UA80111C2 - Pharmaceutically acceptable phosphate-glycerol carrying bodies (variants) - Google Patents

Abstract

This invention relates to three-dimensional synthetic and semi-synthetic compositions having biological activity, and to the uses thereof in the treatment and/or prophylaxis of various disorders in mammalian patients. More particularly it relates to preparations and uses of synthetic and semi-synthetic bodies, such as liposomes, which after introduction into the body of a patient, produce beneficial anti-inflammatory, organ protective and immune regulatory effects. The invention also relates to treatments and compositions for alleviating inflammatory and autoimmune diseases and their symptoms.

Description

Description of the invention

The invention relates to three-dimensional synthetic and semi-synthetic compositions having biological activity, 2 their use in the treatment and/or prevention of various diseases in mammals. In particular, the invention relates to preparations and areas of application of synthetic and semi-synthetic bodies, which after introduction into the patient's body provide favorable anti-inflammatory, organ protective and immunoregulatory effects. The invention also relates to methods of treatment and compositions for alleviating inflammatory and autoimmune diseases and their symptoms.

Professional antigen-presenting cells (APCs), including dendritic cells (DCs) and macrophages (Mp), 70 actively capture and digest antigens (ADs), cell debris, and remove infectious organisms and dying cells, including remnants of dying cells. During this process, ARS can stimulate the production of dominated responses of pro-inflammatory cytokines YOU! (11-12, 1-1, IME-G, TME-A, etc.) or regulatory cytokines TI2/TAZ (such as 1-10, TOR-B, 1/-4, etc.); depending on the nature of the antigen (Ad) or phagocytosed material and the level of maturity/activation of ARS. 12 ARS remove cell debris, some of which are formed from the cell membranes of the body's own body, some from cells of bacterial and parasitic infections and symbiotic organisms, such as intestinal bacteria. While some of these cellular debris initiate a pro-inflammatory response, others initiate a protective and anti-inflammatory response.

A normally functioning immune system is able to distinguish between antigens of foreign introduced organisms (not its own) and antigens of its own tissues or fragments, developing an immune response only against foreign antigens. When the patient's immune system stops distinguishing between "its" and "not its own" antigens, an autoimmune disease begins .

MUO-A-90/11781 describes the use of liposomes as carriers of a medicinal substance for the local treatment of wounds, cuts and wounds, for example, the cornea or the ocular surface (abstract).

M/O-A-95/23592 describes the use of single-layered liposomes having an average diameter in the range from -400 nm to 15 Ohm and which are not bound to the active substance, for the transfer of cholesterol from atherosclerotic He) plaques for the treatment of atherosclerosis, (p .b, lines 6-11).

O5-A-5262405 describes the use of phospholipids of a certain condition for the treatment of airway obstruction in a patient. It follows that the principle of action is based on the effect of a large volume of interaction between phospholipids and the mucosa to reduce the adhesive properties of the mucosa (column 4, lines 47-50). -

This invention is defined at the end in the attached claims. "-

The invention relates to the discovery of the fact that pharmaceutically acceptable bodies, such as liposomes, beads or similar particles, which include phosphate-glycerol groups, after administration to a mammalian patient, cause an anti-inflammatory effect and therefore can be used for the treatment of a number of diseases. These bodies can also include as a minor component an inactive component and/or a component that exhibits activity by

Another mechanism. co

In one embodiment, the invention relates to a composition capable of producing an anti-inflammatory ip mimo response in a mammal, wherein said composition comprises pharmaceutically acceptable bodies having a size of 7520 nanometers (nm) to 500 micrometers (μm) that contain a plurality of phosphate-glycerol groups or groups that can "turn into such groups. Preferably, these bodies are essentially free of non-lipid pharmaceutically active C components. Mostly, phosphate-glycerol groups make up 6095-10095 active groups of these bodies. It is assumed that after administration to a mammal, these bodies interact with the immune system through phosphate-glycerol groups. As a result, after such an introduction, an anti-inflammatory response is produced.

In another embodiment, the invention relates to three-dimensional synthetic or semi-synthetic bodies, also referred to herein as pharmaceutically acceptable bodies, which have a size in the range of 2 nm to 50 µm and are modified in such a way that they contain as a main component at least one anti-inflammatory or promoter ligand, while said ligand includes phosphate-glycerol groups. av | In the following embodiment, the invention relates to three-dimensional synthetic or semi-synthetic bodies, also referred to here as pharmaceutically acceptable bodies, which have a size in the range from 2 nm to 50 μm and - contain phosphate-glycerol groups on their surface. -k 70 The invention facilitates a method of treating a disease mediated by T-cell function, which comprises administering to a mammalian patient an effective amount of a pharmaceutically acceptable body bearing an effective amount of phosphate-glycerin groups to inhibit and/or slow the progression of a disease mediated by T-cell function cells

The invention also facilitates a method of treating an inflammatory disease, which comprises administering to a patient an effective amount of 29 pharmaceutically acceptable bodies carrying an effective amount of phosphate-glycerin

HF) groups, to inhibit and/or slow down the progression of an inflammatory disease. The invention also facilitates a method of treating a disorder of endothelial function which includes administering to a mammalian patient an effective amount of pharmaceutically acceptable bodies carrying an effective amount of phosphate-glycerol groups to inhibit and/or slow the progression of the disorder of endothelial function. 60 The invention also facilitates a method of treating an immune disease characterized by inadequate cytokine expression, which includes administering to a mammalian patient an effective amount of pharmaceutically acceptable bodies carrying an effective amount of phosphate-glycerol groups to inhibit and/or slow the progression of the immune disease.

The invention also facilitates a method of treating or preventing a cardiac disease in mammals, the presence of which or susceptibility to which can be determined based on a prolonged OT-s interval on a patient's electrocardiogram, while the specified method includes administration to a mammalian patient suffering from such a disease or prone to it, a pharmaceutical composition that includes pharmaceutically acceptable biocompatible synthetic or semi-synthetic bodies, also referred to herein as pharmaceutically acceptable bodies, and a pharmaceutically acceptable carrier, wherein at least a portion of said bodies have a size in the range of "2 Ohm to 50 Ohm, and at the same time the surfaces of said bodies modified in such a way that they contain as the main component at least one anti-inflammatory promoter group, and the mentioned group is a phosphate-glycerol group.

In another embodiment, the invention relates to a pharmaceutical composition in a unit dosage form for /o administration to a mammalian patient, which includes pharmaceutically acceptable bodies and a pharmaceutically acceptable carrier, while at least part of said bodies have a size in the range from "20 nm to 500 µm and at the same time the surface of said bodies include phosphate-glycerol groups or groups that can be converted into phosphate-glycerol groups, and said unit dosage form contains from 500 to "2.5 x 109 bodies,

In a further embodiment, the invention relates to a pharmaceutical composition comprising 75 pharmaceutically acceptable biocompatible synthetic or semi-synthetic bodies (also referred to herein as pharmaceutically acceptable bodies) and a pharmaceutically acceptable carrier, wherein at least a portion of said bodies have a size in the range of 2 Ohm to 500 µm and the surfaces of the said bodies are modified in such a way that they contain at least one anti-inflammatory promoter group as the main component, and the mentioned group is a phosphate-glycerin group.

In a further embodiment, the invention relates to a pharmaceutical composition that includes pharmaceutically acceptable biocompatible synthetic or semi-synthetic bodies (which are also referred to herein as pharmaceutically acceptable bodies) and a pharmaceutically acceptable carrier, while at least a portion of said bodies has a size of 2 Ohm to 500 μm and includes cardiolipin.

The bodies described above may additionally include an inactive surface layer and/or a surface layer that

It shows activity through another mechanism, for example, phosphatidylserine. |See, for example, Line ei ai.,

International publication UUO 01/667851. i9)

The invention relates to lyophilized or freeze-dried pharmaceutically acceptable bodies bearing phosphate-glycerol groups or groups that can be converted to phosphate-glycerol groups, and kits including lyophilized or freeze-dried pharmaceutically acceptable bodies containing "h-phosphate-glycerol groups or groups that can be converted to phosphate-glycerol groups, and a pharmaceutically acceptable carrier. --

The invention relates to a method of treating a disease mediated by T-cell function, which comprises administering to a mammalian patient suffering from the disease or at increased risk of a disease mediated by T-cell function, an effective amount of a composition that includes pharmaceutically acceptable bodies of from "2 Ohm to "500 µm, which contain on their surface a plurality of phosphate-glycerin groups or groups that can be converted into said phosphate-glycerin groups, so that after this introduction, the progression of the disease, mediated by the function of T cells, is inhibited and/or slowed down .

The invention relates to a method of treating an endothelial function disorder, which includes administering to a mammalian patient suffering from this disorder or having an increased risk of endothelial function disorder, an effective amount of the composition, which includes pharmaceutically acceptable bodies with a size of from "2Om to U5O0OmMcm, which -sh-th s contain on their surface a number of phosphate-glycerol groups or groups that can be converted into the mentioned phosphate-glycerol groups, so that after this introduction, the progression of the disorder of endothelial function "" is inhibited and/or slowed down.

In a further embodiment, the invention relates to a method of treating an immune disease in a mammalian patient suffering from the disease or at increased risk of the immune disease, which (ee) comprises administering to said mammalian patient an effective amount of a composition comprising pharmaceutically acceptable bodies ranging in size from "2Onm to "50μm, which contain on their surface a number of phosphate-glycerin o groups or groups that can be transformed into the mentioned phosphate-glycerin groups, so that after the introduction - the progression of the immune disease is inhibited and/or slowed down.

