WO2022005339A1

WO2022005339A1 - Therapeutic and diagnostic composition based on antibodies

Info

Publication number: WO2022005339A1
Application number: PCT/RU2021/050193
Authority: WO
Inventors: Петр Иванович НИКИТИН
Original assignee: Петр Иванович НИКИТИН
Priority date: 2020-07-03
Filing date: 2021-07-01
Publication date: 2022-01-06

Abstract

The invention relates to the field of biomedicine and nanomedicine, and more particularly to compositions and methods for treating and diagnosing diseases using lipid-containing medicinal agents, as well as compositions and methods for improving the circulation of said agents in the bloodstream and the delivery thereof to targets, and also improving the therapeutic and/or diagnostic efficiency of lipid-containing medicinal agents. The essence of the invention lies in a composition for use in a method for the therapy or diagnosis of diseases or conditions of the body, which includes at least (i) an antibody against patient blood cells which is in a free molecular form, and (ii) a lipid-containing medicinal agent. The technical result achieved by the invention is that of improving the circulation of an agent in the blood stream, providing more efficient delivery of an agent to a target in the body and improving the efficiency with which diseases are treated and diagnosed with the aid of lipid-containing medicinal agents.

Description

ANTIBODY-BASED COMPOSITION FOR THERAPY AND DIAGNOSTICS

The technical field to which the invention relates

The invention relates to the field of biomedicine and nanomedicine, including compositions and methods for the treatment and diagnosis of diseases based on lipid-containing medicinal agents, as well as compositions and methods for improving circulation in the bloodstream, delivery to targets, as well as therapeutic and / or diagnostic efficacy lipid-containing medicinal agents.

Vehicle tier (Background)

Recently, there has been a steady increase in the use of nanotechnology in biology and medicine, as well as in other fields of science and technology. In the treatment of diseases, nanoparticles have a number of advantages over conventional molecular drugs, for example, they have the ability to carry a large amount of the drug, protecting it from degradation, the ability to carry out targeted delivery using even low-affinity receptors due to their multipoint binding of nanoparticles to targets. In addition, nanoparticles are used for the diagnosis of diseases, as labels in various biosensors, and also as contrast agents.

Of particular interest is the use of various nanoagents (lipid-containing, liposomal, magnetic) for the treatment and diagnosis of diseases such as cancer, stroke, atherosclerosis, infectious diseases (McAteer, M.A., et al. In vivo magnetic resonance imaging of acute brain inflammation using microparticles of iron oxide, Nat Med. 13, 1253-8, 2007), etc. For example, lipid-containing nanoparticles (including liposomes) and magnetic nanoparticles are promising agents for the diagnosis and therapy of diseases. Currently, they are already approved for intravenous injection in humans for the treatment of a number of diseases.

Lipid-containing agents (agents based on lipids and their derivatives), as well as agents based on magnetic nanoparticles, are especially interesting due to their unique properties - biocompatibility, low toxicity of lipids, as well as iron oxides (including those with a low proportion of doping with other elements) , low immunogenicity (in the case of constructing agents from lipids inherent in the patient), the possibility of fusion with the cell membrane and delivery of a tolerable load (payload) directly into the cytoplasm, etc. Such properties allowed lipid-containing agents to occupy a dominant position in the field of nanomedicine, including for gene therapy (for the delivery of genetic information, genome editing systems, etc.).

Therefore, one of the important tasks of modern medicine is the development of approaches to further increase the effectiveness of lipid-containing nanoagents (or simply agents) in terms of drug delivery to targets in the body (patient, subject) for more effective diagnosis and therapy of various diseases and conditions. patients.

A key problem in this area is the short circulation time of agents in the bloodstream due to the rapid clearance of agents by the immune system. Under conditions of short circulation in the bloodstream, nanoagents do not have time to reach their target in sufficient quantities, which leads to a low efficiency of drug delivery and high systemic toxicity of the drug.

To solve this, a similar method is known (W02009065065 A1), in which the agent, which is a liposome, is coated with polyethylene glycol (PEG) molecules, and due to this, the particle surface becomes more hydrophilic and less "noticeable" for cells of the reticuloendothelial system (RES, or as it is also called the system of mononuclear phagocytes). RES cells are extremely efficient at removing nanoparticles from the bloodstream, reducing the circulation time of many types of particles to minutes, and as a consequence, the particles lose their theranostic efficiency, because, for example, from the point of view of targeted drug delivery, most of the particles are removed by the RES cells before they can at least once collide with their target. PEGylation of particles decreases nonspecific interactions of the agent, and also increases the time of its circulation in the bloodstream [Kojima et al., Dendrimer-based MRI contrast agents: the effects of PEGylation on relaxivity and pharmacokinetics. Nanomedicine. 2011], and the efficiency of agent delivery to the target increases, as well as the diagnostic and / or therapeutic effect produced by it.

The disadvantages of this method are that:

1) This approach may not be suitable for complex particles in which PEGylation would impair their essential function. For some agents, surface properties are a key component of their activity. Accordingly, although the circulation time of the agent during its PEGylation in the bloodstream will increase, its useful activity may deteriorate. For example, this can be due to steric hindrances that PEG molecules will create for receptor molecules that must bind to the target, or for particles whose behavior is associated with the variability of the composition of the outer shell [see. M.R. Nikitin, V.O. Shipunova, S.M. Deyev, P.l. Nikitin. Biocomputing based on particle disassembly, Nat Nanotechnol, 9 (9), 716-722 (2014)], or particles whose activity lies in the catalytic properties of the surface, etc.

2) The effect of PEGylation of many particles can be very low, and for some particles, PEGylation not only does not lead to an improvement in characteristics, but can also worsen them.

3) It is known that with repeated introduction of PEGylated particles into the body, the effect of increasing the circulation time of the particles can be significantly reduced [Zhao et al. Repeated injection of PEGylated solid lipid nanoparticles induces accelerated blood clearance in mice and beagles. Int J Nanomedicine. Vol.7, 2891-2900, 2012], and sometimes particles begin to be removed even faster than their non-PEGylated versions.

4) Among the PEGylated particles, it is necessary to distinguish "stele" (the so-called poorly visible by the body) agents, including PEGylated liposomes (for example, the clinically approved drug Kelix for the treatment of cancer), the half-life of which can reach tens of hours. However, this is still significantly less than the circulation time of molecular drugs, for example, monoclonal antibodies. Therefore, prolongation of circulation is required for such agents as well.

The closest method is known (WO2014039874 A2), in which the passive delivery of particles to monocytes (passive - in the sense that it is not mediated by any specific receptors) improves if, in addition to the agent, a fat emulsion (in particular, Intralipid ). Intralipid, when introduced into the bloodstream, saturates the cells of the reticuloendothelial system (in this case, mainly Kupffer's cells of the liver) and leads to their blockage (blockade) in terms of their phagocytic activity. Due to this, the nanoagent introduced after the Intralipid drug circulates in the bloodstream for a longer time and is absorbed by the monocytes / macrophages circulating in the bloodstream much more efficiently than without the administration of the Intralipid drug. Thus, it is possible to improve the delivery of the agent to monocytes, which can be used for the diagnosis and therapy of certain diseases.

The disadvantages of this method are that:

1) To block the RES, a very high dose of the injected emulsion is required - 2 g / kg. Such a dose of a foreign substance can have significant undesirable and negative effects on the body.

2) Even the introduction of such large doses of the emulsion gives only a weak effect of blocking the RES in the sense of increasing the circulation time of particles: by 3.1 and 2.5 times for non-solid nanoparticles and microparticles, respectively. 3) In this method, the agent is delivered only to actively phagocytic blood cells - monocytes and macrophages, and the delivery is due to the specific activity not of agents, but of target cells, whose task in the body itself is, among other things, to absorb foreign particles. Those. in this case, there is no targeting (targeted delivery) of certain cells due to the specific activity of the agent, for example, due to specific receptors (for example, antibodies) conjugated to the agent for surface markers of target cells, but only a change in the biodistribution of particles in organs and cells that foreign particles phagocytose in the body.

4) In this method, the introduction of a drug that blocks RES, significantly changes the composition of the blood of the body, incl. components of the drug - fats, being in the blood in high concentration, can interact with potential targets of the agent. This interaction can have an undesirable effect on the target or interfere with the interaction of the agent with the target, for example, masking them (if the target is cell surface markers), inhibiting phagocytosis of target cells and inhibiting the internalization of the agent into the target cell, etc. This, in turn, can significantly reduce the effectiveness of the agent, despite the increase in the time of its circulation in the bloodstream.

Thus, the required technical result consists in creating a method for increasing the diagnostic and therapeutic efficacy of agents, increasing the efficiency of agent delivery to the body and / or increasing the circulation time of agents (including those based on nano- and microparticles) in the bloodstream of the body (preferably applicable for increasing the effectiveness of the widest possible range of agents) by introducing into the body a minimum amount of potentially foreign and / or toxic objects, if possible, minimally changing the composition of the blood.

Description of the invention

To achieve the specified technical result, a composition (substance / pharmaceutical dosage form / mixture / kit) is proposed for use in a method of therapy or diagnosis of diseases or conditions of the body, including at least i) an antibody in free molecular form against the patient's blood cells, and i) a lipid-containing nanoagent (or - a drug or therapeutic agent, hereinafter, everything that is said about lipid-containing agents where applicable also applies to liposomal agents and vice versa).

In addition, a composition for improving the efficiency and / or reducing the side effects of the magnetic nanoagent, containing at least an antibody against blood cells.

In addition, a composition for improving the efficacy and / or reducing the side effects of a lipid-containing agent, comprising at least an anti-blood cell antibody.

In addition, a composition for improving the effectiveness and / or reducing the side effects of the liposomal agent, containing at least an antibody against blood cells.

In addition, a composition for improving the efficacy and / or reducing the side effects of a liposomal therapeutic agent, comprising at least an antibody against the patient's blood cells.

In addition, a composition for improving the efficacy and / or reducing side effects of a liposomal therapeutic agent administered in a separate dosage form from said composition contains at least an antibody against the patient's blood cells.

In addition, a composition for improving the efficacy and / or reducing the side effects of a liposomal therapeutic agent, comprising at least an antibody against the patient's blood cells. In addition, a collision to improve the effectiveness and / or reduce the side effects of a summer therapy, including the administration of a liposollal therapeutic preparation to a patient, is characterized by the fact that the collapse collapse contains, at least, an antibody against the patient's blood cells.

In addition, a collection for use in therapy or diagnostics, comprising, at least, i) an antibody in free llolecular forlle against the patient's blood cells, and ii) a liposollate therapeutic agent.

In addition, a collection for use in therapy or diagnostics with a HALF liposollal preparation, including at least an antibody in the free llolecular forel against the patient's blood cells.

In addition, the collapse in which the deposited liposollal therapeutic agent is long-circulating in the bloodstream, the vrell half-life of the agent from the bloodstream without administering the collapsed collapse is more than 10 lint, preferably more than 30 lint, preferably more than 1 hour, preferably more than 2 hours, preferably more than 3 hours preferably more than 5 hours, preferably more than 7 hours, preferably more than 10 hours, preferably more than 24 hours.

Moreover, a method for improving the efficacy of a liposollal therapeutic agent, comprising: 1) administering a liposollal therapeutic agent, and 2) administering an antibody against blood cells.

In addition, the method in which the extinguished antibody is administered in a dose effective to improve the efficacy of the extinguished agent.

In addition, the method in which the extinguished antibody is administered in a dose effective to improve the efficacy of the extinguished agent, the extinguished antibody alone does not or will show a therapeutic effect.

In addition, a method for improving the efficacy and / or reducing the side effects of a liposollal therapeutic agent, comprising: 1) administering a liposollal therapeutic agent, and 2) administering an abolished collision.

In addition, a collection for use in therapy or diagnostics, comprising, at least, i) an antibody in free llolecular forel against the patient's blood cells, and ii) a lipid-containing agent.

In addition, a phallacetic collation for use in summer therapy or diagnostics, comprising, at least, i) an antibody against the patient's blood cells, and ii) a lipid-containing agent, characterized by a lipid-containing agent (hereinafter in the number of illusions in the mean in the number of liposollal therapeutic agent) out of bounds with antibodies.

In addition, a collection for use in therapy or diagnostics, including, at least, i) an antibody against the patient's blood cells, and ii) an unbound lipid-containing agent, characterized by the fact that the antibody is (is) a substance in free llolecular forlle. In addition, a collection for use in therapy or diagnostics, comprising, at least, i) an antibody against the patient's blood cells, and ii) a lipid-containing agent, different from the fact that the lipid-containing agent is not associated with the antibodies ...

In addition, the collision, which is presented in the form of a single dosed forlla.

In addition, the collision in which the antibodies against erythrocytes are antibodies.

In addition, there is a collision in which the canceled antibody, when introduced into the body, induces erythrophagocytosis.

In addition, there is a collision in which the antibodies, when introduced into the organism, induce erythrophagocytosis, mediated by Fc-receptoroll.

In addition, a collision in which the antibodies in question is a HIGHLY LOGICAL VIDYLL antibodies organelle (or its variant), including autologous, allogeneic, gullanized or hillernull.

In addition, the collision in which the antibody in question is llonoclonalll antibodies.

In addition, the collision in which the antibody in question is polyclonal antibodies.

In addition, a collision in which the antibody in question is a conglomeration of several llonoclonal antibodies.

In addition, the collision in which the antibodies in question are antibodies isolated from donated blood.

In addition, the collision in which the anti-rhesus RhO (D) antibodies is.

In addition, the collision, in which the dose of the suppressed antibody is selected sufficient for the vrelle circulation in the bloodstream of the suppressed lipid-containing drug to be increased by no more than 30% compared to vrellenella, the circulation in the bloodstream of the suppressed lipid-containing drug without administration of the suppressed antibody ...

In addition, the collision in which the dose of the suppressed antibodies is selected sufficient so that the vrelle circulation in the bloodstream of the suppressed lipid-containing medicinal agent is increased by no more than 3 times compared to vrellenella circulation in the bloodstream of the suppressed lipid-containing drug without the introduction of the suppressed antibody ...

In addition, a collision in which the dose of the suppressed antibody is chosen sufficient so that the delivery efficiency of the suppressed agent to the redundant is increased by no more than 30% compared to the delivery efficiency of the suppressed lipid-containing drug without administration of the suppressed antibody.

In addition, a collision in which the dose of the suppressed antibody is chosen sufficient so that the delivery efficiency of the suppressed agent to the redundant is increased by no more than 3 times compared to the delivery efficiency of the suppressed lipid-containing drug without the administration of the suppressed antibody.

In addition, there is a collision in which the dose of antibodies to be taken does not exceed 5 LH / kg of body weight.

In addition, there is a collision in which the dose of the antibodies taken does not exceed 50 LH / kg body weight.

In addition, there is a collision in which the dose of the antibodies taken does not exceed 500 LH / kg body weight.

In addition, a collision, in which the antibodies and the lipid-containing medicinal agent are administered, are administered to the organism in separate dosed forls.

In addition, the collision, which is presented in the form of a single dosed forlla. In addition, there is a collision, which is characterized by the fact that the injected antibody and the injected lipid-containing medicinal agent are contained in a single dosed forl or in two different dosed forls.

In addition, a collision in which the extinguished antibody and the extinguished lipid-containing drug are introduced into different vrells.

In addition, a collision in which the collapsed antibody is administered prior to administration of the collapsed lipid-containing drug agent.

In addition, the collision in which the lipid-containing drug agent is administered is liposollalll agentoll.

In addition, the collision in which the lipid-containing medicinal agent is dropped is a liposollo.

In addition, the composition in which the lipid-containing drug agent is dropped is a PEGylated liposoll.

In addition, the collision in which the lipid-containing drug agent is administered is liposollalic doxorubicin.

In addition, the collision in which the lipid-containing drug agent is dropped is PEGylated liposollal doxorubicin.

In addition, a collision in which the agent to be copied consists of at least one of the following nano or microparticles: lagnitic, fluorescent, protein (including a cross-linked, polylated or aggregated protein), polylated, in t .h. consisting, at least, of one of the following pollers: polystyrene, dextran, polypeptide, glycolic acid polylactide, or block copolymers) or crystalline (gold, silver, semiconducting, or lettallic) nano- or llicparticles.

In addition, the collision, in which the superimposed agent performs the visualizing function due to the product of the detecting signal, incl. at least one of the following: fluorescent, lullinescent, PET signal, MPT-contrast signal, X-ray contrast, lagnitic signal or signal due to plasllon resonance, or signal due to absorption of light or other electromagnet or acoustic waves.

In addition, a collision in which the agent performs a therapeutic function, for example, due to the delivery of cytostatic or cytotoxic compounds, or medicinal compounds, incl. low molecular weight, fertile, radioactive, heliotherapeutic, substances for photodynallic therapy, hyperterllia.

In addition, the collision is for use in a method of treating cancer.

In addition, the collision is for use in a method of treating a disease or condition selected from the group consisting of: cancer, atherosclerosis, stroke, heart attack.

Kroll, in addition, the colposition for use in the method of conducting angiocontrast.

In addition, the composition for use in a method for the treatment of cancer, characterized by the fact that first the canceled antibody is administered, followed by the administration of the canceled lipid-containing drug agent.

In addition, the collapse, which additionally contains red blood cells.

In addition, there is a collection that additionally contains a collection of donated blood.

In addition, there is a collection that additionally contains autologous blood collectors.

In addition, a collection that additionally contains an erythropoiesis stiller. In addition, collapse, which additionally contains erythropoietin.

Moreover, it is a method of diagnosing or treating diseases or conditions of the organism, into which the collapsed collection is introduced into the organism.

In addition, a method for diagnosing or treating diseases or conditions of the organism, in which i) the collapsed antibody COLLECTION is introduced into the organism, ii) the collapsed lipid-containing drug collision is introduced into the organism.

In addition, a method, characterized by the fact that the agent in question is administered after no more than 10 minutes, preferably not more than 20 minutes, preferably not more than 30 minutes, preferably not more than 45 minutes, preferably not more than 1 hour, preferably not more than 2 hours, preferably not more than 3 hours, preferably not more than 6 hours, preferably not more than 12 hours, after administration of the antibody.

In addition, a method characterized by tell that the agent is administered after no more than 21 days, preferably no more than 14 days, preferably no more than 7 days, preferably no more than 2 days, preferably no more than 24 hours, preferably no more than 18 hours, preferably no more than 12 hours after the injection of the antibodies.

In addition, a method characterized by the fact that the agent is administered no later than 6 hours and no more than 18 hours after the administration of the controlled antibody.

In addition, the method, characterized by the fact that the injection of the canceled antibody into the organism increases the efficiency of the diagnostic or therapeutic action of the canceled agent in comparison with the introduction of the canceled agent without the injection of the canceled antibody.

In addition, a method, characterized by the fact that the administration of a controlled antibody into the organism increases the vrell circulation of the controlled agent in the organism's bloodstream.

Kroll, in addition, a method, which is characterized by the fact that, along with the introduction of an erythrocyte substance and an erythrocyte agent, erythrocytes are introduced into the organism.

In addition, the method is characterized by the fact that, along with the introduction of the expelled substance and the expelled agent, the blood of the sally organelle or its counterparts or donor blood or its counterparts are injected into the organelle.

In addition, a method, characterized by the fact that, along with the introduction of the expelled substance and the expelled agent, a collision is introduced into the organism for the stilling of erythropoiesis.

Kroll addition, a method for the diagnosis or therapy of cancer.

Kroll, in addition, the method of using collapsed collations for the manufacture of drugs for the treatment of cancer.

In addition, it is a method of promoting products to the market, which includes, at least, the promotion for scientific research or diagnostics, monitoring or therapy of diseases or conditions of the organism, at least of the collated collision or method.

In addition, the method of promoting products to the market, including, at least, promoting the delivery or improvement of the delivery of nucleic acids (polynucleotides) (gene therapy, delivery of nucleic acids and, in general, all purposes of COLLECTIONS and methods described in this description), at the latest, collated collision or collated method.

In addition, a llet, characterized by a tell that the advancement is carried out by inserts in a package with a collection collection or its collponentalli (substanceally), containing, at least ller, a collection according to paragraphs. 1-39. In addition, it is a llet, characterized by the tell that the promotion is carried out by written or oral communication to the doctor or medical service provider.

In addition, a business lease that includes, at least, marketing for diagnostics and therapy, or for improving the effectiveness of diagnostics or therapy, or for delivering an agent to an organism, or for increasing the timing of circulation of an agent in the bloodstream of an organism of a given collection or method.

In addition, a business case involving, at least, marketing to deliver or improve the delivery of nucleic acids (polynucleotides) to an organism or to increase the circulation of an agent in the bloodstream of an organism in a given collection or method.

In addition, a collision to increase the diagnostic or therapeutic efficacy of a lipid-containing medicinal agent injected into the organism, which includes, at least, a collponent, which, when a collision is introduced into the organism, promotes the removal of the reticuloendothelial sistella from the bloodstream at the least, of objects, circulating in the bloodstream, but not being artificially created nano- or llylico-particlesalli or opsoninalli, which binds to the collaponentoll nonspecifically, the canceled excretion of the collapsed objects causes, at least, a partial blockage of the reticuloendothelial systtelle.

With etoll, it is podrazulleli that the blockage does not occur due to the saturation of the RES, only the collapse of the collponentol - the collponent only promotes the removal of the collapsed objects from the bloodstream in the body. It is preferable that the collponent is a separate substanceall, which is capable of interfering with an objectalli or objectalli, or, preferably, binding with an objectalli objectalli, or undergoing a chyllic or biochillic, preferably ferventative, reaction in a LLLI.

In this case, facilitating the elimination of the collponent from the bloodstream means that if a collapse was introduced without such a collponent, the elimination of the collapsed objects from the bloodstream would be more glacial, more preferable much more glacial (several times), or even more preferable there would be no blockage of the RES.

