RU2786053C2 - Pharmaceutical composition for use for treatment or prevention of c5-related disease and method for treatment or prevention of c5-related disease - Google Patents

Abstract

FIELD: medicine; therapy.

SUBSTANCE: invention relates to the field of medicine, namely to therapy; it is intended for the treatment or prevention of a C5-related disease. A method for the treatment of prevention of a C5-related disease includes (a) intravenous administration to an individual of a single intravenous dose of an antibody to C5 at a dose of 500 mg, 1000 mg, or 1500 mg of the antibody, as well as (b) subcutaneous administration to an individual of two or more subcutaneous doses of the antibody to C5 (i) at a dose of 340 mg of the antibody once a week or every two weeks, or (ii) from 600 mg to 2000 mg of the antibody every four weeks. The specified antibody to C5 contains a sequence of a variable domain of a heavy chain (VH), consisting of an amino acid sequence SEQ ID NO: 2, a sequence of a variable domain of a light chain (VL), consisting of an amino acid sequence SEQ ID NO: 6, a sequence of a constant domain of a heavy chain (CH), consisting of an amino acid sequence SEQ ID NO: 13, and a sequence of a constant domain of a light chain (CL), consisting of an amino acid sequence SEQ ID NO: 18.

EFFECT: use of the invention allows for an increase in the efficiency and safety of treatment or prevention of C5-related disease.

11 cl, 10 dwg, 3 tbl, 9 ex

Description

The technical field to which the invention belongs

The present invention relates to dosages and schedules for administering an anti-C5 antibody.

Prerequisites for the creation of the invention

The complement system plays a central role in immune complex clearance and immune responses to infectious agents, foreign antigens, virus-infected cells, and tumor cells. Approximately 25-30 complement proteins are known to exist as a complex group of plasma proteins and membrane cofactors. Complement components exercise their immune protective functions by participating in a series of complex enzymatic cleavages and membrane binding events. The resulting cascades of the complement system lead to products with opsonic, immunoregulatory and lytic functions.

It is now generally accepted that the complement system can be activated in three different ways: the classical pathway, the lectin pathway, and the alternative pathway. There are many components involved in these pathways, and although they differ in their initial stages, they then converge and include the same terminal components of the complement system (C5 to C9), which are responsible for the activation and destruction of target cells.

The classical pathway is usually activated by the formation of antigen-antibody complexes. Regardless, the first step in the activation of the lectin pathway is the binding of specific lectins such as mannan-binding lectin (MBL), H-ficolin, M-ficolin, L-ficolin, and C-type lectin CL-11. In contrast, the alternative pathway undergoes spontaneous activation with a low level of turnover, which can easily rise on foreign or other abnormal surfaces (bacteria, yeast, virus-infected cells, or damaged tissue). These pathways converge at a point where complement component C3 is cleaved by an active protease to form C3a and C3b.

C3a is an anaphylatoxin. C3b binds to bacterial and other cells, as well as certain viruses and immune complexes, and marks them for removal from the bloodstream (plays the role of an opsonin). C3b also forms a complex with other components to form C5 convertase, which cleaves C5 to form C5a and C5b.

C5 is a protein with a molecular weight of 190 kDa, which is present in the serum of a healthy individual at a concentration of approximately 80 μg/ml (0.4 μm). C5 is glycosylated, with about 1.5-3% of its mass being carbohydrate. Mature C5 is a heterodimer in which the 115 kDa alpha chain is disulfide-linked to the 75 kDa beta chain. C5 is synthesized as a single chain precursor protein (pro-C5 precursor) consisting of 1676 amino acids (see, for example, PTL1 and PTL2). The pro-C5 precursor is cleaved to release the beta chain as an amino-terminal fragment and the alpha chain as a carboxy-terminal fragment. Polypeptide fragments, representing the alpha chain and the beta chain, are connected to each other through a disulfide bond to form the mature C5 protein.

Mature C5 is cleaved to form a C5a fragment and a C5b fragment during the activation of complement pathways. As a result of the action of C5 convertase, C5a is cleaved from the C5 alpha chain in the form of an amino-terminal fragment containing the first 74 amino acids of the alpha chain. The remainder of the mature C5 is a C5b fragment which contains an alpha chain residue linked by a disulfide to the beta chain. Approximately 20% of the mass of C5a, which is 11 kDa, is accounted for by carbohydrate.

C5a is another anaphylatoxin. C5b combines with C6, C7, C8 and C9 to form a membrane attack complex (MAC, C5b-9, Terminal Complement Component Complex (TCC)) on the surface of the target cell. When sufficient MACs are incorporated into target cell membranes, the pores formed by the MAC mediate rapid osmotic lysis of the target cells.

As stated above, C3a and C5a are anaphylatoxins. They can trigger mast cell degranulation, which causes the release of histamine and other inflammatory mediators, which lead to smooth muscle contraction, increased vascular permeability, leukocyte activation, and other manifestations of inflammation, including cell proliferation leading to increased parenchymal cell counts. C5a also functions as a chemotactic peptide that serves to recruit granulocytes such as neutrophils, eosinophils, basophils, and monocytes to the site of complement activation.

C5a activity is regulated by the plasma enzyme carboxypeptidase N, which removes the carboxy-terminal arginine from C5a, resulting in the formation of the C5a-des-Arg derivative. C5a-des-Arg has only 1% of the anaphylactic activity and polymorphonuclear chemotactic activity inherent in unmodified C5a.

While a properly functioning complement system provides effective protection against infecting organisms, misregulation or activation of the complement system has been implicated in the pathogenesis of various disorders including, for example, rheumatoid arthritis (RA); lupus nephritis; damage caused by ischemia-reperfusion; paroxysmal nocturnal hemoglobinuria (PNH); atypical hemolytic uremic syndrome (aHUS); dense deposit disease (DDD); macular degeneration (eg age-related macular degeneration (AMD)); HELLP syndrome (hemolysis, increased activity of liver enzymes and thrombocytopenia); thrombotic thrombocytopenic purpura (TTP); spontaneous loss of the fetus; Pauci-immune (low-immune) vasculitis (vasculitis with hypocomplementemia); bullous epidermolysis; recurring fetal loss; multiple sclerosis (MS); traumatic brain injury; and damage caused by myocardial infarction, cardiopulmonary bypass, and hemodialysis (see, for example, NPL1). Thus, inhibition of excessive or uncontrolled activation of the complement cascade may have a beneficial clinical effect on patients with these disorders.

Paroxysmal nocturnal hemoglobinuria (PNH) is a rare blood disorder in which red blood cells are defective and therefore destroyed faster than healthy red blood cells. PNH is caused by clonal expansion of hematopoietic stem cells with somatic mutations in the PIG-A (class A phosphatidylinositolglycan) gene located on the X chromosome. Mutations in PIG-A result in early blockade of the synthesis of glycosylphosphatidylinositol (GPI), a molecule required to anchor many proteins to cell surfaces. As a result, in PNH, blood cells are deficient in GPI-anchored proteins, which include the complement-regulating proteins CD55 and CD59. Under normal circumstances, these complement-regulating proteins block the formation of MAC on cell surfaces, thereby preventing erythrocyte lysis. The absence of GPI-anchored proteins causes complement-mediated hemolysis in PNH.

PNH is characterized by hemolytic anemia (low red blood cell count), hemoglobinuria (the presence of hemoglobin in the urine, especially after sleep), and hemoglobinemia (the presence of hemoglobin in the bloodstream). PNH affected individuals are known to have paroxysms, which in this case are cases of dark colored urine. Hemolytic anemia is caused by intravascular destruction of red blood cells by components of the complement system. Other known symptoms include dysplasia, fatigue, erectile dysfunction, thrombosis, and recurring abdominal pain.

Eculizumab is a humanized anti-complement C5 monoclonal antibody that is a first-line therapeutic approved for the treatment of paroxysmal nocturnal hemoglobinuria (PNH) and atypical hemolytic uremic syndrome (aHUS) (see e.g. NPL2). Eculizumab inhibits the cleavage of C5 by C5 convertase with the formation of C5a and C5b, which prevents the formation of the complex of the terminal components of the complement system C5b-9. Both C5a and C5b-9 induce terminal complex mediated events characteristic of PNH and aHUS (see also PTL3, PTL4, PTL5 and PTL6).

Several publications describe antibodies to C5. For example, PTL7 describes an anti-C5 antibody that binds to the C5 alpha chain but does not bind to C5a and blocks C5 activation, and PTL8 describes an anti-C5 monoclonal antibody that inhibits the formation of C5a. On the other hand, PTL9 describes an anti-C5 antibody that recognizes a proteolytic C5 convertase cleavage site on the C5 alpha chain and inhibits the conversion of C5 to C5a and C5b. PTL10 describes an anti-C5 antibody that has an affinity constant of at least 1×10 ⁷ M ^-1 .

Antibodies (IgG) bind to the neonatal Fc receptor (FcRn) and are characterized by long plasma retention times. IgG binding to FcRn typically occurs under acidic conditions (eg, pH 6.0) and rarely occurs under neutral conditions (eg, pH 7.4). Typically, IgGs are non-specifically incorporated into cells via endocytosis and returned to cell surfaces via binding to endosomal FcRn under acidic conditions in endosomes. The IgGs are then separated from the FcRn by dissociation under neutral conditions in the plasma. IgGs that fail to bind to FcRn are cleaved in lysosomes. If the ability of IgG to bind to FcRn under acidic conditions is eliminated by the introduction of mutations in its Fc region, then IgG does not return from endosomes to plasma, which leads to a marked decrease in plasma IgG retention. Published a method to increase its binding to FcRn under acidic conditions, designed to increase the retention of IgG in plasma. When IgG binding to FcRn under acidic conditions is increased by introducing an amino acid substitution into its Fc region, IgG is more efficiently returned from endosomes to plasma, and as a result, its plasma retention is increased. In addition, data have also been published showing that IgG with increased ability to bind to FcRn under neutral conditions is not separated by dissociation from FcRn under neutral conditions in plasma, even if it returns to the cell surface due to its binding to FcRn in acidic conditions. conditions in endosomes, and hence its plasma retention is not altered or rather deteriorated (see, for example, NPL3; NPL4; NPL5).

In recent years, antibodies have been described that bind to antigens in a pH dependent manner (see, for example, PTL11 and PTL12). Said antibodies bind strongly to antigens under neutral conditions in plasma and are separated by dissociation from antigens under acidic conditions which occur in endosomes. After separation by dissociation from antigens, antibodies regain the ability to bind to antigens when they are returned to plasma via FcRn. Thus, one antibody molecule can re-associate with several antigen molecules. In general, the plasma retention time of an antigen is much shorter than that of an antibody that has the above FcRn-mediated recycling mechanism. Thus, when an antigen is bound to an antibody, the antigen normally has a prolonged plasma retention, resulting in an increase in the plasma concentration of the antigen. On the other hand, it has been found that the antibodies described above, which bind to antigens in a pH-dependent manner, allow the elimination of antigens from plasma faster than typical antibodies, since they are separated by dissociation from antigens in endosomes during FcRn-mediated recycling. PTL13 also describes a computer simulation based assay demonstrating that an anti-C5 antibody having pH dependent binding can enhance antigen knockout.