On the other hand, this invention can also be considered as the application of a receptor on the cells of the immune system - mammals, for example, macrophages, which specifically binds to the phosphate-glycerol group. The invention - M relates to bodies that include ligands and groups that can bind to such a receptor and, as a result, produce an anti-inflammatory response. Accordingly, bodies according to the invention can be defined as bodies that include ligands or their active groups that compete with the binding or absorption of bodies expressing phosphate-glycerol groups, described here, by antigen-presenting cells. A person skilled in the art can easily determine whether a particular body is capable of competing in the manner described by conducting simple test studies. For example, the body can be studied using a readily available co-monocytic cell line, such as SHO37. In the first experiment, 0937 cells were incubated only with fluorescently labeled PO liposomes, and in the remaining experiments, 0937 cells were incubated in the presence of fluorescently labeled PO liposomes and various amounts of the test compound. If the absorption of fluorescently labeled PO liposomes in the rest of the experiments is reduced compared to the first experiment, then the test compound is considered to compete for the receptor and is included in the scope of the present invention.

Fig. 1 is a bar chart representing the results of the experiments of the following Example 1 with hypersensitivity mice (CH, a model of acute inflammatory disease mediated by T cells) when using liposomes according to the preferred embodiment of the invention, compared to other liposomes and control experiments.

Figure 2 is a similar graphical representation showing the use of liposomes with different phosphatidylglycerol (PO) content in the CH5 mouse model, Example 2.

Fig. 3 is a similar graphic representation of the results of Example 3, in which different concentrations of 7595 PO-liposomes were used in the model of SNH of mice.

Fig.A4 is a similar graphic presentation of the results of Example 4, in which different concentrations of 10095 PO-liposomes were used in the mouse SNH model.

Fig5 is a similar graphical representation of the results of Example 5, in which liposomes of different sizes were used in the CH5 model. 70 Fig. b6 is a similar graphic representation of the results of Example b, in which the model of hypersensitivity of mice of delayed tin (OH, a model of chronic inflammatory disease mediated by T-cells) was used.

Fig. 7 is a similar graphic representation of the results of Example 7, in which liposomes with cardiolipin were used in the mouse ONZ model.

Fig. 8 is a similar graphic representation of the results of Example 8, in which liposomes with cardiolipin were used in the model of HF in mice.

Fig. 9 shows the percentage change in the decline of the excitatory postsynaptic potential (ERBP) in control and treated mice, which is an indicator of the effect on long-term potentiation (LTP), Example 9.

Figure 10 represents the data shown in Figure 9 in bar chart format, Example 9.

Fig. 11 shows the difference in the concentration of anti-inflammatory cytokine 1-4 in the hippocampus of control and treated animals, Example 10.

Fig. 12 shows the difference in the concentration of the pro-inflammatory cytokine II-IV in the suspension of homogeneous cells of the spleen of control and treated animals, Example 11.

Fig. 13 shows the difference in the concentration of TME-A in the monocytic cell line 0937 treated with different concentrations of 7595 PO-liposomes, Example 12.

Fig. 14 is a graphical representation of the results of Example 13 - the content of endothelin-i in the ears of mice i) treated according to the preferred embodiment of the invention, compared to the control experiment.

Fig. 15 is a graphical representation of the results of Example 14 - ISAM-1-positive cells from cultures

NOMES in the presence and in the absence of compositions according to the preferred embodiment of the invention. According to the invention, patients are injected with pharmaceutically acceptable bodies that carry phosphate-glycerin groups on their surface. Without going into theory, it can be assumed that these bodies interact with the immune system of the patient with accompanying beneficial effects, such as inhibition of pro-inflammatory cytokines and/or promotion of anti-inflammatory cytokines. The cells involved in the reaction can be immune cells, such as professional or non-professional antigen-presenting cells, endothelial cells, o regulatory cells, for example, MK-T cells and others.

These pharmaceutically acceptable bodies include synthetic and semi-synthetic bodies having various shapes, mostly, but not exclusively, spheroidal, cylindrical, ellipsoidal, including flattened and elongated spheroidal, serpentine, kidney-like, etc., and sizes from 720nm to "500 μm in diameter when measured mainly along the longest axis of the body, and contain phosphate-glycerin groups on their surface. Pharmaceutically acceptable bodies contain phosphate-glycerol groups on the surface with predetermined . characteristics. Without getting into the theory, we can assume that these groups are able to interact with the appropriate and? receptor (receptors) (not only with the Re-receptor) on antigen-presenting cells and mimo. The structure of these groups can be changed synthetically and can include all, part or a modified version of the initial phosphate-glycerol group. For example, the negatively charged oxygen of the phosphate group

Go! phosphate-glycerin group can be transformed into a phosphate-ester group (for example, I-OR(IOHOCHHOK"), where Y is the residue o of the phosphate-glycerin group, K' is -CH "CH(OH)СНООН and K" is is an alkyl of 1-4 carbon atoms - or a hydroxy-substituted alkyl of 2-4 carbon atoms and 1-3 hydroxyl groups, provided that K" is more easily pyrolyzed than the K group; on a diphosphate group, including diphosphate ethers (for example , - Г-ОР(ФОХОКЗОР(ОХОК")", where I and K correspond to those defined above, and each K" independently of the others represents hydrogen, an alkyl of 1-4 carbon atoms or a hydroxy-substituted alkyl of 2-4 carbon atoms and 1-3 hydroxyl groups, provided that the second phosphate group I-P(IOXXOK")2| is more easily hydrolyzed by ip mimo than group K; or to a triphosphate group, including triphosphate esters (for example, Г-ОРИЯУОХОК)ОР(ОХОКОР( OKOK")", where | i

KE are as defined above, and each K" independently of the others represents hydrogen, an alkyl of 1-4 carbon atoms or a hydroxy-substituted alkyl of 2-4 carbon atoms and 1-3 hydroxyl groups, provided that the second and third

Ф) phosphate groups are more easily hydrolyzed ip o omimo than the EC group; and top Such synthetically modified ka phosphate-glycerin groups are capable of expressing phosphate-glycerol ip mimo and, accordingly, such modified groups are phosphate-glycerin groups capable of conversion. in Phosphatidylglycerol is a known compound. It can be obtained, for example, by processing the natural dimeric form of phosphatidylglycerol, cardiolipin, with phospholipase Y. It can also be obtained by enzymatic synthesis from phosphatidylcholine using phospholipase O | see, for example, US patent 5,188,951 Tgetriau, ei aiYi. Its chemical structure includes a phosphate-glycerol group and a pair of similar but different chains of fatty acids С.в8-Сго. 65 The term "PO" here refers to phospholipids bearing a phosphate-glycerol group with a wide range of chains of at least one fatty acid, provided that the resulting PO component can act as a structural component of the liposome. Preferably, such POo-compounds can be represented by Formula I: where BC and BC" are independently selected from C.4-Sod hydrocarbon chains, saturated or unsaturated, straight or those containing a limited number of branches, with at least one chain contains from 10 to 24 carbon atoms. Essentially, lipid chains B and 2" form a structural, not an active, 79 component of liposomes. Accordingly, these chains may vary and include one or two such lipid chains, the same or different, provided that they perform a structural function. Preferably, the lipid chains can be from "10 to "24 carbon atoms in length, saturated, monounsaturated or polyunsaturated, straight or with a limited number of branches. Laurate (C12), myristate (C14), palmitate (C16), stearate (C18), arachidate (C20), behenate (C22) and lignocerate (C24) are examples of suitable saturated lipid chains for PO for use in the present invention. Palmitoleate (C16), oleate (C18) are examples of suitable monounsaturated lipid chains. Linoleate (C18), linolenate (C18) and arachidonate (C20) are examples of suitable polyunsaturated lipid chains for use in PO in liposomes according to the invention. Phospholipids with one such lipid chain, also useful in this invention, are known as lysophospholipids. The invention also relates to the use of liposomes, in which the active component is a dimeric form of PO, namely, cardiolipin, but other dimers Formula | with are also suitable. Preferably, such dimers are not synthetically cross-linked with a synthetic d) cross-linking agent such as maleimide, but are cross-linked by removing the glycerol link as described in Ieppideg,

Viospetizig, p. 525 (1970) and is shown below in the reaction scheme:

Kk I - 7

SCHI s ІІ л «c) her

RO d)

And -

Y. banni kinny iy Sanin Ya ts A ;" seam and ! "Enryat: And ! (ee) («c) y - shu 70 saga yiriv

KU

25 where each of B and B independently of each other have the values given above.

Gee! As stated above and, again, without attempting to create any theory, the Po-group and its dimer presumably represent a ligand, since it apparently binds to a specific region of a protein or other molecule where (C of the PO-receptor ), and accordingly this phosphatidylglycerol molecule (and its dimeric form) is sometimes referred to herein as a “ligand” or “linking group.” Such binding apparently occurs through the phosphate-glycerol group because -O-P(HOCHON )-O-CHO-CH(OH)-CHO-OH, which is sometimes referred to herein as a "headgroup," "active group," or "linking group." Accordingly, the terms "linking," "linking "binding group" or "ligand" herein does not refer to any mechanism or principle of action. However, it is suggested that the phosphate-glycerol groups described above are located on the external surfaces of the bodies of the invention to interact with components of the patient's immune system. It should be noted , that this interaction differs from the specific interaction of apoptotic cells with the b5 phosphatidylserine receptor for an antigen-present egg cells.

Examples of "three-dimensional body parts" or "pharmaceutically acceptable bodies" include biocompatible synthetic or semi-synthetic structures such as liposomes, solid spheres, hollow spheres, filled spheres, granules, granules, and microspheres of biocompatible materials, natural or synthetic, commonly used in the pharmaceutical industry. industry Balls can be solid or hollow, or filled with a biocompatible material. The term "biocompatible" refers to substances that, in the quantities used, are either non-toxic or have acceptable toxicity profiles such that their use is acceptable.

Similarly, the expression "pharmaceutically acceptable" in reference to "pharmaceutically acceptable bodies" herein refers to bodies comprising one or more materials that are pharmaceutically acceptable and suitable for intravenous delivery.