In addition, a collision for the delivery of a lipid-containing drug into the body, which includes, at least, a collponent, which, when introduced into the body, promotes the removal of the reticuloendothelial systella from the bloodstream at the end of the body, objects circulating in the bloodstream, but not which are artificially created nano- or llylico-particlesalli or opsoninalli, which bind nonspecifically to canceled collponentoll, the canceled removal of canceled objects causes, at least, partial blockage of the reticuloendothelial sistella.

In addition, a collision is characterized by the tell that the object is collapsed or the object is cells.

In addition, the collapse is characterized by the tell that the object is controlled by the erythrocytes.

In addition, the collision is characterized by troll cells.

In addition, a collision, characterized by the tell that the object is controlled by the leukocytes.

In addition, a collection characterized by the tell that the object is controlled or the object is llolecules. When etoll is treated, it is important that these llolecules do not act exclusively in the role of opsonins, but there is an additional interaction between collponentoll and collapsingly objectally, because it is clear that if a collponent, for example, is some kind of artificial particle, then its introduction will almost always be accompanied by the elimination of opsonins from the bloodstream (various llolecules, such as illunoglobulins, collplent sistella proteins, etc., which nonspecifically bind to these particles and mediate their phagocytosis of the cells of the RES). In addition, there is a collision, which is distinguished by the tell that the collapsed objects are part of the COLLECTION, but are not part of the collateral.

In addition, a collection characterized by the tell that the object is controlled are the cells or llolecules of the sally organism.

In addition, there is a collision, which is characterized by the fact that the collapsed objects are removed from free circulation in the bloodstream.

Under free circulation in the bloodstream, the circulation in the free-flow form decreases, for example, if the erythrocyte is absorbed by the llacrophagoll circulating in the bloodstream, then the red blood cell is considered to be removed from the free circulation in the bloodstream.

In addition, a collision, characterized by the tell that the object is destroyed or the object, is the cells that did not leave the organism.

In addition, there is a collision, which is characterized by the tell that the object is controlled or the cells or molecules that have entered the organism are artificial putells.

In addition, a collection, which is characterized by the tell that the object is controlled or the object, are cells or llolecules that have entered the organelle of the transfusion of blood or blood donor blood collectors.

In addition, a collision, characterized by the tell that the object is controlled or the object, are cells or llolecules that are part of the COLLECTION, BUT are not collponentoll.

In addition, there is a collapse, which is characterized by the fact that the collapsed collponent specifically interacts with the collapse of the object, and this interaction leads to their collapse of the excretion of the reticuloendothelial sistella.

In addition, a collision that is distinguished by the fact that the collated action of the collapsed collateral with the collapsed object is the linking of the collateral collateral with the collapsed collateral object.

In addition, a collision, which is distinguished by the tell that the collapsed collateral acts in a non-covalent manner with the collapsed object.

In addition, a collection characterized by the fact that the collapsed collateral forms a specific collplex with the collapsed object, due to direct or indirect recognition and attachment to the collapsed object.

In addition, a collection characterized by the fact that the collapsed collateral consists, at least ller, of an antibody, which forms a collapsed specific collplex from the collapsed object, due to direct or indirect recognition and attachment to the collapsed object.

In addition, a collision characterized by the fact that the accomplished recognition and attachment of the collapsed or indirectly collapsed collponent to the collapsed object causes predominantly phagocytosis of the collapsed objects of the organism's cell, mediated by the Fc receptoroll.

In addition, there is a collision, characterized by the fact that the antibodies are targeted against the organelle's erythrocytes or troll cells or their allogeneic variants.

In addition, a collision that is distinguished by the fact that the collateralized collponent consists, at least ller, of a highly colological type of antibodies of the organism of an antibody (or its variant), incl. autologous, allogeneic, gullanized or chiller antibodies.

In addition, a collision is characterized by the fact that the collapsed collponent consists, at least ller, of a lonoclonal antibody.

In addition, a collision, characterized by the fact that the collapsed collateral promotes the elimination of collapsed objects, which are the cells that make up the COLLECTION, by recognizing the collapsed cells and promoting their phagocytosis by the reticuloendothelial sistella. In addition, a collision is characterized by the fact that the collateralized collponent consists, at least, of several antibodies targeting several different types of cells or llolecules of the organism.

In addition, a collision, which is distinguished by the fact that the collapsed collponent consists, at least of several, at least of lonoclonal, or at least of highly colological types of antibodies of antibodies (or of their collisions and variants), incl. autologous, allogeneic, gullanized or chiller antibodies, against organelle cells or llolecules or their allogeneic analogs.

In addition, a collision, which is characterized by the fact that the collapsed component consists, at least of all, of an antibody combined with another million of substances, including, in the number of molecules, particles, cells or their associations.

In addition, there is a collision, characterized by the fact that the antibodies are targeted against the organell's erythrocytes.

In addition, there is a collision, characterized by the fact that the antibodies are targeted against the organell's troll cells.

In addition, there is a collision, characterized by the fact that the suppressed antibodies are directed against the organelle's leukocytes.

In addition, a collision, which is characterized by the fact that the accomplished recognition and attachment of the collapsed or indirectly collapsed collateral to the collapsed object causes phagocytosis of the collapsed objects in other cells of the organism.

In addition, a collision, characterized by the fact that the controlled excretion of the reticuloendothelial sistella of the collapsed objects is caused by their agglutination under the action of the collapsed collponent of the collapse.

In addition, a collision, characterized by the fact that the controlled excretion of the reticuloendothelial sistella of the collapsed objects is caused by their aging under the action of the collapsed collponent of the collapse.

In addition, a collision, characterized by the fact that the controlled excretion of the reticuloendothelial sistella of the collapsed objects is caused by their damage to the cells under the action of the collapsed collateral of the collapse.

In addition, a collision, characterized by the fact that the controlled excretion of the reticuloendothelial sistella of the collapsed objects is caused by their llodification under the action of the collapsed collponent of the collapse.

In addition, a collision, characterized by the fact that the collapsed collateral causes an increase in the phagocytic activity of the cells of the reticuloendothelial sistella of the organism, which remove the collapsed objects from the bloodstream.

In addition, a collision, which is characterized by the fact that the collapsed component is a hyllicell compound, llolecule, particle or cell (including a bacterium, leukocytol, etc.) or a combination of such objects, but does not contain erythrocytes.

In addition, a collision, characterized by the fact that the collapsed component is a hyllicell compound, a molecule, a particle or a cell (including a bacterium, leukocytol, etc.) or a combination of such objects, but does not contain objects obtained by any or lmodification of erythrocytes.

Moreover, there is a collapse that is different from the fact that the collapsed collponent is different from the erythrocyte or lodified erythrocyte.

In addition, collapse, the introduction of which into the organism leads to a reduced blockage of the reticuloendothelial sistella, so that the vrell circulation of the reduced lipid-containing drug in the bloodstream increases. In addition, a collision, the introduction of which into the organism leads to a reduced blockage of the reticuloendothelial sistella, so that the vrelle elimination half-life (or vrelle elimination) of the expelled agent from the circulation in the bloodstream increases no more than the number of cells by 1.2, preferably by 1.5, preferably by 1.75, preferably by 2, more preferably 3, more preferably 5, more preferably 7, more preferably 10, more preferably 15, more preferably 20, and even more preferably 25 times the elimination half-life of the eliminated agent without collision.

In addition, collapse, the introduction of which into the organism leads to a canceled blockage of the reticuloendothelial sistella, so that the vrelle elimination half-life of the eliminated agent from the circulation in the bloodstream increases no more than 5 times compared to the vrellenella elimination half-life of the eliminated agent without the introduction of collapse.

In addition, there is a collapse for which the canceled blockage of the reticuloendothelial sistella is of a permanent nature.

In addition, there is a collision that is characterized by the fact that the class of the collateral does not exceed 20% of the class of objects that cause blockage of the reticuloendothelial sistella.

In addition, a collection characterized by a tell that the class of the collector does not exceed 75%, preferably 50%, even more preferably 25%, even more preferably 10%, even more preferably 1%, even more preferably 0.1%, even more preferably 0.01% and even more preferably 0.001% of Classes of objects excreted by the reticuloendothelial sistella from the bloodstream to ensure blocking of the RES. When ETHOLL is under a cell, for example, it is difficult to lower the cell with or without the amount of water they contain.

In addition, a collision characterized by the fact that in its composition the dose of active colponents, the presence of which causes a blockage of the reticuloendothelial sistella, does not exceed 5 lg / kg of body weight.

In addition, a collision that is characterized by tellthat in its composition the dose of active colponents, the presence of which causes blockage of the reticuloendothelial sistella, does not exceed, at least one of the following doses: 5, 2.5, 1, 0.5, 0.25, 0.1, 0.05 , 0.025, 0.01, 0.005, 0.0025, 0.0015 g / kg body weight.

In addition, a collision, the introduction of which into the organism increases the efficiency of passive or directed delivery of the canceled agent to the target, at least 2 times, more preferably 3, more preferably 5, more preferable 7, and even more preferable 10 times in comparison with cases delivery of the collapsed agent to the client without collision.

In addition, a collision that is distinguished by the fact that the collateralized collponent consists, at least ller, of llonoclonal or, at least ller, highly colological type of antibodies organelle antibodies, incl. autologous, allogeneic, gullanized, or hiller antibodies (or their variants) capable of specifically recognizing and binding spontaneously or indirectly to cells or llolecules circulating in the bloodstream (naturally circulating or introduced from outside).

In addition, there is a collision, characterized by the fact that the antibodies are combined with other objects, including, in the number of molecules, particles, cells or their associations.

In addition, there is a collision, which is characterized by the fact that the administration of antibodies causes agglutination of cells or llolecules.

In addition, a collision, characterized by the fact that the accomplished recognition and attachment of the suppressed antibodies, either indirectly or indirectly to the cells, causes the phagocytosis of the suppressed cells.

In addition, a collision characterized by the fact that the controlled recognition and attachment either directly or indirectly to the cells of the collapsed antibodies causes predominantly Fc-mediated phagocytosis of the collapsed cells. Moreover, a collision, the introduction of which into the organism increases the diagnostic or therapeutic efficacy of the extinguished agent by at least 1.3 times as compared to the case of delivery of the expelled agent to the organelle without the introduction of a collision.

In addition, the collision for which the lipid-containing medicinal agent is reduced in number of at least one of the following nano or microparticles: lagnitic, fluorescent, protein (including a cross-linked, polymerized or aggregated protein), polylactic (including polystyrene, dextran, polypeptide, polylactide glycolic acid or other polylactic and block copolymers, etc.) or crystalline (gold, silver, semiconductor, etc.) nano- or llic-particles.

In addition, the collision for which the agent is nano- or microparticles (including particles including nano- or microparticles).

In addition, a collision, characterized by a tell that the agent being controlled is a nano- or microparticle with an average value of one roll of radiosity (i.e., taking into account the variance) exceeding 10 nll, or 19 NLL, or 29 NLL, or 29 NLL, or 49 NLL, or 74 NLL, or 99 NLL, or 199 NLL, or 299 NLL, or 399 NLL, or 499 NLL, or 599 NLL, or 699 NLL, or 799 NLL, or 899 NLL, or 999 NLL, or 1299 NLL, or 1499 NLL, or 1999 NLL, or 2499 NLL, or 2999 NLL.

In addition, a collision, characterized by a tell that the agent is a nano- or microparticle with an average depletion in one-roll of eradication (i.e., taking into account the variance) not exceeding 10 nll, or 19 NLL, or 29 NLL, or 29 NLL, or 49 NLL, or 74 NLL, or 99 NLL, or 199 NLL, or 299 NLL, or 399 NLL, or 499 NLL, or 599 NLL, or 699 NLL, or 799 NLL, or 899 NLL, or 999 NLL, or 1299 NLL , or 1499 NLL, or 1999 NLL, or 2499 NLL, or 2999 NLL.

In addition, a collision, characterized by the fact that the agent used is used to diagnose or treat diseases or conditions of the organism, including one or more diseases from the following list: cancer, atherosclerosis, stroke, heart attack, incl. including diagnostics of diseases or conditions of the organism using angiocontrast.

In addition, a collision, which is distinguished by the tell that the agent under control performs a visualizing function due to the product of a detecting signal, incl. at least one of the following list: fluorescent, lullinescent, PET signal, MPT-contrasting signal, ultrasound-contrasting, X-ray contrast, lagnitny signal or signal due to plasllon resonance or another method based on the absorption of agentoll light, electrolagnet, acoustic waves (signals), etc.

In addition, a collision, characterized by the fact that the agent in question performs a therapeutic function, for example, by delivering agents for hyperterllia (lagnitic or gold particles), cytostatic or cytotoxic compounds, or medicinal compounds, incl. low molecular weight, fertile, radioactive, heliotherapeutic, substances for photodynallic therapy.

In addition, a collision to increase the diagnostic or therapeutic effect of INSERTED into the organism of the agent includes, at least, a collponent, which, when introduced into the organism, causes, at least, a partial blockage of the reticuloendothelial sistella of the cell.

In addition, there is a collision for the delivery of an agent into the organism, which includes, at least, a collponent, which, when introduced into the organism, causes, at least, a partial blockage of the reticuloendothelial sistella of the cell.

In this case, under the blocking of RES, the cellcalli podrazulleli that the organs of the RES excrete a significant amount of cells that have been "blocked", with ETOLL it is clear that a poll of these RES cells displays other objects as well (since this is one of the main functions of cells RES). With ETOLL, for example, in a one-roll embodiment of the invention, the introduction of COLLECTION significantly increases the number of REMOVED cells (in comparison with cases without the introduction of composition), and their removal plays a significant role in the saturation of the RES. In an advantageous embodiment of the invention, the number of cells removed from the bloodstream in a certain period of time (10 minutes, 1 hour, 3 hours, 6 hours, 9 hours, 12 hours, 15 hours, 18 hours, 24 hours, 2 days, 3 days, 4 days , 5 days, 7 days, or 10 days) significantly increases (by 1.5, 2, 3, 4, 5, 7, 10, 20, 30, 40, 50, 75 or more times) compared to the number of RES cells removed from the bloodstream cells without the introduction of the composition.

In addition, a composition characterized in that said component consists at least of antibodies highly homologous to the specific antibodies of the body against said cells, which provide said blocking of the reticuloendothelial system, incl. an autologous, allogeneic, humanized, or chimeric antibody.

In addition, the composition, characterized in that the said component leads to increased excretion of cells circulating in the bloodstream by the reticuloendothelial system, thereby achieving the aforementioned at least partial blocking of the reticuloendothelial system by cells.

In addition, a composition characterized in that said component contains said cells, which block, at least partially, the reticuloendothelial system, due to their rapid removal from the circulation in the bloodstream, sufficient to ensure said blocking of the reticuloendothelial system.

In addition, a composition characterized in that said cells causing said blockage of the reticuloendothelial system are erythrocytes.

In addition, a composition characterized in that said cells causing said blockage of the reticuloendothelial system are platelets.

In addition, a composition characterized in that said cells that cause said blockage of the reticuloendothelial system are leukocytes.

In addition, a composition characterized in that said cells are cells introduced into an organism by transfusion of a donor's or organism's material.

In addition, a composition characterized in that said cells are treated variants of body cells or allogeneic cells.

In addition, a composition characterized in that said treated variants of body cells or allogeneic cells are cells, including those incubated with specific antibodies, that recognize and bind directly or indirectly to said variants of body cells or allogeneic cells.

In addition, the composition, characterized in that the said specific antibodies are highly homologous to the specific antibodies of the body, antibodies (or their variants), incl. autologous, allogeneic, humanized or chimeric antibodies.

In addition, a composition characterized in that said specific antibodies are monoclonal antibodies.

In addition, a composition characterized in that said cells are part of said component.

In addition, a composition characterized in that said cells are cells that have not left the body.

In addition, a composition characterized in that the said component specifically interacts with the cells circulating in the bloodstream, and this interaction leads to their excretion from the bloodstream by the reticuloendothelial system, which leads to the aforementioned blockage of the reticuloendothelial system. In addition, a collision, which is characterized by the fact that the suppressed interaction of the collapsed component with the cells circulating in the bloodstream is the binding of the collateral cells with the collapsed cells.

In addition, there is a collapse, which is characterized by the fact that the collapsed collponent interacts with the cells circulating in the bloodstream non-covalently.

In addition, a collision characterized by the tell that the collapsed collponent forms a specific collplex with the collapsed cage due to direct or indirect recognition and adhesion to the collapsed cage.

In addition, a collision characterized by the fact that the collapsed collponent consists, at least, of an antibody that forms a collapsed specific collplex from the collapsed cell due to direct or indirect recognition and attachment to the collapsed cell.

In addition, there is a collision, characterized by the fact that the target antibody is directed against the organelle's red blood cells.

In addition, a collision characterized by the fact that the accomplished recognition and attachment of the collapsed or indirectly collapsed collateral to the cells causes phagocytosis of the collapsed cells of the other cells of the cells of the organism.

In addition, a collision, characterized by the fact that the accomplished recognition and attachment of a proliferated or indirectly collapsed collponent to the cells, causes predominantly phagocytosis of the collapsed cells of other cells of the cells of the organism, mediated by the Fc receptoroll.

In addition, a collision, characterized by the fact that the collapsed cells are agglutinated cells, and either are part of the collapsed collateral, or agglutinated under the action of the collapsed collateral.

In addition, a collision, characterized by the fact that the collapsed cells are damaged cells, and either are part of the collapsed collateral or are damaged under the action of the collapsed collateral.

Moreover, there is a collision, which is characterized by the fact that the destroyed cells are lodified cells and either are part of the canceled colponent, or are llodified under the action of the canceled colponent.

Moreover, there is a collision that is characterized by the fact that the canceled lodified cells are fertilely lodified, incl. with a band of neurallinidase, trypsin, galactose oxidase, etc.

Moreover, there is a collection that is characterized by the fact that the destroyed lodified cells are chyllically modified, incl. with a mixture of glutaraldehyde, hydrogen peroxide.

In addition, there is a collision that is characterized by the fact that the collapsed cells are old cells and either are part of the collapsed collateral or age under the influence of the collapsed collateral.

In addition, a collision is characterized by the fact that the collapsed collateral consists, at least, of several antibodies targeting several different types of organelle cells or their allogeneic variants.

In addition, a collection that is characterized by the fact that the collateralized collateral consists, at least of several, of several, at least of, lonoclonal, or at least of high-level, of antibodies of the body of antibodies (or their combinations and variants), incl. autologous, allogeneic, humanized or chimeric antibodies against body cells or their allogeneic variants.

In addition, a composition characterized in that said component is different from an erythrocyte or a modified erythrocyte.

In addition, a composition characterized in that said agent is nano- or microparticles (including particles including nano- or microparticles).

In addition, a composition characterized in that said agent is a nano- or microparticle with a size in one of the dimensions exceeding 10 nm.

In addition, a composition, the introduction of which into the body leads to the said blocking of the reticuloendothelial system so that the half-life (or elimination time) of the said agent from circulation in the bloodstream increases by at least 1.2, preferably 1.5, preferably 1.75, more preferably 2, more preferably 3, more preferably 5, more preferably 7, more preferably 10, more preferably 15, more preferably 20, and even more preferably 25 times the half-life of said agent without administration of the composition.

In addition, the composition, characterized in that the dose of active components in its composition, the presence of which causes blockage (or one way or another leads to blockage) of the reticuloendothelial system, does not exceed 5 mg / kg of body weight.

In addition, a composition characterized in that the dose of active components in its composition, the presence of which causes blockage (or one way or another leads to blockage) of the reticuloendothelial system, does not exceed at least one of the following doses: 5, 2.5, 1, 0.5, 0.25, 0.1, 0.05, 0.025, 0.01, 0.005, 0.0025, 0.0015 g / kg of body weight.

In addition, the composition, the introduction of which into the body increases the efficiency of delivery (passive or directed) of the said agent to the target, at least 2 times, more preferably 3, more preferably 5, more preferably 7, and even more preferable 10 times compared to with the case of delivery of the said agent to the target without administration of the composition.

In addition, a composition, the introduction of which into the body increases the diagnostic or therapeutic efficacy of the said agent, at least 1.3 times in comparison with the case of delivery of the agent to the target without administration of the composition.

In addition, a composition characterized in that said agent consists of at least one of the following nano or microparticles: magnetic, fluorescent, protein (including a cross-linked, polymerized or aggregated protein), polymeric (from polystyrene, dextran, polypeptide, polylactide glycolic acid, etc.) or crystalline (gold, silver, semiconductor, etc.) nano- or microparticles.

In addition, a composition characterized in that said agent is used for the diagnosis or therapy of diseases or conditions of the body, including one or more diseases from the following list: cancer, atherosclerosis, stroke, heart attack, incl. including diagnostics of diseases or conditions of the body using angiocontrast.

In addition, a composition characterized in that said agent performs a visualizing function by producing a detected signal, incl. at least one of the following: fluorescent, fluorescent, PET, MPT contrast, radiopaque, magnetic, or plasmon resonance

In addition, a composition characterized in that said agent performs a therapeutic function, for example, by delivering cytostatic or cytotoxic compounds or medicinal compounds to targets, incl. low molecular weight, enzymatic, radioactive, chemotherapeutic, substances for photodynamic therapy, hyperthermia. In addition, a collision, introduced into the organism in order to increase the diagnostic or therapeutic effect of the agent and containing, at least, highly colologic vidyll antibodies of the organism antibodies against erythrocytes or trollbocytes (or their variants), incl. autologous, allogeneic, gullanized, or chiller antibodies.

In addition, a collision containing, in number, erythrocytes or trollbocytes.

In addition, there is a collision, characterized by the fact that the antibodies are associated with erythrocytals or trollbocytals of the organelle or their allogennally variantally.

In addition, a collision introduced into the organism in order to increase the diagnostic or therapeutic effect of the agent or to deliver the agent or to increase the circulation time of the agent in the bloodstream, and containing, at least, highly colologic VIDOVYLL antibodies of the organism, antibodies against erythrocytes or trollbocytes (or their variants) , incl. autologous, allogeneic, gullanized, or chiller antibodies.