Link List

Patent Literature

[PTL1] US Pat. No. 6,355,245,

[PTL2] US Pat. No. 7,432,356,

[PTL3] W02005/074607,

[PTL4] W02007/106585,

[PTL5] W02008/069889,

[PTL6] W02010/054403,

[PTL7] WO 95/29697,

[PTL8] W02002/30985,

[PTL9] W02004/007553,

[PTL10] W02010/015608,

[PTL11] WO2009/125825,

[PTL12] WO 2011/122011,

[PTL13] WO 2011/111007.

Non-Patent Literature

[NPL1] Holers et al., Immunol. Rev. 223:300-316, 2008,

[NPL2] Dmytrijuk et al., The Oncologist 13(9):993-1000, 2008,

[NPL3] Yeung et al., J Immunol. 182(12):7663-7671, 2009,

[NPL4] Datta-Mannan et al., J Biol. Chem. 282(3): 1709-1717, 2007,

[NPL5] Dall'Acqua et al., J. Immunol. 169(9):5171-5180, 2002.

Brief summary of the invention

The present invention provides pharmaceutical compositions for use in the treatment or prevention of a C5 related disease and methods of treating or preventing a C5 related disease. The present invention also provides doses and schedules for administering an anti-C5 antibody or a pharmaceutical composition containing an anti-C5 antibody.

When creating the present invention, the effective doses and regimens of administration of antibodies to C5 intended for use in the treatment and/or prevention of C5-related diseases have been studied. As a result, the present invention has found that when an antibody is administered intravenously, the plasma concentration of anti-C5 antibody rapidly increases beyond the therapeutically effective level, and the concentration is maintained at a certain level by subcutaneous administration of the antibody after intravenous administration.

Appropriate dosing conditions have also been studied in the present invention, including antibody dose, intravenous or subcutaneous frequency and intervals, interval between intravenous and subcutaneous administration, and transition from treatment with other drugs.

In particular, the present invention provides:

[1] A pharmaceutical composition for use in the treatment or prevention of a C5-related disease, wherein the composition is in a subcutaneous injection form, contains an anti-C5 antibody, and is administered subcutaneously, wherein the initial dose of the composition is administered after a dose of another pharmaceutical composition that is administered intravenously, wherein another pharmaceutical composition is formulated for intravenous injection and contains an anti-C5 antibody.

[2] A pharmaceutical composition for use in the treatment or prevention of a C5-related disease, wherein the composition is in the form of an intravenous injection, contains an anti-C5 antibody, and is administered intravenously, wherein the dose of the composition is administered before an initial dose of another pharmaceutical composition that is administered subcutaneously, wherein another pharmaceutical composition is formulated for subcutaneous injection and contains an anti-C5 antibody.

[3] The pharmaceutical composition according to [1] or [2], wherein the initial dose of the subcutaneous injection composition is administered on the same day or 1 to 28 days after the intravenous injection dose of the composition.

[4] The pharmaceutical composition according to any one of [1] to [3], wherein the subcutaneous injection composition is administered once every 3 to 42 days.

[5] The pharmaceutical composition according to any one of [1] to [4], wherein the intravenous injection composition is administered two or more times and from once per hour to once every 14 days.

[6] The pharmaceutical composition according to any one of [1] to [5], wherein the subcutaneous injection composition is administered at a dose of 17 to 6000 mg of the antibody.

[7] The pharmaceutical composition according to any one of [1] to [6], wherein the composition for intravenous injection is administered at a dose of 50 to 5000 mg of the antibody.

[8] A pharmaceutical composition according to any one of [1] to [7], wherein the composition is for treating or preventing a C5-related disease in an individual who has been treated with at least one pharmacological product intended for use in treating or preventing the disease , wherein the initial dose of the composition for intravenous injection is administered after the final dose of the pharmacological product.

[9] The pharmaceutical composition according to [8], wherein the initial dose of the composition for intravenous injection is administered on the third day or 3 days after the final dose of the pharmacological product.

[10] The pharmaceutical composition according to [8] or [9], wherein the pharmaceutical product contains an siRNA targeting C5 mRNA, or an anti-C5 antibody other than the anti-C5 antibody contained in the composition.

[11] The pharmaceutical composition according to any one of [8]-[10], wherein the pharmacological product contains eculizumab or a derivative thereof.

[12] A pharmaceutical composition for use in the treatment or prevention of a C5-related disease, wherein the composition is in the form of a subcutaneous injection, contains an anti-C5 antibody, and is administered subcutaneously at a dose of 17 to 6000 mg of the antibody.

[13] A pharmaceutical composition for use in the treatment or prevention of a C5-related disease, wherein the composition is formulated for intravenous injection, contains an anti-C5 antibody, and is administered intravenously at a dose of 50 to 5000 mg of the antibody.

[14] The pharmaceutical composition according to any one of [1]-[13], wherein the C5-related disease is any disease selected from the group consisting of rheumatoid arthritis (RA); lupus nephritis; damage caused by ischemia-reperfusion; paroxysmal nocturnal hemoglobinuria (PNH); atypical hemolytic uremic syndrome (aHUS); dense deposit disease (DDD); macular degeneration; HELLP syndrome (hemolysis, increased activity of liver enzymes and thrombocytopenia); thrombotic thrombocytopenic purpura (TTP); spontaneous loss of the fetus; Pauci-immune vasculitis; bullous epidermolysis; recurring fetal loss; multiple sclerosis (MS); traumatic brain injury; and damage caused by myocardial infarction, cardiopulmonary bypass, or hemodialysis.

[15] A method for treating or preventing a C5-related disease, comprising intravenously administering to an individual a first pharmaceutical composition, which is formulated for intravenous injection and contains an anti-C5 antibody, and subcutaneously administering to an individual a second pharmaceutical composition, which is formulated for subcutaneous injection and contains antibody to C5, after administration of the first composition.

[16] The method according to [15], wherein the initial dose of the second composition is administered on the same day or 1-28 days after the final dose of the first composition.

[17] The method according to [15] or [16], wherein the second composition is administered once every 3-42 days.

[18] The method according to any one of [15]-[17], wherein the first composition is administered two or more times and from once an hour to once every 14 days.

[19] The method according to any one of [15]-[18], wherein the second composition is administered at a dose of 17 to 6000 mg of antibody.

[20] The method according to any one of [15]-[19], wherein the first composition is administered at a dose of 50 to 5000 mg of the antibody.

[21] The method according to any one of [15]-[20], which is for the treatment or prevention of a C5-related disease in an individual who has been treated with at least one pharmacological product intended for use in the treatment or prevention of this disease, in which the initial the dose of the first composition is administered after the administration of the final dose of the pharmacological product.

[22] The method according to [21], wherein the initial dose of the first composition is administered on the third day or 3 days after the administration of the final dose of the pharmacological product.

[23] The method according to [21] or [22], wherein the pharmacological product contains an siRNA targeting C5 mRNA, or an anti-C5 antibody other than the anti-C5 antibody contained in the first composition.

[24] The method according to any one of [21]-[23], wherein the pharmacological product contains eculizumab or a derivative thereof.

[25] A method for treating or preventing a C5-related disease, comprising administering to an individual a pharmaceutical composition that contains an anti-C5 antibody by subcutaneous injection at a dose of 17 to 6000 mg of the antibody.

[26] A method for treating or preventing a C5-related disease, comprising administering to an individual a pharmaceutical composition that contains an anti-C5 antibody by intravenous injection at a dose of 50 to 5000 mg of the antibody.

[27] The method according to any one of [15]-[26], wherein the C5-related disease is any disease selected from the group consisting of rheumatoid arthritis (RA); lupus nephritis; damage caused by ischemia-reperfusion; paroxysmal nocturnal hemoglobinuria (PNH); atypical hemolytic uremic syndrome (aHUS); dense deposit disease (DDD); macular degeneration; HELLP syndrome (hemolysis, increased activity of liver enzymes and thrombocytopenia); thrombotic thrombocytopenic purpura (TTP); spontaneous loss of the fetus; Pauci-immune vasculitis; bullous epidermolysis; recurring fetal loss; multiple sclerosis (MS); traumatic brain injury; and damage caused by myocardial infarction, cardiopulmonary bypass, or hemodialysis.

[28] The use of an anti-C5 antibody for the preparation of a pharmaceutical composition for the treatment or prevention of a C5-related disease, wherein the composition is in a subcutaneous injection form, contains the C5 antibody and is administered subcutaneously, and the initial dose of this composition is administered after a dose of another pharmaceutical composition , which is administered intravenously, and where another pharmaceutical composition is prepared in the form for intravenous injection and contains an antibody to C5.

[29] The use of an antibody to C5 for the preparation of a pharmaceutical composition intended for the treatment or prevention of a C5-related disease, where the composition is prepared in the form for intravenous injection, contains the antibody C5 and is administered intravenously, while the dose of this composition is administered before the initial dose of another pharmaceutical composition , which is administered subcutaneously, and where the other pharmaceutical composition is prepared in the form for subcutaneous injection and contains an antibody to C5.

[30] Use according to [28] or [29], wherein the initial dose of the subcutaneous injection composition is administered on the same day or 1-28 days after the dose of the intravenous injection composition.

[31] The use according to any one of [28]-[30], wherein the subcutaneous injection composition is administered once every 3 to 42 days.

[32] The use according to any one of [28]-[31], wherein the composition for intravenous injection is administered two or more times and from once an hour to once every 14 days.

[33] The use according to any one of [28]-[32], wherein the subcutaneous injection composition is administered at a dose of 17 to 6000 mg of antibody.

[34] The use according to any one of [28]-[33], wherein the composition for intravenous injection is administered at a dose of 50 to 5000 mg of the antibody.

[35] The use according to any one of [28]-[34], wherein the composition for intravenous injection and the composition for subcutaneous injection are for the treatment or prevention of a C5-related disease in an individual who has been treated with at least one pharmacological product intended for the treatment or prevention of this disease, wherein the initial dose of the composition for intravenous injection is administered after the final dose of the pharmacological product.

[36] The use according to [35], where the initial dose of the composition for intravenous injection is administered on the third day or 3 days after the final dose of the pharmacological product.

[37] Use according to [35] or [36], wherein the pharmacological product contains an siRNA targeting C5 mRNA, or an anti-C5 antibody other than the anti-C5 antibody contained in the composition.

[38] Use according to any one of [35]-[37], wherein the pharmacological product contains eculizumab or a derivative thereof.

[39] The use of an anti-C5 antibody for the preparation of first and second pharmaceutical compositions for the treatment or prevention of a C5-related disease, wherein the first pharmaceutical composition is formulated for intravenous injection, contains an anti-C5 antibody, and is administered intravenously, wherein the second pharmaceutical composition is formulated in form for subcutaneous injection, contains an antibody to C5 and is administered subcutaneously, and the introduction of the first and second pharmaceutical compositions should be carried out in the order, including the use of one or more intravenous doses of the first pharmaceutical composition and one or more subcutaneous doses of the second pharmaceutical composition.

[40] The use of an anti-C5 antibody for the preparation of a pharmaceutical composition for the treatment or prevention of a C5-related disease, wherein the composition is formulated for subcutaneous injection, contains an anti-C5 antibody, and is administered subcutaneously at a dose of 17 to 6000 mg of the antibody.

[41] The use of an anti-C5 antibody for the preparation of a pharmaceutical composition for the treatment or prevention of a C5-related disease, wherein the composition is formulated for intravenous injection, contains an anti-C5 antibody, and is administered intravenously at a dose of 50 to 5000 mg of the antibody.