Such bodies may include liposomes formed from lipids, one of which is PO. Alternatively, the 7/0 pharmaceutically acceptable bodies may be solid spheres, hollow spheres, filled spheres, granules, granules and microspheres of biocompatible materials comprising one or more biocompatible materials such as polyethylene glycol, poly(methyl methacrylate), polyvinylpyrrolidone, polystyrene and a wide range of other natural, semi-synthetic and synthetic materials with phosphate-glycerin groups attached to them.

As noted above, phosphatidylglycerol analogs with modified active groups that also /5 interact with PoO receptors on antigen-presenting cells, through the same receptor pathway as PO, or otherwise, to produce an anti-inflammatory response in the recipient's body, are considered included within the expression "phosphatidylglycerol". This term includes, without limitation, compounds in which one or more hydroxyl groups and/or phosphate groups are derivatized or in salt form. Many such compounds form free hydroxyl groups immediately after administration or later and, accordingly, include

RoO corpses capable of transformation.

Preferred compositions according to the invention are liposomes, which can consist of various lipids. However, mostly none of the lipids is positively charged. In the case of liposomes, phosphatidylglycerol PO may make up almost all or the entire layer(s) or wall(s) of the liposome, with its phosphate-glycerol group facing outward to act as a linking group, and the lipid chain or chains forming structural wall.

Liposomes, or lipid vesicles, are closed bags with a size in the micron or submicron i) range, the walls (monolayers or multilayers) of which include suitable amphiphilic substances. Usually they contain an aqueous environment, although for the present invention the internal content is not important, and they are generally inactive.

Accordingly, in a preferred embodiment, liposomes, as well as other pharmaceutically acceptable bodies, essentially "do not contain non-lipid pharmaceutically active components (for example, contain "190) and more preferably, do not contain non-lipid pharmaceutically active components at all. Such liposomes are obtained and processed in such a way that the active groups are located outside the liposome body. Thus, PO in liposomes of the preferred variant -" where the implementation of the invention serves as both a ligand and a structural component of the liposome itself.

Thus, a preferred embodiment of the invention relates to liposome bodies that have on their surfaces, or can be processed or induced to have on their surfaces one or more phosphate-glycerol groups acting as binding groups. Phosphatidylglycerol is preferred

PO-ligand, and such lipids should be from 1095 to 10095 liposomes, and the rest - an inactive component, for example, phosphatidylcholine RS, or a component with a different mechanism of action, for example, phosphatidylserine P, or their mixtures. Inactive ingredients such as RS are preferred. "

At least 109595 kg. such a liposome consists of PO, preferably at least 5095, more preferably from 60 to 10095, and most preferably from 70 to 9095, and in one of the most preferred embodiments, the liposome contains 7595 wt. RO. ;" Mixtures of PO-liposomes with inactive liposomes and/or with liposomes from phospholipids having a different mechanism of action can also be used, provided that the total amount of PO in the mixture remains above Minimum 71095 and preferably above 60905. o Regarding non-liposomal bodies for use in the invention, as indicated above, they include biocompatible solid or hollow beads of suitable size. Biocompatible non-liposomal synthetic or o semi-synthetic bodies can be selected from polyethylene glycol, poly(methyl methacrylate), polyvinylpyrrolidone, - polystyrene and a wide range of other natural, semi-synthetic and synthetic materials, with phosphate-glycerol groups attached to them. Such materials include biodegradable polymers such as those described in U.S. Patent No. 4,938,763, incorporated herein by reference in its entirety. as Biodegradable polymers are described in the literature and include, for example, straight chain polymers such as polylactides, polyglycolides, polycaprolactones, polyanhydrides, polyamides, polyurethanes, polyetheramides, polyorthoesters, polydioxanones, polyacetals, polyketals, polycarbonates, polyorthocarbonates, polyphosphazenes, polyhydroxybutyrates, polyhydroxyvalerates, polyalkylene oxalates, polyalkylenesuccinates, poly(malic acid), poly(amino acids), polyvinylpyrrolidone, polyethylene glycol, polyhydroxycellulose,

F) chitin, chitosan and their copolymers, terpolymers and combinations. Other biodegradable polymers include, for example, gelatin, collagen, etc.

Suitable substances for derivatization in order to attach the phospholipid(s) or its parts with bo groups or binding groups to three-dimensional bodies are produced serially, for example, by the company |Roiuzsiepsez

Ips., 400 Maieu Coide, UMagipdiop, RA 18976, or Zidta Alagispy Ripe SNetisaI5). Methods of their derivatization are known in chemical technology. Certain preferred examples of such methods are described in International Patent Application

PCT/САО2/01398 Mazodetp Igeiiapa I itpied, which is included in this application in the form of a reference.

The invention provides that the patient can be a mammal, including, but not limited to, humans and domestic animals, such as cows, horses, pigs, dogs, cats, etc.

Phospholipids are amphiphilic molecules (ie, amphiphiles), meaning that the compound includes molecules that contain a polar water-soluble group attached to a water-insoluble hydrocarbon chain.

Amphiphiles serving as matrix layers have defined polar and apolar regions. Amphiphiles may include, in addition to the PO according to the invention, other natural lipids used together with a phospholipid carrying an active group, alone or in admixture with another lipid. Amphiphiles serving as a liposome layer(s) can be inert, structure-forming synthetic compounds, such as polyoxyethylene alkyl ethers (simple), polyoxyethylene alkyl ethers (complex) and sucrose diesters.

Methods of obtaining liposomes of the appropriate size are known in science and do not form part of the present invention.

References can be made to various manuals and literature on this subject, for example, the review article 7/0 GMirozotez az RNagtaseciisai Oozade Grogtz", Bu UespegKe! Vageppoi2 and Ovap 9. A. Spgotteip, and the literary sources listed here, for example Mem, K .S. "Irozotev: And Rgasiisa! Arrgoasi", IC. Rgezz ai Ohio Opimegeiu

Rgezz (1990)|.

The diameter of the liposome, as well as other pharmaceutically acceptable bodies, in the preferred embodiment of the invention is from 20nm to 7500μm, more preferably from 7200nm to 1000nm, even more preferably from 7500nm to 7/5. 790 nm, and most preferably from 8 nm to 120 nm (preferably when measured along the longest axis of the body).

Pharmaceutically acceptable bodies can be suspended in a pharmaceutically acceptable carrier, such as sterile saline, sterile water, pyrogen water, isotonic saline, and phosphate-buffered solutions (eg, sterile aqueous solutions containing phosphate buffer), together with other non-toxic, biocompatible substances used in pharmaceutical formulations, such as auxiliary pharmaceuticals, buffers, preservatives, etc. Preferably, the pharmaceutically acceptable bodies are prepared as a liquid suspension in a sterile biocompatible fluid, such as buffered saline, and administered to the patient by any suitable route that allows them to interact with one or more components of the immune system, for example, intra-arterially, intravenously, or most preferably, by intramuscular or subcutaneous injection.

The invention provides that pharmaceutically acceptable bodies can be freeze-dried or lyophilized i) so that they can later be resuspended for administration. The invention also relates to a kit of parts comprising lyophilized or freeze-dried binding-bearing bodies and a pharmaceutically acceptable carrier such as sterile saline, sterile water, pyrogen-free water, isotonic saline, and phosphate-buffered saline ( for example, sterile aqueous solutions containing phosphate buffer), as well as other non-toxic biocompatible substances used in pharmaceutical formulations, such as pharmaceutical excipients, buffers, preservatives, etc. The composition may also include protective "- agents for freeze-drying, known in pharmaceutical technology, for example, lactose or sucrose.

The preferred method of administration of pharmaceutically acceptable bodies to a patient is by injection, which is administered daily, several times a week, once a week, or once a month, for a period of one week to several months. co

The frequency and duration of the treatment course will vary depending on the characteristics of the specific patient and the specific condition being treated, its severity, and whether the treatment is preventive, curative, or curative. Planning and optimization of treatment are familiar to experienced doctors. Intramuscular injection, especially in the gluteal muscle, is the most preferred. One of the separate injection courses, at least for some indications according to the invention, is an injection into the gluteal muscle of the corresponding number of bodies on day 1, the next injection on day c 2, the next injection on day 14 , and then "booster" injections at monthly intervals, if appropriate. . By implication, in many embodiments of the invention, pharmaceutically acceptable bodies containing on its surface RO groups as binding groups, act as modifiers of the patient's immune system, similar to the effect of a vaccine. Accordingly, they are used in such quantities and by such methods of introduction as to ensure a sufficient localized concentration of bodies at the site of introduction. Quantities of such bodies suitable for modification

Go! of the immune system, may not directly correlate with the recipient's body size and therefore may differ markedly from the doses of drugs designed to provide therapeutic levels of active substances in the patient's bloodstream and tissues. Accordingly, doses of medicines should be much higher than doses of immune system modifiers.

The correlation between the weight of liposomes and the number of liposomes can be determined by an experienced specialist in liposome compositions based on the information that a bilayer vesicle with a diameter of 10 Ohm contains 81,230 lipid molecules per 1 vesicle, distributed approximately 50:50 between the layers | see Kispaga Naitidap - 1992 Opimegeyu oi VpyizA

SoIstria RIO Tpeziz "Ttapvtetbrgape pH agadiepiv ip IProzotevs (ptisgoiogt): agid-mevisie ipiegasiope and rgoiop

Pih", ribiiznei ru Mayopa! (ibgagu oi Sapada, ONChauzsha, Sapada (1993); MOpimegeyu Misgoiitv ogdeg po.