In addition, a collision introduced into the organism in order to increase the diagnostic or therapeutic effect of the agent and containing, at least, highly colologic VIDOVYLL antibodies of the organism antibodies against erythrocytes or troll cells (or their variants), incl. autologous, allogeneic, gullanized, or chiller antibodies.

In addition, a collision for the delivery of an agent and containing, at least ller, highly colologic VIDOVILL antibodies or antibodies against erythrocytes or troblocytes (or their variant), incl. autologous, allogeneic, gullanized, or chiller antibodies.

In addition, a collision to increase the timing of the circulation of the agent in the bloodstream and containing, at least, highly colologic VIDOVILL antibodies of the organelle antibodies against erythrocytes or troll cells (or their variantalli), incl. autologous, allogeneic, gullanized, or chiller antibodies.

In addition, a method for diagnosing or treating diseases or conditions of the organism, in which: i) one of the above COLLISIONS is introduced into the organism, at least, ii) before or after the introduction of the collapsed collision, or simultaneously with the collapse of the collision, an agent is introduced mediating the diagnostic or therapeutic action.

Kroll of the method, which is characterized by tell, is that the agent is administered no later than 6 hours and no more than 18 hours after the administration of the copied collection.

Kroll of the method, characterized by the fact that the agent is injected after no more than 11 hours and no more than 13 hours after the introduction of the collapse.

Kroll of the method, characterized by tell that the agent to be administered is administered no later than 20 days, preferably 15 days, preferably 10 days, preferably 7 days, preferably 5 days, preferably 4 days, preferably 3 days, preferably 2 days, preferably 1 day, and even more preferably 15 hours after the introduction of the collated collection.

Kroll of the method, characterized by the fact that the controlled introduction into the organism of the canceled collision increases the effectiveness of the diagnostic or therapeutic action of the canceled agent by increasing the timing of the circulation of the canceled agent in the organism's bloodstream.

Kroll is a method characterized by the fact that, along with the introduction of the canceled collapse and the canceled agent, a reducing agent is introduced into the organism, which causes an increase (in comparison with the cases without the administration of the canceled reducing agent) in the level of cells or llolecules, the removal of which from the bloodstream causes the canceled collapse of the canceled collapse.

Kroll of the method, characterized by the fact that the reducing agent used is the blood of the Sally organelle or its colponentally or donated blood or its colponentally. In addition, a method characterized by the fact that the reducing agent used is erythropoietinol.

In addition, a collposition for delivery of a lipid-containing drug to the organism and / or increasing the diagnostic or therapeutic efficacy of a lipid-containing drug, and including, at least, a collponent or colponents, which i), when introduced into the organism, lead to augmentednoll the elimination of cells or llolecules of the organism from the circulation in the bloodstream, or ii) when introduced into the organism, lead to an increased excretion from the circulation in the bloodstream of cells or llolecules introduced into the organism, or iii) sally are cells, which are rapidly eliminated from the circulation after the introduction of COLLPOSITION into the organism in the bloodstream, in these cases, the increased or rapid elimination of the canceled cells or llolecules from the circulation in the bloodstream leads to an increase in the timing of the circulation of the canceled agent in the bloodstream.

In addition, a collposition for delivery of a lipid-containing drug to the organism and / or facilitating the delivery of a lipid-containing drug to the organism and / or for increasing the diagnostic or therapeutic efficacy of the agent, and including, at least, a collponent or colponents that i) when introduced into the organism lead to enhanced Nolla excretion of cells or llolecules of the organism from the circulation in the bloodstream, or ii) when introduced into the organism lead to enhanced Nolla excretion of cells or llolecules introduced into the body from circulation in the bloodstream, or iii) Sally are cells or llolecules after the introduction of COLLPOSITIONS in the body, they are quickly removed from the circulation in the bloodstream, and in these cases, the increased or rapid removal of the canceled cells or llolecules from the circulation in the bloodstream leads to an increase in the timing of the circulation of the canceled agent in the bloodstream.

In addition, the method of delivery of a lipid-containing drug into the organism, into which: i) inject into the organism, at least one of the above COLLECTIONS, ii) before or after the introduction of the collapsed collection or simultaneously with the introduction of the collapsed collision, the collapsed agent is administered, if it is is not a collponentoll collection.

Kroll is a method, which is characterized by the fact that the agent to be suppressed mediates a diagnostic or therapeutic effect.

Kroll of the method, which is characterized by the fact that the agent is injected after no more than 4, 7, 13, 30, 50, 70, 100, 125, 150, 200, 300, 400, 500, 600, 700, 800, 900 or 1000 hours after the introduction of the canceled collection.

Kroll of the method, characterized by the fact that, along with the introduction of the canceled collapse and the canceled agent, a reducing agent is introduced into the organism, which causes an increase (in comparison with the cases without the introduction of the canceled reducing agent) the level of cells or llolecules, the enhanced or rapid removal of which from the bloodstream causes the canceled collagen collapsed.

Kroll of the method, characterized by the fact that the reduced reducing agent is the blood (or its colponentally) of a sally organelle or donor.

In addition, a method characterized by the fact that the reducing agent used is erythropoietinol.

In addition, a collision introduced into the organism in order to increase the circulation of the lipid-containing drug in the bloodstream of the organism, containing at least: i) component 1, which, when introduced into the body, leads to an increased elimination from circulation in the bloodstream of erythrocytes or platelets that have entered the bloodstream either naturally or artificially, compared to the rate of their removal in the case without the introduction of the said component, or ii) component 2 , consisting at least of an antibody (or its variants) highly homologous to the specific antibodies of the organism, incl. an autologous, allogeneic, humanized or chimeric antibody capable of forming, directly or indirectly, a specific complex with objects that are naturally in the body or that have entered the body artificially.

In this case, the mentioned objects can be molecules, particles, cells, as well as their various variants or their combinations, incl. complexes.

In addition, a composition characterized in that said antibody of said component 2 is a gamma immunoglobulin.

In one embodiment of the invention, said objects may be cells. Cells that entered the body (or the bloodstream) naturally mean cells of the body itself, which were naturally created in the body and did not leave the body, while cells that entered the body artificially mean cells that were introduced into the body, including ... cells of the body that were isolated from the body and then injected back into it. The same is true for other objects, such as molecules.

In addition, a composition characterized in that said erythrocytes or platelets that have entered the bloodstream artificially, or said objects that have entered the body artificially, have been introduced into the body by transfusion of material from a donor or body material.

In one embodiment of the method, said material introduced into the body by transfusion is the blood of a donor or the body itself or its components.

In addition, a composition characterized in that said erythrocytes or platelets that have entered the bloodstream artificially, or said objects that have entered the body artificially, are a component of said composition.

In addition, a composition characterized in that said erythrocytes or platelets that have entered the bloodstream by artificial means are part of said component 1 of the composition.

In addition, the composition, characterized in that the said erythrocytes or platelets, which are intensively removed from the circulation in the bloodstream, are treated variants of body cells or allogeneic cells, either being one of the components of the composition, or entering the body artificially.

In addition, a composition characterized in that said treatment of variants of body cells or allogeneic cells includes incubation with specific antibodies that recognize and bind directly or indirectly to said variants of body cells or allogeneic cells.

In addition, a composition characterized in that said objects are erythrocytes.

In addition, a composition characterized in that said objects are leukocytes.

In addition, a composition characterized in that said objects are platelets.

In addition, a composition characterized in that said objects are albumin.

In addition, a composition characterized in that said objects are immunoglobulins. In addition, a collection that is characterized by the tell that the object is killed and the object has entered the bloodstream of the artificial putell are the object that has been introduced into the organism.

In addition, a collision, characterized by the fact that the collapsed collateral 1 or the collapsed collponent 2 leads mainly to the enhanced collapse of the removal of erythrocytes or trolbocytes from the collapsed circulation, which did not leave the organism.

In addition, a collision characterized by the tell that the collapsed collateral 1 or collapsed collponent 2 specifically interacts with the collapsed erythrocytal or trollbocytalli or object, and this interaction leads to the reinforced noll removal of the collapsed or erythrocyte objects from the circulation.

In addition, a collision, characterized by the fact that the collapsed action of the collapsed collateral 1 or the collapsed collponent 2 from the collapsed erythrocytalli or trollbocytalli or objectalli is the binding of the collponent ells with the collapsed or trollbocytalli object.

In addition, there is a collision, characterized by the fact that the collapsed collateral 1 or the collapsed collponent 2 interacts with the erythrocytals or trollbocytals or objects circulating in the bloodstream non-covalently.

In addition, a collision characterized by the tell that the collapsed binding of the collapsed collateral 1 or collapsed collateral 2 with the collapsed erythrocytal or trollbocytalli or objectalli is non-covalent.

Moreover, there is a collision, which is characterized by the fact that the agent in question is nano- or microparticles (including particles including nano- or microparticles).

In addition, a collision, characterized by the fact that the agent being tested is a nano- or microparticle with a single roll of radiation exceeding 10 nill.

In addition, there is a collapse, characterized by the fact that the increased excretion of erythrocytes or trollbocytes is caused by their aging under the action of the canceled colponent 1.

In addition, a collision, characterized by the fact that the suppressed increased excretion of erythrocytes or troll cells or objects is caused by their agglutination under the action of the collapsed collateral 1 or collapsed collateral 2 of the collision.

In addition, there is a collapse, which is characterized by the fact that the increased excretion of erythrocytes or trollbocytes is caused by their damage to the cells under the action of the canceled colponent 1.

In addition, collapse, characterized by the fact that the collapsed collponent 1 or collponent 2 causes an increase in the phagocytic activity of the organelle cells, which leads to an enhanced collapse of the collapsed erythrocytes or trolbocytes or objects from the circulation.

In addition, a collapse characterized by the fact that an increase in the time of circulation of the extinguished agent is caused, at least, by a partial blockage of the reticuloendothelial sistella of the organism due to the removal of erythrocytes or trolley cells or objects from the circulation in the bloodstream.

In addition, a collection that is characterized by the fact that the collateralized collponent 1 consists, at least of all, of a highly colological type of antibodies of the organism of antibodies (or its variants), incl. an autologous, allogeneic, gullanized, or hillerized antibody capable of specifically recognizing and binding spindle or indirectly to erythrocytal or trollbocytal.

In addition, a collision, characterized by the fact that the collapsed collponent 1 forms a specific collplex with erythrocytal or trollbocytal due to direct or indirect recognition and attachment of erythrocytal or trollbocytall to the collapse. In addition, a collision, which is distinguished by the fact that the copied antibody of the collapsed collateral 2 forms a collapsed specific collplex with the collapsed object, due to direct or indirect recognition and attachment to the collapsed object.

In addition, there is a collision, characterized by the fact that the antibodies are targeted against the organelle's erythrocytes or their allogeneic counterparts.

In addition, there is a collision, characterized by the fact that the antibodies are targeted against the organell's troll cells or their allogeneic counterparts.

In addition, there is a collision, characterized by the fact that the antibodies are targeted against the organelle's leukocytes or their allogeneic counterparts.

In addition, there is a collision, which is characterized by the fact that the introduction of the collapsed antibodies causes the agglutination of the collapsed objects.

In addition, a collision characterized by the tell that the collapsed recognition and attachment of the collapsed or indirectly collapsed collponent 1 or collapsed collponent 2 to the erythrocytal or trollbocytall or objectall causes phagocytosis of the collapsed erythrocytes or trollbocyte objects.

In addition, a collision characterized by the tell that the predicted recognition and attachment of the proliferated or indirectly collapsed collponent 1 or the collapsed collponent 2 to the erythrocytal or trollbocytal or objectall causes predominantly phagocytosis of the objects collapsed to the erythrocytal-mediated cells or organisms of the receptor-collapsed cells or organisms

In addition, a collision characterized by the tell that the collapsed collateral 1 or the collapsed collponent 2 is a hyllicell compound, llolecule, particle or cell (including bacteria, leukocytoll, etc.) or a combination of such objects, but does not contain erythrocytes.

In addition, a collision characterized by the tell that the collapsed collateral 1 or the collapsed collponent 2 is a hyllicell compound, llolecule, particle or cell (including bacteria, leukocytoll, etc.) or a combination of such objects, but does not contain objects obtained by any modification of erythrocytes.

In addition, a collection that is distinguished by the fact that the collapsed collateral 1 or the collapsed collponent 2 consists, at least ller, of several highly collological types of antibodies of the organism of antibodies (or their variants), incl. autologous, allogeneic, gullanized, or chiller antibodies.

In addition, a collision, which is characterized by the fact that the collapsed antibodies of the collapsed collateral 2 are llonoclonal antibodies, or the collapsed collponent 1 consists, at least, of the collapsed collponent antibodies.

In addition, a collision, characterized by the fact that the collapsed antibody of collapsed collateral 2 is combined with other substances, including, in the number of lololecules, particles, cells or their associations, or in which collapsed collponent 1 or collapsed collponent 2 consists, at least , from an antibody combined with another million substances, including, in the number of molecules, particles, cells, or their combinations.

In addition, a collection that is distinguished by the fact that the collapsed collateral 1 or the collapsed collponent 2 consists, at least ller, of several highly collological types of antibodies of the organism of antibodies (or their variants), incl. autologous, allogeneic, gullanized or chiller antibodies directed against organelle cells or llolecules or their allogeneic variants.

In addition, a collision characterized by the tell that the collapsed collponent 1 or the collapsed collponent 2 consists, at least, of a collplex of a substance (including llolecules, cells, particles) with a cell or lloleculally isolated from the organism or their allogeneically analogous. In addition, a collection that is distinguished by the fact that the collapsed collateral 1 or the collapsed collponent 2 consists, at least, of the collplex of llolecules or cells (or their allogeneic variants) isolated from the organelle and associated spindle or indirectly with LLLI antibodies.

In addition, a collection that is distinguished by the fact that the collapsed collponent 1 or the collapsed collponent 2 consists, at least, of a collplex of llolecules or cells (or their allogeneic variants) isolated from the organelle and associated pryallo or indirectly with NILLI highly gallially vidyll vidyll antitellar or their variants), incl. autologous, allogenic, gullanized, or hillernally antibodies.

In addition, a collection characterized by the fact that the collapsed collponent 1 or the collapsed collponent 2 consists, at least, of a collplex of llolecules or cells (or their allogeneic variants) isolated from the organelle and highly-logical types of antibodies of the organism of antibodies (or their variants), .h. autologous, allogeneic, gullanized or chiller antibodies, against data of all llolecules or cells (or their variants).

In addition, a collection characterized by the telltale of collponentoll 1 or collaponentoll 2 are collapses of cells or llolecules isolated from the organelle or their allogeneic analogs specifically associated with antibodies against erythrocytes or trollbocytes.

In addition, a collision characterized by the fact that the collapsed collponent 1 or the collapsed collponent 2 consists, at least, of a collplex of a substance (llolecules, cells, particles, etc.) non-covalently connected by a specific biololecular bond with the erythrocyltol ( or their allogenic analogue).

In addition, a collection that is distinguished by the fact that the collapsed collateral 1 or the collapsed collponent 2 consists, at least ller, of a highly colloquial type of antibodies of the organism of antibodies (or its variants), incl. an autologous, allogeneic, gullanized or hillerized antibody capable of specifically recognizing and binding spindle or indirectly to cells or llolecules circulating in the bloodstream (naturally circulating or artificially introduced).

In addition, the collision, which is characterized by the fact that the antibodies that are capable of specifically recognizing and binding either indirectly or indirectly to the cells or llolecules circulating in the bloodstream, is the llonoclonalnyl antibodies.

In addition, a collision, characterized by the fact that the antibodies (or variants thereof), capable of specifically recognizing and adhering spontaneously or indirectly to cells or llolecules circulating in the bloodstream, are connected to other millions of substances, including, in toll, llolecules, particles, cells or their associations.

In addition, there is a collision, characterized by the fact that the suppressed antibodies specifically interacts with the cellalli or llolekulalli of the organelle, and this interaction leads to the enhanced excretion of the expelled cells or llolecules of the organelle from the circulation.

In addition, a collision, which is distinguished by the fact that the collapsed collateral 1 or the collapsed collponent 2 consists, at least, of several, at least lonoclonal, or at least highly colological types of antibodies of the organism of antibodies (or their collbations and variants), in t. h. autologous, allogeneic, gullanized or chiller antibodies, against organelle cells or llolecules or their allogeneic analogs. Naturally, here and in other similar cases, it is meant that the antibody lays down to be directed against, for example, cellular llarkers of these cells, certain epitopes of cellular llarkers, etc.

In addition, a collision that is distinguished by the fact that the collapsed collateral 1 or the collapsed collponent 2 consists at least of several, at least lonoclonal, or at least vysokogolologicheskogo VIDOVILL antibodies of organelle antibodies (or their collections and variants). h. autologous, allogeneic, gullanized or chiller antibodies against different cells or llolecules of the organelle or their allogeneic analogs. In addition, the collapse, which is different, is that the collapsed collponent 1 is different from the erythrocyte or lodified erythrocyte.

In addition, collapse, the introduction of which into the organism leads to an increase in the circulation time of the extinguished agent in the bloodstream, so that the vrelle elimination half-life of the expelled agent from the expelled circulation increases no more than 3 times compared to the vrellenella elimination half-life of the expelled agent without the introduction of the collapse.

Moreover, a collision, the introduction of which into the organism leads to an increase in the rate of circulation of the extinguished agent in the bloodstream so that the vrelle elimination half-life (or vrelle elimination) of the agent from the circulation in the bloodstream increases no more than the number of people by 1.2, preferably 1.5, preferably 1.75, more preferable 2, more preferably 3, more preferably 5, more preferably 10, more preferably 12.5, more preferably 15, more preferably 20, and even more preferably 25 times the half-life of the agent without administration

COLLPOSITIONS.

In addition, collapse, the introduction of which into the organism leads to an increase in the circulation time of the eliminated agent in the bloodstream, while phenollene increases the circulation time of the eliminated agent is of a permanent nature.

Moreover, a collision, the introduction of which into the organism increases the vrelle circulation of the agent, the introduction of which, in turn, is used for the diagnosis or monitoring or therapy of a disease, while an increase in the circulation time of the canceled agent increases the efficiency of the diagnosis or therapy of the disease.

In addition, a method for increasing the circulation of a lipid-containing drug in the organelle's bloodstream, into which: i) is injected into the organelle, at least one of the above COLLEGES, ii) is injected into the expelled organelle.

In addition to the way in which the administration of the agent is used for diagnostics or monitoring or therapy, an increase in the timing of the circulation of the suppressed agent in the bloodstream of the organism increases the efficiency of the diagnosis or therapy.

Kroll of the method, which differs from the tell, that the dose of the reduced colposition is selected when it is introduced into the body to increase the reduced circulation rate by the required number of times, optillizing the beneficial effect of increasing the circulation rate, and POSSIBLE negative effects (e.g., lowering the concentration of llolecules or blood cells, withdrawal organally RES).

Kroll of the method, which is characterized by tell, is that the dose of the reduced collapse is selected when it is introduced into the body to increase the reduced circulation rate by the necessary number of times, optimizing the beneficial effect of increasing the circulation time, and POSSIBLE negative effects.

Kroll of the method, differing tell, that the introduction of the collapsed collision or its variants is repeated many times.

Kroll of the method, differing tell, that the introduction of the collapsed collision or its variants is repeated many times. Under a variantoll of a collision, a collision is subdivided that is different in composition from the collated one, but it is also a pre-bulletin of the present invention by its own indications.

Kroll of the method, which is characterized by the fact that, along with the introduction of the canceled agent and the canceled collision, a reducing agent is introduced into the organism, causing an increase in the level of cells or llolecules, which are intensively removed from the circulation in the bloodstream under the action of the canceled collapse, in comparison with Trowells without the introduction of the canceled reducing agent ...

Kroll of the method, characterized by the fact that, along with the introduction of the canceled agent and the canceled collision, a reducing agent is introduced into the organism, causing an increase in the level erythrocytes or platelets in the blood compared with the level without the administration of the said reducing agent.

In addition, a method characterized in that said reducing agent is the blood of the organism itself or its components or donor blood or its components.

In addition, a method characterized in that said reducing agent is erythropoietin.

In addition, a composition introduced into the body in order to increase the diagnostic or therapeutic effect of the agent introduced into the body, and including at least a component that, when introduced into the body, leads to at least partial blockage of the reticuloendothelial system due to active excretion it from the circulation in the bloodstream of at least one of the following substances: i) cells or molecules of the organism itself, or ii) cells or molecules that have entered the body artificially, but not included in the composition, or iii) cells included in the composition compositions, incl. included in the mentioned component.

In addition, a composition for increasing the diagnostic or therapeutic effect of a lipid-containing drug agent includes at least a component that, when the composition is administered to the body, promotes the removal of the reticulo-endothelial system from the circulation in the bloodstream, at least of objects that are cells or molecules, if the mentioned molecules are not excreted from the bloodstream mainly as nonspecifically interacting with the aforementioned component of opsonins, and the said removal of the mentioned objects from the circulation in the bloodstream leads to at least partial blockage of the reticuloendothelial system.

In addition, a composition introduced into the body in order to increase the diagnostic or therapeutic effect of a lipid-containing medicinal agent, and including at least a component that, when introduced into the body, causes active excretion (promotes excretion) from circulation in the bloodstream of body objects or objects introduced into the body regardless of the introduction of the composition, or the cells that make up said component,

In addition, a composition introduced into the body in order to increase the diagnostic or therapeutic effect of a lipid-containing medicinal agent, and including at least a component that, when introduced into the body, causes active elimination (promotes elimination) from circulation in the bloodstream (cells organism or cells that make up the said composition, or objects introduced into the body, regardless of the introduction of the composition)

In addition, a composition introduced into the body in order to increase the diagnostic or therapeutic effect of a lipid-containing drug agent, and comprising at least a component that, when introduced into the body, causes at least partial blockage of the reticuloendothelial system, at least , one of the following substances: cells or molecules capable of forming a specific complex with the said component.