[42] Use according to any one of [28]-[41], wherein the C5-related disease is any disease selected from the group consisting of rheumatoid arthritis (RA); lupus nephritis; damage caused by ischemia-reperfusion; paroxysmal nocturnal hemoglobinuria (PNH); atypical hemolytic uremic syndrome (aHUS); dense deposit disease (DDD); macular degeneration; HELLP syndrome (hemolysis, increased activity of liver enzymes and thrombocytopenia); thrombotic thrombocytopenic purpura (TTP); spontaneous loss of the fetus; Pauci-immune vasculitis; bullous epidermolysis; recurring fetal loss; multiple sclerosis (MS); traumatic brain injury; and damage caused by myocardial infarction, cardiopulmonary bypass, or hemodialysis.

[43] A product intended for the treatment or prevention of a C5-related disease, which contains (i) a container; (ii) a pharmaceutical composition in a container, wherein the pharmaceutical composition is formulated for subcutaneous injection, contains an anti-C5 antibody and is intended for subcutaneous administration; and (iii) a document providing instructions that the pharmaceutical composition is administered after a dose of another pharmaceutical composition that is administered intravenously, wherein the other pharmaceutical composition is formulated for intravenous injection and contains an anti-C5 antibody.

[44] A product intended for the treatment or prevention of a C5-related disease, which contains (i) a container; (ii) a pharmaceutical composition in a container, wherein the pharmaceutical composition is formulated for intravenous injection, contains an anti-C5 antibody and is intended for intravenous administration; and (iii) a document providing instructions that the pharmaceutical composition is administered prior to an initial dose of another pharmaceutical composition that is administered subcutaneously, wherein the other pharmaceutical composition is formulated for subcutaneous injection and contains an anti-C5 antibody.

[45] A product intended for the treatment or prevention of a C5-related disease, which contains (i) a first container; (ii) the first pharmaceutical composition in the first container, where the first pharmaceutical composition is prepared in the form for intravenous injection, contains an antibody to C5 and is intended for intravenous administration; (iii) a second container; (iv) a second pharmaceutical composition in a second container, wherein the second pharmaceutical composition is formulated for subcutaneous injection, contains an anti-C5 antibody, and is intended to be administered subcutaneously; and (v) a document providing instructions that the first pharmaceutical composition and the second pharmaceutical composition are to be administered in an order involving the administration of one or more intravenous doses of the first pharmaceutical composition for intravenous injection and one or more subcutaneous doses of the second pharmaceutical composition for subcutaneous injections.

[46] A product according to any one of [43]-[45], wherein the product is for the treatment or prevention of a C5-related disease in an individual who has been treated with at least one pharmacological product intended for use in the treatment or prevention of that disease, where the document also contains instructions that the initial dose of any of the pharmaceutical compositions is administered after the final dose of the pharmacological product.

[47] A product intended for the treatment or prevention of a C5-related disease, which contains (i) a container; (ii) a pharmaceutical composition in a container, wherein the pharmaceutical composition is in a subcutaneous injection form and contains an anti-C5 antibody; and (iii) a document providing instructions that the pharmaceutical composition is administered subcutaneously and at a dose of 17 to 6000 mg of the antibody.

[48] A product intended for the treatment or prevention of a C5-related disease, which contains (i) a container; (ii) a pharmaceutical composition in a container, wherein the pharmaceutical composition is formulated for intravenous injection and contains an anti-C5 antibody; and (iii) a document providing instructions that the pharmaceutical composition is administered intravenously and at a dose of 50 to 5000 mg of the antibody.

[49] The product according to any one of [43]-[48], wherein the C5-related disease is any disease selected from the group consisting of rheumatoid arthritis (RA); lupus nephritis; damage caused by ischemia-reperfusion; paroxysmal nocturnal hemoglobinuria (PNH); atypical hemolytic uremic syndrome (aHUS); dense deposit disease (DDD); macular degeneration; HELLP syndrome (hemolysis, increased activity of liver enzymes and thrombocytopenia); thrombotic thrombocytopenic purpura (TTP); spontaneous loss of the fetus; Pauci-immune vasculitis; bullous epidermolysis; recurring fetal loss; multiple sclerosis (MS); traumatic brain injury; and damage caused by myocardial infarction, cardiopulmonary bypass, or hemodialysis.

Brief description of the drawings

On FIG. 1 shows the time course of 305LO15 concentration in plasma of cynomolgus monkeys following subcutaneous injection (a) or intravenous injection (b) of the 305LO15 antibody. Plasma 305LO15 concentration was assessed by ELISA. Each dot corresponds in Fig. 1a the average value for three males or three females, and in FIG. 16 average for six males or six females.

On FIG. 2 shows the time course of 305LO15 concentration in the plasma of cynomolgus monkeys when the monkeys were treated with an initial intravenous injection and subsequent subcutaneous maintenance injections of the 305LO15 antibody. Plasma 305LO15 concentration was assessed by ELISA. Each point corresponds to the average value for five males or five females.

On FIG. 3a is a graph showing plasma concentrations of free C5 at each plasma antibody concentration for each indicated dosage amount and route of administration (IV—intravenous injection; SC—subcutaneous injection).

On FIG. 3b is a graph showing the levels of measured complement activity at each plasma antibody concentration for each indicated dosage amount and route of administration (IV—intravenous injection; SC—subcutaneous injection).

On FIG. 4a shows simulated plasma concentrations of 305LO15 over time for the dosing regimens planned for part 1 of the clinical study (IV - intravenous injection; SC - subcutaneous injection),

On FIG. 4b shows simulated plasma concentrations of 305LO15 over time for the dosing regimens planned for part 2 of the clinical study (SC - subcutaneous injection; QW - once every week administration).

On FIG. 4c shows simulated plasma concentrations of 305LO15 over time for the dosing regimens planned for part 3 of the clinical study (SC - subcutaneous injection; QW - once every week; Q2W - once every two weeks; Q4W - once every four weeks).

On FIG. 5 shows the profile of immunocomplexes formed by eculizumab (ECZ), human C5 (hC5) and/or 305LO15 analyzed in vitro by gel filtration at pH 7.4 (upper panel) and at pH 6.0 (lower panel).

On FIG. 6 shows human measured and simulated plasma concentration versus time profiles of 305LO15 in healthy individuals who received 75 mg by IV infusion (over 60 minutes). The shaded gray area indicates the 95% prediction interval of the simulated 305LO15 concentration versus time profile. The solid line indicated by an arrow corresponds to the simulated time profile of the median 305LO15 concentration. The dotted line indicates the PD cut-off dose of 40 μg/g.

On FIG. 7 shows human measured and simulated plasma concentration versus time profiles of 305LO15 in healthy individuals who received 125 mg by IV infusion (over 60 minutes). The shaded gray area indicates the 95% prediction interval of the simulated 305LO15 concentration versus time profile. The solid line indicated by an arrow corresponds to the simulated time profile of the median 305LO15 concentration. The dotted line indicates the PD cut-off dose of 40 μg/g.

On FIG. 8 shows human measured and simulated plasma concentration versus time profiles of 305LO15 in healthy individuals who received 100 mg SC. The shaded gray area indicates the 95% prediction interval of the simulated 305LO15 concentration versus time profile. For simulations performed with a single 100 mg SC dose, a bioavailability of 90% is expected. The dotted line indicates the cut-off dose PD of 40 μg/g.

On FIG. 9 shows the relationship between plasma concentrations of 305LO15 and hemolytic activity (LIA) after a single IV infusion or SC injection in healthy individuals. An ex vivo LIA dose-response curve was generated using human plasma samples treated with various concentrations of 305LO15.

On FIG. 10a shows the time profile of LDH (lactate dehydrogenase) serum levels in a PNH patient, patient X, who received 375mg IV dose of 305LO15 on day 1, 500mg IV dose of 305LO15 on day 8, 1000mg IV dose of 305LO15 on day 22 , 170 mg SC dose of 305LO15 on day 36, and 170 mg SC dose of 305LO15 on day 43 in part 2 of the study.

On FIG. 10b shows the time profile of hemolytic activity (LIA) in a PNH patient, patient X, who received 375 mg IV dose of 305LO15 on day 1, 500 mg IV dose of 305LO15 on day 8, 1000 mg IV dose of 305LO15 on day 22, SC -dose 170 mg 305LO15 on day 36 and SC dose 170 mg 305LO15 on day 43 in part 2 of the study.

Description of embodiments of the invention

The techniques and procedures described or referenced herein are generally well known to those skilled in the art and widely used using conventional techniques such as those described in Sambrook et al., Molecular Cloning: A Laboratory. Manual, 3rd ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. 2001; Current Protocols in Molecular Biology, ed. F.M. Ausubel et al., 2003; Methods in Enzymology series, Academic Press, Inc.: PCR 2: A Practical Approach, ed. M.J. MacPherson, B.D. Hames and G.R. Taylor, 1995, Antibodies, A Laboratory Manual, ed. Harlow and Lane, 1988 and Animal Cell Culture, ed. R.I. Freshney, 1987; Oligonucleotide Synthesis, ed. M.J. Gait, 1984; Methods in Molecular Biology, Humana Press; Cell Biology: A Laboratory Notebook, ed. J.E. Cellis, 1998, from Academic Press; Animal Cell Culture, ed. R.I. Freshney, 1987; J.P. Mather and P.E. Roberts, Introduction to Cell and Tissue Culture, Plenum Press, 1998; Cell and Tissue Culture: Laboratory Procedures, ed. A. Doyle, J.B. Griffiths and D.G. Newell, J. Wiley and Sons, 1993-1998; Handbook of Experimental Immunology, ed. D.M. Weir and C.C. Blackwell; Gene Transfer Vectors for Mammalian Cells, ed. J.M. Miller and M.P. Calos, 1987; PCR: The Polymerase Chain Reaction, ed. Mullis et al., 1994; Current Protocols in Immunology, ed. J.E. Coligan et al., 1991; Short Protocols in Molecular Biology, Wiley and Sons, 1999; C.A. Janeway and P. Travers, Immunobiology, 1997; P. Finch, Antibodies, 1997; Antibodies: A Practical Approach, ed. D. Catty, IRL Press, 1988-1989; Monoclonal Antibodies: A Practical Approach, ed. P. Shepherd and C. Dean, Oxford University Press, 2000; E. Harlow and D. Lane, Using Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory Press, 1999; The Antibodies, ed. M. Zanetti and J. D. Capra, Harwood Academic Publishers, 1995; and Cancer: Principles and Practice of Oncology, ed. V.T. DeVita et al., J.B. Lippincott Company, 1993.

I Definition

"Dosage interval" (interval between separate administrations) means the interval between administration of the nth dose (n is an integer of 1 or more) and administration of the (n+1)th dose.

II. Pharmaceutical composition for subcutaneous injection

The pharmaceutical composition formulated for subcutaneous injection according to the present invention is for the treatment or prevention of a C5 related disease, contains an anti-C5 antibody, and is administered subcutaneously. Subcutaneous injection can be performed using conventional devices and methods for introducing the pharmaceutical composition into the subcutaneous tissue by injection. You can also choose specific devices and methods for subcutaneous injection.

Examples of anti-C5 antibodies

The anti-C5 antibody is not limited to a specific variant, and can be selected accordingly from the antibodies known as anti-C5 antibodies. The term "antibody to C5" or "antibody that binds to C5" refers to an antibody that has the ability to bind to C5 with sufficient affinity so that the antibody can be used as a C5 targeting therapeutic agent.