OMIO0406756; Sapadiapa by. 942042220, IBVM 0315796936). From here you can calculate, for example, that the dose

ХХ108 vesicles (the order of the dose used ip mimo in the individual examples below) is equivalent to 4.06x1073 lipid molecules. Using Avogadro's number for the number of lipid molecules in a gram-molecule o (mol), 6.023x1023, we can determine that this is 6.74x1077! mol, which at the molecular weight of PO 729 is approximately 3.83x10 8 g, or 38.Zng PO for such a dose. 60 The amounts of pharmaceutically acceptable bodies to be administered will vary depending on the nature of the mammalian disease to be treated and the nature and characteristics of the patient. Preferably, an effective amount of pharmaceutically acceptable bodies is non-toxic to the patient, and is not so great as to harm the immune system. When using intra-arterial, intravenous, subcutaneous or intramuscular administration of a sterile aqueous suspension of pharmaceutically acceptable bodies, it is preferable to administer, for each dose, from 70.1 to 50 ml of liquid, which contains the number of bodies, in general, equivalent to 1095-100090 of the number of leukocytes, which is usually contained in an equivalent volume of whole blood. Preferably, the number of bodies administered per delivery to a human patient is from 500 to -2.5X10 9 ("25Ong bodies in the case of a liposome and proportionally to the difference in density for other body variants), more preferably from 1000 to 71500000000, even more preferably from 10,000 to "5,000,000 and most preferably from 7,200,000 to "2,000,000.

Because the pharmaceutically acceptable bodies in the method of the invention act as immune system modifiers, similar to a vaccine, the number of such bodies injected at the injection site per administration may represent a more meaningful quantification than the number or weight of bodies per unit body weight the patient For the same reason, it can be argued that effective amounts or body numbers used for a small animal may not translate directly on a weight basis to effective amounts for larger mammals (ie, greater than 5 kg).

This invention is suitable for use in the prevention and/or treatment of a wide range of mammalian diseases involving T-cell function, inflammation, endothelial dysfunction and inappropriate cytokine expression. A patient suffering from such a disease or prone to it may be selected for treatment. 75 The term "treating" means reducing symptoms, such as, but not limited to, reducing the severity or number of symptoms of a particular disease or limiting further progression of symptoms.

With respect to disorders of T cell function (diseases mediated by T cells), these disorders include all and any diseases mediated at least in part by T cells, and include, for example, ulcers, wounds, and autoimmune diseases, including but not limited to not only, diabetes, scleroderma, psoriasis and rheumatoid arthritis.

The invention is suitable for use in inflammatory allergic reactions, organ and cell transplant reactions, and microbial infections causing inflammatory reactions. The invention is also suitable for prophylactic use against oxidative stress and/or ischemic reperfusion injury, ingestion of poisons, exposure to toxic chemicals, radiation damage, and exposure to airborne or water-borne irritants, such as those that cause inflammation. The invention is also suitable for use in inflammatory, allergic and T-cell-mediated diseases of internal organs, such as kidneys, liver, heart, etc. i9)

Diseases involving inappropriate cytokine expression for which the present invention is applicable include any and all diseases involving inappropriate cytokine expression, such as neurodegenerative diseases. Neurodegenerative diseases, including Down's syndrome, Alzheimer's disease and Parkinson's disease, are associated with increased levels of certain cytokines, including interleukin-1B (1-18) (see Sgiyip MUZT ei aiYi. (1989); Mody M. ei ai (1996)). It was also shown that 1-18 inhibits long-term potentiation in the hippocampus. (1998)). Long-term potentiation in the hippocampus represents a certain form of synaptic plasticity and is usually considered a suitable model of memory and learning. (1993)). Thus, inappropriate expression of cytokines in the brain is currently considered to be involved in the development and progression of neurodegenerative diseases and neuroinflammatory diseases.

Thus, the invention is suitable for the treatment and prevention of a wide range of neurodegenerative and other neurological diseases of mammals, including Down's syndrome, Alzheimer's disease, "

Parkinson's disease, senile dementia, depression, Huntington's disease, peripheral neuropathies, syndrome

Juliana-Barra, spinal cord diseases, neuropathic joint diseases, chronic inflammatory demyelinating disease, neuropathies including mononeuropathy, polyneuropathy, symmetric distal sensory neuropathy, neuromuscular synapse disease, myasthenia gravis, and amyotrophic lateral sclerosis (ALS). "» Treatment and prevention of these neurodegenerative diseases is a particularly preferred embodiment of the invention, with the treatment of Alzheimer's disease, Parkinson's disease and ALS being the most preferred. (ee) With respect to diseases involving endothelial dysfunction, the invention is suitable for the treatment and prevention of a wide range of a range of such mammalian diseases, including all and any diseases mediated at least in part by endothelial dysfunction, for example, cardiovascular diseases - such as atherosclerosis, peripheral arterial disease or arterial occlusive disease, congestive heart failure, cerebrovascular disease (stroke), myocardial infarction , angina pectoris, hypertension, etc., vasospastic diseases such as Raynaud's disease, cardiac syndrome X, migraine, etc., and - M damage as a result of ischemia (ischemic damage or ischemic reperfusion damage). In general, this can be essentially any disease, the pathology of which in an inappropriately functioning endothelium is keyed.

Other indications for the compositions and methods of the invention include administration to patients to accelerate the healing of wounds and ulcers, as well as administration to patients before surgery to accelerate the speed of their recovery after operations, including the speed of healing of surgical wounds and sutures.

F. With respect to "cardiac diseases", the present invention is suitable for the treatment and prevention of a wide range of such mammalian diseases, including all and any diseases related to the heart, for example, ventricular arrhythmias (ventricular tachycardia or fibrillation) and sudden cardiac death. death from heart disease. Because the predisposition of patients to heart diseases, such as arrhythmias and sudden death from heart disease, can often be determined by prolonged intervals of OT-c in the rhythm of the heartbeat. Administration of the compositions according to preferred embodiments of the invention should reduce OT-s intervals in mammalian patients, which will be an indicator of reduced susceptibility to arrhythmia and sudden death from heart disease.

Next, the invention is described, for illustrative purposes, by means of non-limiting examples. 65 Examples

In the following examples, abbreviations have the meanings defined below. If the abbreviation is not defined, it has the generally accepted meaning. μg - microgram μl - microliter

MKM - micromolar

СНнЗ - contact hypersensitivity cm - centimeter

OM5O - dimethyl sulfoxide

OMEV - 2,4-dinitrofluorobenzene 70 rnz - hypersensitivity of delayed tin

EUN - ethanol he gram h. - an hour

NO - Hertz v.m. - intramuscularly i.p. - intraperitoneally kg - kilogram

And RZ - lipopolysaccharide

G TR - long-term potentiation mg - milligram min. - minute ml - milliliter

MM - millimolar mes - millisecond ng - nanogram schi nm - nanometer i)

NM - nanomolar

РВ5 - saline solution, buffered with phosphate

PCR - polymerase chain reaction "- with RORZ - 1-palmitoyl-2-oleoyl-zp-glycero-3-I(phospho-1| -serine, here it is called P5

POPO - 1-palmitoyl-2-oleoyl-zp-glycero-3-Iphospho-rac-(I -glycerin)|), here it has the name PO --

RORSO - 1-palmitoyl-2-oleoyl-zp-glycero-3-phosphocholine, here it has the name RS rpm - revolutions per minute "- s - second

Unless otherwise specified, the exact form of lipids used in the experiments was РОРБ5, РОРО and o

RORS defined above. co

Example 1

Liposomes with an average diameter of 100--20 nm were obtained according to standard methods known in science and had the following composition:

Group A-100965 P5 «

Group B-10095 PO 8 s Group C - control, without liposomes. The total suspension of each liposome composition containing 4.8 x 1077 liposomes/ml was diluted with RVZ to obtain an injection suspension containing x 10 9 particles/ml. Liposome suspensions were used for injections into female mice of VAI V/s (duskgop and arogaiyugiez) aged 6-8 weeks and weighing 19-23g, to determine the effect on ear swelling in the model of contact hypersensitivity (CH5) of mice. The CHO model was tested for (ee) Tp1-mediated inflammatory reactions. o The animals were divided into three groups, 5 animals in each group. Groups A and B received about 3 x 105 of the above-described liposomes (that is, 10095 RS and 10095 PO, respectively), in a volume of about 5 Ωcl. Group C was the control group that did not receive liposomes. -l 20 Protocol

The following experiments were performed: -

Ф) име) 60 b5 ptn father-in-law his eto dent her her n-x t is set ; - Petu ZHE; - MEbyna Gyaishoe (deky! deno |Denea ldenei den IDeNKYE Gdevu, z | I se ! | | ovo |. E - tk | zay z st sla p 7 sleep: ee vu fs yn geo t te I ask Perlknyaa set; y y From oderdensya U I 2 g, I SU Y I. : : zeeenoMVi" : churovoe: |, ; pdv. ; . | Kenya -Sh and .

I ssh Di sho EE, M In KOMA DAO KA u tya Zodi i styu sovet vimy t otv tt UllyuTITNCAT pache until I I - Mon ankotig I

On days 1-6, the mice of Groups A and B were injected with the corresponding drug liposomes. About 300,000 liposomes were administered in a 50-μl volume by intramuscular (IM) injection, for a total of about 1,800,000 liposomes over the entire test period. Mice in the control group (Group C) were not injected with liposomes, but were sensitized, challenged and tested in the same manner as Groups A and B, as described below. era ay kohi r shk sii i ke siv sonya pinini inii by the wall opitittunntrnj i Ddsi isnntt 4 bin nni bin ptn lunch - ini int iii: tanya Yan pe Na pre r sa ee y r ovyychevde i ve nen ah SE EV r DE , What Shk zhi wai g

I - g Ts fan con - y t : you 7 y y opnykryaaut U |! and with ! " : namznnann | and 7 all HER, I h- : Ж "-NIZaTsi ; . ; SCHI acCE Her : and - ШИ I and vych in | ET B M son k Eh SHU sh Mech 1 I av) lav Do i Klevan | Teiseokee | Teibsshnnann | You Ben 7 rennia | Breektt en zya KVT 0b56 | back; | TSEKTSi | Tek | LI'Ovii: | I | PREKSHYA, DYING s : Щ Her doc IE Ч cha t : : . Mizatsa! ХХ жая: 1 ; ш DI t0 Sensitization 8 s On day 1, after liposome injection on this day, mice were anesthetized with 0.2 ml of a solution of B5mg/ml sodium pentobarbital by i.p. injection. The abdominal skin of the mouse was sprayed with 7095 EN and a blade was used ". . - . scalpel to remove an area of hair about 1 inch in diameter from the abdomen. Then, 25 μl of 0.595 2,4-dinitrofluorobenzene (OMEV) in a 4:1 mixture of acetone and olive oil was applied to the shaved area using a pipette.