In addition, a composition for increasing the diagnostic or therapeutic effect of a lipid-containing drug agent, and comprising at least a component that, when the composition is introduced into the body, promotes the removal of objects from the circulation in the bloodstream, including cells or molecules, if the said molecules are not excreted mainly as opsonins that interact nonspecifically with the mentioned component, and said excretion leads to at least partial blockage of the reticuloendothelial system

In addition, a composition introduced into the body in order to increase the diagnostic or therapeutic effect of a lipid-containing drug or to deliver the agent or to increase the circulation time of the agent in the bloodstream, and containing at least highly homologous to the species antibodies of the body antibodies against erythrocytes or thrombocytes (or their options), incl. autologous, allogeneic, humanized or chimeric antibodies.

In addition, a composition introduced into the body in order to increase the diagnostic or therapeutic effect of a lipid-containing medicinal agent and containing at least highly homologous to the specific antibodies of the body antibodies against erythrocytes or thrombocytes (or their variants), incl. autologous, allogeneic, humanized or chimeric antibodies.

In addition, a composition for delivery of a lipid-containing drug into the body and containing at least highly homologous to the specific antibodies of the body antibodies against erythrocytes or thrombocytes (or their variants), incl. autologous, allogeneic, humanized or chimeric antibodies.

In addition, a composition for increasing the circulation time of a lipid-containing medicinal agent in the bloodstream and containing at least highly homologous to the specific antibodies of the body antibodies against erythrocytes or thrombocytes (or their variants), incl. autologous, allogeneic, humanized or chimeric antibodies.

In addition, a method for increasing the circulation time of lipid-containing medicinal agents in the bloodstream of the body due to the introduction into the said living organism of the said agent along with the composition, the introduction of which leads to increased removal of erythrocytes or platelets from the circulation in the bloodstream in comparison with the rate of their removal in the case of without the introduction of the composition, and the composition is different from erythrocytes.

In addition, a method for increasing the circulation time of a lipid-containing drug in the bloodstream of an organism due to the introduction into said living organism of the said agent along with a composition, the introduction of which leads to an increased removal of erythrocytes or platelets from the circulation in the bloodstream in comparison with the rate of their removal in the case of without the introduction of the composition, and the composition is different from modified in any way erythrocytes, incl. destroyed erythrocytes.

In addition, a method of promoting a product to the market, including at least promotion for the diagnosis, monitoring or therapy of diseases or conditions of the body, or for scientific research, at least one of the mentioned compositions or methods.

In addition, a method characterized in that the promotion is carried out by an insert in a package with a commercial composition containing at least one of the mentioned compositions.

In addition, a method characterized in that the promotion is carried out by written or oral communication to the doctor or health care provider.

In addition, the method, characterized in that the promotion is carried out by posting information on the Internet.

In addition, the method, characterized in that the promotion is carried out by publication on the Internet.

In addition, a method, characterized in that the promotion is carried out by publishing on a corporate page on the Internet.

In addition, a method of promoting products to the market, including at least promotion for the diagnosis, monitoring or therapy of diseases or conditions of the body, or for scientific research of an agent, nanoagent, nanoparticle, substance, composition, device, device, method (and etc.) as products that improve their properties (or show improved characteristics / possessing improved or advanced characteristics) when introduced into an organism or as part of one of the completed collections or in rallies of one of the above methods.

In addition, a business llet that includes, at least, marketing for diagnostics and therapy, or to improve the effectiveness of diagnostics or therapy, or to deliver an agent to the organism, or to increase the timing of circulation of an agent in the organism's bloodstream, at least one of the collapsed collections and ways.

In addition, a business lease that includes, at least, marketing for the diagnosis, monitoring or therapy of diseases or conditions of the organism, or for the scientific research of an agent, nanoagent, nanoparticle, substance, COLLECTION, device, device, summer medicine (etc. .) as a product that improves its properties (or shows improved characteristics / has improved or advanced characteristics) when introduced into an organism or as part of one of the approved collections or in rallies of one of the approved methods.

In addition, a composition for improving the magnetic delivery of a therapeutic agent, characterized in that said composition contains at least an antibody against the patient's blood cells.

In addition, a collision for improving the lagnitic delivery of a therapeutic agent, characterized by the fact that the collated collation contains, at least, an antibody against the patient's blood cells.

In addition, a collision for improving the effectiveness and / or reducing the side effects of a therapeutic agent containing lagnitic nanoparticles differs in that the collated collation contains, at least, an antibody against the patient's blood cells.

In addition, a collision for improving the effectiveness of an agent containing lagnitic nanoparticles, characterized by the fact that the collated collation contains, at least, an antibody against the patient's blood cells.

In addition, there is a collection for improving the effectiveness of a lagnitic nanoagent, which is distinguished by the fact that the collated collection contains, at least, an antibody against the patient's blood cells.

In addition, a collection for use in therapy or diagnostics, comprising, at least, i) an antibody in free llolecular forel against the patient's blood cells, and ii) a therapeutic agent containing at least a lagnitic nanoparticle.

In addition, the collision in which the agent to be used contains a lagnitic particle from the following group of laterals: iron oxide; iron oxide; doped one of the following elements: nickel, cobalt, zinc, llarganese, lignium; iron oxide, doped with ONE or several other or several elelentally; iron oxide coated with a gold shell.

In addition, a method for diagnosing or treating diseases or conditions of the organism, in which: i) the collapsed collapse is introduced into the organism, the extinguished therapeutic agent is introduced into the organism, a gradient of the lagnitic field is created in the area of the extinguished therapeutic agent.

In addition, a method of lactic delivery of a therapeutic agent (therapeutic lagnitic agent / lagnitic particle / agent containing a lagnitic nanoparticle) to lagnitism inside the organism, which consists in: i) introducing an antibody against the patient's blood cells into the organism (subject / patient), ii) introducing into organelle of the expelled therapeutic agent, iii) creation of a lagnitic field in the area of the expelled organelle.

In addition, the way in which the antibodies in question are located (is the antibodies) in the free llolecular forlle. Moreover, a method for diagnosing or treating diseases or conditions of the organism, in which: i) the collapsed collapse is introduced into the organism, ii) the extinguished therapeutic agent is introduced into the organism, iii) a lagnitic field is created in the region of the collapsed organism.

In addition, a method for diagnosing or treating diseases or conditions of the organism, in which: i) the extinguished COLLECTION antibody is introduced into the organism, ii) the extinguished therapeutic agent of the collision is introduced into the organism, iii) a lagnitic field is created in the region of the extinguished organism.

In addition, the way in which the antibodies in question are located (is the antibodies) in the free llolecular forlle.

In addition, a composition in which said antibody is an anti-CD47 antibody.

In addition, there is a collision in which the antibody in question recognizes (binds, specifically to) the CD47 receptor.

In addition, the collision further comprises an anti-CD47 antibody.

In addition, the collection additionally includes a P (ab) -fragment anti-CD47 antibody.

In addition, the collection additionally includes a P (ab) 2 fragment of an anti-CD47 antibody.

In addition, the collection additionally includes an anti-CD47 antibody fragment without an Fc fragment.

In addition, the collision further comprises an anti-CD47 antibody lacking an Fc fragment.

In addition, the collision is an additional inhibitor of the interaction of the CD47 receptor with the SIRPa ligandol.

In addition, the collision additionally includes a llolecule that blocks the interaction of CD47 with the SIRPa ligandol.

In addition, the collection additionally includes blood cells associated with ONLY or several killer compounds from the group: anti-CD47 antibody, P (ab) -fraglent antibodies against CD47, P (ab) 2-fragment of antibodies against CD47, fragment of anti-CD47 antibodies without Fc-fragment, anti-CD47 antibody fragment without Fc-fragment, anti-CD47 antibody lacking Fc-fragment, inhibitor of CD47 receptor interaction with SIRPa ligandol, llolecule, blocking CD47 interaction with SIRPa ligandoll.

In addition, a method further comprising introducing into the organism (patient) one or more compounds (substances) from the group: anti-CD47 antibody, P (ab) -fragment antibodies against CD47, P (ab) 2 -fragment of anti-CD47 antibodies, fragment of anti-CD47 antibody without Fc-fragment, fragment of anti-CD47 antibody without Fc-fragment, anti-CD47 antibody devoid of Fc-fragment, inhibitor of the interaction of the CD47 receptor with the SIRPa ligandol, llolecule that blocks the interaction of CD47 with the SIRP ligandoll.

In addition, a composition for improving the efficacy and / or reducing the side effects of a therapeutic agent used to deliver a heterologous polynucleotide, containing at least an antibody against the patient's blood cells.

In addition, a collision to improve the efficiency of delivery of genetic information into an organism with an agent containing, at least, an antibody against the patient's blood cells. In addition, there is a collision for which the predicted improvement in delivery efficiency is an increase in delivery efficiency to a certain extent.

In addition, a collision to improve the efficiency of delivery of genetic information into an organism with an agent containing, at least, an antibody against the patient's blood cells.

In addition, a collision for delivery of a polynucleotide to an organism (or delivery of a polynucleotide encoding a therapeutic gene product to an organism) (or targeted delivery of a gene / polynucleotide) (or delivery of a polynucleotide encoding a therapeutic gene product to an organism), containing, at least, an antibody against the organism's blood cells.

In addition, a collision for delivery of a polynucleotide to an organism (or delivery of a polynucleotide encoding a therapeutic gene product to an organism) (or targeted delivery of a gene / polynucleotide) (or delivery of a polynucleotide encoding a therapeutic gene product to an organism), containing at least:

1) an antibody against the organism's blood cells,

2) an agent containing the polynucleotide.

In addition, a collection for the treatment of a disease (or for the treatment of a disease or condition of the organism, caused by a decrease in the disease or an increase in the level of expression, functional activity or lutation of the protein), containing, at least, an antibody against the blood cells of the organism.

In addition, a collision for the treatment of a disease (or for the treatment of a disease or condition of the organism caused by a decrease in the disease or an increase in the level of expression, functional activity or lutation of the protein), containing at least:

1) an antibody against the organism's blood cells,

2) an agent containing a polynucleotide, which is transcribed into a nucleic acid or encodes a collapsed protein or its analogue (including a peptide), which therapeutically provides the functional activity of the collapsed protein, restores the activity level of the collapsed protein, or brings the activity level of the collapsed protein back to normal.

In addition, a collision in which the agent being dropped is a liposolle containing the dropped heterologous polynucleotide.

In addition, the collision in which the agent to be deposited includes a lipid.

In addition, a collision in which the collapsed agent is a collplex that includes the collapsed heterologous polynucleotide and a cationic lipid.

In addition, a collision in which the collapsed agent is a collplex, which includes the collapsed heterologous polynucleotide and a positively charged poller.

In addition, the collision in which the agent used includes polyethylenillin.

In addition, a collection in which the agent to be used contains a dendriller.

In addition, a collection in which the agent to be copied contains at least one of the following nano or microparticles: lagnitic, fluorescent, protein (including a cross-linked, polymerized or aggregated protein), polylated, incl. consisting, at least, of one of the following pollers: polystyrene, dextran, polypeptide, glycolic acid polylactide, or block copolymers) or crystalline (gold, silver, semiconductor, or letallic) nano- or llic-particles.

In addition, the collision in which the abolished agent includes a virus.

In addition, a collision in which the agent to be dropped includes a viral vector from the group: adeno-associated virus, adenovirus, lentivirus, herpesvirus, alphavirus. In addition, the collision in which the agent being collapsed comprises an adeno-associated virus vector that includes a capsid protein, a gollologous capsid protein sequence selected from the group consisting of: AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9 , AAV10, AAV-DJ.

In addition, a collision, in which the agent being dropped, additionally performs a visualizing function due to the product of a detecting signal, incl. at least one of the following: fluorescent, lullinescent, PET signal, MPT-contrast signal, X-ray contrast, lagnitic signal or signal due to plasllon resonance, or signal due to absorption of light or other electromagnet or acoustic waves.

In addition, the collision, in which the agent used includes additional therapeutic substances from the group: cytostatic compounds, cytotoxic compounds, low molecular weight medicinal compounds, ferlents, radioactive compounds, heliotherapeutic compounds, substances for photodynamic therapy, a substance for hypertellia.

In addition, a collision in which the overlaid heterologous polynucleotide encodes a protein.

In addition, a collision in which a heterologous polynucleotide encodes a nucleic acid that suppresses protein synthesis in the body.

In addition, a collision in which the heterologous polynucleotide is an antisense oligonucleotidol.

In addition, a collision in which the heterologous polynucleotide is RNA.

In addition, there is a collision in which the heterologous polynucleotide is the llal of the interfering RNA.

In addition, a collision in which the heterologous polynucleotide is llycroRNA.

In addition, a collision in which the predicted delivery of a heterologous polynucleotide is used to treat a disease selected from the group: cancer, lung, kidney, liver disease, blood clotting, diabetes, stroke, heart attack, Alzheiller's disease, Parkinson's disease, Huntington's disease, hellophilia, alliotrophilic lateral sclerosis, Gaucher's disease, Pollpe's disease, impaired lledi accumulation, impaired iron accumulation, infectious disease, viral disease, bacterial infection, sepsis, hereditary disease, orphan disease.

In addition, a collision in which a heterologous polynucleotide encodes a protein or peptide selected from the group: insulin, hormones, including growth hormone, luteinizing throat, angiopoietins, angiostatin, erythropoietin, growth factors, trollbopoietin, interleukins, blood factors, histocompatibility receptors.

In addition, a collision in which a heterologous polynucleotide encodes, at least ller, a genoll editing fertile.

In addition, a collision in which a heterologous polynucleotide encodes, at least ller, a genoll editing nuclease.

In addition, a collision in which the heterologous polynucleotide encodes, at least, the Cas9 nuclease.

In addition, there is a collision in which the antibodies, when introduced into the organism, induce erythrophagocytosis, mediated by Fc-receptoroll. In addition, a collision in which the antibodies in question is a HIGHLY LOGICAL VIDYLL antibodies organelle (or its variant), including autologous, allogeneic, gullanized or hillernull.

In addition, the collision in which the anti-rhesus RhO (D) antibodies is.

In addition, a collision in which the dose of the suppressed antibody is chosen sufficient so that the vrelle circulation in the bloodstream of the suppressed therapeutic agent is increased by 30% compared to the vrellenella circulation in the bloodstream of the suppressed therapeutic agent without the introduction of the suppressed antibody.

In addition, the collision in which the dose of the suppressed antibody is chosen sufficient so that the vrelle circulation in the bloodstream of the suppressed therapeutic agent is increased no more than three times compared to the vrellenella circulation in the bloodstream of the suppressed therapeutic agent without the introduction of the suppressed antibody.

In addition, a collision in which the dose of the suppressed antibody is chosen sufficient so that the delivery efficiency of the suppressed agent to the target is no less increased by 30% compared to the delivery efficiency of the suppressed therapeutic agent without the administration of the suppressed antibody.

In addition, the collision, in which the dose of the suppressed antibody is selected sufficient to increase the delivery efficiency of the suppressed agent to the target is no less than 3 times compared to the delivery efficiency of the suppressed therapeutic agent without the administration of the suppressed antibody.

In addition, there is a collision in which the dose of antibodies taken up does not exceed 0.1 LH / kg of body weight.

In addition, there is a collision in which the dose of antibodies to be taken does not exceed 1 LH / kg of body weight.

In addition, there is a collision in which the dose of the antibodies taken up does not exceed 10 LH / kg of body weight.

In addition, there is a collision in which the dose of antibodies to be consumed does not exceed 100 LH / kg body weight.

In addition, there is a collision in which the dose of the antibodies consumed does not exceed 1000 LH / kg body weight.

In addition, a collision in which the extinguished antibody and the extinguished therapeutic agent are administered to the organism in separate dosed forls.

In addition, the collision, in which it is presented in the form of a single dosed forlla.

In addition, the collation, which is presented in the form of a single dosed forlla, is in the free llolecular forlla.

In addition, there is a collision, differing in the tell that the injected antibody and the injected therapeutic agent are contained in a single dosed forl or in two different dosed forls.

In addition, a collision in which the suppressed antibody and the suppressed therapeutic agent are injected into a different vrell.

In addition, a collision in which the extinguished antibody is administered prior to the administration of the extinguished therapeutic agent. In addition, the collision in which the therapeutic agent is administered is a PEGylated liposollo.

In addition, the collapse, which additionally contains red blood cells.

In addition, a collection that additionally contains an erythropoiesis stiller.

In addition, collapse, which additionally contains erythropoietin.

In addition, the collision is for use in a method of treating cancer.

In addition, the composition for use in a method for the treatment of cancer, characterized by the fact that first the collapsed antibody is administered, followed by the administration of the collapsed lipid-containing drug agent.

In addition, a method for improving the efficacy and / or reducing the side effects of a therapeutic agent used (intended) for delivery of a heterologous polynucleotide, comprising:

1) administration of the controlled therapeutic agent, and

2) the introduction of a collapsed collection.

In addition, the method in which the extinguished antibody is administered in a dose effective to improve the effectiveness of the extinguished agent, the extinguished antibody alone does not or will show a therapeutic effect.

In addition, a method characterized by the fact that the agent is administered no later than 6 hours after the administration of the antibody.

In addition, the method, characterized by the fact that the agent is administered no more than 18 hours after the administration of the antibody.

In addition, a method characterized by the fact that, along with the introduction of the antibodies and the antibodies, erythrocytes are injected into the organism.

In addition, a method characterized by the fact that, along with the introduction of the antibodies and the antibodies, the organelle is injected with the blood of the organelle or its counterparts or donated blood or its counterparts.

In addition, a method characterized by the fact that, along with the introduction of the antibodies and the antibodies in the body, a collision is introduced into the organism for the stilling of erythropoiesis.

Kroll addition, a method for the diagnosis or therapy of cancer.

In addition, the method further includes the step of quantifying the blockage of the reticuloendothelial sistella, which includes administering a controlling agent and irradiating its concentration in the bloodstream after a specified time interval to calculate the blockage characteristic of the reticuloendothelial systella. In addition, the method further includes the step of radiating a quantitative characteristic of the blockage of the reticuloendothelial sistella, which includes: a) the step of administering a controlling agent before the controlled administration of the suppressed antibodies and irradiating its concentration in the bloodstream after a certain time interval to calculate the characteristic of blocking the reticulo- endothelial sistella. b) the step of administering the controlling agent after the controlled administration of the suppressed antibodies and eradication of its concentration in the bloodstream after a certain time interval to calculate the characteristic of blocking the reticuloendothelial sistella.

In addition, a method of delivering a polynucleotide to an organism (or a method of delivering a polynucleotide encoding a therapeutic gene product to an organism) to an organism:

1) injecting an antibody against blood cells,

2) an agent containing the polynucleotide is introduced.

In addition, a method of directed delivery of a gene (polynucleotide) to an organism (or a method of delivering a polynucleotide encoding a therapeutic gene product to an organism), into which:

1) an antibody against blood cells is introduced into the organism,

2) the agent containing the polynucleotide is introduced into the organism.

In addition, a method of treating a disease (or for treating a disease or condition of the organism caused by a decrease in the disease or an increase in the level of expression, functional activity or lutation of a protein), in which:

1) an antibody against the organelle's blood cells is injected into the organism,

Kroll, in order to achieve the specified technical result, COLLECTIONS and methods are proposed that have a collapse of the above characteristics. In addition, it should be noted that in all cases of collapse, for example, cells or llolecules can be subranged, for example, both the organelle's cells and llolecules, and allogeneic cells, or variants of organelle cells or allogeneic cells.

The characteristics and advantages of the collations and methods according to this invention will be further illustrated in the following detailed description. While the manufacture and use of various embodiments of the present invention are discussed in detail below, it should be understood that the present invention provides a variety of illustrative inventive concepts that can be embodied in a wide range of specific contexts. The specific embodiments discussed herein merely illustrate various ways of making and using the invention and do not limit the scope of the invention. All content provided herein (hereinafter and hereinafter) is incorporated herein by reference in its entirety for all purposes.

Dictionary

To facilitate understanding of the essence of this invention, here (earlier and further in the ETOLL text) the following terllins are used:

Terllin "nanoparticles" and "nanoagents" subranges supra-molecular structures, which consist of more than one llolecule. With ETHOLL, although nanoparticles often denote objects up to 100 nll, in the descriptions of the present invention, objects with large 100 nll people are also reframed - preferably up to 5LLKLL, but in some cases the razller was allowed up to UOnll. Taquill scrambled, under nanoparticles include both nanoparticles and microparticles, microspheres, and the like. In addition, particles with the mentioned dimensions are meant, both in all dimensions, and in at least one.

Free molecular form - a state of a molecule in which it is predominantly not bound (does not form a complex / is not associated) with other molecules or objects (cells / nanoparticles), preferably not covalently bound. In particular, within the framework of the present invention, it is meant that said antibody is not associated with said lipid-containing agent. In the case of non-covalent interactions, in a statistical sense, the standard understanding of "binding" is implied, i.e. the presence of a sufficiently high affinity constant. Within the framework of this invention, it is understood that an increase in the efficiency of delivery of an agent to a target or an increase in circulation of an agent in the bloodstream is not a consequence of the formation of an associate of said antibody with said agent (and the hitch-hiking agent expected in such a case on blood cells).

Therapeutic (or drug) molecules / agents. Therapeutic molecules / agents can be understood as substances, substances, molecules, supramolecular agents, nanoparticles, microparticles, nanoagents, microagents, formulations, dosage forms, etc., which are used both for therapy and diagnosis of various diseases or conditions of the body. or a patient, incl. include labels, contrast agents, controlled release components, paramagnetic and magnetic agents, cytotoxic agents, and the like. In addition, in one embodiment of the invention, the said composition is used to improve the effectiveness of agents, nanoparticles and microparticles for carrying out various scientific studies, for example, for biovimaging of tumors, angiocontrast, MRI and CT contrasting of various organs, structures, delivery of various probes, toxins and other known problems using nano- and micro-sized objects. In this case, the “therapeutic” function of the agent is interpreted not from the point of view of clinical significance, but also from the point of view of research activities using nanoagents (etc.) as means of nanomedicine. In this case, an improvement in the effectiveness of the agent can be the achievement of the same result, but with a lower dose of the agent, higher quantitative indicators (contrast enhancement, detection limit, etc.) are achieved.