In one aspect of the invention, anti-C5 antibodies inhibit C5 activation. In some embodiments, anti-C5 antibodies that prevent C5 cleavage resulting in C5a and C5b, thereby preventing C5a-associated anaphylactic activity and also preventing assembly of the C5b-9 membrane attack complex (MAC) associated with C5b. In some embodiments, anti-C5 antibodies block the conversion of C5 to C5a and C5b by C5 convertase. In some embodiments, anti-C5 antibodies block the access of the C5 convertase to the C5 cleavage site. In some embodiments, anti-C5 antibodies block hemolytic activity due to C5 activation. In other embodiments, anti-C5 antibodies inhibit C5 activation in a classical and/or alternative manner.

According to one aspect of the invention, the pharmaceutical compositions of the present invention can be used to treat or prevent a C5 related disease based at least in part on the aforementioned anti-C5 antibody activities, such as inhibition of C5 activation.

In some embodiments of the invention, the activity of C5 can be assessed by its ability to carry out cell lysis in the total water of the individual. The ability of C5 to carry out cell lysis or its reduction can be assessed using methods well known in this field, for example, using a standard hemolytic assay, such as the hemolytic assay described in Experimental Immunochemistry, ed. Kabat and Mayer, 2nd ed., 135-240, CC Thomas, Springfield, IL, 135-139, 1961, or a standard variant of this assay, such as the hen erythrocyte hemolysis method described, for example, in Hillman et al, N. Engl. J. Med. 350(6): 552-559, 2004. In some embodiments, C5 activity or its inhibition is quantified using the CH50eq assay. The CH50eq assay is a method for measuring the total activity of the classical complement pathway in serum. This test is a lytic activity assay that uses antibody-sensitized erythrocytes as an activator of the classical complement pathway and uses various dilutions of test serum to determine the amount needed to achieve 50% lysis (CH50). The percentage of hemolysis can be determined, for example, using a spectrophotometer. The CH50eq assay is an indirect measure of terminal complement component complex (TCC) formation, since the TCC itself is directly responsible for the measured hemolysis. Inhibition of C5 activation can be detected and/or measured using the methods set forth and illustrated in the Experimental Examples. Using assays of these or other suitable types, candidate antibodies having the ability to inhibit C5 activation can be screened. In some embodiments, the inhibition of C5 activation comprises an assayed decrease in C5 activation of at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, or 40%, or more, compared to exposure to a negative control under similar conditions. In some embodiments, it refers to inhibition of C5 activation by at least 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95%, or more .

In preferred embodiments, the epitope of the anti-C5 antibody is different from that of eculizumab.

In some embodiments, an anti-C5 antibody of the present invention binds to an epitope in the C5 beta chain. In some embodiments, an anti-C5 antibody binds to an epitope in the MG1-MG2 domain of the C5 beta chain. In some embodiments, an anti-C5 antibody binds to an epitope in a fragment consisting of amino acids 19-180 of the C5 beta chain. In some embodiments, an anti-C5 antibody binds to an epitope in the MG1 domain (amino acids 20-124 of SEQ ID NO: 1) of the C5 beta chain. In some embodiments, an anti-C5 antibody binds to an epitope in a fragment consisting of amino acids 33-124 of the C5 beta chain (SEQ ID NO: 1).

In some embodiments, an anti-C5 antibody of the present invention binds to an epitope in the C5 beta chain that consists of the MG1 domain. In some embodiments, an anti-C5 antibody binds to an epitope in the beta chain (SEQ ID NO: 1) of C5 that contains at least one fragment selected from the group consisting of amino acids 47-57, 70-76, and 107- 110. In some embodiments, an anti-C5 antibody binds to an epitope in the beta chain fragment (SEQ ID NO: 1) of C5 that contains at least one amino acid selected from the group consisting of Thr47, Glu48, Ala49, Phe50, Asp51, Ala52, Thr53, Lys57, His70, Val71, His72, Ser74, Glu76, Val107, Ser108, Lys109 and His110. In some embodiments, an anti-C5 antibody binds to an epitope in the beta chain fragment (SEQ ID NO: 1) of C5 that contains at least one amino acid selected from the group consisting of Glu48, Asp51, His70, His72, Lys109, and His110. In some embodiments, the binding of an anti-C5 antibody to a C5 mutant is reduced compared to its binding to wild-type C5, where the C5 mutant has at least one amino acid substitution at a position selected from the group consisting of Glu48, Asp51, His72, and Lys109 . In another embodiment, the pH-dependent binding (described below) of an anti-C5 antibody to a C5 mutant is reduced compared to its pH-dependent binding to wild-type C5, where the C5 mutant has at least one amino acid substitution at a position selected from the group, which consists of His70, His72 and His110. In a further embodiment of the invention, in the C5 mutant, the amino acid at a position selected from Glu48, Asp51 and Lys109 is changed to alanine and the amino acid at a position selected from His70, His72 and His110 is changed to tyrosine.

In another aspect of the invention, the anti-C5 antibodies of the present invention may have pH-dependent binding characteristics. In the context of the present description, the term "pH-dependent binding" means that the antibody has a "reduced ability to bind to C5 at acidic pH compared to its binding at neutral pH" (both terms can be used interchangeably for the purposes of this description). For example, antibodies with "pH dependent binding characteristics" include antibodies that bind to C5 with higher affinity at neutral pH than at acidic pH. In some embodiments, antibodies bind to C5 with an affinity of at least 2, 3, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 200, 400, 1000, 10,000 or more times higher at neutral pH than at acidic pH. In some embodiments, antibodies bind to C5 with higher affinity at pH 7.4 than at pH 5.8. In other embodiments, the antibodies bind to C5 with an affinity of at least 2, 3, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 200, 400, 1000, 10,000 or more times higher at pH 7.4 than at pH 5.8.

For the purposes of this description, the "affinity" of an antibody to C5 is expressed in terms of the KD of the antibody. The KD of an antibody means the dissociation equilibrium constant of the antibody-antigen interaction. The higher the KD value, which characterizes the binding of an antibody to its antigen, the weaker its binding affinity for the specified specific antigen. Thus, in the context of the present description, the concept of "higher affinity at neutral pH than at acidic pH" (or the equivalent concept of "pH-dependent binding") means that the KD, which characterizes the binding of an antibody to C5 at acidic pH, is higher than the KD , which characterizes the binding of the antibody to C5 at neutral pH. For example, in the context of the present invention, an antibody is considered to bind to C5 with higher affinity at neutral pH than at acidic pH if the KD characterizing the binding of the antibody to C5 at acidic pH is at least 2 times higher than the KD characterizing binding of the antibody to C5 at neutral pH. Thus, anti-C5 antibodies of the present invention include antibodies whose binding to C5 at acidic pH is characterized by a KD value of at least 2, 3, 5, 10, 15, 20, 25, 30, 35, 40 , 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 200, 400, 1000, 10,000 or more times the KD characterizing binding to C5 at neutral pH. In another embodiment, the KD value of the antibody at neutral pH may be 10 ^-7 , 10 ^-8 , 10 ^-9 , 10 ^-10 , 10 ^-11 , 10 ^-12 M or less. In another embodiment, the KD value of the antibody at acidic pH may be 10 ^-9 M, 10 ^-8 M, 10 ^-7 M, 10 ^-6 M or more.

In other embodiments, an antibody is said to bind to C5 with a higher affinity at neutral pH than at acidic pH if the KD value characterizing the binding of the antibody to C5 at pH 5.8 is at least 2-fold higher than the KD , characterizing the binding of the antibody to C5 at pH 7.4. In some embodiments, antibody binding to C5 at pH 5.8 is characterized by a KD value that is at least 3, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65 , 70, 75, 80, 85, 90, 95, 100, 200, 400, 1000, 10000 or more times the KD characterizing the binding of the antibody to C5 at pH 7.4. In another embodiment, the KD value of the antibody at pH 7.4 may be 10 ^-7 , 10 ^-8 , 10 ^-9 , 10 ^-10 , 10 ^-11 , 10 ^-12 M or less. In another embodiment, the KD value of the antibody at pH 5.8 may be 10 ^-9 , 10 ^-8 , 10 ^-7 , 10 ^-6 M or more.

The binding characteristics of an antibody to a particular antigen can also be expressed as the kd of an antibody. The kd value of an antibody denotes the dissociation rate constant of an antibody relative to a particular antigen and is expressed as reciprocals of a second (ie, s ⁻¹ ). An increased kd value means weaker binding of the antibody to its antigen. Thus, the present invention includes antibodies whose binding to C5 has a higher kd value at acidic pH than at neutral pH. The antibodies of the present invention include antibodies whose binding to C5 at acidic pH is characterized by a kd value of at least 2, 3, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 200, 400, 1000, 10000 or more times the kd characterizing the binding of the antibody to C5 at neutral pH. In another embodiment, the kd value of the antibody at neutral pH may be 10 ^-2 , 10 ^-3 , 10 ^-4 , 10 ^-5 , 10 ^-6 1/s or less. In another embodiment, the kd value of the antibody at acidic pH may be 10 ^-3 , 10 ^-2 , 10 ^-1 1/s or more. The antibodies of the present invention also include antibodies whose binding to C5 has a higher kd at pH 5.8 than at pH 7.4. The antibodies of the present invention include antibodies whose binding to C5 at pH 5.8 has a kd value of at least 3, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 200, 400, 1000, 10000 or more times the kd characterizing the binding of the antibody to C5 at pH 7.4. In another embodiment, the kd value of the antibody at pH 7.4 may be 10 ^-2 , 10 ^-3 , 10 ^-4 , 10 ^-5 , 10 ^-6 1/s or less. In another embodiment, the kd value of the antibody at pH 5.8 may be 10 ^-3 , 10 ^-2 , 10 ^-1 1/s or more.

In some cases, "reduced ability to bind to C5 at acidic pH compared to the ability to bind at neutral pH" is expressed as the ratio of the KD value, which characterizes the binding of the antibody to C5 at acidic pH, and the KD value, which characterizes the binding of the antibody to C5 at neutral pH ( or vice versa). For example, an antibody may be considered to have "reduced ability to bind to C5 at acidic pH compared to the ability to bind at neutral pH" for the purposes of the present invention if the antibody's acid/neutral pH KD ratio is 2 or more. In some embodiments, the KD ratio at pH 5.8/pH 7.4 for the antibody is 2 or more. In some embodiments, the acid/neutral pH KD ratio for an antibody may be 2, 3, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 200, 400, 1000, 10000 or more. In another embodiment, the KD value of the antibody at neutral pH may be 10 ^-7 , 10 ^-8 , 10 ^-9 , 10 ^-10 , 10 ^-11 , 10 ^-12 M or less. In another embodiment, the KD value of the antibody at acidic pH may be 10 ^-9 ,10 ^-8 , 10 ^-7 , 10 ^-6 M or more. In other cases, an antibody may be considered to have "reduced ability to bind to C5 at acidic pH compared to the ability to bind at neutral pH" if the antibody has a KD ratio at pH 5.8/pH 7.4 of 2 or more. In some exemplary embodiments of the invention, the KD ratio at pH 5.8/pH 7.4 for an antibody may be 3, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60 , 65, 70, 75, 80, 85, 90, 95, 100, 200, 400, 1000, 10000 or more. In another embodiment, the KD value of the antibody at pH 7.4 may be 10 ^-7 , 10 ^-8 , 10 ^-9 , 10 ^-10 , 10 ^-11 , 10 ^-12 M or less. In another embodiment, the KD value of the antibody at pH 5.8 may be 10 ^-9 , 10 ^-8 , 10 ^-7 , 10 ^-6 M or more.

