the sequence, select the NUU of SEQ ID NO: 28, 32 and 34.

49. The method according to item 45, wherein the specified number of analog constatine is in the range from 2 mg/ml to 20 mg/ml

50. The method according to item 45, wherein from 100 μg to 2,000 mcg analog constatine, preferably from 250 micrograms up to 1,500 µg analog constatine, more preferably from 400 mg to 1,200 mg of analog constatine injected into the eye.

51. The method according to item 45, characterized in that the liquid composition further comprises an inhibitor of angiogenesis.

52. A liquid composition for the formation of a gel-like structure in the extravascular space of the subject, containing analogue of constatine as the gel-forming substance and water, characterized in that the concentration of analog constatine is in the range from 3 to 50 mg/ml

53. The liquid composition according to paragraph 52, wherein the concentration of analog constatine is in the range from 5 to 30 mg/ml, preferably in the range from 8 to 25 mg/ml

54. The liquid composition according to paragraph 52, wherein the specified analog constatine contains a peptide selected from the group of sequences SEQ ID NO: 3,4, 5, 6 and 7.

55. The liquid composition according to paragraph 52, wherein the specified analog constatine has an activity of at least 100 times greater activity of SEQ ID NO: 8, preferably specified analog constatine has an activity of at least 200 times greater activity of SEQ ID NO: 8.

56. The liquid composition according to paragraph 52, wherein the specified analog constatine contains a peptide selected from the following group: SEQ ID NO: 14, 21, 28, 29, 30, 31, 32, 33, 34 and 36, or preferably from the following group: SEQ ID NO: 28, 32 and 34.

57. The liquid composition according to paragraph 52, wherein the composition consists essentially of analog constatine and water.

58. The liquid composition according to paragraph 52, wherein the composition further comprises excipients selected from amino acids and sugar alcohols.



 

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2 cl

FIELD: medicine, pharmaceutics.

SUBSTANCE: what is presented is a pharmaceutical composition in the form of a spray for treating a damage by non-lethal irritants (e.g. pelargonic acid morpholide), containing pediphene in the following proportions, wt/volume %: pediphene hydrochloride 0.01-10.0; sodium chloride 0.1-10.0; water for injections up to 100 ml. What is shown is the efficacy of pediphene in compliance with the declared application ensured by the local anaesthetic effect of the drugs.

EFFECT: drug preparation has no allergenic and immunotoxic properties.

3 dwg, 40 tbl

FIELD: medicine.

SUBSTANCE: invention relates to medicine, namely to ophthalmology and can be applied for treatment of traumatic and dystrophic injuries of eye cornea. For this purpose peptide fragments of protein S100b - SP2 and/or SP3 in concentrations 10-6 M are introduced in conjunctive cavity or in endonasal way. Introduction is realised daily two times per day during 10 days.

EFFECT: method ensures fast and high-quality restoration of injured zone of cornea due to stimulation of regenerative-reparative processes in cornea.

3 ex, 2 cl

FIELD: medicine.

SUBSTANCE: invention refers to ophthalmology and may be used for treating degenerative and dystrophic retinal diseases. That is ensured by subcutaneous introduction of the preparation Coenzyme compositum 0.5 ml within the mastoid process, the preparation Lymphomyosot - within the temporal fossa, as well as by parabulbar introduction of the preparation Placenta compositum. The course is 10 sessions every second day 1-2 days a year. The introductions are double-sided if the retina in both eyes is involved.

EFFECT: invention provides improving the peripheral circulation in the eyes, increasing visual acuity due to the integrated effect of the drugs administered.

2 cl, 5 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: group of inventions refers to pharmaceutical formulations and methods for preparing them of lipids for ophthalmological application containing a phospholipid ingredient containing natural zwitterion phospholipids, and an oil ingredient containing natural oil-in-water emulsions. The oil ingredient and phospholipid ingredient are preferentially related as 3:1; the phospholipid ingredient is found in the amount of 0.1%-5%, and the oil ingredient is found in the amount of 0.3%-15%.

EFFECT: group of inventions provides ocular drug delivery applicable for treating dry eye syndrome; it has an ability to recover the lipid lachrymal film layer, inhibits the inflammatory element observed.

37 cl, 19 tbl, 4 ex, 7 dwg

FIELD: medicine.

SUBSTANCE: invention relates to medicine, in particular to ophthalmology, and is intended for treatment of eye burns. For this purpose instillations of eye drops of 2% of mexidol solution in conjunctival cavity is carried out 2-4 times per day immediately after burn and during the following two weeks.