Provocation bo After the liposome injection on day b, mice were given a provocation test of OMEV by applying 1 omcl 0.290 av | OMEV on the dorsal surface of the right ear with a pipette and application of 1 omcl carrier on the left ear with a pipette. with Results

On day 7, 24 hours after challenge, each animal was anesthetized with halothane and ear thickness was measured using a Reasosk spring-loaded micrometer. The results were presented as the difference between the thickness of the OMEV-treated right ear and the thickness of the vehicle-treated left ear. The experiments were repeated three times 7 on similar animals. The increase in ear swelling was used as a measure of the SNOZ-response. The significance of the results was determined on the basis of a two-sample Student's χ-test. A P value of 0.05 was considered significant.

The results are presented in Fig. 1, in the form of a bar chart showing the average swelling values of 25 ears from three experiments, in μm.

GF) Fig. 1 shows that with the help of injection of liposomes according to the invention, a significant reduction in the swelling of the ears was achieved. The reduction achieved with 100906 Rho-liposomes is significantly higher than for 10095 Re-liposomes.

Example 2

Liposomes with an average diameter of 100-20 nm were obtained according to standard methods known in science, and 60 had the following composition:

Group A-10095 RO

Group B - 7595 RO, 2595 RUS

Group C - 5095 RO, 5095 RS

Group O - 2595 RO, 75965 RS because Group E - only RV5

Group E - without injections

The total suspension of each liposome composition containing 4.8 x 1077 liposomes/ml was diluted with PB5 to obtain an injection suspension containing 12 x 106 liposomes/ml. Liposome suspensions were used for injection into mice to determine the effect on ear swelling in the murine model of CHD, a biological system useful for studying TP1-mediated inflammatory responses. Female mice were used for these experiments

VAI V/s (dasKvgop I arogaigiez) aged 6-8 weeks and weighing 19-23Hg.

The animals were divided into 6 groups (Groups A-E, described above) with 10 animals in each group. Control groups that did not receive injections (Group E) or received injections of RV5 without liposomes (Group 70. E) were also included in the study. Animals of Groups A-O were injected with 10 ml of the above-described liposome suspensions, each of which contained about 6 x 105 liposomes.

Protocol

The test involved sensitization (Zepv) with a potentially inflammatory substance, injection of liposomes (Ipi) into test animals or PB5 into control animals, and provocation (Spa!) with a potentially inflammatory substance followed by ear measurements (Meavs) to determine whether , whether liposome injection is effective against the development of provocation-induced inflammation.

The following experiments were performed:

Soil | Liposome | Deme 7 | Dan-?. Day I am Day; | Denv5 | Devvb | Day? ( 201 Tk Ch. I I i ! | her;

AND! in yeRO colors and |) oi 1 ba) Me 1/9

Eat more ! | And Shchi | ! , CI TO, : ke Y i

Here is her mouth NEO I Ks NN of ethics nev en as on EK et sv skit of landslides: DRINK

OV yr benn | Зб | Z o) o u ba; Me o

I Ж I : І і Id) І і ! Ka | | 7 Sho with | | What is it and what is it. and. 1. And | With co

Era evddvns nkvstasn NN: NOT UT J same kn st SV as penis. pyt Ta SHE TIVNE pis NOT with UNN of your Ar, HER | And | Her " ! t signs: with Z and; |!

Ki V.E. ; g, ! : Y t sh ! B Plural. ! Her ; І І i 45. MK Her i і p sha Mch RN YN Da t -zhk lyk x klyny lyh tivy pi to spost t Sv, rt. On days 1-6, mice were injected with the corresponding liposomes as indicated above. Liposomes were administered by i.m. injections in

And av | volume of 5Omcl, that is, 600,000 liposomes per injection, to the total amount for the entire test period of about 3,600,000 liposomes. Mice in the control group were not injected with liposomes, but were sensitized, challenged and tested in the same manner as the other groups of mice as described below. - 70 Sensitization (Zepv) ha On day 1, after liposome injection on this day, mice were anesthetized with 0.2 ml of B5 mg/ml sodium phenobarbital solution by i.p. injections. The abdominal skin of the mouse was sprayed with 7095 EUN and a scalpel was used to remove an area of hair about 1 inch in diameter from the abdomen. Then, 25 µl of 0.595 2,4-dinitrofluorobenzene (OMEV) in a mixture of 41 acetone:olive oil was applied to the shaved area using a pipette. 99 Provocation (Spai)

GF) On day 6, after liposome injection on this day, mice were challenged (Spa!) with OMEV as follows: from 10 µl 0.296 OMEV was pipetted onto the dorsal surface of the right ear, and 1 µl of the vehicle was pipetted onto the left ear.

Results 60 On day 7, 24 hours after challenge, each animal was anesthetized with halothane and ear thickness (Meaz) was measured using a Reasosc spring-loaded micrometer. The increase in ear swelling was used as a measure of the CH53 response. Results were presented as the difference between the thickness of the treated right ear and the thickness of the vehicle-treated left ear. The significance of the results between the two groups was determined on the basis of a two-sample Student's i-test. A P value of 0.05 was considered significant. The results are presented graphically in Fig. 2, in the form of a bar chart showing the swelling of the ears in microns. When constructing the diagram, the average value from the corresponding experiments was used.

Fig. 2 shows that with the help of 100 and 7595 PO, a significant reduction in ear swelling was achieved, which indicates that both of these concentrations provide protection against the development of inflammation from contact with the allergen

Ingredient, OMEV. 5095 and 2595 PO-liposomes also showed a decrease compared to both control experiments, but the difference in this experiment did not reach statistical significance.

Example C

Liposomes with an average diameter of 100-20nm were obtained according to standard methods known in science, and had a composition of 75956 РО, 25956 РОС. A total suspension containing 4.8 x 1014 liposomes/ml was used as described above and diluted in PB5 to obtain an injectable suspension containing the following concentrations of liposomes:

Group y y ; o ce tera i tia "IN tia o so M na; roya i ko Her game "Y. I : | And injection and I

F | in 4! : :

ХО | 7596 RO, goy Yes and a! in 250 iinnvtentvinayatinnh i- annia nn ti guilty: ns tt a PN pi t nn

Cedar inv i NE te tin PE enie 5 pe Rove t YOU EE E ev slia t Re E i

And Bai: sha

E notable jan Ko o E anger of the left ZK i vya dpovzhvnyakh nt --2 t --- in che:

TE "Bealiposom. | | I 16; es

VAiIV-C mice were divided into six groups (Groups A-P), including a control group that did not receive liposomes, but received injections of bOmcl RVZ (Group P). Mice were sensitized on the side, the selected dose of liposomes was injected intramuscularly into the right thigh muscle on the same day, but after sensitization (day 1) and on days 2, 3, 4, and 5. On day b, mice were injection and provocation on the ear as described in Example 1. The thickness of the ear -o s was measured as described 24 hours after the provocation. and The results (Fig. 3) show a significant difference between the control group (Group E) and Group C (12 x 108 "» liposomes/ml), between the control group and Group O (12 x 107 liposomes/ml), as well as between the control group and

Group E (12 x 109 liposomes/ml). Between the control group and Groups A or B (12 x 1071 and 12 x 109 liposomes/ml,

Accordingly) there was very little difference, suggesting the existence of an optimal concentration range (ee) of liposomes, above which the beneficial effect may decrease. In other experiments, a decrease in the effect was also observed when the concentration of liposomes was reduced below 12 x 107 liposomes/ml.

Example 4 - Liposomes of composition 10095 RO with an average size of 100-20 nm were obtained according to standard methods. Four -l 20 groups of 10 mice (Groups A-0) were sensitized, injected and challenged according to the method and schedule described in Example 3, with the following amounts of 10095 PO-liposomes in 50 μl of suspension: Group -6 x 107

Group B - 6 x 109

Group C - 6 x 109 52 Group O - 6 x 107

GF! The results, together with the control experiment with RVZ5 from Example 4, are presented in the form of a bar chart in Fig.4. A significant reduction in ear swelling compared to the control group can be seen for each test group, but the difference between the different groups is very small.

Example 5 Liposomes of the composition 7590 РО, 25906 РОС with an average size of 50, 100, 200 and 400 nm were obtained according to standard methods. They were tested in a murine model of CKD as described in Examples 3 and 4, using 6 x 10? liposomes in

BOmcl of suspensions for each injection and using the "sensitization-injection-challenge" methodology and schedule described in Example 3. Groups were formed as follows:

Group A - 50-nm liposomes, while Group B - 100-nm liposomes

Group C - 200-nm liposomes

Group 0 - 400-nm liposomes

Group E - without liposomes

The results are presented in Fig.5. The results for Group 0, using liposomes with a diameter of 40 Ohm, were little different from the control group (Group E), indicating the possibility of a critical size range in this model.