In some cases, "reduced ability to bind to C5 at acidic pH compared to the ability to bind at neutral pH" is expressed as the ratio of the kd value characterizing the binding of the antibody to C5 at acidic pH, and the kd value characterizing the binding of the antibody to C5 at neutral pH ( or vice versa). For example, an antibody may be considered to have "reduced ability to bind to C5 at acidic pH compared to the ability to bind at neutral pH" for the purposes of the present invention if the antibody's acid/neutral pH kd ratio is 2 or more. In some embodiments, the kd ratio at pH 5.8/pH 7.4 for an anti-C5 antibody is 2 or more. In some exemplary embodiments of the invention, the acid/neutral pH kd ratio for an antibody may be 2, 3, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 200, 400, 1000, 10000 or more. In other exemplary embodiments of the invention, the ratio kd at pH 5.8/pH 7.4 for an antibody can be 2, 3, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55 , 60, 65, 70, 75, 80, 85, 90, 95, 100, 200, 400, 1000, 10000 or more. In another embodiment, the kd value of the antibody at neutral pH may be 10 ^-2 , 10 ^-3 , 10 ^-4 , 10 ^-5 , 10 ^-6 1/s or less. In another embodiment, the kd value of the antibody at acidic pH may be 10 ^-3 , 10 ^-2 , 10 ^-1 1/s or more. In some exemplary embodiments of the invention, the ratio kd at pH 5.8/pH 7.4 for an antibody may be 2, 3, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55 , 60, 65, 70, 75, 80, 85, 90, 95, 100, 200, 400, 1000, 10000 or more. In another embodiment, the kd value of the antibody at pH 7.4 may be 10 ^-2 , 10 ^-3 , 10 ^-4 , 10 ^-5 , 10 ^-6 1/s or less. In another embodiment, the kd value of the antibody at pH 5.8 may be 10 ^-3 , 10 ^-2 , 10 ^-1 1/s or more.

In the context of the present description, the term "acidic pH" means a pH from 4.0 to 6.5. The term "acidic pH" includes one of the following pH values 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9 , 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6 .2, 6.3, 6.4 and 6.5. In specific aspects of the invention, "acidic pH" corresponds to 5.8.

In the context of the present description, the term "neutral pH" means a pH from 6.7 to about 10.0. The term "neutral pH" includes one of the following pH values 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6 , 7.7, 7.8, 7.9, 8.0, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8 .9, 9.0, 9.1, 9.2, 9.3, 9.4, 9.5, 9.6, 9.7, 9.8, 9.9, and 10.0. In specific aspects of the invention, "neutral pH" corresponds to 7.4.

The KD and kd values described herein can be determined using a biosensor based surface plasmon resonance method to characterize antibody-antigen interactions. The KD values and kd values can be determined at 25°C or 37°C.

In another aspect of the invention, the anti-C5 antibody comprises a heavy chain variable domain (VH) sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99 % or 100% identical to the amino acid sequence of SEQ ID NO: 2. In some embodiments, the VH sequence is the amino acid sequence of SEQ ID NO: 2. In some embodiments, the VH sequence is at least 90%, 91%, 92 identical %, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, contains substitutions (e.g., conservative substitutions), insertions, or deletions relative to the reference sequence, but an anti-C5 antibody containing the indicated sequence retains ability to bind to C5. In some embodiments, a total of 1 to 10 amino acids are changed, inserted, and/or deleted by deletion in SEQ ID NO: 2. In some embodiments, the substitutions, insertions, or deletions affect regions outside the HVR (i.e., FR). Optionally, the anti-C5 antibody contains the VH sequence shown in SEQ ID NO: 2, including post-translational modifications of said sequence. In a specific embodiment, the VH contains one, two, or three HVRs selected from: (a) HVR-H1, which contains the amino acid sequence of SEQ ID NO: 3, (b) HVR-H2, which contains the amino acid sequence of SEQ ID NO: 4 , and (c) HVR-H3 which contains the amino acid sequence of SEQ ID NO: 5.

In the context of the present description, the term "HVR" means "hypervariable region", and the term "FR" means "framework region".

The term "framework region" or "FR" refers to variable domain residues other than hypervariable region (HVR) residues. The variable domain FR typically consists of four FR domains: FR1, FR2, FR3, and FR4. Thus, the HVR and FR sequences typically have the following location in the VH (or VL): FR1-H1(L1)-FR2-H2(L2)-FR3-H3(L3)-FR4.

The term "hypervariable region" or "HVR" in the context of the present description refers to each of the regions of the variable domain of an antibody, the sequences of which are hypervariable ("complementarity determining regions" or "CDRs") and/or form loops of a certain structure ("hypervariable loops") , and/or contain antigen-contacting residues ("antigen-contacts"). Typically, antibodies contain six HVRs; three in VH (H1, H2, H3) and three in VL (L1, L2, L3). As used herein, examples of HVRs include (a) hypervariable loops comprising amino acid residues 26-32 (L1), 50-52 (L2), 91-96 (L3), 26-32 (H1), 53-55 (H2), and 96-101 (H3) (Chothia and Lesk, J. Mol. Biol. 196: 901-917, 1987); (6) CDRs including amino acid residues 24-34 (L1), 50-56 (L2), 89-97 (L3), 31-35b (H1), 50-65 (H2) and 95-102 (H3) ( Kabat et al., Sequences of Proteins of Immunological Interest, 5th ed., Public Health Service, National Institutes of Health, Bethesda, MD, 1991, NIH publication 91-3242); (c) antigen contact regions comprising amino acid residues 27 c-36 (L1), 46-55 (L2), 89-96 (L3), 30-35b (H1), 47-58 (H2) and 93-101 (H3) (MacCallum et al., J. Mol. Biol. 262: 732-745, 1996); and (d) combinations of residues specified in subparagraphs (a), (b) and/or (c), including amino acid residues HVR 46-56 (L2), 47-56 (L2), 48-56 (L2), 49 -56 (L2), 26-35 (H1), 26-35b (H1), 49-65 (H2), 93-102 (H3) and 94-102 (H3). Residues in HVR and other residues in the variable domain (eg, residues in FR) are numbered according to Kabat et al., supra.

In another aspect of the invention, the anti-C5 antibody comprises a light chain variable domain sequence ((VL) that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% 99% or 100% identical to the amino acid sequence of SEQ ID NO: 6. In some embodiments, the VL sequence is the amino acid sequence of SEQ ID NO: 6. In some embodiments, the VL sequence is at least 90%, 91% identical, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, contains substitutions (e.g., conservative substitutions), insertions, or deletions relative to the reference sequence, but an anti-C5 antibody containing the specified sequence, retains the ability to bind to C5 In some embodiments, a total of 1 to 10 amino acids are changed, inserted, and/or deleted by deletion in SEQ ID NO: 6. In some embodiments, the substitutions, insertions, or deletions affect regions outside HVR (i.e. FR). Optionally, the anti-C5 antibody contains the VL sequence shown in SEQ ID NO: 6, including post-translational modifications of said sequence. In a specific embodiment, the VL contains one, two, or three HVRs selected from: (a) HVR-L1, which contains the amino acid sequence of SEQ ID NO: 7, (b) HVR-L2, which contains the amino acid sequence of SEQ ID NO: 8 , and (c) HVR-L3 which contains the amino acid sequence of SEQ ID NO: 9.

In another embodiment, the antibody is a full-length antibody, such as an intact IgG1-, IgG2-, IgG3-, or IgG4-subclass antibody, or an antibody engineered to have regions of two or more IgGs selected from IgG1, IgG2 , IgG3 and IgG4, in the region(s) which/which are homologous among IgG subclasses, by recombination. The antibody may comprise a suitable Fc region containing the sequence of a human Fc region (eg, human IgG1, IgG2, IgG3, or IgG4 Fc regions).

Fc-region variants

In some embodiments, an anti-C5 antibody of the present invention comprises an Fc region variant in which one or more amino acid modification(s) are introduced into the antibody's native Fc region sequence. A variant Fc region may comprise a human Fc region sequence (eg, human IgG1, IgG2, IgG3, or IgG4 Fc regions) comprising an amino acid modification (eg, substitution) at one or more amino acid positions.

Some embodiments of the invention provide a variant of an antibody that has some, but not all, effector functions, making it a promising candidate for applications where the in vivo half-life of the antibody is important, but some effector functions (such as complement-dependent cytotoxicity (CDC) and ADCC) are unnecessary or harmful. In vitro and/or in vivo cytotoxicity assays can be performed to confirm the reduction/depletion of CDC and/or ADCC activity. For example, Fc receptor (FcR) binding assays can be performed to ensure that the antibody lacks the ability to bind to Fc-gamma R (and therefore likely lacks ADCC activity), but retains the ability to bind to FcRn. Primary cells mediating ADCC, ie. NK cells express only Fc-gamma RIII while monocytes express Fc-gamma RI, Fc-gamma RII and Fc-gamma RIII. FcR expression data on hematopoietic cells are summarized in Table 3 on p. 464 by Ravetch and Kinet, Annu. Rev. Immunol. 9: 457-492, 1991. Examples of in vitro assays for ADCC activity of a molecule of interest are, but are not limited to, those described in US 5,500,362 (see, for example, Hellstrom et al., Proc. Natl. Acad. Sci. USA 83: 7059-7063, 1986 and Hellstrom et al., Proc. Nat'l Acad. Sci. USA 82: 1499-1502, 1985); US 5821337 (see Bruggemann et al., J. Exp. Med. 166: 1351-1361, 1987). Alternatively, non-radioactive assays can be used (see, for example, non-radioactive flow cytometry-based ACTI™ cytotoxicity assay (CellTechnology, Inc. Mountain View, Calif.); and non-radioactive CytoTox 96 cytotoxicity assay (Promega, Madison, CA). Wisconsin. Suitable effector cells for such assays are peripheral blood mononuclear cells (PBMCs) and natural killer (NK) cells. Alternatively, or in addition, the ADCC activity of a molecule of interest can be assessed in vivo, e.g., in an animal model, e.g. as described in Clynes et al., Proc. Natl. Acad. Sci. USA 95: 652-656, 1998. C1q binding assays can also be performed to confirm that the antibody cannot bind C1q and therefore lacks CDC activity. (See WO 2006/029879 and WO 2005/100402 for an ELISA for assessing C1q and C3c binding.) lysed by the CDC (see for example, Gazzano-Santoro et al., J. Immunol. Methods 202: 163-171, 1996; Cragg et al., Blood 101: 1045-1052, 2003; and Cragg, Blood 103: 2738-2743, 2004). Determination of FcRn binding and in vivo clearance/half-life can also be performed using methods known in the art (see, for example, Petkova et al., Int'l. Immunol. 18(12): 1759-1769, 2006).

Antibodies with reduced effector function include antibodies with the replacement of one or more of the following residues of the Fc region, such as 238, 265, 269, 270, 297, 327 and 329 (see, for example, US 6737056). These Fc mutants include Fc mutants with substitutions at two or more of the following amino acid positions: 265, 269, 270, 297, and 327, including the so-called "DANA" Fc region mutant carrying the substitution of residues 265 and 297 with alanine (US 7332581).

Several antibody variants with increased or decreased ability to bind to FcR have been described (see, for example, US 6737056; WO 2004/056312 and Shields et al., J. Biol. Chem. 276: 6591-6604, 2001).