A "label" or "detectable element" is a composition detectable by spectroscopic, photochemical, biochemical, immunochemical, chemical, or other physical means. For example, commonly used labels include 32P, fluorescent dyes, electron dense reagents, enzymes (for example, as commonly used in ELISA), biotin, digoxigenin, or haptens, and proteins or other entities that can be detected, for example, by incorporating radioactive peptide or antibody tags that specifically react with the target peptide. Any method known in the art for conjugating a labeled antibody can be used, for example, using the methods described in (Hermanson, G. T., Bioconjugate Techniques, Academic press, 2013).

As used herein, the term "pharmaceutical" refers to a composition that is useful in treating a disease or symptom of a disease.

The term "treatment", "therapy", "diagnosis", "treatment with" (treating, treatment, therapy) refers to any indication of success in the treatment or improvement of a disease, injury, pathology or condition, including any objective or subjective parameters such as decrease; remission; reducing symptoms or slowing down the deterioration of the patient's condition, pathology or patient's condition; slowing down the rate of degeneration; what makes the final stage of the disease less painful; improving the physical or mental well-being of the patient. Treatment or improvement of symptoms can be based on objective or subjective parameters; including results of physical examination, neuropsychiatric examinations, and / or psychiatric evaluation. For example, certain methods presented here can successfully treat cancer by reducing cancer incidence and inducing cancer remission. The term "treatment" and its continuation includes the prevention of injury, pathology, condition or disease. In this case, the term "therapy" also includes "diagnosis", and also if a therapeutic agent is used for research purposes (as described above), then the use of a research agent is considered as a "therapy" of a condition or (treatment of a condition). destination.

"Disease", "Disease" or "condition" refers to the state of health of a patient or subject to be treated with various drugs, dispersions, or other methods provided herein.

As used herein, the term "contrast agent" refers to a composition that, when administered to a subject, improves the detection limit or detection capability of a physical method, technique, or medical imaging device (e.g., X-ray instrument, X-ray, CT, PET, MPT (MRI), ultrasound, and others methods). The contrast agent can enhance the magnitude of signals associated with various structures or fluids within a subject.

"Patient" or "subject in need thereof" refers to a living organism suffering from or (possibly) prone to a disease or condition that can be treated by administering a drug, pharmaceutical composition, or agent as defined herein. Non-limiting examples include humans, other mammals, bulls, rats, mice, dogs, monkeys, goats, sheep, cows, deer, and other non-mammalian animals. In particular, the patient can be human.

As used herein, the term “controlled release component” or “externally activated agent” refers to a compound that, when combined with a composition described herein (including embodiments), releases, under the conditions of use (in a patient), the composition at a controlled rate. Such compounds include high molecular weight, anionic mucomimetic polymers, gelling polysaccharides and fine drug carrier substrates, and others. These components are discussed in more detail in US patents 4911920; 5403841; 5212162; and 4,861,760. The entire contents of these patents are incorporated herein by reference in their entirety for all purposes. In some embodiments, the implementation of the "controlled release component" can be a sustained release, sustained release, sustained release, time release, timed release, controlled release, modified release, or sustained release compound. In some embodiments, the compound is degraded at the site of administration (eg, subcutaneous, intravenous) or within the digestive tract (eg, stomach, intestines) when the compound and composition are administered orally. In some embodiments, the controlled release component is a polymer and may be referred to as a “controlled release polymer”. In addition, an agent activated by an external effect can be activated (for example, release a therapeutic agent or bind to a target) as a result of exposure to an external magnetic / electric field, ultrasound, light, an electron beam, as a result of the radioactive decay of any atoms, as a result of interaction with chemical microenvironment, etc.

As used herein, the term "paramagnetic agent" refers to a paramagnetic compound useful in diagnostic imaging techniques (eg, magnetic resonance imaging) as a contrast agent. In some embodiments, the paramagnetic agent includes gadolinium, iron oxide, iron platinum, or manganese.

Variants of cytotoxic agents (or, toxins) include, but are not limited to, the following substances: ricin, doxorubicin, daunorubicin, taxol, ethidium bromide, mitomycin, etoposide, tenoposide, vincristine, vinblastine, colchicine, dihydroxyanthracindione Dion, actinomycinoma PE) A, PE40, abrin and glucocorticoid and other chemotherapeutic agents, as well as radioisotopes. Suitable detectable markers include, but are not limited to the following substances, a radioisotope, a fluorescent compound, a bioluminescent compound, a chemiluminescent compound, a metal chelator, or an enzyme.

The term "lipid" is used broadly herein to include triglycerides, liglycerides, monoglycerides, fatty acids, steroids, waxes, and various classes of emulsifiers that are used for emulsion stabilization.

The term "referred to" refers to both the above-described entities and those described below.

It should be noted that throughout the application, alternatives are written in Markush groups, for example, each position of a therapeutic agent may contain more than one candidate therapeutic agent. In particular, it is contemplated that each member of the Markush group should be considered separately, thus containing a different embodiment, and the Markush group should not be read as a whole. In addition, all cited references are incorporated herein by reference.

Detailed description of the embodiments of the invention.

We found that the introduction of antibodies against blood cells (for example, erythrocytes) into the body can induce phagocytosis of these blood cells by cells of the reticuloendothelial system (RES, mononuclear phagocytic system - MFS), which at a certain dose of antibodies leads to saturation of the phagocytic activity of RES. Such saturation leads to the fact that, with the simultaneous or subsequent administration of nanosized agents, the circulation time of such agents in the bloodstream increases. An increase in the circulation time of drugs in the bloodstream, in turn, leads to an increase in the efficiency of delivery of such agents to targets and, as a result, to an increase in their therapeutic or diagnostic efficiency.

It is especially surprising that in this way we were able to extend the circulation time not only for agents circulating shortly in the bloodstream, but also for the so-called stealth agents, the half-life of which from the bloodstream is several tens of hours. In particular, the prolongation of circulation (area under the curve) for clinically approved liposomal doxorubicin (Kelix preparation) was 42% as shown in Example 6. This unexpected result significantly improved cancer therapy with Kelix - 10 days after treatment, tumor volume in mice injected with the drug after administration of anti-erythrocyte antibodies was 2.1 times less than in mice injected with the drug without antibodies. A similar improvement effect was observed for non-stealth liposomal and lipid-containing agents, including for the treatment of other tumors - Lewis carcinoma, breast tumors, etc.

Accordingly, we called a similar method of blocking RES (MFS) by RES-cytoblockade or RES-immunoblokada (MFS-cytoblockade or MFS-immunoblokada). In the case of using antibodies against erythrocytes, this method can also be called RES-erythroblockade (MFS-erythroblockade).

Blockage (blockade) of the reticuloendothelial system is understood as a condition in which the ability of the reticuloendothelial system to remove any agents from the bloodstream decreases, incl. this can occur as a result of active phagocytosis by RES cells of any objects. In this case, in particular, a decrease in their phagocytic activity and, as a result, an increase in the circulation time of the administered agents in the bloodstream can be observed. In this case, a significant increase in the circulation time of the said agent in the bloodstream is preferable, however, an increase in the circulation time of the agent may not occur if, for example, the agent is able to quickly recognize its target in the body and bind to it. In this case, blocking the RES will allow the agents to avoid absorption by the cells of the RES and to bind to the target, but due to its rapid attachment to the target, the agent will be eliminated from the bloodstream almost as quickly. In this case, there will be a change in the biodistribution of the agent between the RES cells and the target. This invention makes it possible to increase the circulation time of therapeutic agents by reducing the rate of their absorption by the organs of the reticuloendothelial system as a result of saturation of RES with blood cells, the elimination of which is facilitated by the said antibodies of the composition. Therefore, the present invention is fundamentally different from the phenomenon of hitchhiking (RBC-hitchhiking, etc.), in which the agent, by binding to an anti-cell antibody, "clings" to long-circulating blood cells. Thus, the essence of the present invention is to prolong the circulation of freely circulating agents in the bloodstream (ie agents not associated with cells), or in free circulation within the blood plasma compartment rather than the blood cell compartment. Circulation in a cell-unbound state provides agents with additional therapeutic options that bound agents do not.

This invention improves the delivery of the agent to various targets in the body. In this sense, the improvement of delivery (implementation of more efficient targeted delivery) in some cases is meant in the sense of the effect of the mentioned antibodies on the biodistribution of the agent in organs, tissues, and cells of the body. In some embodiments of the invention, the induced cytoblockade reduces the accumulation of the agent in the liver, therefore, an increase in the concentration of agents in other tissues is observed, depending on the properties of the agent, the properties of antibodies and their dosage. In this case, it is possible to achieve various changes in biodistribution, which can be used for therapeutic purposes. Thus, in the mentioned embodiments, the hepatoprotective function of the method is achieved. At the same time, in some embodiments of the invention, the accumulation of the agent in the bone marrow is increased, which can be used to treat various diseases of the bone marrow. In addition, a significant decrease in the accumulation of agents in the liver leads to a significant increase in the accumulation, for example, in tumors. Tk. the size of the liver is often much larger than the volume of other targeted organs; even insignificant changes in the accumulation of agents in the liver lead to a significant increase in accumulation in smaller organs and targets.

It is important to note that this phenomenon is temporary, that is, it does not have a long-term compromise of the immune system and the body's defense systems.

In another embodiment, the compositions of this invention (including embodiments) can be delivered using liposomes that fuse with the cell membrane or endocytosis, as well as using ligands attached to the liposome, or attached directly to an oligonucleotide that bind to surface membrane receptors. proteins of the cell, which leads to endocytosis. Using liposomes, especially where the liposome surface carries target cell specific ligands, or is otherwise preferentially targeted to a specific organ, specific delivery of the compositions of the present invention to target cells in vivo can be accomplished. (See, for example, Al-Muhammed, J. Microencapsul, 13: 293-306, 1996; Chonn, Curr. Opin. Biotechnol. 6: 698-708, 1995; Ostro, Am. J. Hosp., Pharm., 46: 1576-1587, 1989).

Pharmaceutical compositions include those compositions in which the active ingredient is contained in a therapeutically effective amount, that is, in an amount effective to achieve its intended purpose. The actual amount effective for a particular application will depend, in part, on the condition being treated. When incorporated into a treatment protocol for a particular disease, such compositions will contain an amount of the active ingredient effective to achieve the desired result, for example, modulating the activity of the target molecule and / or reducing, eliminating or slowing the progression of disease symptoms. Determination of the therapeutically effective amount of a compound in this invention is well within the capabilities and skill of those skilled in the art, especially in light of the detailed description of the present invention.

The dosage and frequency (one or more doses) administered to a mammal can vary depending on many factors, for example, whether the mammal is suffering from another disease, the type of concurrent treatment, the mode of administration of the drug, size, age, sex, general health, body weight, body mass index, diet, nature and degree of symptoms of the disease, from complications of the disease or other health problems. Other therapeutic regimens or agents can be used in combination with the methods and compositions described herein (including embodiments). Adjustment and manipulation of established dosages (eg, frequency and duration) is well within the skill of those skilled in the art.

Dosages can vary depending on patient requirements and the compound used. The dose administered to the patient should be sufficient to produce a beneficial therapeutic response in the patient over time. The size of the dose will also be determined by the presence, nature and extent of any adverse side effects. Determining the correct dosage for a particular situation is within the skill of the art. Typically, treatment begins with lower doses that are less than the optimal dose of the compound. Thereafter, the dosage is increased in small increments until the optimal effect for the specific conditions is achieved.

The dosage and intervals can be adjusted individually to provide levels of compound administered that are effective for the particular clinical condition being treated. This will provide a therapeutic regimen that is commensurate with the severity of the individual's disease condition.

Using the methods described herein, an effective prophylactic or therapeutic regimen can be planned that does not cause significant toxicity and at the same time is effective for treating the clinical symptoms of a particular patient. This planning should include careful selection of the active compound by considering factors such as the pharmacological activity and toxicity of the compound, relative bioavailability, patient weight, the presence and severity of adverse side effects, and the preferred route of administration of the selected agent.

In addition, when the disease is a treatable cancer by topical administration of a pharmacologically active agent (s), an additional aspect of this embodiment of the invention comprises administering to a patient a suitable pharmacologically active agent (s) in the present invention capable of treating cancer in a formulation of the invention.

The introduction can be carried out in various ways, for example, intra-arterial and / or by injection. In an example of this aspect, a suitable pharmacologically active agent (s) comprises an antineoplastic agent (eg, a taxane (paclitaxel, docetaxel, and the like) and the like) or the like.

In some embodiments, the subject has cancer from the group of primary and secondary liver tumors (e.g., hepatocellular carcinoma, multifocal hepatoma, etc.), solid tumors with local or regional manifestation (e.g., metastatic breast cancer, prostate cancer, pancreatic cancer, non-small cell lung cancer (squamous cell carcinoma), colon cancer, renal cancer, bowel sarcoma, esophageal cancer, melanoma, ependymoma, head and / or neck cancer, and the like) or the like.

In one embodiment of the invention, a lipid-containing drug is used as said agent, which is liposomes, nano-sized lipid droplets, emulsions, solid lipid particles and the like, including those filled with various drugs, for example, molecular drugs - doxorubicin, paclitaxel etc., or nanoparticles - magnetic, gold, etc.

In one embodiment of the invention, an increase in the circulation time of the agent in the bloodstream of the body or blocking of the RES is achieved by introducing antibodies into said living organism, the introduction of which leads to an increased removal of erythrocytes or platelets (including native or intact) from the circulation in the bloodstream of the body as compared to the rate of their removal in the case without the introduction of antibodies. Under native erythrocytes or platelets are meant unchanged cells that have not undergone hemolysis or any physical, chemical or biochemical treatment (such treatment can, for example, be ultrasonic exposure, osmotic lysis, formalin fixation, etc.). Accordingly, in one embodiment of the invention, the clearance of native cells is mediated by antibodies that interact non-covalently with blood cells.

In one embodiment of the invention, the composition is chosen such that it does not contain erythrocytes, or more generally, modified erythrocytes. In another embodiment of the method, said antibody composition interact (bind, recognize due to its antigen-recognition site) with epitopes of erythrocytes or platelets that are located directly in the body, i.e. which are not introduced into the body within the framework of a given effect on the body (i.e., if the body has been previously transfused or transplanted with material that is not associated with the need to modulate the behavior of the agent in the body, then we can talk about such cells and molecules). Modified erythrocytes are understood as substances obtained by any physical, chemical or biochemical treatment of erythrocytes or blood components containing erythrocytes. Such treatment may, for example, be ultrasound, osmotic lysis, formalin fixation, etc. Such modifications can lead to negative toxic effects of these agents.

In addition, in one embodiment of the invention, the composition may contain complexes of preselected erythrocytes or other cells or molecules of the body with antibodies (or their variants) complementary to homologous species antibodies of the body, incl. autologous, allogeneic, humanized or chimeric antibodies. A composition containing such complexes will not cause, for example, anemia, and the injected cells or molecules will be excreted by the RES organs, which will lead to blocking of the RES organs. The collection of cells or molecules of the body can be carried out in advance so that the body has time to restore the levels of these substances at the time of administration of the composition. In addition, the restoration of the level of these substances can be carried out not only while waiting for the body to restore their content, but also actively contribute to this by introducing reducing agents. Such a reducing agent, for example, can be erythropoietin, which accelerates the maturation of erythrocytes. Another option for administering a reducing agent is donated blood transfusion. The introduction of the reducing agent can be carried out both before and during or after the introduction of the agent and the composition.

In one embodiment of the invention, antibodies non-covalently bind to cells, i. E. in this case, new covalent bonds are not formed, which could lead to undesirable toxic effects of drugs.

The term "anti-cell antibody" means that the antibody binds or is capable of binding to cells, preferably capable of binding specifically (in the traditional sense of molecular and cellular biology) - i.e. specifically form a complex with cells. In one embodiment of the invention, the antibody recognizes a specific receptor, receptor epitope, or antigenic determinant of a receptor on a cell.

In one embodiment of the invention, said anti-blood cell antibody is selected to be highly specific for a particular population of blood cells (eg, erythrocytes). In this case, it is preferable that this antibody binds only to this population of blood cells and does not bind to other cells of the body, for example, vascular epithelial cells. For example, in one embodiment of the invention, allogeneic antibodies against erythrocytes are used, isolated from the repertoire of anti-erythrocyte antibodies of an organism susceptible to autoimmune hemolytic anemia (see Examples with antibody 34-3C) - or from the repertoire of similar autoimmune patients for other blood cells. In another embodiment of the invention, antibodies are used against antigenic determinants or receptors that are characteristic not only of a certain population of blood cells, but also on other cells of the body, for example, antibodies against the CD47 receptor. In such a case, it is preferable to select a receptor that is sufficiently expressed on blood cells, and also select an appropriate monoclonal or polyclonal antibody or a mixture of different antibodies, so that the administration of antibodies effectively induces phagocytosis of blood cells by the reticuloendothelial system, while minimizing negative effects on other cells expressing this receptor.

In addition, the specificity of antibodies to blood cells is determined according to standard concepts in the field of molecular and cellular biology. Namely, it is important that the affinity of these antibodies for the selected population is such that the constant of association with a receptor specific for a given cell population is higher than 10 ^L 6 1 / M, more preferably 10 ^L 7 1 / M, more preferably 10 ^L 8 1 / M, more preferably 10 ^L 9 1 / M, preferably 10 ^L 10 1 / M, preferably 10 ^L 11 1 / M.

The specificity of an antigenic determinant is also determined according to the standards of cell biology, namely, in the presence of a significantly higher level of expression of this receptor in a given cell population compared to other cell populations.

In addition, to classify antibodies as antibodies against erythrocytes, the characterization of the number of antibody molecules bound to an erythrocyte after incubation of cells with a solution of antibodies can be used. The determination of such an indicator can be carried out using flow cytometry or other type of analysis. So, for example, after 1-hour incubation 1 mln. cells in a volume of 10 μg / ml with a solution of antibodies with a concentration of 10 μg / ml, this can be 25 molecules / cell, preferably 50 molecules / cell, preferably 100 molecules / cell, preferably 200 molecules / cell, preferably 300 molecules / cell, preferably 500 molecules / cell, more preferably 750 molecules / cell, more preferably 1000 molecules / cell. These numbers and protocol can be changed according to accepted principles in cell biology.

In addition, in one embodiment of the invention, the hematocrit, hemolysis, and / or the level of the said cells (against which the said antibody is directed) is assessed to verify the phagocytosis of said cells by the reticuloendothelial system in order to possibly adjust the dose of the administered antibody and / or said lipid-containing agent.

In one embodiment of the invention, the elimination of cells from the bloodstream is facilitated by additional treatment with enzymes. For example, the treatment of erythrocytes with an enzyme, for example, neuraminidase, trypsin, galactose oxidase, etc. or a combination of enzymes leads to an acceleration of their excretion by the reticulo-endothelial system from the circulation in the bloodstream. This occurs due to, for example, a change in the glycosylation profile of the erythrocyte surface, which can recognize RES cells and remove "obsolete" erythrocytes, or due to the appearance of new antigenic determinants on the erythrocyte, which will cause it to bind to the said antibodies and lead to elimination from the bloodstream for example, by Fc receptor-mediated phagocytosis. It is clear that although the treatment of cells with enzymes has certain advantages due to the availability of enzymes, the treatment of cells only with antibodies is a milder effect on the body (in terms of toxicity - due to the possibility of the appearance of new antigenic determinants, reproducibility, etc.).

In any of the above embodiments of the invention, the active component or components of the composition can be composed of a combination of different objects, including those that possess the described properties and those that do not. Those. the composition may contain not only, for example, an allogeneic antibody against erythrocytes, but also various auxiliary components such as buffer salts and the like, and in addition, such an antibody may, for example, be conjugated to other molecules, particles, etc. ...

The mentioned lipid-containing agent can be a wide variety of substances containing lipids, for example, it can be nano- and microparticles in the usual sense of colloidal chemistry, bacteria or cells (in the preferred embodiment of the invention), various molecules (in the preferred embodiment, macromolecules), macromolecular complexes, etc. Agents can be, inter alia, diagnostic and / or therapeutic (including theranostic) agents or substances that in one way or another mediate diagnostic and / or therapeutic functions. For example, an agent can perform an imaging function from a diagnostic point of view. or monitoring a condition / disease of the body, so, for example, it can be an agent that directly produces a detectable signal (fluorescent, luminescent or radioactive agent), or changes the signal detected from other substances (for example, from water protons for MPT contrast agents). In addition to physical signals, the agent can participate in the production of biochemical signals, which, for example, allow the body to be tested for the presence of a disease and, if necessary, signal it. In addition, the agent can perform other functions (including therapeutic ones), for example, for the targeted delivery of drugs, the implementation of a hyperthermic effect on the body (if, for example, it contains magnetic nanoparticles), photodynamic therapy (in one way or another, causing, for example, cytotoxicity due to external radiation), and the like. The said lipid-containing agent may be or may contain one or more substances / elements from the following: therapeutic molecules / agents, labels or other detectable elements, contrast agents, controlled release components, agents activated by external influences, paramagnetic agents, cytotoxic agents ...

In one embodiment of the invention, the effect of increased circulation in the bloodstream of the lipid-containing agent, as well as the effect of increasing the efficiency of its delivery to the target, allows non-diagnosis of diseases. Thus, Example 11 shows how an increased accumulation of Agent in a tumor is recorded non-invasively using an MPQ detector (see Nikitin et al. Nat Nanotechnol, 2014). In addition, agents are imaged with magnetic resonance imaging or CT diagnostics. In addition to the diagnostic nature, the increased delivery of the agent to the target can be used for therapeutic purposes or simultaneously diagnostic and therapeutic (so-called theranostic) purposes.