Example 6

The total suspension of 75,956 PO-liposomes with an average diameter of 100-2 Ohm, containing 4.8 x 10 liposomes/ml, was diluted 70 to obtain an injection suspension containing 6 x 10 5 liposomes/ml. The liposome suspension was used to administer to mice to determine the effect on ear swelling in the OH5 mouse model. As in

In example 1, we used female mice VAY V/s (daskzgop and arogayogiev) aged 6-8 weeks and weighing 19-23 kg.

The animals were divided into 3 groups of 10 animals in each group. The control group (Group C) received only injections of RV5. Animals of Groups A and B received injections of HOmcl suspension containing 6 x 109 liposomes.

Protocol

On days 13-18, mice were injected with 7595 PO-liposomes as indicated below. Liposomes were administered in a 50-μl volume by i.m. injection, i.e., 600,000 liposomes per injection, for a total of about 3,600,000 liposomes during the entire test period. Sensitization and provocation were carried out as described in Example 2.

The results of

The results are graphically presented in Fig. b and show that 7595 RO is effective in the ONZ model on day 16,

24 hours after the third injection made after the second provocation. (uh)

Example 7

Liposomes with a composition of 10095 cardiolipin (SI) and an average diameter of 100-2O0nm were obtained according to standard methods. they were used at a dose of 6 x 107 liposomes per 50-μl injection in the mice ONZ model described in "

Example 6. Data obtained for animals that were injected with C-liposomes (Group A; 10 animals) were compared with data obtained for animals that were injected with only PB5 (Group B; 10 animals). The methods of sensitization, injections and provocations corresponded to those described in Example 2. The results of measuring the thickness of the ears on day 19, 24 hours after the sixth injection, are presented in Fig. 7. The results showed a significant decrease in swelling of the ears in the test group with SI injections. (Group A).

Example 8

Liposomes with an average diameter of 100 nm, which included 10095 cardiolipin or 7595 cardiolipin and 2595 ROS, were obtained according to standard methods. Three groups (Groups A-C) of 10 mice were sensitized on day 1. o The control group received RVZ injections on days 1, 2 and 6 (Group C). The other two groups received injections of liposomes: b x 105 10095 cardiolipin (Group A) or 6 x 105 7595 cardiolipin (Group B), in a volume of BOmcl, according to the same schedule. Mice were challenged on day 7 and ear thickness was measured as described in previous examples. - 70 Fig. 8 shows the average measurements in each group. Both groups receiving C-liposomes showed a statistically significant suppression of CH5 compared to the control group.

Example 9

An animal model of long-term potentiation (LTP) was used to study the cellular and molecular mechanisms of cognitive function. TR is a certain form of synaptic plasticity that occurs in the hippocampal formation, which is considered the biological substrate of learning and memory.

GF) AI. Mayte 361: 31-39 (1990)). Conduct an electrophysiological study of TR in rats by methods well known to experts. Then the animals are killed to study biochemical changes in the hippocampal tissues.

Comparison of electrophysiological data with biochemical hippocampal changes is useful for determining how the cellular events underlying TR may be altered in animals suffering from diseases or 60 disorders associated with neuroinflammation, such as aging, stress, Alzheimer's disease and bacterial infection.

Systemic administration of lipopolysaccharide (II P5), a component of the cell wall of gram-negative bacteria, provokes activation of the immune system by inducing an increase in pro-inflammatory cytokines, such as II-18. As mentioned above, one of the examples of neuronal deficits induced by ARZ and 1/-18 is the deterioration of I TR in the hippocampus.

The TR indicator is the average slope of the post-synaptic excitatory potential (ERP). After tetanic stimulation, the slope of erzr (95) sharply increases, which shows increased synaptic activity.

I P5-induced inhibition of I TR reduces the increase in slope and/or causes a faster return of the erzr slope to the baseline, which means that the increased synaptic activity is short-lived. Appropriate measurements of the slope of erzr (95) at certain time intervals after tetanic stimulation can be used to reflect memory and its loss after an inflammatory stimulus, as well as inflammation in the hippocampus of the brain.

Liposomes with an average diameter of 100--20 nm were obtained according to standard methods known in science, and had a composition of 7595 RO and 2595 ROS. The total liposome suspension containing 2.9 x 1074 liposomes/ml was diluted with PB5 to obtain an injection suspension containing 1.2 x 107 liposomes/ml. it was used for administration to rats in order to determine the effect on I P5-induced deterioration of I TR. For these experiments, 70 male M/iziag rats (Viokezoigsez Opii, Thipku Sopede, Yuibiip) weighing about ZOOg were used.

The animals were divided into four groups of 8 animals in each group for the following treatment:

Group A - saline solution and control experiment

Group B - salt solution и-РО

Group C - IGBR is the control experiment

Group C-IRB - RO 150 μl of each of the above drugs was administered by i.m. injections on days 1, 13, and 14. VioY groups received a total of 5,400,000 liposomes (1,800,000 liposomes per injection). The ITR procedure and the tissue preparation procedure were performed on day 0.

Procedure I TR

Rats were anesthetized by i.p. urethane injections (1.5 g/k). Rats received either I RB5 (10 Ωkg/kg) or saline intraperitoneally. Three hours later, a bipolar stimulating electrode and a unipolar recording electrode were placed in the perforant path and in the cell body of the dorsal region of the dentate gyrus, respectively. Test discharges with a frequency of 0.033 N?i were applied and responses were recorded for 1 Ohv. before and after 45Xmin. after high-frequency stimulation (S series of stimuli was performed with 30-second intervals, 250 H2 for 200 Ohms). Ge

Rats were killed by decapitation. The hippocampus, tetanized and non-tetanized dentate gyrus, cortex and entorhinal cortex were prepared on ice, cut into samples and frozen in ml of Krebs solution (composition

Krebs in mm: Mass 136, KSI 2.54, KNoRO, 1.18, Ma5O,-7H.O 1.18, MansSo» 16, glucose 10, CaSi» 1.13), which contained 1095 ЮМ5О.

Results -

The results are presented in Fig.9. The diagram shows the difference in the excitatory postsynaptic potential "- (erzr) recorded in the cell bodies of labrocytes. The data represent the average values of 7-8 observations in each treatment group and are presented in the form of the average percentage change in the slope of erzr per Zbs, - normalized in relation to the average value 5 minutes before tetanic stimulation. at

Fig. 9 shows that I RoZ-induced inhibition of I TR in the synapses of the perforant path of labrocytes was inhibited by pretreatment with Roz liposomes. Black triangles represent Group A (salt solution and (ee) control experiment), white triangles represent Group B (saline solution and РО), black squares represent Soil C (I RB5 and control experiment) and white squares represent Soil О (IRZ Ro ).

Fig. 10 presents an analysis of average values 40-45 minutes after tetanic stimulation, which shows that the population erzr-slope decreased in the control-I P5 group (white columns) and that PO liposomes (shaded columns) significantly reversed this effect direction ("p" 0.01). As an indicator of the functionality of memory and learning, the improvement in TR slowing demonstrated in this Example means the suitability of such a treatment in the case of dementias, for example, Alzheimer's disease and memory impairment. is" Example 10

I-4 is one of several cytokines produced by the T2 subclass of lymphocytes and known for their anti-inflammatory effects. Fig. 11 shows that the concentration of II-4 in the hippocampus significantly increased in the 1st Po-group, (ee) supplemented with PO-liposomes ("p" 0.05). The white columns represent the control group (Group E), and the shaded columns represent the Ro-treated group (Group P). II-4 was measured by EG ISA and the results were presented in pg (picgrams) II-4/mg of total protein. Such an upregulation of anti-inflammatory cytokine 1-4 in the brain is an indicator of the possibility of using the method and A preferred embodiment of the composition of the invention is 90 in the treatment of a wide range of neuroinflammatory diseases, including Parkinson's disease, ALS, chronic inflammatory demyelinating disease SIRO and Julian Barr syndrome - Example 11

I-18 is one of several cytokines produced by the TI1 subclass of lymphocytes and known for their pro-inflammatory effects. Spleens of animals treated as described in Example 9, groups C and 0, were removed and spleen cells were collected. they were prepared as follows: o Fig. 12 shows that the concentration of I./-1B8 in the cells of the spleen was significantly reduced in the I P5-group, which was reformulated with PO-liposomes ("p" 0.05). I-18 was measured by EGISA and presented in pg 1I/-18/MG of the total protein. This shows the systemic anti-inflammatory effect of the method and compositions according to the preferred embodiment of the invention. Because Example 12

ShC937 is a leukemic monocytic cell line that can be differentiated into macrophages by using phorbol esters. Treatment with lipopolysaccharide (I P5), a component of the cell wall of gram-negative bacteria, stimulates an inflammatory response in SHO937 cells, with upregulation of the expression of a number of inflammatory molecules, including TMEo. This model is an experimental system for evaluating 65 anti-inflammatory therapies. Macrophages can be grown in a culture medium in the presence of an anti-inflammatory composition under investigation, and TMEco expression can be measured.

Liposomes with an average diameter of 100--20 nm were obtained according to standard methods known in science and had a composition of 7595 phosphatidylglycerol (PO) and 2595 phosphatidylcholine (ROS). The total concentration of liposomes was 40 mM of lipids and was diluted to the following final concentrations for analysis: 100 µM phosphatidylglycerol (PO) 40 µM РО 10 µM РО 4.0 µM РО 1 mM µM РО 70 SHUO37 cells were cultured by growing in KRMI medium (C5IVSO VK) supplemented with 1090 embryonic bovine serum (EB) and 195 penicillin/streptomycin, and grown at 372C in an atmosphere containing 595 CO. 5 x 10 cells were seeded into the cells of b-h-arun plates and caused differentiation into macrophages by treatment with 150 nm of phorbol myristate acetate (PMA) for 2-3 days. The cell medium was replaced with complete medium after SHOZ7 cells were differentiated into macrophages. Then the cells were incubated for another 75 24 hours. to minimize pleotropic effects due to PMA treatment.