EFFECT: chosen concentration of mexidol and mode of introduction in claimed method ensure effective treatment of burns at early terms after trauma, including by normalisation of activity of enzymes and proteins, which take part in processes of reparation and regeneration, fast growth of vessels and recovery of microcirculation, as well as prevention of formation of deep ulcers of cornea.

9 dwg, 6 ex

FIELD: medicine.

SUBSTANCE: invention relates to medicine, in particular to ophthalmology, and can be applied for cell therapy in case of different ophthalmopathologies, accompanied by dystrophic and atrophic processes as well. Three-component complex for cell therapy contains mesenchymal stem cells, labeled by magnetic microparticles. Cells are translocated into biological or synthetic fine-pore material, which in its turn is strongly fastened with polymer magnetic material with induction of constant magnetic field 1.5 mT, with multipolar reverse magnetisation.

EFFECT: invention ensures directed supply of stem cells to pathological nidus and holding stem cells for specified time with creation of possibility of giving complex any form, size and space configuration, suitable for extrascleral implantation to any area of eyeball or visual.

2 cl, 1 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to pharmaceutical compositions containing (2-hydroxyethoxy)amide 6-(4-brom-2-chlorophenylamino)-7-fluor-3-methyl-3H-benzoimidazole-5-carboxylic acid hydrosulphate and solvates, crystalline forms and amorphous forms thereof, to using the above compositions as a drug; and to methods for preparing the above compositions.

EFFECT: preparing the new pharmaceutical compositions.

20 cl, 7 tbl, 7 ex, 5 dwg

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to a pharmaceutical composition for outpatient treatment and prevention of the cardiovascular diseases, containing therapeutic amounts of a vasodilator, a renin-angiotensin system inhibitor, a thrombocyte aggregation inhibitor, a cholesterol-lowering agent, and an antihypoxic agent. As a vasodilator, the declared composition contains an agent possessing α-adrenergic receptor antagonist action, and a thrombocyte aggregation inhibitor is presented by an ADP-dependent thrombocyte activation mechanism blocking agent.

EFFECT: invention provides the integrated therapeutic effect on the cardiovascular system after acute administration that improves the compliance with treatment regimen by the patient.

20 cl, 1 tbl, 15 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: microparticle contains an agglomerate of particles containing a hydrophilic active substance, wherein the particle contains an amphiphilic polymer composed of a hydrophobic segment of polyhydroxy acid and a hydrophilic segment of polysaccharide or polyethylene glycol, and a hydrophilic active substance. What is also disclosed is a method of producing the agglomerated microparticles, which involves (a) a stage of preparing a reverse phase emulsion, (b) a stage of preparing a solid residue containing the hydrophilic active substance, and (c) a stage of introducing the solid residue into a liquid phase containing a surface modifier.

EFFECT: agglomerated microparticles provide the effective encapsulation of the hydrophilic active substance and the release of the hydrophilic active substance at an appropriate speed.

14 cl, 22 dwg, 4 tbl, 31 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to medicine and represents gel-forming mixed dextran esters containing phosphate and carbamate groups of general formula: {C6H7O2(OH)3-x-y{[(OP(O)ONa)mONa)]xl[(O2P(O)ONa)k]x2}x(OCONH2)y}n, wherein x=x1+x2 is a degree of substitution in phosphate groups (mono- and diesters), x=0.47-1.09; X1 is a degree of substitution in monoesters, X1=0.01-0.48; m is a number of phosphates in monoesters, m=1-2; x2 is a degree of substitution in diesters, x2=0.01-1.09; k is a number of phosphates in diesters, k=1-2; y is a degree of substitution in carbamate groups, y=0.39-1.23; n is a degree of polymerisation, 20≥n≤1000.

EFFECT: invention provides producing low-toxic low- and high-substituted dextran phosphates in the form of hydrogels containing additionally carbamate groups and possessing antiproliferative activity with respect to cancer cells.