In specific embodiments, the antibody variant comprises an Fc region with one or more amino acid substitutions that increase ADCC, such as substitutions at positions 298, 333, and/or 334 of the Fc region (EU residue numbering).

In some embodiments, changes are made in the Fc region that result in an altered (i.e., either increased or decreased) C1q binding ability and/or complement dependent cytotoxicity (CDC), as described, for example, in US 6194551, WO 1999 /51642 and Idusogie et al., J. Immunol. 164: 4178-4184, 2000.

Antibodies with extended half-life and increased ability to bind to the neonatal Fc receptor (FcRn), which is responsible for the transfer of maternal IgG to the embryo (Guyer et al., J. Immunol. 117: 587-593, 1976 and Kim et al., J Immunol 24: 2429-2434, 1994) are described in US 2005/0014934A1 (Hinton et al.). These antibodies contain an Fc region with one or more substitutions that increase the binding of the Fc region to FcRn. These Fc region variants include variants with substitutions of one or more of the following Fc region residues: 378, 380, 382, 413, 424 or 434, for example, a substitution of residue 434 in the Fc region (US 7371826).

Additional examples regarding other variants of the Fc region are also described in Duncan, Nature 322: 738-740, 1988; in US Pat. Nos. 5,648,260 and US 5,624,821 and in WO 1994/29351.

In some embodiments, an anti-C5 antibody of the present invention comprises the VH shown above as one of the embodiments of the invention and a heavy chain constant region containing the amino acid sequence shown in one of SEQ ID NOs: 10, 11, 12 , 13, 14, and 15. In some embodiments, the anti-C5 antibody comprises the VL shown above as one of the embodiments of the invention and a light chain constant region containing the amino acid sequence shown in one of SEQ ID NOs: 16, 17 and eighteen.

In some embodiments, an anti-C5 antibody of the present invention is any of the antibodies described in WO 2016/098356, WO 2017/123636 and WO 2017/132259.

In some embodiments of the invention, in the case where the pharmaceutical composition is administered by subcutaneous injection to the individual repeatedly, the composition is administered every 3-42 days. A suitable interval between subcutaneous injections can be determined within an acceptable range depending on the condition of the individual or patient. The specific interval between subcutaneous injections is 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, 15 days, 16 days, 17 days, 18 days, 19 days, 20 days, 21 days, 22 days, 23 days, 24 days, 25 days, 26 days, 27 days, 28 days, 29 days, 30 days, 31 days, 32 days, 33 days, 34 days, 35 days, 36 days, 37 days, 38 days, 39 days, 40 days, 41 days or 42 days. In preferred embodiments of the present invention, the interval between injections is 4-35 days. In other preferred embodiments of the invention, the interval between injections is 5-31 days. In the most preferred embodiments of the invention, the interval between injections is 7, 14 or 28 days or 1 month. The pharmaceutical composition can be administered once every week (weekly), once every 2 weeks (every two weeks or at two week intervals) or once every 4 weeks (every four weeks or monthly). A longer interval allows to reduce the (drug) burden (burden) on the patient or his suffering.

In some embodiments, the pharmaceutical composition is administered subcutaneously at a dose of 17 to 6000 mg of the antibody. A suitable dose can be determined within an acceptable range depending on the condition of the individual or patient. In the case where the pharmaceutical composition is administered by subcutaneous injection to an individual multiple times, the doses do not have to be the same all the time and can be determined arbitrarily if the doses are within an acceptable range. For example, the dose can be gradually reduced. In preferred embodiments, the dosage range is from 25 to 4000 mg of the antibody. In other preferred embodiments of the invention, the dosage range is from 50 to 2000 mg of the antibody. In the most preferred embodiments of the invention, the dose range is from 75 to 1000 mg of antibody. Specific preferred doses are 100, 170, 340, 600 and 680 mg. Other preferred doses are 170, 340, 600 and 680 mg. When the pharmaceutical composition is administered weekly (about 7 days apart), a dose of 170 mg is preferred. When the pharmaceutical composition is administered once every 2 weeks (about 14 days apart), a dose of 340 mg is preferred. When the pharmaceutical composition is administered once every 4 weeks (with an interval of about 28 days), a dose of 600 mg or 680 mg is preferred. A dosage regimen of 600 mg or 680 mg once every 4 weeks may be particularly preferred as it reduces patient burden or suffering.

In a specific embodiment of the invention, a pharmaceutically acceptable carrier suitable for a conventional subcutaneous injection composition may be included in the pharmaceutical composition. The type of carrier for subcutaneous injection may be selected from carriers generally known in the art.

In a specific embodiment of the invention, the pharmaceutical composition may be any one selected from the group that includes liquid, semi-solid and solid composition. The solid composition is obtained, as a rule, by lyophilization from a liquid formulation. The lyophilized composition is typically reconstituted with water or saline immediately prior to subcutaneous injection.

When the composition is administered subcutaneously, the concentration of the anti-C5 antibody in the liquid formulation of the composition is within the generally accepted range known in the art. In a specific embodiment of the invention, the volume of subcutaneous injection is determined by the usual range known in this field.

In some embodiments, the C5-associated disease is a complement-mediated disease or condition that involves excessive or uncontrolled C5 activation. In some embodiments, the C5-related disease is at least one selected from the group consisting of rheumatoid arthritis (RA); lupus nephritis; damage caused by ischemia-reperfusion; paroxysmal nocturnal hemoglobinuria (PNH); atypical hemolytic uremic syndrome (aHUS); dense deposit disease (DDD); macular degeneration; HELLP syndrome (hemolysis, increased activity of liver enzymes and thrombocytopenia); thrombotic thrombocytopenic purpura (TTP); spontaneous loss of the fetus; Pauci-immune vasculitis; bullous epidermolysis; recurring fetal loss; multiple sclerosis (MS); traumatic brain injury; and damage caused by myocardial infarction, cardiopulmonary bypass or hemodialysis; resistant generalized myasthenia gravis (gMG); neuromyelitis of the optic nerve (NMO). In a preferred embodiment, the C5-related disease is at least one selected from the group consisting of PNH, aHUS, gMG and NMO.

In some aspects of the invention, the initial dose of the composition for subcutaneous injection is administered after a dose of another pharmaceutical composition that is administered intravenously. The pharmaceutical composition for intravenous injection is the composition described above in the present description.

In some embodiments of the present invention, the subcutaneous injection composition is administered subcutaneously on the same day or one or more days after the dose of the intravenous injection composition that is administered intravenously. One or more doses of the composition for intravenous injection may be administered to an individual or patient prior to administration of the initial dose of the composition for subcutaneous injection. In a preferred embodiment of the present invention, the initial dose of the subcutaneous injection composition is administered after the final dose of the intravenous injection composition.

In some embodiments, implementation of the present invention, the initial dose of the composition for subcutaneous injection is administered on the same day or 1-28 days after the dose of the composition for intravenous injection. The specific interval between administration of a dose of the composition for intravenous injection and the initial dose of the composition for subcutaneous injection is 0 days (i.e. within 24 hours), 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, 15 days, 16 days, 17 days, 18 days, 19 days, 20 days, 21 days, 22 days, 23 days, 24 days, 25 days, 26 days, 27 days or 28 days. In preferred embodiments of the present invention, the interval is 1-14 days. In other preferred embodiments of the present invention, the interval is 3-10 days. In the most preferred embodiments of the invention, the interval is 5-8 days.

III. Pharmaceutical composition for intravenous injection

The pharmaceutical composition formulated for intravenous injection according to the present invention is for use in the treatment or prevention of a C5 related disease, contains an anti-C5 antibody and is administered intravenously. Intravenous injection can be performed using conventional devices and methods for introducing the pharmaceutical composition into a vein by injection. You can also choose specific devices and methods for intravenous injection.

The anti-C5 antibody used for intravenous injection may be any of the antibodies described above for pharmaceutical compositions for subcutaneous injection. The pharmaceutical composition for intravenous injection and the pharmaceutical composition for subcutaneous injection may contain the same anti-C5 antibody or may contain a different anti-C5 antibody. In preferred embodiments, the anti-C5 antibody used for intravenous injection is the same antibody as described above for subcutaneous injection.

In some embodiments, implementation of the present invention, the pharmaceutical composition is administered intravenously at a dose of 50-5000 mg of the antibody. A suitable dose can be determined within an acceptable range depending on the condition of the individual or patient. In the case where the pharmaceutical composition is administered intravenously to an individual multiple times, the doses do not have to be the same all the time and can be determined arbitrarily if the doses are within an acceptable range. For example, the dose can be gradually reduced. In preferred embodiments, implementation of the present invention, the dose range is from 55 to 4000 mg of antibody. In other preferred embodiments of the present invention, the dosage range is from 60 to 2500 mg of the antibody. In the most preferred embodiments of the present invention, the dosage range is from 70 to 1500 mg of antibody. Specific preferred doses are doses containing 75, 125, 150, 300, 375, 500 and 1000 mg. Other preferred doses are 37, 500 and 1000 mg, and among them, 1000 mg is the most preferred.

In some embodiments of the present invention, the number of administrations of the pharmaceutical composition that is administered by intravenous injection is not specifically limited and may be one or more administrations. In preferred embodiments of the invention, the number of injections may be one, two or three injections. In other preferred embodiments of the invention, the number of injections is one or two injections. In the most preferred embodiment of the invention, the number of injections is one injection. Reducing the number of injections allows you to reduce the burden on the patient or his suffering.

In some embodiments, implementation of the present invention, in the case where the pharmaceutical composition is administered repeatedly intravenously to the individual or patient, the composition is administered once an hour or every 14 days. A suitable interval between intravenous injections can be determined in an acceptable range depending on the condition of the individual or patient. The specific interval between intravenous injections is 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours. , 16h, 17h, 18h, 19h, 20h, 21h, 22h, 23h, 24h, 25h, 26h, 27h, 28h, 29h, 30h, 31h, 32 h, 33 h, 34 h, 35 h, 36 h, 37 h, 38 h, 39 h, 40 h, 41 h, 42 h, 43 h, 44 h, 45 h, 46 h, 47 h, 48 h, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days or 14 days. In preferred embodiments of the invention, the interval between injections is from 24 hours to 10 days. In other preferred embodiments of the invention, the interval between injections is from 48 hours to 7 days. In the most preferred embodiments of the invention, the interval between injections is from 3 to 5 days.

In a specific embodiment of the invention, a pharmaceutically acceptable carrier suitable for a conventional composition for intravenous injection may be included in the pharmaceutical composition. The type of carrier for intravenous injection may be selected from carriers generally known in the art.

In some embodiments, the formulation of the pharmaceutical composition may be any one selected from the group that includes liquid, semi-solid, and solid formulations. The solid composition is obtained, as a rule, by lyophilization from a liquid preparation. The lyophilized composition is typically reconstituted with water or saline immediately prior to intravenous injection. The formulation of the pharmaceutical composition for intravenous injection may be the same as or different from that of the pharmaceutical composition for subcutaneous injection. In preferred embodiments of the invention, the formulation of the pharmaceutical composition for intravenous injection is the same as the formulation of the pharmaceutical composition for subcutaneous injection, which helps to reduce the cost of production. When the composition is administered intravenously, the concentration of anti-C5 antibody in the liquid formulation of the composition is within the generally accepted range known in the art. In a specific embodiment of the invention, the volume of intravenous injection is determined by the usual range known in this field.