In one embodiment of the invention, the requirement of said antibody to be an antibody against red blood cells may be considered broader than the standard understanding of affinity / avidity for red blood cells, but in the ability of said antibody to form, directly or indirectly, a specific complex with an red blood cell. The ability of a substance to form, directly or indirectly, a specific complex with something, for example, with the cells of the body, means that either the given substance (for example, an antibody) can directly recognize and bind to any cell or molecule, or recognize and bind to other additional molecules , which in turn (also directly or through other molecules) can bind to these cells of the body. Such additional molecules can be created by the body itself, or also introduced into the body, including as part of the composition. In addition, indirect recognition and binding means that, for example, an antibody may not bind directly to a cell, but rather bind to another molecule that has bound to the cell. For example, a humanized anti-rat immunoglobulin G antibody can recognize and bind to a rat anti-human erythrocyte antibody associated with erythrocytes.

In all of the above and below listed cases, circulation in the bloodstream can be understood as circulation in the bloodstream in general (in the "spatial" sense, that is, inside the bloodstream, vessels, capillaries, etc.), and free circulation in the bloodstream ( i.e. circulation in free form without association or binding with cells, other agents or objects, for example, if an erythrocyte is absorbed by a macrophage circulating in the bloodstream, then the red blood cell is considered to be removed from free circulation in the bloodstream, although in a spatial sense it is still there in the bed of a blood vessel). In an advantageous embodiment of the invention, withdrawal from circulation is understood to mean withdrawal from free circulation.

In one embodiment of the invention, a reversible short-term retargeting of RES to the body's own cells / molecules is achieved in order to prevent an "attack" on the drug, which is fundamentally different from the known methods, since the dose of the introduced foreign blocking RES agent can be reduced 10-100 times compared to existing methods to achieve a 1000% increase in the half-life of the drug (compared to the circulation time of the drug without the introduction of any additional compositions). This Possibly due to the fact that most of the mass / volume of the total complexes excreted by the RES organs and causing blockage of the RES organs belongs to its own blood components (components of the organism itself) and is a natural material for it. It also allows to minimize the toxicity of the method as much as possible. only a small amount of a foreign (and therefore potentially toxic) blocking agent (in this case, said antibodies) is required to create the desired effect.

In one of the embodiments of the invention, a composition is used that contains at least an antibody (or variants thereof) highly homologous to the antibodies of the organism, incl. an autologous, allogeneic, humanized or chimeric antibody capable of forming, directly or indirectly, a specific complex with cells or molecules that are naturally in the body, or that have entered the body artificially. In another embodiment of the invention, antibodies (or their variants), homologous (mainly highly homologous) to the specific antibodies of the organism, are administered to block the organs of RES, incl. autologous, allogeneic, humanized or chimeric antibodies against the patient's blood cells, for example, his erythrocytes, platelets or leukocytes. In this case, the organs of RES actively absorb blood cells that have bound with antibodies, after which their phagocytic activity against the agent decreases, in particular, the half-life of the agent from the bloodstream may increase. After the introduction of antibodies, the effect increases for some time, then subsides. In one embodiment of the invention, if the agent is administered after administration of the antibodies, their elimination half-life will increase over time and then decline to the interval that was observed before the administration of the antibodies (see Examples 4-10). In the implementation of the method with antibodies against erythrocytes, caused by anemia (due to a decrease in hematocrit) is short-lived and after a certain time the level of erythrocytes in the bloodstream, as a rule, is restored to the level observed before the introduction of antibodies.

The introduction of antibodies (or their variants) highly homologous to the specific antibodies of the body, incl. autologous, allogeneic, humanized or chimeric antibodies, and not, for example, heterologous anti-lymphocyte serum, for blocking RES, allows you to use only the natural mechanism of removing cells from the blood due to phagocytosis by Fc-receptor-mediated RES cells and does not introduce unnecessary danger (toxicity) due, for example, to the "attack" of phagocytes with anti-lymphocyte serum proteins. In a preferred embodiment of the invention, the Fc portion of the antibody is identical to the Fc portion of the antibodies of the organism itself. In addition, the high homology to the body's antibodies will allow avoiding or minimizing the immune response to the components of the composition, which will allow it to be administered multiple times to achieve the effect. In addition, there will be no various side effects from the immune response, up to such as the inability to carry out immunoassays of blood in which certain antibodies are used (for example, if a mouse antibody against human erythrocytes is introduced into a person, and as a result of the immune response, antibodies against murine immunoglobulins, the use of many tests for carrying out immunoassays of blood, in which mouse antibodies are involved, will be difficult due to the possibility of errors in the test results).

In addition, since the number of erythrocytes in the blood is many times higher than the number of leukocytes and platelets, the use of compositions for removing erythrocytes by RES cells to block RES, incl. to increase the circulation time of the agent, preferable in view of fewer negative effects on the health of the body. In particular, after removing a certain part of erythrocytes from the bloodstream by the RES cells, in one embodiment of the invention, the hematocrit is restored by blood transfusion. This avoids the negative consequences of anemia associated with low hematocrit.

One of the embodiments of the invention (when the components of their own blood are removed by the RES cells) is characterized by a low toxicity of the approach, since The absorption of blood components by the RES organs (including erythrocytes) occurs throughout the life of mammals, here this process is only accelerated for a short time. So, for example, during the day under natural conditions, the body processes about 1% of erythrocytes, and to achieve blocking of RES, it is required to remove from blood flow of less than 10% of erythrocytes (in Example 4, the maximum drop in hematocrit was by 10-15%, i.e., from 50% to 45%, and then, the minimum hematocrit was observed on the fourth day), while the blocking effect lasts about 5 -10 days, during which approximately the same number of red blood cells would be processed. Therefore, this method is characterized by low health risks. In addition, for example, in the case of using antibodies against blood cells to increase the circulation time of the agent, a natural mechanism is used to remove cells from the bloodstream by RES cells. Namely, the antibody non-covalently binds to the cell and signals to the RES cells that this complex must be removed from the bloodstream. An important advantage of the method is that there is no chemical modification of blood molecules / cells, no new covalent bonds are formed, or new chemical compounds, etc., which could cause any toxic effects.

In one embodiment of the invention, an antibody (and / or dose of antibody) is selected that does not or leads to little activation of the complement system, preferentially promoting Fc receptor mediated phagocytosis by RES cells.

In addition, in one embodiment of the invention, the composition (or said antibody) does not cause agglutination of blood cells. In this case, the dose of the agent administered can be varied over wide ranges without fear of vascular embolization.

In addition, in one embodiment of the invention, the components of the composition, conversely, cause agglutination of blood cells. In such a case, the choice of the dose of the composition is required to prevent thrombus formation, which leads to vascular embolism.

In addition, in one embodiment of the invention, the dose of the injected antibodies is selected to increase the effect of blocking RES or to increase the circulation time of the agent in the bloodstream by the required number of times, optimizing the beneficial effect of increasing the circulation time, and the possible negative effects of lowering the concentration of blood molecules / cells. excreted by the RES cells.

This approach can also be used to block phagocytic RES cells in other natural fluids, as well as in body tissues. With the introduction of appropriate agents that cause the phagocytic cells of the RES to actively absorb the cells and / or molecules of the body or other objects, their ability to phagocytose the agent decreases, which leads to an increase in the target efficiency of the agent.

This invention can be used for diagnostic and / or therapeutic purposes. Including for the purpose of improving the targeted delivery of various lipid-containing agents to various targets in the body. The nature and type of targets can be very diverse, for example, it can be specific cells, tissues, areas of the body (including malignant neoplasms and foci of inflammation).

The application of the present invention can be, for example, as follows. An antibody that blocks RES (for example, humanized antibodies against erythrocytes) is intravenously injected into the body, and then lipid-containing agents containing toxic molecules (for example, the anticancer drug doxorubicin or cisplatin) are injected intravenously. The agents, either through the enhanced permeability and retention effect or through specific anti-cancer antibodies, accumulate in the tumor, where doxorubicin or cisplatin is desorbed and causes the death of tumor cells (as shown in Examples 14-15).

In one embodiment of the invention, a monoclonal antibody is used, preferably against erythrocytes or platelets, even more preferably associated with allogeneic erythrocytes or platelets, preferably with erythrocytes or platelets of the body or allogeneic variants thereof. In this case, in one embodiment of the method, the monoclonal antibody is selected according to the type and isotype so as to achieve controlled removal of cells from the circulation, predominantly by a certain mechanism, for example, by phagocytosis of cells to which the antibodies have bound, by an Fc-receptor-mediated mechanism (to block, for example, the body's Kupffer cells), agglutination of erythrocytes to filter aggregates by the spleen, due to complement-mediated hemolysis of cells to load the kidneys, etc. In this case, in one embodiment of the invention, a combination of different antibodies is used to influence both the biodistribution of excreted cells or other objects that block RES cells, and the biodistribution of the agent throughout the tissues of the body. Those. for example, if the liver is preferentially blocked, the drug will preferentially accumulate in the target (eg, tumor) as well as spleen, lungs, and the like. In addition, the use of monoclonal antibodies makes it possible to create a composition with an accurate and well reproducible composition, which is useful for quality control and minimizing the harmful effects of such a composition.

In another embodiment of the invention, polyclonal antibodies are used. For example, xenogenic or xenogenic mixed with allogeneic (see Example 13) or allogenic, isolated from donated blood, for example, anti-rhesus RhO (D) antibodies.

In one embodiment of the invention, a humanized or chimeric antibody is used, preferably against erythrocytes or platelets, even more preferably associated with allogeneic erythrocytes or platelets, preferably with erythrocytes or platelets of the body or allogeneic variants thereof. In addition, the preparation of a monoclonal antibody, for example, murine or rat, against, for example, human erythrocytes, and its subsequent humanization, allows you to obtain a high affinity antibody of the desired isotype, with the desired Fc fragment and the like. It is also possible to obtain the desired antibody variant using phage display technology.

AHTM-CD47 antibodies

In one embodiment of the invention, antibodies against the CD47 receptor are used as antibodies against erythrocytes. This cell surface marker is a glycoprotein that plays an important role in cell-cell communication. Among other things, the function of this receptor on red blood cells is to prevent the capture of normal red blood cells by macrophages and other phagocytic cells involved in erythrophagocytosis. Receptor ligand - SIRPA (SIRPa, CD172a). In one embodiment of the invention, an anti-CD47 antibody (see Examples 18-23) is used to block the interaction of the receptor with the natural ligand of SIRPa. In this case, the introduction of this antibody leads to the elimination of erythrocytes by the reticuloendothelial system, which leads to its temporary blockage. This, in turn, leads to an increase in the circulation time of various nano- and micro-agents in the bloodstream, which are administered simultaneously or after the administration of these antibodies. These antibodies have a twofold effect. First, these antibodies mediate antibody-dependent phagocytosis of erythrocytes. Secondly, these antibodies screen the CD47 receptor, which signal to macrophages about the "normality" of a given erythrocyte. This allows you to enhance erythrophagocytosis, "deceiving" the recognition system between erythrocytes and macrophages. That is, when the CD47 receptor is blocked / shielded, it is no longer able to perform its function of inhibiting erythrocyte phagocytosis by the macrophage. Such a violation of the natural signaling pathway between the erythrocyte and macrophages makes it possible to enhance erythrophagocytosis, due to which the blockage of the reticuloendothelial system is realized. In one embodiment of the invention, not a full-length antibody is used, but its antigen-binding fragment (Fab, Fab2 or the like), or other inhibitors of the CD47-SIRPa interaction, for example, peptides. Accordingly, to increase the effect of prolonging the circulation of therapeutic agents in the bloodstream and their therapeutic efficacy, it is possible to use various antibodies synergistically, for example, isolated from patients suffering from autoimmune hemolytic anemia and antibodies that “interfere” / disrupt / use integral natural pathways for the regulation of erythrocytosis, as well as elimination of other blood cells.

Moreover, a composition comprising an anti-CD47 antibody has an additional anti-tumor effect by shielding the CD47 receptor on cancer cells, which prevent phagocytosis by cells of the immune system. Thus, in one embodiment of the invention, antibodies against blood cells are used, which have both the functions of prolonging the circulation of nanoagents introduced into the bloodstream and other functions and activities from the viewpoint of combating target diseases.

In one embodiment of the invention, the composition further comprises a component that results in enhanced clearance of body molecules, preferably macromolecules or macromolecular complexes, from circulation. This is important, including for the comprehensive blocking of cells capable of removing agents from the circulation in the bloodstream, mainly for blocking all types of such cells. This makes it possible to significantly reduce the dose of the agent that must be administered in order to deliver a certain amount of it to the target and to increase the specificity of delivery, i.e. deliver as much of the administered dose of the agent to the target as possible.

In one embodiment of the invention, an antibody or a mixture of antibodies is used simultaneously against several types of blood cells - erythrocytes, platelets and / or leukocytes (including those associated with erythrocytes, leukocytes and / or platelets), more preferably together with components that promote enhanced excretion various molecules (mainly macromolecules or macromolecular complexes) from the body. This allows you to block the largest part of the cells that are able to remove the agent from the circulation in the bloodstream, and therefore allows you to reduce the required dose of the agent.

Various aspects regarding the said antibody

Note that the mechanism and behavior of the present invention to prolong the circulation of various nanoagents in the bloodstream is very similar to the mechanism of anti-erythrocyte antibodies in the treatment of immunothrombocytopenia. Sequestration of autoantibody-sensitized platelets during immune thrombocytopenia (ITP) can be suppressed by regular injections of intravenous immunoglobulin (IVIG) or anti-RhD (in Rh-positive patients). Although the mechanisms of their action are still unclear, most researchers consider the inhibition of IFS to be one of the most probable mechanisms. On toxicity issues, the FDA revised the anti-RhD package insert in 2009 to highlight the following warning: "Intravascular hemolysis (IVH) leading to death has been reported in patients with ITP." All rare cases of mortality were caused by intravascular hemolysis. However, based on extensive 20 years of experience with the drug, the FDA considers the drug to be safe.

In this regard, it should be noted that in certain embodiments of the present invention, when using anti-erythrocyte antibodies, additional tests are performed to identify patients at risk of developing I VH, which can significantly reduce the risk of side effects of antibodies against red blood cells (RBC) and make such therapy much more secure;

In certain embodiments of the present invention, anti-RBC antibodies are obtained from a murine model of autoimmune hemolytic anemia (AIHA). A clone obtained in a similar way from a patient with AIHA in humans, especially if it does not fix complement (for example, lgG4), can also help reduce toxicity.

The polyclonal nature of anti-RhD derived from donor serum makes it much more difficult to control the reproducibility of the composition. In turn, side effects for a given patient may be inconsistent from batch to batch, and their causes can be very difficult to investigate. Accordingly, the proposed monoclonal antibody (obtained from an AIHA patient) will be a much more reproducible agent, free from such unexpected batch-specific toxicity problems.

In a study by Song et al. Blood (2003), the authors showed that treatment of ITP in a mouse model requires much higher doses of antibodies than those used in humans. Specifically, they showed that an anti-mouse erythrocyte anti-TER-119 monoclonal antibody was rather unproductive for the treatment of ITP at a dose of 5 μg and effective at a dose of 50 μg. At the same time, our new experiments show that 25 MKr TER-119 (the same dose that we used with 34- ЗС) successfully prolongs the circulation of nanoparticles. Thus, in a mouse model, ITP treatment and prolongation of nanoparticle circulation are achieved using similar doses of the anti-RBC TER-119 monoclonal antibody.

Accordingly, it appears that: i) mice generally require more antibodies for RES cytoblockade, or ii) polyclonal anti-RhD is more effective than 34-3C or TER-119, for example, due to different levels of antigen expression. In both cases, there is some possibility that anti-RhD itself will be effective in improving nanoparticle circulation even at already approved doses of 50-75 mcg / kg.

In any event, we should note that these anti-RhD doses have been approved for the treatment of ITP based on the following considerations: 1) there are several safe alternatives such as IVIG; 2) ITP itself is not a life-threatening disease; 3) treatment must be safe for repeated injections (once a month) over a long period of time. Therefore, in some embodiments of the invention, the use of the proposed cytoblockade in combination with nanomedicine would be the treatment of life-threatening conditions (such as cancer) that have different thresholds of efficacy / toxicity. Therefore, with careful control of the risk of intravascular hemolysis, the dose of anti-RhD or anti-RBC clones obtained from AIHA can be significantly increased with an overall increase in the effectiveness of treatment of the disease.

Magnetic delivery

In one embodiment of the invention, administration of said anti-blood cell antibody is used to increase magnetic delivery of a magnetic agent to a target. With this method of delivery, a specially generated magnetic field is used to hold or direct an agent with increased magnetic susceptibility (magnetic nanoagent, superparamagnetic, ferro- and ferri-magnetic agent) to its target. For example, the agent can contain magnetic nanoparticles, superparamagnetic nanoparticles, ferromagnetic particles, and the like. In this case, the external field can be both variable and constant. The increase in the circulation time of the agent in the blood, achieved by the introduction of the above antibodies (as described above), leads to a greater accumulation of the agent in the focusing area of the magnetic field. The mentioned focusing of the magnetic field means the formation of such a magnetic field, the strength of which in the target area, or the magnitude of the field gradient in the target area, or the product of these values is greater than in any other parts of the body.

In this case, it is necessary to mean the focusing of the magnetic field on a target with a restriction according to Earnshaw's theorem, namely, to understand that not an absolute maximum of the field strength is created in the target, but only a greater value than in some other parts of the body.

In addition, in one embodiment of the invention, an alternating magnetic field is used to actively direct the circulating magnetic agents into the target area. In another embodiment of the method, a magnetic field is used as a means of long-term retention of magnetic agents in the target area. It is known that many nanoagents (including lipid-containing drugs), spreading well by the bloodstream in the vessels of the body, quickly enter the target, for example, a tumor. And then, within a fairly short time (minutes, tens of minutes, hours), their concentration quickly decreases, which reduces the therapeutic effect. In this case, in one embodiment of the invention, the magnetic field makes it possible to retain such agents in the target area for much longer, and therefore to implement the desorption of the drug (for example, cytotoxic drugs) to affect the target, for example, to kill tumor cells.

Examples 14-15 demonstrate an increase in the effectiveness of antitumor magnetic agents when they are magnetically delivered using said antibodies.

One of the ways to generate a magnetic field is to place a permanent magnet, preferably neodymium, near (in the area) of the target. Another way is to use electromagnets, which are also located near the target. Also shown methods of magnetic delivery using gradient coils of an MRI tomograph, as well as other more complex systems consisting of several permanent magnets or electromagnets. When forming a focusing system for magnetically controlled delivery, try to achieve the distribution of the force field acting on the magnetic inoagents, which ensures maximum efficient accumulation in the target.

In general, the essence of the present invention in one embodiment is to reduce the rate of elimination of magnetic nanoagents from the bloodstream by the immune system. This allows the invention to be used in conjunction with virtually any magnetic focusing (delivery) system that allows an increase in the accumulation of magnetic particles in the target area with traditional systemic delivery of the agent (i.e., without the use of said antibodies).

Polynucleotide delivery and other aspects

The compositions and methods of in vivo transfection according to this invention will significantly advance in the creation of drugs for gene therapy, namely, the treatment of diseases by delivering genetic material directly to certain cells of a patient, in order, for example, to increase the expression of specific genes or reduce the production of a desired protein. Since the limiting factor of such therapy is direct delivery of nucleic acids to the target cells, the selection of a reliable and efficient viral or non-viral delivery vector is of particular importance. The well-established, traditionally used viral vectors based on adenoviruses, lentiviruses and adeno-associated viruses, however, have a number of serious disadvantages, such as immunogenicity and carcinogenicity. Non-viral vectors are safer. These include polyplexes based on cationic or neutral biodegradable polymers, lipoplexes (cationic liposomes, niosomes), complexes of DNA with dendrimers or peptides, as well as combinations of all of the above vectors with other nanoparticles, for example, magnetic, gold, etc.

In accordance with the present invention, it is possible to use a combination of different delivery methods for transgenes in a single delivery vehicle. For example, non-viral and viral transgene delivery methods can be combined by encapsulating or complexing viral particles with liposomes, nanoparticles, lipid nanoparticles, polymers, microparticles, microcapsules, micelles, extracellular vesicles, etc.

Currently, lipid vectors are one of the most commonly used carriers of nucleic acids (NK) for transfection for gene therapy. In one embodiment of the invention, liposomes are used that bind NKs on the surface or encapsulate them in the internal cavity, formed with cationic lipids (eg DOTMA, DOSPA, DOTAP and DMRIE) used as precursors. Cationic lipids in liposomes are diluted with neutral lipids (eg DOPE) to increase the stability and transfection activity of liposomal vectors.

In one embodiment of the invention, cationic liposomes are formed using cationic cholesterol derivatives.

In one embodiment of the invention, cationic liposomes are formed through the use of lipidoids.

In one embodiment of the invention, NK delivery is carried out using formally neutral liposomes constructed from zwitterionic lipids.

Liposomes (phospholipid vesicles) used for gene therapy (lipofection) compatible with this invention may vary in size, lipid composition, charge, dispersion, method of preparation, membrane fluidity, fusogenicity (tendency to fusion with membranes), phospholipidic capacity components for micelle formation and complexation with DNA. The vesicle charge is determined by the presence of polyions or cationic lipids; membrane fluidity negatively correlates with the amount of cholesterol and depends on the degree of "unsaturated ™" and the length of the fatty acid residues of phospholipids.

In one embodiment, the efficiency of complexation with DNA is enhanced by the use of cone lipids (e.g. dioleyltrimethylammonium (DOTMA)) which have a high spontaneous micelle capacity. In one embodiment, substituted tertiary amines such as DOTMA and DOTPA are used as cationic lipids. In one embodiment, as cationic lipids, positively charged lipids with an ether bond in the glycerol part with different ratios of hydrophobic and hydrophilic parts in the molecule, as well as different distances between them, are used, which provides a wide variety of structures of DNA-liposome complexes.In one embodiment, dioleylphosphatidylcholine ( DOPC) and dioleyl phosphatidylethanolamine (DOPE) In some embodiments, liposome fusogenicity is reduced by the use of dioleyl phosphatidylethanolamine as phospholipid components of liposomes, which causes increased hydration of lysosomes. Absorption DNA-liposomal complexes are prepared on the basis of cationic liposomes. In one embodiment, the DNA-liposomal complexes are prepared from neutral (zwitterionic) liposomes.