The cells were then incubated with the following compounds:

Group A Saline solution, buffered with phosphate (РВ5) - negative control experiment,

Group B 1Ong/ml IR - positive control experiment,

Group C 1Ong/ml GR - 100μM RO,

Group O 1Ong/ml GRB5 - 40μM RO,

Group E TIOng/ml RB - 10μM RO,

Group E 1Ong/ml I R and 4.0 μM PO, or

Group OS 1Ong/ml I R y- 1MmkM RO.

Cells were incubated as described above at 372°C to 590°C. After 18 hours Supernatants from each treatment group were collected and analyzed on TME-5 using a standard analytical kit of OpapiyaKipe o

Epgute-i ipkei Ittipozogrepi Azzau (EGIZA) (KO zuvietv, Mippearoiv, OBA).

The results

Fig. 13 shows the amount of produced TME-A in PO per ml. These results demonstrate that i937-differentiated macrophages express very low levels of TME-5 under normal conditions. However, after /P5 treatment, they secrete large amounts of TME-5 into the environment, which is an indicator of cellular stress. Incubation of cells with PO-liposomes inhibits the secretion of TME-5 in a dose-dependent manner, while the highest concentration of 100 μM provides a 98905 decrease, and even the lowest concentration of iμM causes a 5895 decrease in TME-o expression. at

Example 13 3o In order to determine the effect of PO-liposomes according to the preferred variant of the invention on endothelial function, the content of endothelin-y (ET-1) was determined in the ears of mice that participated in the SNZ studies described in

Examples 3. Endothelium is a powerful vasoconstrictor, exhibits inotropic and mitogenic effects, modulates salt and water homeostasis and plays an important role in maintaining vascular tone and blood pressure. "

A variety of evidence supports that endogenous ET-1 may be involved in the pathophysiology of conditions associated with slow vasoconstriction, such as heart failure. In heart failure, elevated levels of circulating ET-1 and rBid-ET-1 are observed |Ciappezzi Yu, Oye! Ku 5, Miaie KI. "Tpe goYe ohi

Thus, ET-1 is a marker of endothelial function, and the increased production of ET-1 in the tissue is an indicator of the deterioration of endothelial function.

To determine the expression of ET-1, the ears of mice (right provoked ear) were collected after 24 hours. after provocation in bo CH3-experiments. Ears were taken from mice receiving i.m. injections of RV5 for 6 days (Group A) and mice that av! received in.m. injection of liposomes 75956 РО/2595 РОС (600000 liposomes/injection; Group B). Ears were stored in KMAIaieg at -202C until RNA extraction. RNA was extracted and cDNA was generated using reverse transcriptase (RT) together with ET-1-specific primers, as an internal control experiment, and PCR was performed with - 70 using shrachtin-specific primers. PCR products were separated on a 1,595 agarose gel and DNA-M bands were counted by densitometric analysis. The ET-1/B-actin ratio was calculated.

PCR composition:

PCR mixture (ET-1) PCR mixture (B-actin) bml of PCR buffer (10x) bml of PCR buffer (10x) (F. 1, Bml. Masig(BOMM) 1.5μcl. Masig(5OMM) co imcl aMTR (10MM) imcl aMTR (10MM)

0.1 µl of primer 1 (25 µM) 1 µl of primer 1 (10 µM) in 0.5 µl of primer 2 (25 µM) µl of primer 2 (10 µM)

O.2Bmcl TAY O.2Bmcl TAY 2.5mcl kKDdNK 2.5mk kKDdNK

ZVmcl of water 37.75mcl of water

Total 5µl Total 5Omcl b5

Primers: |As described above - see, for example, Mapo, !; Nizaip, M; and Ziemzhmat. 0. in "Sopaiopai! sagaias omegehrgesviop oi epdoipeiiip-1 ip (gapzdepis tise), EABEV U 15(53:А1138-А1138 Ragi 2, MAKV 20011.

ET-1 () 5-САС САС TTS TO Test t Too-3

ET-1 (FD) 5-SSA DOS AOS TSS ASA dDAS dO-3!

P-actin (P) 5-STO BOS SOS TST ASO SAC SAA-3

P-actin (g) 5-STS TTT SAT TS ASO SAS SAT TTO-3

PCR parameters: 9420-5 minutes 9490-30 s 602C-30s ZO cycles 729060 -60s 7220-10 minutes 420 - exposure time

After six daily injections of 7595 PO-liposomes, the ET-1 level decreased by 3695 compared to a control group of 75 mice receiving PB5 in the same injection regimen. The results are graphically presented in Fig. 14. This reduction means a beneficial effect of liposome injections according to the preferred embodiment of the invention on the endothelial function of the mammalian patient, due to the reduction of Tp1-mediated inflammation.

Example 14

Intercellular adhesion molecule-1 (ISAM-1) is a cell surface molecule expressed by several types of cells, including leukocytes and endothelial cells. It takes part in the adhesion of monocytes to endothelial cells and plays a certain role in inflammatory processes and in the system of immune protection mediated by T cells. ISAM-1 expression is believed to be involved in the clinical manifestations of various diseases, mostly by interfering with normal immune function. Among these diseases are malignant formations (for example, melanoma and lymphomas), many inflammatory diseases (for example, asthma and autoimmune diseases), atherosclerosis, ischemia, some neurological diseases and allogeneic transplants of He) organs (Map de BioiIre A, map deg 5aaod RT , "IpiegseiIshag adpeviop toiescie-1" in). My. Honey. (1996) 74:1 13-33).

Human umbilical vein endothelial cells (HOMESv) are the primary endothelial cell line isolated from umbilical cords in this manner.

775 flasks were prepared by coating with 0.296 gelatin (5-7 ml/flask) for a minimum of 15/20 minutes or overnight. The remainder was removed. The umbilical cord was sprayed with ethanol before the 7095 procedure and the remnants of the placenta that were on the umbilical cord were cut off. The umbilical cord was then cut to an approximate length of 5-6 inches. The umbilical cord contains two thick-walled arteries and one larger, thin-walled vein. A vein was found and the grooved edge of the cork was inserted into it. Then about 20 cm of rope was used to tie the umbilical cord to the cork.

Then the umbilical cord was washed with phosphate-buffered saline (PB5) several times until the washing PB5 looked clean. After that, 15-20 ml of collagenase solution was placed on the umbilical cord, wrapped in tin foil and incubated for 15 minutes at 372C. After incubation, the tied end of the umbilical cord was cut off and the collagenase was poured into a 50-ml test tube. Collagenase was then passed through the umbilical cord again, the umbilical cord was massaged to release endothelial cells, and then PB5 was passed through the umbilical cord and collected in the same tube containing the collagenase solution. it was centrifuged at 16,000 rpm, the supernatant was removed, and a drop of C from the sediment was resuspended in 10-12 ml of complete medium M199. The medium containing the cells was then added to the gelatinized flasks. Liposomes with an average diameter of 100-20 nm were obtained by standard techniques known in the art and had a composition of 7595 phosphatidylglycerol (PO) and 2595 phosphatidylcholine (ROS). The total concentration of liposomes was 40 mM and was diluted to 100 mM for analysis. because the NOMES cells were distributed among flasks for tissue culture, allowed to attach to the surface of the flasks, and then aw! processed as follows: with Group A - RV5 - negative control experiment,

Group B - 50Ong/ml I R - positive control experiment, -k 70 Group C - 50Ong/ml I RZ i- 100MCM RO sh Group 0 - 5O0Ong/ml I RB5 i- 100μM ROS 7" Cells were incubated at 372С, 5956 СО After 18 h, the supernatants of each treatment group were collected and analyzed for ET-1 using a standard EGISA kit (from Azzau Oezidpv), and the cells were collected for analysis for ISAM-1 as follows: 99 Cells were first washed with RVZ and then incubated with cell culture buffer dissociation at 372C for 25-300 minutes.

GF) Cells were washed by centrifugation and incubated with anti-CO54 (ISAM-1) antibody for 30 minutes. 7 The secondary RITS antibody was then added and incubated with the cells as before. Then they were resuspended in them

RVZ and analyzed for fluorescence on a flow cytometer.

Results 60 The results are shown in Fig. 15, which is a graphical representation of the percentage of cells positively stained in the ISAM-1 assay in the respective cultures. It should be noted that the number of positively stained cells in the culture containing PO-liposomes is reduced to the level of the negative control experiment, and is much lower than the level of the positive control experiment.

Example 15 because Microglial cells (macrophages of the brain) were cultivated and their production of TME-o, an inflammatory cytokine, was measured. Cells were stimulated with immunoglobulin (Id) from patients suffering from ALS, resulting in an 800-fold increase in TME-o production. When the same cells were grown in the presence of ALS-IdO and liposomes

PO, the production of TME-o, decreased by "75905, which shows the potential of the preferred embodiments of the invention for the treatment of ALS. The results are presented in Fig.16.