In some aspects of the invention, the dose of the composition for intravenous injection is administered before the initial dose of another pharmaceutical composition, which is administered subcutaneously. The pharmaceutical composition for subcutaneous injection is one of the compositions described above.

In some embodiments of the invention, the composition for intravenous injection is administered intravenously on the same day or one or more days before the initial dose of the composition for subcutaneous injection, which is administered subcutaneously. One or more doses of the composition for intravenous injection may be administered to an individual or patient prior to administration of the initial dose of the composition for subcutaneous injection. In a preferred embodiment of the invention, the final dose of the composition for intravenous injection is administered before the initial dose of the composition for subcutaneous injection.

In some embodiments of the present invention, the dose of the composition for intravenous injection is administered on the same day or 1-28 days prior to the initial dose of the composition for subcutaneous injection. The specific interval between administration of a dose of the composition for intravenous injection and the initial dose of the composition for subcutaneous injection is 0 days (i.e. within 24 hours), 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, 15 days, 16 days, 17 days, 18 days, 19 days, 20 days, 21 days, 22 days, 23 days, 24 days, 25 days, 26 days, 27 days or 28 days. In preferred embodiments of the invention, the interval is 1-14 days. In other preferred embodiments of the invention, the interval is 3-10 days. In the most preferred embodiments of the invention, the interval is 5-8 days.

IV. Switching From Another Pharmaceutical Product The pharmaceutical compositions for subcutaneous or intravenous injection of the present invention may be used to treat or prevent a C5-related disease in an individual who has been treated one or more times with at least one pharmacological product intended for use in the treatment or prevention of this disease. For example, the pharmaceutical compositions of the present invention can be used to treat a C5-related disease in a patient who has previously been treated with at least one pharmacological product intended for the treatment of this disease, but for whom an improved response to treatment with pharmaceuticals is expected. compositions of the present invention. In such cases, drug treatment can be switched from the pharmacological product to the pharmaceutical composition of the present invention. In preferred embodiments of the invention, the initial dose of the composition for intravenous injection proposed in the present invention is administered after the final dose of the pharmacological product used for the previous treatment.

In some embodiments, the pharmacological product contains an active substance that is different from the anti-C5 antibody used in the subcutaneous and intravenous injection composition described above. In some embodiments, the active ingredient of the pharmaceutical product is an siRNA targeting C5 mRNA, or an anti-C5 antibody other than the anti-C5 antibody contained in the subcutaneous and intravenous injection composition described above. In preferred embodiments, the pharmacological product contains an antibody that is different from the antibodies in the subcutaneous and intravenous injection composition described above. In the most preferred embodiment of the invention, the antibody included in the pharmacological product that was used for the previous treatment is eculizumab or its derivative.

In some embodiments of the invention, the initial dose of the composition for intravenous injection proposed in the present invention is administered on the same day or one or more days after administration of the final dose of the pharmacological product. The specific interval between administration of the final dose of the pharmacological product and the initial dose of the intravenous injection composition of the present invention is 0 days (meaning that the initial dose of the intravenous injection composition is administered on the same day as the final dose of the pharmacological product), 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, 15 days, 16 days, 17 days, 18 days, 19 days, 20 days or 21 days. In preferred embodiments of the invention, the composition for intravenous injection proposed in the present invention is administered 3 or more days after the final dose of the pharmacological product. In more preferred embodiments, the intravenous injection composition of the present invention is administered 7 or more days after the final dose of the pharmacological product. In other preferred embodiments of the invention, the composition for intravenous injection proposed in the present invention is administered 14 or more days after the final dose of the pharmacological product. In the most preferred embodiments of the invention, the composition for intravenous injection proposed in the present invention is administered 21 or more days after the final dose of the pharmacological product.

V. Method of treatment or prevention

Other objects of the present invention are methods for the treatment or prevention of C5-related disease.

In one of the embodiments of the invention, the method includes intravenous administration of a first pharmaceutical composition, which is formulated for intravenous injection and contains an antibody to C5, and subcutaneous administration of a second pharmaceutical composition, which is formulated for subcutaneous injection and contains an antibody to C5, after administration of the first compositions.

The conditions and diseases targeted by the pharmaceutical composition for subcutaneous or intravenous injection in said method are the same as those mentioned above in sections "II. Pharmaceutical composition for subcutaneous injection" and "III. Pharmaceutical composition for intravenous injection.

VI. Vaccination

Infectious diseases, such as meningococcal infections, may occur in an individual administered with an anti-C5 antibody. To prevent these infectious diseases, the subject may be immunized with a vaccine known to be able to treat or prevent the disease before, after, or at the time of administration of the composition for subcutaneous and intravenous injection.

VII. Product

Another object of the invention is an article or product containing/containing materials that can be used to treat or prevent associated with C5 diseases mentioned above. The article or product contains one or more containers and a label or package insert, which are placed on or enclosed in the container. Acceptable containers are, for example, jars, vials, syringes, IV solution bags, SC solution syringes, etc. Containers can be made from various materials such as glass or plastic. The container contains a composition that, alone or in combination with another composition, is effective in treating or preventing the condition and may have a sterile access port (for example, the container may be an intravenous solution bag or a vial fitted with a stopper that can be pierced with a needle to subcutaneous injections). At least one active ingredient in the composition is an anti-C5 antibody as described above. The label, package insert, or similar document indicates that the composition can be used to treat the selected condition.

In some embodiments of the invention, the composition in the container of the article or product is in the form of a subcutaneous injection. According to this embodiment of the invention, the article or product may further include a package insert leaflet with information that the composition is administered after a dose of another pharmaceutical composition intended for intravenous administration. According to this embodiment of the present invention, another pharmaceutical composition contains an anti-C5 antibody and is formulated for intravenous injection.

In some embodiments of the invention, the composition contained in the container of the article or product is in the form of an intravenous injection. According to this embodiment of the present invention, the article or product may further include a package insert leaflet stating that the composition is to be administered prior to the initial dose of another pharmaceutical composition intended for subcutaneous administration. According to this embodiment, another pharmaceutical composition contains an anti-C5 antibody and is formulated for subcutaneous injection.

According to some other embodiments of the invention, the article or product may contain (a) a first container with a first composition, where the composition contains an antibody to C5 and is prepared in the form for intravenous injection, and (b) a second container with a second composition, where the composition contains an antibody to C5 and formulated for subcutaneous injection. In this embodiment of the invention, the product may also contain a package insert leaflet with information that the compositions can be used to treat a specific condition, such as a C5-related disease, and additional information that the administration of the first pharmaceutical composition and the second pharmaceutical composition should be carried out in the order of one or more intravenous injections of the first pharmaceutical composition for intravenous injection and one or more subcutaneous injections of the second pharmaceutical composition for subcutaneous injection.

In addition, the article or product may contain an additional container containing a composition that contains an additional cytotoxic or other therapeutic agent. According to this embodiment of the invention, the product may further comprise a package insert leaflet with information that the compositions can be used to treat a particular condition. In an alternative or additional embodiment, the product may further include another additional container with a pharmaceutically acceptable buffer, such as bacteriostatic water for injection (BWFI), phosphate buffered saline, Ringer's solution and dextrose solution. In addition, it may include other products required from a commercial and consumer point of view, in particular other buffers, diluents, filters, needles and syringes.

Examples

Below are examples of methods and compositions proposed in the invention. As should be apparent, various other embodiments of the invention may be practiced based on the general description of the invention provided above.

Example 1

Creation of an antibody to C5

Antibody to C5 305LO15 described in WO 2016/098356 was obtained by a standard method. In general, the method was as follows. The genes encoding the 305LO15 heavy chain variable (VH) were fused with the genes encoding the SG115 modified variant (SEQ ID NO: 13) of the heavy chain constant domain (CH) of human IgG1. The genes encoding the light chain variable domain (VL) 305LO15 were combined with the genes encoding the human light chain constant domain (CL) (SKI, SEQ ID NO: 18). Antibodies were expressed in HEK293 cells co-transfected with a combination of heavy and light chain expression vectors and purified using protein A.

Example 2

Protein-based pharmaceuticals, such as antibody-based drugs, are typically administered intravenously because proteins are difficult to concentrate. Thus, eculizumab, which is currently the only antibody approved for treatment, is administered intravenously. If it becomes possible to use subcutaneous injection, then this should ease the burden on the patient, for example, in terms of outpatient treatment and the length of time spent in the clinic for intravenous injection, since it will be possible to self-inject.

The 305LO15 antibody described in WO 2016/098356 was chosen to study the feasibility of subcutaneous administration of an anti-C5 antibody due to its high solubility. It was expected that the higher solubility would allow the administration of the protein-based pharmaceutical agent in a smaller volume, allowing the use of subcutaneous injection.

To compare the pharmacokinetic characteristics of 305LO15 after subcutaneous injection or intravenous injection of monkeys, cynomolgus was divided into two groups, a subcutaneous injection group and an intravenous injection group. 305LO15 was administered subcutaneously to the animals in the first group (three males and three females) once every week for a total of 22 times, each time administering the antibody at a dose of 40 mg/kg. The plasma concentration of 305LO15 in these animals was measured by ELISA at the time points indicated in FIG. 1a. 305LO15 was administered intravenously to the animals in the second group (six males and six females) once every week for a total of five times, each time administering the antibody at a dose of 40 mg/kg.

Animal plasma concentration of 305LO15 was measured by ELISA at the time points shown in FIG. 1b.

As shown in FIG. 1a, the plasma concentration of 305LO15 in cynomolgus monkeys was increased by subcutaneous injections, which confirms that 305LO15 can be administered by subcutaneous injection for therapeutic purposes. The rate of initial increase in plasma concentration with subcutaneous injection (Fig. 1a) was lower than with intravenous injection (Fig. 1b).

Example 3

A study was then made as to whether the relatively low rate of increase in plasma concentration after the initial subcutaneous injection could be compensated for by intravenous injection of 305LO15. Five male and five female cynomolgus monkeys received a single intravenous injection with an initial dose of 100 mg/kg 305LO15 antibody. One week after the initial intravenous administration, a subcutaneous injection of 305LO15 was started. Subcutaneous injection was performed a total of 26 times at a frequency of once every week, administering at each dosing antibody at a dose of 40 mg/kg.

As shown in FIG. 2, the rate of increase in plasma concentration of 305LO15 was greater when the initial dose of the antibody was administered by intravenous injection before maintenance doses by subcutaneous injection than when the initial dose was not administered by intravenous injection. The plasma concentration of 305LO15 reached a level comparable to that shown in FIG. 1a and maintained at this level. It can be assumed that this faster increase in plasma concentration of 305LO15 contributes to a more rapid achievement of the therapeutic effect of the drug.

Example 4

To assess the effective concentration of 305LO15 in plasma, the ratio between the concentration of free antigen (C5) in plasma or complement activity and the concentration of antibody (305LO15) in plasma in cynomolgus monkeys was determined. 305LO15 was administered intravenously (0.8, 4 or 20 mg/kg) or subcutaneously (4 mg/kg). Plasma antibody and free C5 concentrations were measured by ELISA. Complement activity was measured using a erythrocyte hemolytic assay (RBC). As shown in FIG. 3, the free C5 concentration decreased to less than a few μg/ml when the plasma concentration of 305LO15 was maintained above 40 μg 305EO15/ml (Fig. 3a) and the plasma 305LO15 concentration was 40 μg/ml or higher, inhibited complement activity (hemolysis) to less than 20% of baseline (ie, in the absence of antibody) (FIG. 3b). Complement activity was found to be suppressed to less than 20% of baseline (ie, in the absence of antibody injection) when the plasma concentration of 305LO15 was maintained at or above 40 μg/mL.