In some implementations, native plasmid DNA is enclosed in a phospholipid bilayer and protected from the action of DNases, topoisomerase 1 and fluorescent dyes (ethidium bromide and DAPI), which does not affect its ability to internalize and interact with other membranes.

In one embodiment of the invention, solid lipid nanoparticles [47, 48] with a diameter of about 100 nm, which are aggregates of low molecular weight lipids (fatty acids, cholesterol and their derivatives) suspended in the presence of a surfactant (for example Tween-80, Pluronic). Such lipid nanoparticles bind NC on the surface through electrostatic interactions.

In some embodiments, dendrimers are used as gene therapy vectors, which are highly symmetric branched macromolecules with low polydispersity.

In one embodiment of the invention, the agents comprise polycations such as L-polylysine (3000-10,000 Da), polyethyleneimine (5 kDa-100 kDa), as well as complexes of polyethyleneimine (hexamer) with mono-, di- and tricholesterol derivatives, lipospermine (cationic polypeptide sperm with hydrophobic groups). Lipospermin has a high affinity for DNA, while forming non-micellar hydrophobic structures similar to liposomes. The agents can also contain cationic polypeptides that are metabolically active and can block ion channels and inhibit the permeability of mitochondrial membranes to calcium and phosphate ions.

In addition, the agent may include poly- (alkylacrylic acid) polymers, including non-cytotoxic polyacrylic acid (PAA), which are considered endosomolytic polymers and therefore increase gene expression as well as reduce toxicity in vivo.

In one embodiment, PLGA-PEI nanoparticles are used as a non-viral vector for the delivery of genetic material, which are capable of self-assembly of DNA and provide higher stability, ease of manipulation and an economical option than cationic liposomes. In addition, surface modification of composite PLGA-PEI / DNA nanoparticles by treatment with polyethylene glycol (PEGylation) reduces their cytotoxicity, prolongs their in vivo existence and expression, for example, of plasmid DNA.

In one embodiment, polyesters (poly-b-amino ester, PBAE) are used to protect NA from enimatic degradation.

In one embodiment, gold nanoparticles, carbon nanotubes, quantum dots, silicon nanoparticles are used as a vector for delivery of DNA and RNA. In one embodiment of the invention, an extended gene-encoding dsDNA, mRNA, non-coding RNAs that regulate the expression of target genes (siRNA, microRNA) were used as the transported material. In one embodiment of the invention, nucleic acids are delivered from the group: plasmid DNA (pRNA), also small interfering RNA (siRNA), short hairpin RNA (shRNA) and antisense oligonucleotides.

In one embodiment, viral vectors for the delivery of genetic material were obtained from native viral particles by replacing the virus's own genes with genetically engineered constructs. Protein components of the capsid protect NK from degradation in lysosomes and facilitate the delivery of genetic material to the nucleus.

In one embodiment, adenoviral vectors were used, which are non-enveloped viral particles with a diameter of 80-100 nm containing double-stranded DNA (dsDNA); adeno-associated vectors (AAV), which usually have a small, non-enveloped virion with a diameter of about 20 nm and a genome of 4.7 thousand bases (thus), represented by single-stranded DNA (ssDNA); gammaretroviruses (mouse leukemia virus) and lentiviruses (HIV-1) (however, the low efficiency of transduction of nondividing cells with gamma retroviral vectors limits their use by transferring genes into hematopoietic cells); lentiviral vectors, which are a protein capsid with a diameter of 80-100 nm, packed in a lipid envelope; herpesvirus vectors, which have great potential for use in the clinic, obtained on the basis of herpes simplex virus type 1 (HSV-1); alphavirus vectors obtained from viruses of the Togaviridae family were used to deliver siRNA-coding constructs.

In some embodiments, targeting molecules (eg, cell receptor ligand peptides) have been introduced onto the surface of viral vectors. In one embodiment, to allow co-delivery of genetic material and small molecule drugs, viral vectors have been modified by mixing fragments of different viruses or introducing peptide inserts so that the genome of the virus is altered to form chimeric or mosaic capsids.

In some embodiments, viral vectors are rendered properties uncharacteristic of viruses (eg, the presence of reporter groups) by incorporating synthetic polymers or inorganic nanoparticles into viral particles.

In one embodiment of the invention, the agent is or contains a viral vector. As used herein, the term "viral vector" has a common meaning in the art and refers to a vector derived from various serotypes of gammaretroviruses, lentiviruses, adeno- and adeno-associated viruses, herpes simplex viruses, rhinoviruses, etc., capable of infecting humans, monkeys or others. mammalian species, as well as those obtained from wild-type viruses or by various manipulations with their genes, including rearrangements, insertions, mutations, etc. Such vectors can have one or more wild-type genes, as well as completely or partially deleted sequences, provided that functional flanking regions are preserved, which are responsible for the preservation, replication and correct packaging of the virion. Expression vectors (expression cassettes) are constructed using known methods to ensure functionality and integrity of its elements, including a transcription initiation region, a nucleic acid molecule of a given sequence, and a terminal region. The composition of these elements is selected in such a way as to ensure their functionality in mammalian cells. Any viral strains or serotypes can be used as a "viral vector" in the present invention. For example, the genome or particle of an rAAV vector (with capsids VP1, VP2 and / or VP3) can include any serotype, for example, AAV-1, -2, -3, -4, -5, -6, -7, -8, -9, -10, -1 1, -12, -rh74, -rhlO, or AAV-2i8, where vectors may be based on the same strain or serotype (or subgroup and variant) or be different from each other.

An immune response, such as the humoral immunity of the host organism, can develop against virtually any vector and component of the delivery system, including viruses, transport proteins, polymer structures, nanoparticles, etc., as well as the polynucleotides themselves or proteins or peptides encoded by this polynucleotide. This can potentially lead to partial or complete inhibition of vector functions and / or disrupt the processes of cell transduction, expression or functions of both the heterologous polynucleotide itself and the functions or activity of the encoded proteins or peptides.

In some embodiments of the present invention, the heterologous polynucleotide used for delivery encodes a protein that can be selected from the following group: insulin, glucagon, growth hormone (GH), parathyroid hormone (PTH), growth hormone release factor (GRF), follicle stimulating hormone (FSH) , luteinizing hormone (LH), human chorionic gonadotropin (hCG), vascular endothelial growth factor (VEGF), angiopoietins, angiostatin, granulocyte colony stimulating factor (GCSF), erythropoietin (EPO), connective tissue growth factor (CTGF) (basic growth factor) bFGF), acidic fibroblast growth factor (aFGF), acidic a-glucosidase (GAA), epidermal growth factor (EGF), transforming growth factor a (TGFa), platelet growth factor (PDGF), insulin growth factors I and II (IGF- I and IGF-II)), TGF, activins, inhibitors, bone morphogenic protein (BMP), nerve growth factor (NGF), brain neurotrophic factor (BDNF), neurotrophins NT-3 and T4 / 5, ciliary neurotrophic i factor (CNTF), glial cell line neurotrophic factor (GDNF), neuroturin, agrin, netrin-1 and netrin-2, hepatocyte growth factor (HG F), ephrins, noggin and tyrosine hydroxylase.

In certain embodiments of this invention, the expected effect can be obtained without the specific administration of drugs that suppress the immune response (immunosuppressants). In other embodiments of the invention, at least one immunosuppressive agent may be added prior to, concurrently with, or after administration of the vector to the subject. In certain embodiments of the invention, an anti-inflammatory agent can be used as such an immunosuppressive agent.

The compositions and vectors of this invention can be administered in any sterile, biocompatible pharmaceutical carrier, including, but not limited to, saline, phosphate-saline solution, dextrose solution, or water, as long as the therapeutic and pharmacological properties of the composition are preserved. The composition can be administered to a subject alone or in combination with other agents that affect dose size, frequency of administration, and / or therapeutic efficacy of the drug.

According to this invention, the introduction of the drug can be carried out systemically, regionally, locally or in any other way, for example, by injection or infusion. Delivery of pharmaceutical compositions in vivo, as a rule, can be carried out both by injection using a conventional syringe and other known methods of drug administration. For example, the compositions can be delivered subcutaneously, epidermally, intradermally, intrathecally, intrabitally, intramucosally, intraperitoneally, intravenously, intrapleurally, intraarterially, orally, intrahepatically, through the portal vein, or intramuscularly.

Reducing side effects, reducing systemic toxicity of the therapeutic agent due to the administration of antibodies

The disclosed compositions and methods, in some embodiments of the invention, can reduce the side effects of agent administration. This means that with the introduction of the above antibodies, the side effects of the agent are reduced compared to the administration of agents based on magnetic particles, polyethyleneimine and polynucleotides (from Examples 27-29) in 12 hours after the administration of antibodies, the toxicity of these complexes decreases, namely, significantly reduces the lethality ... So, for example, if, 12 hours after the administration of antibodies, a dose of such an agent is administered twice as high as the LD50 for the control experiment without the administration of antibodies, then all animals in the experiment survive. In some embodiments of the invention, this allows for significantly higher doses of agents to be administered and to achieve significantly greater delivery of the payload of an agent, for example, a polynucleotide (which in turn leads to a significant increase in the expression of a protein or targeted nucleic acid).

The effect of reducing the toxicity of a therapeutic agent (in particular, said agent for delivering a polynucleotide) in some embodiments of the method (for example, in which said antibody is an antibody against erythrocytes) is explained by an increase in the level of a number of cytokines, in particular IL-10, as was shown in [Robak, Blood , 2012]. Among other functions, this interleukin has protective properties, prevents excessive activation of inflammation, inhibits the cytotoxic activity of NK cells, etc. In addition, due to this effect, in some embodiments of the invention, an additional level of control over the specificity of the therapeutic effect is achieved. So, for example, the mentioned antibody with effector functions is selected, which is personalized for the patient (including with a selected dose) or applied to a certain group of patients based on the analysis of their genome or determination of the probability of higher expression or susceptibility to IL-10, or with by control measurement of the level of IL-10 in response to the introduction of a small dose of this antibody to the patient before therapy (see [Robak, Blood, 2012]). Moreover, in some embodiments of the inventions, the introduction of such antibodies allows to reduce the systemic activity of the drug delivered by the agent (for example, the expression of a protein or nucleic acid from a therapeutic polynucleotide agent), especially in macrophages and other cells of the reticuloendothelial system, but to increase or maintain such activity in target cells.

Control agent

In one embodiment of the invention, the state of the mononuclear phagocytic system and the degree of its blockage are additionally controlled depending on the administration of antibodies using an additional controlling agent. Administration of a controlling agent allows the rate of elimination of nanoagents from the bloodstream of organisms to be measured as a control measure. For example, in one embodiment of the invention, for such a measurement, a fluorescently labeled liposomal monitoring agent is administered to the body, and then measurements of the level of this monitoring agent in the bloodstream are carried out either non-invasively by measuring the fluorescent signal from the monitoring agent, or invasively using blood sampling at certain time intervals. intervals and also measurements of the fluorescent signal in the given samples. Then, the characteristic of the circulation time or other quantitative parameter determining the rate of excretion of the controlling agent from the bloodstream is calculated based on the obtained data. It can be a characteristic of half-life, the area under the circulation curve, and based on measurements over a relatively short period of time, for example 1 hour, 2 hours, 3 hours, 4 hours, 6 hours, 8 hours, 12 hours, 24 hours or more. In addition, such a quantitative characteristic can be a percentage of the administered dose over a certain period of time. In addition, several such reference points can be used to refine a given quantitative characteristic.

Moreover, in one embodiment of the invention, such a quantitative time characteristic is measured before the administration of the said antibodies, as well as after the administration. At the same time, comparing these values determine the degree of blocking of the mononuclear phagocytic system in quantitative terms. This procedure allows you to evaluate and measure the effectiveness of these antibodies in a particular patient, as well as to evaluate the effectiveness of dosage of these antibodies to induce cytolacade. This allows, if necessary, to adjust either the dose or the specificity of antibodies (that is, choose the optimal antibody for therapy or use an antibody of a different specificity) individually (personalized) for a particular patient.

This procedure also avoids the negative effects of administering said polynucleotide delivery agent if the patient is unresponsive to said antibodies. In this case, an important characteristic of the controlling agent is low toxicity and the ability to produce a strong and pronounced detectable signal for its highly sensitive registration and reduction of the administered dose.

Any nano- and micro-sized objects can be a controlling agent. Preferably, the colloidal and other physicochemical characteristics of the controlling agent are close to or similar to said polynucleotide delivery agent. This allows you to increase the effectiveness and information content of such a test. These measurements allow predicting the effect of the antibody on prolonging the circulation time of the agent or its elimination by the phagocytic system.

The controlling agent can be various inorganic, organic nano- and microparticles, including magnetic, fluorescent, protein (including those that are cross-linked, polymerized or aggregated protein), polymeric (from polystyrene, dextran, polypeptide, glycolic acid polylactide and etc.) or metallic (gold, silver, semiconductor, etc.) quantum dots, carbon quantum dots, radioactive particles, and other particles.

The controlling agent can be detected by the product of the signal, incl. at least one of the following: fluorescent, fluorescent, PET, MRI contrast, radiopaque, magnetic, or plasmon resonance.

The operation of the proposed method is illustrated by the drawings of FIG. 1-4.

List of drawing figures.

Fig. 1. The dynamics of the circulation of magnetic particles introduced into the bloodstream of the mouse, measured as indicated in Example 3. In addition, an exponential approximation of the dynamics of magnetic particles is shown to determine the half-life of particles from the bloodstream.

Fig. 2. Increase in the circulation time of liposomes loaded with magnetite in the case of administration after injection of a solution of monoclonal mouse antibodies against mouse erythrocytes (clone 34-3C) as described in example 4.

Fig.Z. Deceleration of tumor growth upon administration of a composition containing antibodies against erythrocytes and a lipid-containing agent as described in Example 14. Black triangle, solid line — tumor growth in a test group of mice injected with a lipid-containing agent after administration of anti-erythrocyte antibodies; black circle, dashed line — tumor growth in the control group of mice, which were injected with a lipid-containing agent without administration of anti-erythrocyte antibodies.

Fig. 4. Fluorescent signal of organ homogenates in mice administered with AAV-DJ virus with and without administration of 34-3C antibodies (as described in Example 24).

EXAMPLES

The embodiments of the invention are varied. Here are various examples. The following examples are given to illustrate the present invention and do not limit its application.

Example 1. Synthesis of liposomes loaded with magnetite.

A solution of 22 mg of cholesterol and 42 mg of fosatidylcholine in 6 ml of a mixture of chloroform / methanol (in a ratio of 6: 1) was evaporated from a round bottom flask in a rotary evaporator. Then, 1.5 ml of a 1% solution of magnetite nanoparticles coated with citrate in 125 mM ammonium sulfate was added to the flask. Then, the liposomes are processed in an ultrasonic bath and then passed through an extruder with a membrane installed with a pore size of 100 nm. Then, the liposomes are magnetically separated from the not included magnetite and transferred to physical. solution. The final concentration of liposomes is adjusted to 200 μg magnetite / 100 μl suspension.

Example 2. Synthesis of liposomes loaded with magnetite and doxorubicin.

A solution of 22 mg of cholesterol and 42 mg of fosatidylcholine in 6 ml of a mixture of chloroform / methanol (in a ratio of 6: 1) was evaporated from a round bottom flask in a rotary evaporator. Then, 1.5 ml of a 1% solution of magnetite nanoparticles coated with citrate in 125 mM ammonium sulfate was added to the flask. Then, the liposomes are processed in an ultrasonic bath and then passed through an extruder with a membrane installed with a pore size of 100 nm. Then, the liposomes are magnetically separated from the unencumbered magnetite and transferred into a doxorubicin hydrochloride solution at the rate of 33 μg of doxorubicin hydrochloride per 1 mg of lipid. Then the resulting mixture is incubated at 60 ° C for 15 minutes, after which the liposomes are magnetically separated from the non-included doxorubicin and transferred to physical. solution. The final concentration of liposomes is adjusted to 200 μg magnetite / 100 μl suspension.

Example 3. Measurement of the circulation time of liposomes loaded with magnetite in the bloodstream of a mouse.

The tail of a Balb / c mouse was placed in the coil of the MPQ magnetic nanoparticle detector created by the inventor and described earlier in [Nikitin PI, Vetoshko PM. Magnetic susceptibility meter. RF patent 2177611 (2000), see also Nikitin et al. Nat Nanotechnol, 2014], and fixed. Then, 100 μl of a suspension of liposomes loaded with magnetite was injected into the retroorbital sinus, and the dynamics of the content of magnetite in the bloodstream of the tail veins and arteries of the mouse was recorded (see Fig. 1). The half-life of particles from the bloodstream was calculated either by approximating the obtained data by an exponential dependence, or as the average time required for a 2-fold decrease in the signal in the middle section of the curve.

Example 4. Increasing the circulation time of liposomes loaded with magnetite.

A solution of monoclonal mouse antibodies against mouse erythrocytes (clone 34-3C) (or an equal volume of 100 μl of saline in the control experiment) was injected into Balb / c mice at a dosage of 1.3-3 mg / kg intraperitoneally or into the retroorbital sinus. Then, 10-15 hours later, liposomes loaded with magnetite were injected into the retroorbital sinus, and the dynamics of the concentration of particles in the bloodstream was measured as described in example 3. The half-life of particles with the introduction of antibodies was on average 6.3 times higher than in control experiments.

Example 5. Increasing the circulation time of liposomes loaded with doxorubicin.

A solution of monoclonal mouse antibodies against mouse erythrocytes (clone 34-3C) (or an equal volume of 100 μl of saline in the control experiment) was injected into Balb / c mice at a dosage of 1.3-3 mg / kg intraperitoneally or into the retroorbital sinus. Then, 10-15 hours later, liposomes loaded with magnetite were injected into the retroorbital sinus, and the dynamics of the concentration of particles in the bloodstream was measured by measuring the fluorescent signal of doxorubicin in selected blood samples every 3 hours. The circulation time of the particles was defined as “the area under curve "(area under curve - AUC). It turned out to be on average 7.1 times higher with the introduction of antibodies than in the control experiments.

Example 6. Increasing the circulation time of PEGylated liposomes loaded with doxorubicin.

A solution of monoclonal mouse antibodies against mouse erythrocytes (clone 34-3C) (or an equal volume of 100 μl of saline in the control experiment) was injected into Balb / c mice at a dosage of 1.3-3 mg / kg intraperitoneally or into the retroorbital sinus. Then, 10-15 hours later, PEGylated liposomes loaded with doxorubicin were injected into the retroorbital sinus, and the dynamics of the concentration of particles in the bloodstream was measured by measuring the fluorescent signal of doxorubicin in collected blood samples every 3 hours. Particle circulation time was defined as the area under curve (AUC). It turned out to be on average 42% higher with the introduction of antibodies than in the control experiments.

Example 7. Increasing the circulation time of liposomes loaded with magnetite and doxorubicin.

A solution of monoclonal mouse antibodies against mouse erythrocytes (clone 34-3C) (or an equal volume of 100 μl of saline in the control experiment) was injected into Balb / c mice at a dosage of 1.3-3 mg / kg intraperitoneally or into the retroorbital sinus. Then, 10-15 hours later, liposomes loaded with magnetite and doxorubicin were injected into the retroorbital sinus, and the dynamics of the concentration of particles in the bloodstream was measured as described in example 3. The half-life of particles with the introduction of antibodies was on average 4.4 times higher than in control experiments.

Example 8. Increasing the circulation time of liposomes loaded with magnetite in the bloodstream using a complex of cells and antibodies.

A solution of monoclonal mouse antibodies against mouse erythrocytes (clone 34-3C) (or an equal volume of saline in the control experiment) was incubated for 1 hour with erythrocytes isolated from 100-200 μl of mouse blood (three times centrifuged with phosphate buffered saline). The resulting complexes of erythrocytes and antibodies were injected into the retroorbital sinus in Balb / c mice. Then, 10 hours later, liposomes loaded with magnetite were injected into the retroorbital sinus, and the dynamics of the concentration of particles in the bloodstream was measured as described in example 3. The half-life of particles with the introduction of a complex of antibodies and cells was on average 2.5 times higher than in control experiments.

Example 9. Reversibility of the effect of increasing circulation time.

A solution of monoclonal mouse antibodies against mouse erythrocytes (clone 34-3c) (or an equal volume of saline in the control experiment) was injected into Balb / c mice at a dosage of 1.5 mg / kg into the retroorbital sinus. Then, 10 days later, liposomes loaded with magnetite were injected into the retroorbital sinus, and the dynamics of the concentration of particles in the bloodstream was measured as described in example 3. The half-life of particles in the case of administration of antibodies did not differ from the half-life in the control experiment (within the experimental error).

Example 10 The effectiveness of increasing circulation time with repeated administrations of a liposome-based composition loaded with magnetite.

A solution of monoclonal mouse antibodies against mouse erythrocytes (clone 34-3c) (or an equal volume of saline in the control experiment) was injected into Balb / c mice at a dosage of 1.5 mg / kg into the retroorbital sinus, then after 12 hours liposomes loaded with magnetite were injected and measured the kinetics of elimination as described in example 3. The same animals were carried out the same procedure after 10 days and again after 10 days. The ratio of the half-life of particles in the case of administration of antibodies to the half-life of particles in the case without administration of antibodies practically did not change for repeated injections and amounted to 6-6.5 times.