2 cl, 3 dwg, 14 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention relates to field of pharmacy and represents microsphere with controlled release, which has covering layer and contains core, which contains exendin as active ingredient and biodegradable polymer, and covering layer, which covers core with covering material, exendin being exendin-4 (SEQ ID NO:2), biodegradable polymer represents polymer, selected from group, consisting of polylactide (PLA), polyglycolide (PGA), lactide and glycolide copolymer (PLGA), polyorthoester, polyanhydride, polyhydroxybutyric acid, polycaprolactone and polyalkylcarbonate; copolymer or simple mixture of two or more polymers, selected from said group of polymers; copolymer of said polymer and polyethylene glycol (PEG); or polymer-sugar complex, in which sugar is bound with said polymer or said copolymer, covering material is selected from group, consisting of essential amino acids, polypeptides and organic nitrogenous compounds, essential amino acid being one or more, selected from group, consisting of arginine, lysine and histidine; polypeptide represents L-Lys-L-Thr-L-Thr-L-Lys-L-Ser; and organic nitrogenous compound is selected from group, consisting of creatine, creatinine and urea, content of covering layer constitutes from 0.01 to 5 wt fractions in terms per 100 wt fractions of microsphere.

EFFECT: invention ensures increase of bioaccessability and reduction of initial peak of exendin for prevention of such side effects as vomiting, nausea, headache.

10 cl, 7 ex, 5 tbl, 7 dwg

FIELD: medicine, pharmaceutics.

SUBSTANCE: dosage form contains oxycodone hydrochloride as a physiologically active substance (A), optionally one or more physiologically combined excipients (B), a synthetic or natural polymer (C) and optionally natural, semisynthetic or synthetic wax (D).

EFFECT: dosage form of oxycodone hydrochloride possess degradation resistance of at least 400 N to less than 500 N, and releases max 99% of oxycodone hydrochloride in the physiological conditions after 5 h.

14 cl, 7 dwg, 17 ex

FIELD: medicine.

SUBSTANCE: declared invention refers to medicine. What is declared is a delayed release complex pharmaceutical composition containing a delayed release part and an immediate release part. The delayed release part contains AII-receptor blocker as an 'active ingredient'. The immediate release part contains HMG-CoA reductase inhibitor as an active ingredient.

EFFECT: declared composition is effective for treating hypertension and preventing complications in the patients suffering metabolic syndromes, such as diabetes, obesity, hyperlipidemia, coronary vessel disease, etc.

10 dwg, 13 bl, 8 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to pharmaceutical industry and represents a pharmaceutical preparation for controlled release of a contained active ingredient, differing by the fact that said pharmaceutical preparation contains medically acceptable yeast able for alcoholic fermentation.

EFFECT: invention provides release of the active ingredient from the pharmaceutical preparation regardless of the environmental conditions.

13 cl, 4 ex, 3 dwg

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to medicine and may be used for treating patients with hypertension and lipid storage disease. There is applied a combined drug containing dihydropyridine, calcium canal blockers, and statin, a hypolipidemic agent. The drug is prepared in such a manner that release rate of said ingredients can be controlled with respect to each other.

EFFECT: method allows higher clinical effectiveness and compliance, prevented antagonist and side effects of the combined therapy.

27 cl, 12 tbl, 10 dwg, 7 ex

FIELD: medicine.

SUBSTANCE: there are offered an oral pharmaceutical composition containing a) a testosterone ester and a fatty acid ester of a medium chain size; and b) two or more lipid ingredients, at least first of which contains a hydrophilic surfactant and at least second of which contains a lipophilic surfactant; said lipid ingredients together provide solubilisation of said testosterone ester in amount 10-20 % of weight of a pharmaceutical composition; a method for prevention or release of testosterone deficiency symptoms in mammal subjects and a method for maintenance of prolonged oral testosterone release.

EFFECT: invention provides medium-size testosterone and fatty acid ester delivery of intensified and prolonged adsorption, desired testosterone levels which are detected in people who do not suffer testosterone deficiency.

30 cl, 15 dwg, 4 tbl

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to pharmacology, pharmaceutics and medicine, more specifically to a new generation of high-stable dosage forms prepared with using the process of sublimation in a specific mode with a composition containing no stabilising agents reducing the width of therapeutic action of finished dosage form substantially. The invention concerns the pharmaceutical composition for injections and infusions containing 3-oxy- and methylpyridine derivatives and pharmaceutically acceptable salts thereof as an active ingredient, sodium chloride or potassium chloride as an additive agent, in the form a lyophilisate. The process involves the sublimation followed by the vacuum dewatering for at least 64 hours.

EFFECT: pharmaceutical composition possesses high stability for the whole shelf-life as opposed to all known pharmaceutical formulations of these compounds; it preserves pharmacological activity and enables dissolving the composition immediately before use and reducing a risk of the negative effect of the thermal sterilisation of aqueous solutions.

2 cl, 7 ex, 2 tbl

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