Claims (59)

Formulation of the invention 70 1. A pharmaceutical composition, in a unit dosage form, for administration to a mammalian patient, which contains pharmaceutically acceptable bodies and a pharmaceutically acceptable carrier, characterized in that at least a part of said bodies has a size in the range of 20 nm to 500 μm, and while the surfaces of said bodies include phosphate-glycerol groups or groups that can be converted into phosphate-glycerol groups, and said single dosage form contains from 500 to 2.5 x 109 bodies, 2. Pharmaceutical composition according to claim 1, which differs in that the bodies are liposomes. C. The pharmaceutical composition according to claim 1 or 2, which is characterized by the fact that it does not essentially contain non-lipid pharmaceutically active components. 4. The pharmaceutical composition according to item C, which is characterized by the fact that it does not contain non-lipid pharmaceutically active components. 5. Pharmaceutical composition according to any of claims 2-4, which is characterized by the fact that the liposomes include from 60 to 100 96 phosphatidylglycerol, while their phosphate-glycerol end groups are located on the outer surfaces of the liposomes. b. Pharmaceutical composition according to claim 5, which differs in that the liposomes include 70-90 90 phosphatidylglycerol. with 7. Pharmaceutical composition according to claim 5 or b, which differs in that the rest of the liposomes is phosphatidylcholine. at 8. Pharmaceutical composition according to claim 1, which is characterized by the fact that said bodies include surface groups that specifically bind to receptors on cells of the immune system of a mammal, specific for binding to phosphate-glycerol groups. "- 9. Pharmaceutical composition according to any one of claims 1-8, which is characterized in that the single dosage form contains from 10,000 to 5,000,000 bodies. - 10. A pharmaceutical composition according to any of claims 1-9, which differs in that it is used as "a medicine." 11. A composition that contains pharmaceutically acceptable biocompatible bodies with a size of 20 nm to 500 μm, which o Contain a plurality of expressed on the surface or such that can be expressed on the surface, ee) phosphate-glycerin active groups, while the said composition is essentially not contains pharmaceutically active non-lipid components, for the manufacture of a medicinal product for the treatment or prevention of a disease selected from a disease mediated by T-cell function, an inflammatory disease and an immune "disease associated with inappropriate expression of cytokines." 12. The composition according to claim 10, where the phosphate-glycerol groups are expressed on the surface of the bodies. - with 13. The composition according to claim 11 or 12, where the phosphate-glycerin active groups make up from 60 to 100 95 active c groups on the surface of the bodies. "» 14. The composition according to any one of claims 11-13, where the phosphate-glycerol groups represent end groups of phosphatidylglycerol (Po) ligands. 15. The composition according to any one of claims 11-14, where the bodies are liposomes. (uh) 16. The composition according to claim 15, where the liposomes consist of 50-100 95 wt. phosphatidylglycerol. 17. The composition according to claim 15 or 16, where the liposomes consist of 65-90 95 wt. phosphatidylglycerol. at 18. The composition according to any one of claims 15-17, where the liposomes have a diameter of 20 nm to 1000 nm. - 19. The composition according to any one of claims 11-18, where a single dosage form of the mentioned composition contains from 500 shu 20 to 25x 109 bodies. 20. The composition according to claim 19, where a single dosage form of said composition contains from 10,000 to 50 million bodies. -. and 21. The composition according to any one of claims 11-20, wherein the disease is a disease mediated by T-cell function selected from ulcers and autoimmune diseases. 22. The composition according to any one of claims 11-20, wherein the disease is an autoimmune disease selected from scleroderma, psoriasis and rheumatoid arthritis. at 23. The composition according to any one of claims 11-20, where the disease is an inflammatory allergic reaction. 24. The composition according to any one of claims 11-20, where the disease is contact hypersensitivity. eat) 25. The composition according to any one of claims 11-20, wherein the disease is delayed hypersensitivity. 26. A composition containing pharmaceutically acceptable biocompatible bodies in the size range from 20 nm to 500 μm, which contain active groups on the surface that consist essentially of phosphate-glycerin groups or groups that can be converted to phosphate-glycerin groups, wherein said composition is substantially free of pharmaceutically active non-lipid components, for the manufacture of a medicament for the treatment or prevention of a disease associated with endothelial dysfunction selected from peripheral arterial occlusive disease, congestive heart failure, ventricular arrhythmia, sudden cardiac death b5, stroke, myocardial infarction, angina pectoris, hypertension, vasospastic disease, ischemic injury and ischemic reperfusion injury. 27. The composition according to claim 26, where the treatment or prevention is carried out due to the therapeutic interaction of said bodies with the patient's immune system to suppress pro-inflammatory cytokines and/or promote anti-inflammatory cytokines. 28. The composition according to claim 26 or 27, where a single dosage form of said composition contains from 500 to 2.5 x 109 bodies. 29. The composition according to any one of claims 26-28, where the bodies are liposomes. 30. Pharmaceutical composition according to any one of claims 26-29, where the composition does not contain non-lipid pharmaceutically active components. 76 31. The composition according to claim 29 or 30, where the liposomes contain from 60 to 100,956 phosphatidylglycerol, while their phosphate-glycerol end groups are located on the outer surfaces of the liposomes. 32. The composition according to item ZO, where the liposomes contain from 70 to 90 96 phosphatidylglycerol. 33. The composition according to claim 31 or 32, where the rest of the liposomes is phosphatidylcholine. 34. The composition according to any one of claims 26-33, where said bodies include surface groups that specifically bind to receptors on cells of the mammalian immune system specific for binding to phosphate-glycerol groups. 35. The composition according to any one of claims 28-34, where a single dosage form of said composition contains from 10,000 to 50 million bodies. 36. The composition according to any one of claims 11-20, wherein the disease is associated with inappropriate expression of cytokines. 37. The composition according to claim 36, where the disease is a neurodegenerative disease. 38. The composition of claim 37, wherein the neurodegenerative disease is selected from Down's syndrome, Alzheimer's disease, Parkinson's disease, senile dementia, depression, Huntington's disease, peripheral neuropathies, Julian Barr syndrome, spinal cord diseases, neuropathic joint diseases, chronic inflammatory demyelinating disease, mononeuropathy, polyneuropathy, symmetric distal sensory neuropathy, diseases of neuromuscular synapses, myasthenia gravis and amyotrophic lateral sclerosis (ALS). 39. The composition according to claim 38, where the neurodegenerative disease is Alzheimer's disease, i) Parkinson's disease or ALS. 40. A single dose of a composition containing pharmaceutically acceptable biocompatible bodies with a size of 20 nm to 500 μm in an amount of 500 to 2.5 x 109 bodies, said bodies contain active groups on the surface, which consist "-- essentially of phosphate-glycerin groups or groups that can be converted into phosphate-glycerin groups, and said composition essentially does not contain pharmaceutically active non-lipid components, for the treatment or prevention of atherosclerosis. "h- 41. The composition according to claim 40, which differs in that the bodies are liposomes containing phosphatidylglycerol. at 42. The composition according to claim 41, which differs in that the liposomes consist of 60-100 96 phosphatidylglycerol. ee) 43. The composition according to claim 42, which differs in that the liposomes consist of 50-90 96 phosphatidylglycerol. 44. A composition of a substance capable of exerting an anti-inflammatory effect in a mammal, wherein said composition contains pharmaceutically acceptable liposomes with a size of 20 nm to 500 μm, the liposomes containing 60-100 95 wt. " of phosphatidylglycerol with phosphate-glycerol groups oriented outward, and liposomes essentially do not contain non-lipid pharmaceutically active components. - with 45. The composition according to claim 44, which differs in that the liposomes contain at least 90 wt. phosphatidylglycerol. "» 46. The composition according to claim 44 or 45, which differs in that the rest of the liposome is an inactive component. 47. The composition according to claim 46, which differs in that the inactive component is phosphatidylcholine. 48. The composition according to any one of claims 44-47, which is characterized in that said composition essentially does not contain (ee) non-lipid pharmaceutically active components. 49. The composition according to any one of claims 44-48, characterized in that the liposomes have a size of 50 to 500 nm. at 50. The composition according to claim 49, which differs in that the liposomes have a size from 80 to 20 nm. - 51. The composition according to any of claims 44-50, which is characterized by the fact that it contains a mixture of said liposomes containing phosphatidylglycerol with inactive liposomes and/or liposomes from phospholipids that act by a different mechanism, while the PO-liposomes are at least 10 95 of the total mixture. - M 52. Use of a composition containing pharmaceutically acceptable biocompatible bodies with a size of 20 nm to 500 μm, containing a plurality of surface-expressed or surface-expressable phosphate-glycerin active groups, while said composition essentially does not contain pharmaceutically active Non-lipid components, in the manufacture of a medicinal product for the treatment or prevention of a disease selected from a disease mediated by the function of T cells, an inflammatory disease and an immune and F) disease associated with inappropriate expression of cytokines. co 53. Application according to claim 52, where the composition is a composition according to any of claims 12-25 and 36-39. 54. Use of a composition containing pharmaceutically acceptable biocompatible bodies with a size in the range from about 20 nm to 500 μm, which contain active groups on the surface, which consist essentially of phosphate-glycerol groups or groups that can be converted into phosphate-glycerol groups , wherein said composition is substantially free of pharmaceutically active non-lipid components, in the manufacture of a medicament for the treatment or prevention of a disease associated with endothelial dysfunction selected from peripheral arterial occlusive disease, congestive heart failure, ventricular arrhythmia, sudden death 65 From heart disease , stroke, myocardial infarction, angina pectoris, hypertension, vasospastic disease, ischemic injury and ischemic reperfusion injury. 55. Use according to claim 54, wherein the composition is a composition according to any of claims 27-35. 56. The use of a composition containing pharmaceutically acceptable biocompatible bodies with a size in the range from 20 nm to 500 μm, which contain active groups on the surface, which consist essentially of phosphate-glycerol groups or groups that can be converted into phosphate-glycerol groups, where said composition essentially does not contain pharmaceutically active non-lipid components, in the manufacture of a medicinal product for the treatment or prevention of atherosclerosis. 57. Use according to claim 56, where the composition is a composition according to any of claims 41-43. 58. Use of a composition containing pharmaceutically acceptable liposomes with a size from 20 nm to 500 μm, 7/0 Liposomes contain 60-100 90 wt. of phosphatidylglycerol with outward-facing phosphate-glycerol groups, and liposomes essentially free of non-lipid pharmaceutically active components, for the preparation of a medicament capable of exerting an anti-inflammatory effect ip mimo in a mammal. 59. Use according to claim 58, where the composition is a composition according to any of claims 45-51. c) 6) "- "- "- "c) d) - and? (ee) («c) - - 70 - ime) 60 b5