Example 11. Increasing the efficiency of delivery of liposomes loaded with magnetite and doxorubicin into a tumor. Mice were injected subcutaneously with 10 ^{L of} 6 B16F10 melanoma cells. After the development of tumors, mice with a tumor in the size of 150-200 cubic meters. mm with a neodymium magnet attached to the tumor, liposomes loaded with magnetite and doxorubicin were injected as described in Example 4 (after and without administration of a solution of monoclonal mouse antibodies against mouse erythrocytes). Organs and tumor were extracted 5 hours after particle injection. The content of magnetic particles in the tumor was measured using a magnetic nanoparticle detector created by the author of the invention and described earlier in [Nikitin PI, Vetoshko PM. Magnetic susceptibility meter. RF patent 2177611 (2000)]. The content of particles (per gram of tissue) in tumors of mice in the case of administration of anti-erythrocyte antibodies was, on average, 7.3 times higher than the content of particles in tumors of mice in the case of no administration of anti-erythrocyte antibodies. In addition, the non-invasive detection of these liposomes in the tumor using the MPQ method (the detected signal in the tumor is 2.3 times higher in the experimental group than in the control group) makes it possible to diagnose the presence of a tumor.

The same experiment was repeated with other tumors. The efficiency of delivery to the tumor when the agent was injected after antibodies increased 3.4 times for the BT-474 human breast tumor xenograft, and 2.5 times for Lewis lung carcinoma.

Example 12. Increasing the efficiency of delivery of liposomes loaded with magnetite and doxorubicin into a tumor by preliminary administration of xenogeneic antibodies against erythrocytes.

Mice were injected subcutaneously with 10 ^{L of} 6 B16F10 melanoma cells. After the development of tumors, mice with a tumor measuring 50-200 cubic meters. mm, liposomes loaded with magnetite and doxorubicin were injected 12 hours after administration of a solution of polyclonal rabbit gamma immunoglobulins against mouse erythrocytes at a dosage of 50 mg / kg or without the introduction of such antibodies. Organs and tumor were extracted 5 hours after particle injection. The content of magnetic particles in the tumor was measured using a magnetic nanoparticle detector, as in Example 11. The content of particles (per gram of tissue) in tumors of mice in the case of administration of anti-erythrocyte antibodies was on average 2 times greater than the content of particles in tumors of mice in the case of without the introduction of anti-erythrocyte antibodies.

Example 13. Increasing the circulation time of liposomes loaded with magnetite in the bloodstream of a mouse by the introduction of polyclonal xenogeneic antibodies and secondary allogeneic antibodies.

A solution of polyclonal rabbit antibodies (gamma-immunoglobulins) against mouse erythrocytes (or an equal volume of 100 μl of saline in the control experiment) was intravenously injected into Balb / c mice at a dosage of 10 mg / kg. After 1 hour, the mice were intravenously injected with a solution of polyclonal mouse antibodies against rabbit gamma-immunoglobulins (or an equal volume of 100 μl of physiological saline in the control experiment) at a dosage of 1.3-3 mg / kg. Then, 10-15 hours later, liposomes loaded with magnetite were injected into the retroorbital sinus, and the dynamics of the concentration of particles in the bloodstream was measured as described in example 3. The half-life of particles with the introduction of rabbit and mouse antibodies was on average 2.3 times longer than in control experiments (without the introduction of antibodies).

Example 14. Slowdown of tumor growth upon administration of a composition containing antibodies against erythrocytes and a lipid-containing agent.

Mice were injected subcutaneously with 10 ^{L of} 6 B16F10 melanoma cells. After the development of tumors, mice with a tumor of about 50-200 cubic meters. mm was first injected with a solution of monoclonal mouse antibodies against mouse erythrocytes (as described in Example 3) and nothing was administered to the mice in the control group. Then, 12 hours later, mice of both groups were injected with liposomes loaded with magnetite and doxorubicin (with a neodymium magnet attached to the tumor). Tumor growth in both cases is shown in Fig. 3. As can be seen from Fig. 3, the tumor size 10 days after the injection of liposomes after the injection with the antibody was 3.5 times smaller than after the injection of liposomes.

Example 15. Extension of the lifetime of animals with melanoma upon administration of a composition containing antibodies against erythrocytes and a lipid-containing agent. Mice were injected subcutaneously with 10 ^{L of} 6 B16F10 melanoma cells. After the development of tumors, mice with a tumor of about 50-200 cubic meters. mm was first introduced a solution of monoclonal mouse antibodies against mouse erythrocytes (as described in Example 3) or physical. solution in the control group, and then 12 hours later liposomes loaded with magnetite and doxorubicin (with a neodymium magnet attached to the tumor). The average survival time of animals in the group to which the agent with doxorubicin was administered after administration of monoclonal mouse antibodies against mouse erythrocytes turned out to be longer than the average life time in the group of animals to which only the agent was administered by 30%.

Example 16. Inhibition of tumor growth with the introduction of antibodies against erythrocytes and PEGylated liposomal doxorubicin.

Mice were injected subcutaneously with 10 ^{L of} 6 B16F10 melanoma cells. After the development of tumors, mice with a tumor of about 50-200 cubic meters. mm was first injected with a solution of monoclonal mouse antibodies against mouse erythrocytes (as described in Example 3) and nothing was administered to the mice in the control group. Then, 12 hours later, mice of both groups were injected with the preparation of PEGylated liposomal doxorubicin Kelix. The tumor size 10 days after the injection of liposomes after the injection with the antibody was 1.9 times smaller than after the injection of liposomes alone.

Example 17. Extension of the survival time of animals with melanoma with the introduction of antibodies against erythrocytes and PEGylated liposomal doxorubicin.

Mice were injected subcutaneously with 10 ^{L of} 6 B16F10 melanoma cells. After the development of tumors, mice with a tumor of about 50-200 cubic meters. mm was first introduced a solution of monoclonal mouse antibodies against mouse erythrocytes (as described in Example 3) or physical. solution in the control group, and then, 12 hours later, the preparation of PEGylated liposomal doxorubicin Kelix was administered. The average survival time of animals in the group to which Kelix was injected after the administration of monoclonal mouse antibodies against mouse erythrocytes turned out to be longer than the average lifetime in the group of animals to which only the agent was administered, by 23%.

Example 18. Increasing the circulation time of magnetic nanoparticles by the introduction of antibodies against CD47.

A solution of monoclonal mouse antibodies against mouse / rat / human CD47 (clone MIAP410) (or equal volume - 100 μl - saline in the control experiment) was injected into Balb / c mice at a dosage of 1.3-3 mg / kg intraperitoneally or into the retroorbital sinus. Then, 10-15 hours later, magnetic nanoparticles Chemicell fluidMAG-ARA-lOOnm were injected into the retroorbital sinus, and the dynamics of the concentration of particles in the bloodstream was measured as described in example 3. The half-life of particles with the introduction of antibodies was on average 3.2 times higher than in the control experiments.

Example 19. Increasing the circulation time of magnetic nanoparticles by introducing a mixture of antibodies 34-3C and MIAP410.

A solution of monoclonal mouse antibodies against mouse / rat / human CD47 (clone MIAP410) at a dose of 1.3 mg / kg and monoclonal mouse antibodies against erythrocytes clone 34-3C at a dose of 1.3 mg / kg (or an equal volume - 100 μl - saline in a control experiment ) was injected into Balb / c mice in the retroorbital sinus. Then, 12 hours later, magnetic nanoparticles Chemicell fluidMAG-ARA-100nm were injected into the retroorbital sinus, and the dynamics of the concentration of particles in the bloodstream was measured as described in example 3. The half-life of particles with the introduction of antibodies was on average 12.5 times higher than in control experiments.

Example 20. Increasing the circulation time of magnetic nanoparticles by introducing a mixture of antibodies 34-3C and Fab-fragments of antibodies against CD47.

A solution of Fab fragments of monoclonal mouse antibodies against mouse / rat / human CD47 at a dose of 1.3 mg / kg and monoclonal mouse antibodies against erythrocytes clone 34-3C at a dose of 1.3 mg / kg (or an equal volume of 100 μL of saline in control experiment) was injected into Balb / c mice in the retroorbital sinus. Then, 12 hours later, magnetic nanoparticles Chemicell fluidMAG-ARA-lOOnm were injected into the retroorbital sinus, and the dynamics of the concentration of particles in the bloodstream was measured as described in example 3. The half-life of particles with the introduction of antibodies was on average 11.3 times higher than in control experiments.

Example 21. Increasing the circulation time of magnetic liposomes by the introduction of antibodies against CD47.

A solution of monoclonal mouse antibodies against mouse / rat / human CD47 (clone MIAP410) (or equal volume - 100 μl - saline in the control experiment) was injected into Balb / c mice at a dosage of 1.25 mg / kg intraperitoneally or into the retroorbital sinus. Then, 10-15 hours later, liposomes loaded with magnetite were injected into the retroorbital sinus, and the dynamics of the concentration of particles in the bloodstream was measured as described in example 3. The half-life of particles with the introduction of antibodies was on average 2.5 times higher than in control experiments.

Example 22. Increasing the circulation time of liposomes loaded with magnetite in the bloodstream using a complex of cells and antibodies against CD47.

A solution of monoclonal mouse antibodies against mouse / rat / human CD47 (clone MIAP410) (or an equal volume of saline in the control experiment) was incubated for 1 hour with erythrocytes isolated from 100-200 μl of mouse blood (three times centrifuged with phosphate buffered saline ). The resulting complexes of erythrocytes and antibodies were injected into the retroorbital sinus in Balb / c mice. Then, 10 hours later, liposomes loaded with magnetite were injected into the retroorbital sinus, and the dynamics of the concentration of particles in the bloodstream was measured as described in example 3. The half-life of particles with the introduction of a complex of antibodies and cells was on average 2.1 times higher than in control experiments.

Example 23. Increasing the circulation time of liposomes loaded with magnetite in the bloodstream using a complex of cells and a mixture of antibodies 34-3C and MIAP410.

A solution of monoclonal mouse antibodies against mouse / rat / human CD47 (clone MIAP410) at a dose of 1.3 mg / kg and monoclonal mouse antibodies against erythrocytes clone 34-3C at a dose of 1.3 mg / kg (or an equal volume of saline in the control experiment) was incubated for 1 hour with erythrocytes isolated from 100-200 μl of mouse blood (three times purified in a centrifuge with phosphate-buffered saline). The resulting complexes of erythrocytes and antibodies were injected into the retroorbital sinus in Balb / c mice. Then, 10 hours later, liposomes loaded with magnetite were injected into the retroorbital sinus, and the dynamics of the concentration of particles in the bloodstream was measured as described in example 3. The half-life of particles with the introduction of a complex of antibodies and cells was on average 3.2 times higher than in control experiments.

Example 24. Increasing the efficiency of delivery of a gene (polynucleotide) encoding a protein using a virus.

The AAV-DJ virus with an inserted polynucleotide expressing the fluorescent protein GFP, at a dose of 5 * 10 ^{L of} 12 viral particles per mouse, is injected into Balb / c mice without the administration of 34-3C antibodies or 12 hours after the administration of 34-3C (as described in Example 4). After 3 weeks, the organs of the animal are extracted, homogenized, centrifuged, and the expression level of the reporter protein is measured using a fluorometer. Figure 4 shows significantly greater protein expression in mice treated with 34-3C antibody as compared to controls.

Example 25 Synthesis of a polynucleotide delivery agent using a cationic lipid agent DOTAP (lipid containing agent).

A solution of DOTAP and cholesterol in chloroform in a ratio of 3 to 1 by weight is evaporated on a rotary vacuum evaporator at 45 degrees (until a thin lipid film is formed on the walls of the flask). Then, a 5% glucose solution is added, treated with ultrasound, and the resulting suspension is pushed through 100-nm polycarbonate membranes. Then, mix the liposome suspension with plasmid DNA (encoding Nanoluc luciferase) in a ratio of 4: 1 by weight (4 mg of liposomes are added to 1 mg of DNA). Then incubated for 20 minutes.

Example 26. Increasing the efficiency of delivery to the lungs of a gene (polynucleotide) encoding a protein using a cationic lipid-based agent DOTAP (lipid-containing agent).

The obtained lipoplexes from Example 25 (in an amount of 100 μg by plasmid DNA) were injected into Balb / c mice without the administration of 34-3C antibodies or 12 hours after the administration of 34-3C (as described in Example 4). After 24 hours, the lungs are extracted, homogenized, centrifuged and the substrate coelenterazine-h is added to the aliquot and the level of luciferase expression is measured using a luminometer. The expression of luciferase (by the level of luminescence) in the case of mice injected with the 34-3C antibody is 76% higher than in the control.

Example 27. Synthesis of magnetic nanoagents for delivery of heterologous polynucleotide.

Magnetite particles are synthesized as described in Nikitin et al. Nat Nanotechnol. 2014 coated with sodium citrate (as in Example 1). Then, to 1 mg of this particle add 15 mg of branched polyethyleneimine (average molecular weight 25 kDa), incubate for 30 minutes at room temperature, sonicating every 5 minutes. The particles are washed three times with water, separating the particles on a magnet. Finally, resuspend the particles in phosphate buffered saline using an ultrasonic bath. Then add 0.15 mg of plasmid DNA (encoding nanoLuc luciferase), incubate at room temperature for 1 hour, periodically treating the suspension in an ultrasonic bath, and then wash the particles three times with water, removing unbound DNA. Finally, the particles are resuspended in a 5% glucose solution.

Example 28. Increasing the efficiency of delivery to the lungs of a gene (polynucleotide) using magnetic agents and the TER-119 antibody.

The obtained magnetic nanoagents from Example 27 (in an amount of 100 μg for plasmid DNA) were injected into Balb / c mice without the administration of TER-119 antibodies or 12 hours after the administration of TER-119 (as described in Example 4). After 24 hours, the lungs are extracted, homogenized, centrifuged and the substrate coelenterazine-h is added to the aliquot and the level of luciferase expression is measured using a luminometer. The expression of luciferase (by the level of luminescence) in the case of mice with the injected antibody 34-3C is 1.5 times higher than in the control (without the introduction of 34-3C).

Example 29. Increasing the efficiency of delivery to the lungs of a gene (polynucleotide) using magnetic agents and antibody 34-3C.

The obtained magnetic nanoagents from Example 27 (in an amount of 100 μg for plasmid DNA) were injected into Balb / c mice without the administration of TER-119 antibodies or 12 hours after the administration of TER-119 (as described in Example 4). After 24 hours, the lungs are extracted, homogenized, centrifuged and the substrate coelenterazine-h is added to the aliquot and the level of luciferase expression is measured using a luminometer. The expression of luciferase (by the level of luminescence) in the case of mice with the injected antibody 34-3C is 1.9 times higher than in the control (without the introduction of 34-3C).

Claims

Claim

1) A composition for use in a therapeutic or diagnostic method comprising at least i) an antibody in free molecular form against the patient's blood cells, and i) a lipid-containing drug agent.

2) A composition according to claim 1, wherein said antibody is an anti-erythrocyte antibody.

3) A composition according to claim 1, wherein said antibody, when administered to an organism, induces erythrophagocytosis.

4) A composition according to claim 1, wherein said antibody, when administered to an organism, induces Fc receptor-mediated erythrophagocytosis.

5) A composition according to claim 1, wherein said antibody is highly homologous to the species antibodies of the organism (or a variant thereof), including autologous, allogeneic, humanized or chimeric.

6) A composition according to claim 1, wherein said antibody is a monoclonal antibody.

7) The composition of claim 1, wherein said antibody is a polyclonal antibody.

8) A composition according to claim 1, wherein said antibody is a mixture of several monoclonal antibodies.

9) A composition according to claim 1, wherein said antibody is an antibody isolated from donated blood.

10) A composition according to claim 1, wherein said antibody is an anti-rhesus RhO (D) antibody.

11) The composition according to claim 1, in which the dose of said antibody is selected sufficient so that the circulation time in the bloodstream of said lipid-containing drug is increased by at least 30% compared to the circulation time in the bloodstream of said lipid-containing drug without introduction of the said antibody.

12) A composition according to claim 1, in which the dose of said antibody is selected sufficient so that the circulation time in the bloodstream of said lipid-containing drug is increased by at least 3 times compared to the circulation time in the bloodstream of said lipid-containing drug without introduction of the said antibody.

13) A composition according to claim 1, wherein the dose of said antibody is selected sufficient so that the efficiency of delivery of said agent to the target is increased by at least 30% compared to the efficiency of delivery of said lipid-containing drug without administration of said antibody.

14) A composition according to claim 1, wherein the dose of said antibody is selected sufficient so that the efficiency of delivery of said agent to the target is increased by at least 3 times as compared to the efficiency of delivery of said lipid-containing drug without administration of said antibody.

15) A composition according to claim 1, wherein the dose of said antibody does not exceed 5 mg / kg body weight.

16) A composition according to claim 1, wherein the dose of said antibody does not exceed 50 mg / kg body weight.

17) A composition according to claim 1, wherein the dose of said antibody does not exceed 500 mg / kg body weight.

18) The composition of claim 1, wherein said antibody and said lipid-containing drug are administered to the body in separate dosage forms.

19) The composition according to claim 1, which is presented in the form of a unit dosage form. 20) A composition according to claim 1, wherein said antibody and said lipid-containing drug are contained together in a single dosage form or in two different dosage forms.

21) The composition of claim 1, wherein said antibody and said lipid-containing drug are administered at different times.

22) A composition according to claim 1, wherein said antibody is administered prior to administration of said lipid-containing drug.

23) A composition according to claim 1, wherein said lipid-containing drug agent is a liposomal agent.

24) A composition according to claim 1, wherein said lipid-containing drug agent is a liposome.

25) The composition of claim 1, wherein said lipid-containing drug agent is a pegylated liposome.

26) The composition of claim 1, wherein said lipid-containing drug agent is liposomal doxorubicin.

27) The composition of claim 1, wherein said lipid-containing drug agent is pegylated liposomal doxorubicin.

28) A composition according to claim 1, in which said agent consists of at least one of the following nano or microparticles: magnetic, fluorescent, protein (including a cross-linked, polymerized or aggregated protein), polymeric , incl. consisting of at least one of the following polymers: polystyrene, dextran, polypeptide, glycolic acid polylactide, or block copolymers) or crystalline (gold, silver, semiconducting, or metal) nano- or microparticles.

29) A composition according to claim 1, in which said agent performs a visualizing function by producing a detected signal, incl. at least one of the following: fluorescent, fluorescent, PET signal, MPT contrast signal, X-ray contrast signal, magnetic signal, or plasmon resonance signal, or signal due to absorption of light or other electromagnetic or acoustic waves.

30) A composition according to claim 1, wherein said agent performs a therapeutic function, for example, by delivering cytostatic or cytotoxic compounds or medicinal compounds, incl. low molecular weight, enzymatic, radioactive, chemotherapeutic, substances for photodynamic therapy, hyperthermia.

31) The composition of claim 1, which further comprises erythrocytes.

32) The composition according to claim 1, which further contains components of donated blood.

33) The composition of claim 1, which further comprises autologous blood components.

34) The composition of claim 1, which further comprises an erythropoiesis stimulant.

35) The composition of claim 1, which further comprises erythropoietin.

36) Composition according to PP. 1-35 for use in a method of treating cancer.

37) Composition according to PP. 1-35 for use in a method of treating a disease or condition selected from the group consisting of: cancer, atherosclerosis, stroke, heart attack.

38) Composition according to PP. 1-35 for use in an angiocontrast method. 39) Composition according to PP. 1-35 for use in a method of treating an oncological disease, characterized in that said antibody is first administered, followed by said lipid-containing drug.

40) A method for diagnosing or treating diseases or conditions of the body, in which the composition according to claims 1-35 is introduced into the body.

41) A method for diagnosing or treating diseases or conditions of an organism, in which: i) the said antibody of the composition according to claims 1-35 is administered to the organism, and) said lipid-containing medicinal agent of the composition according to claims 1-35 is administered to the organism.

42) The method according to claim 41, wherein said agent is administered at least 6 hours and not more than 18 hours after administration of said antibody.

43) The method according to claim 41, wherein said administration of said antibody to the body increases the diagnostic or therapeutic effect of said agent as compared to administration of said agent without administration of said antibody.

44) The method according to claim 41, wherein said administration of said antibody to the body increases the circulation time of said agent in the bloodstream of the body.

45) The method according to claim 41, characterized in that, along with the introduction of said antibody and said agent, erythrocytes are introduced into the body.

46) The method according to claim 41, characterized in that along with the introduction of said antibody and said agent, the blood of the organism itself or its components or donor blood or its components are introduced into the body.

47) The method according to claim 41, characterized in that, along with the introduction of said antibody and said agent, a composition for stimulating erythropoiesis is administered to the body.

48) The method according to claim 41 for the diagnosis or therapy of cancer.

50) The method of claim 40 further comprising the step of quantifying blockage of the reticuloendothelial system, which includes administering a controlling agent and measuring its concentration in the bloodstream after a specified time interval to calculate a blockage characteristic of the reticuloendothelial system.

51) The method according to claim 40, further comprising the step of measuring the quantitative characteristic of the blockage of the reticuloendothelial system, which includes: a) the step of administering a controlling agent prior to said administration of said antibodies and measuring its concentration in the bloodstream after a certain time interval to calculate the characteristic blocking of the reticulo-endothelial system. b) a step of introducing a controlling agent after said administration of said antibodies and measuring its concentration in the bloodstream after a certain time interval to calculate the characteristic of blocking the reticuloendothelial system.

49) Method of using compositions according to claims 1-35. for the manufacture of drugs for the treatment of cancer.

50) A method of promoting products to the market, including at least promotion for research, diagnostics, monitoring or therapy of diseases or conditions of the body, at least i) a composition according to claims 1-35, or i) a method according to claims ... 40-49. 51) A method according to claim 50, characterized in that the promotion is carried out by an insert in a package with a commercial composition containing at least the composition according to claims. 1-35.

52) The method according to claim 50, characterized in that the promotion is carried out by written or oral communication to the doctor or health care provider.

53) A business method, including at least marketing for diagnostics and therapy, or to improve the effectiveness of diagnostics or therapy, or to deliver an agent to the body, or to increase the circulation time of the agent in the bloodstream of the body i) a composition according to claims. 1-35, or i) the method according to PP. 40-49.