Example 5

To determine doses and administration schedules of 305LO15 suitable for maintaining an effective concentration of 305LO15 in clinical trials, the dependence of the concentration of 305LO15 in human plasma was predicted by modeling. First, plasma concentrations of 305LO15 were measured as a function of time after antibody administration to cynomolgus monkeys at doses of 0.8, 4, 20 μg antibody/kg at each dose. PK parameters in the case of cynomolgus monkeys were assessed by data analysis using a 2-compartment model. Human PK parameters were estimated from Cintomolgus monkey PK parameters using allometric scaling. Evaluated PK parameters for humans are presented in Table 1 (abbreviations used to denote parameters in Table 1: BW - body weight; CL - total clearance of the drug; F - bioavailability or the relative amount of the administered drug that entered the bloodstream; Ka - absorption rate constant; MAb - monoclonal antibody; PK - pharmacokinetic parameter; Q - intercompartmental clearance; SC - subcutaneously; Vc - volume of distribution in the central compartment; Vp - volume of distribution in the peripheral compartment).

The human PK profile was assessed for each cohort participating in the Phase 1/2 study (FIG. 4). Part 1 of the study was designed to include three groups of individuals. The first group was a group of individuals who were administered 305LO15 intravenously once at a dose of 75 mg/person. The second group was a group of individuals who were administered 305LO15 intravenously once at a dose of 150 mg/person. The third group was a group of individuals who were administered 305LO15 subcutaneously once at a dose of 170 mg/person. It was predicted that the plasma concentration of 305LO15 in individuals administered any one of the above doses would not be maintained above the threshold (40 μg/ml) for more than one week (FIG. 4a).

Part 2 of the study was designed to include a group of individuals who were administered 305LO15 intravenously three times (first at 300 mg/person, then 500 mg/person one week after the first dose, and finally 1000 mg/person two weeks after second administration) and, two weeks after the final intravenous administration, 305LO15 was administered subcutaneously once a week at a dose of 170 mg/person. It was predicted that when using the dosing regimen planned for part 2 of the study, the concentration of 305LO15 will be maintained at a level above the cut-off PK level (40 μg/ml) (Fig. 4b).

Part 3 of the study was designed to include three groups of individuals. First, 305LO15 was administered to individuals in all groups intravenously once at a dose of 500 mg/person. Starting from the day following the day of initial administration, 305LO15 was administered subcutaneously to individuals in the first group once every week at a dose of 170 mg/person, to individuals in the second group once every two weeks at a dose of 340 mg/person, and to individuals in the third group once every four weeks at a dose of 600 mg/person. It was predicted that the concentration of 305LO15 will be maintained above the PD threshold (40 μg/ml) in all three groups participating in part 3 of the study (Fig. 4c).

Example 6

Eculizumab has been used to treat patients with PNH or aHUS. In the event that 305LO15 was predicted to be more beneficial than eculizumab in a patient with PNH or aHUS, the patient could be switched from eculizumab drug therapy to 305LO15 drug therapy. It should be taken into account that there is a possibility that 305LO15 forms an immune complex with C5 and eculizumab in the body, since the epitope for 305LO15 is different from the epitope for eculizumab, which means that 305LO15 binds to the C5 beta chain, while eculizumab binds to the C5 alpha chain. The formation of such an immune complex can cause abnormal activation and therefore should be avoided as much as possible.

To investigate whether 305LO15 forms an immune complex with eculizumab (ECZ, heavy chain sequence shown in SEQ ID NO: 19 and light chain sequence shown in SEQ ID NO: 20) and human C5 (hC5, SEQ ID NO: 21), in vitro analysis was performed using gel filtration (SEC). hC5 was obtained according to the method described in WO 2016/098356. ECZ, hC5 and 305LO15 were dialyzed against D-PBS, pH 7.4, to exchange buffer. Then ECZ and hC5 were mixed and incubated at 37°C for 3-4 hours. After incubation, 305LO15 was added to the mixture containing ECZ and hC5. The final concentration of ECZ and hC5 was adjusted to 200 μg/ml, and the concentration of 305LO15 was adjusted to 0, 25, 125, 250, or 1250 μg/ml. After incubation of these mixtures at 37° C. for 3-4 hours, SEC analysis was performed at pH 7.4 and pH 6.0. The specific conditions for the SEC are listed below:

HPLC system: Waters Alliance e2695 HPLC system

column: TSKgel G4000SWXL column temperature: 25°C

eluent: 50 mM Na-PB/300 mM NaCl, pH 7.4 or pH 6.0

flow rate: 0.5 ml/min

detection: UV absorption at 220 nm

injection: 10 µl.

As a result, it was found that 305LO15 at concentrations from 125 to 1250 μg/ml had no effect on the SEC profile at pH 6.0, but had an effect on the profile at pH 7.4. More specifically, when 305LO15 was mixed with ECZ and hC5, peaks of large ECZ/hC5/305LO15 immune complexes that were not detected at pH 6.0 were detected at pH 7.4 (see peaks indicated by arrows in Fig. 5, top panel), in addition to the peaks corresponding to small complexes (see peaks labeled '2Ag+Ab' and 'Ag+Ab' in Fig. 5, top panel). At pH 6.0, only peaks corresponding to small complexes were detected (see peaks presented in Fig. 5, lower panel, corresponding to '2Ag+Ab' 'Ag+Ab' in Fig. 5, upper panel). This means that 305LO15 binds to the ECZ/hC5 complex at pH 7.4 but not at pH 6.0, which is consistent with the binding characteristics of 305LO15 (binding to hC5 at pH 7.4 but not at pH 6.0) . Based on these results, it was suggested that the simultaneous administration of 305LO15 and ECZ to an individual may result in the formation of large plasma immune complexes.

Example 7

Study BP39144 is an adaptive phase I/II study to evaluate the safety, efficacy, pharmacokinetic characteristics (PK) and pharmacodynamic characteristics (PD) of 305LO15 in healthy volunteers and patients with paroxysmal nocturnal hemoglobinuria (PNH).

The BP39144 Part 1 study (randomized, investigator/individual blinded, adaptive, placebo-controlled, parallel group study) assessed the safety and tolerability of single doses of 305LO15 in healthy volunteers. At each dose level in each cohort, a total of 5 healthy volunteers were randomly assigned to receive a single dose of 305LO15 or placebo intravenously (IV; cohorts 1 and 2) or subcutaneously (SC; cohort 3). The study had an adaptive design, according to which sufficient safety, tolerability, PK and PD were constantly assessed before starting the next administration. The target dose levels for cohorts 1, 2, and 3 were 75 mg IV, 150 mg IV, and 170 mg SC, respectively (FIG. 4a). Based on a preliminary analysis of PK and PD data obtained for cohort 1, doses for cohorts 2 and 3 were reduced to 125 mg IV (cohort 2) and 100 mg SC (cohort 3).

When performing an IV infusion, blood samples were collected before dosing, after the end of the IV infusion (1 h), at 2, 6, 12, 24, 48, 72, 96, and 144 h after dosing, and on days 14, 21, 28 , 35, 42, 56, 84, and 91. For SC administration, blood samples were collected pre-dose, 12, 24, 48, 72, 96, and 144 hours post-dose, and on days 14, 21, 28, 35, 42, 56, 84 and 91.

Preliminary safety results from the BP39144 Part 1 study showed that 305LO15 was well tolerated at all dose levels studied (75 and 125 mg IV and 100 mg SC). Available clinical safety information did not raise significant safety concerns, and no evidence was found to suggest adverse events or abnormalities in laboratory testing. There were no noteworthy data or trends in vital signs or 12-lead electrocardiograms in all treatment groups.

Preliminary serum PK results and simulated profiles: 305bO15 concentration-time are shown in FIG. 6, FIG. 7 and FIG. 8. These data indicate that the 305LO15 FC in healthy individuals was consistent with the human FC previously predicted from cynomolgus FA data (Example 5). It appears that after IV dosing, exposure should be dose proportional in the range of 75 to 125 mg. The end half-life (tm) for a typical 70 kg patient is estimated to be approximately 25 days. Preliminary PK results after SC administrations indicate that 305LO15 exposure peaked around day 7 and bioavailability was approximately 90%. After absorption, t _1/2 was comparable to t _1/2 in the case of IV infusion.

Preliminary estimates of the exposure-response relationship confirmed the previously obtained prediction, namely, that approximately 40 μg of 305LO15 per milliliter of blood is required to ensure complete complement inhibition (Fig. 9).

Example 8

Part 2 of the BP39144 study (an open-label, multi-dose, global, multicenter, in-patient, dose escalation study) assessed safety and tolerability over a total of 5 months, and the pharmacodynamic effects of multiple doses of 305LO15 on complement activity in the treatment of previously untreated patients with PNH. This study included six patients with PNH. Study patients were not treated with any complement inhibitor or were previously treated but discontinued due to lack of efficacy due to a single heterozygous C5 missense mutation and had elevated serum LDH levels (>1.5 x ULN) at screening (ULN: upper limit of normal range).

One patient (Patient X) of the six patients was a PNH patient and was a candidate for complement inhibitor treatment. Patient X was administered IV three single incremental doses; by a single infusion of 375 mg of 305LO15 on day 1, then by a single infusion of 500 mg of 305LO15 on day 8, and then by a single infusion of 1000 mg of 305LO15 on day 22. The first SC administration (170 mg) was started on day 36, then weekly ( QW) SC injections (170 mg) of 305LO15, which were to be continued throughout the treatment period of 5 months.

Preliminary results of pharmacodynamic evaluation for patient X are presented in Table 2 and in FIG. 10. Levels of LDH, which serves as a pharmacodynamic marker of hemolysis, decreased to levels within the normal range by day 15, continued to decrease further until day 36, and were maintained at a normalized level until day 43 (Table 2 and Fig. 10a). As shown in Table 2 and in FIG. 106, results from a liposomal immunoassay (LIA) indicated that complement activity was completely inhibited at the end of the infusion on study entry day, day 1. The 375 mg dose of 305LO15 given on day 1 maintained complete complement inhibition until the next dose was administered. 305LO15 dose of 500 mg on day 8. The 305LO15 dose of 500 mg given on day 8 maintained complete complement inhibition until the next dose of 305LO15 of 1000 mg on day 22. The 305LO15 dose of 1000 mg, which administered on day 22 maintained complete complement inhibition until the next 170 mg SC dose of 305LO15 was administered on day 36. The SC dose administered on day 36 maintained complete complement inhibition until day 43.

305LO15 was well tolerated with only mild abdominal pain unrelated to treatment.

Example 9

Part 3 of the BP39144 study (an open-label, multi-dose, global, multi-centre study) assessed the safety and tolerability over a total of 5 months, and the pharmacodynamic effects of multiple doses of 305LO15 on complement activity, in PNH patients treated with eculizumab during the considered time period. This study included 18 patients with PNH. Patients participating in the study were treated continuously with eculizumab for at least 3 months prior to entry into the study and the patients were regularly infused with eculizumab.

One patient (Patient Y) of 18 patients was a PNH patient who was treated with eculizumab, with eculizumab administered last 14 days prior to the first IV infusion of 305LO15. Patient Y was given a loading dose by single infusion; administered a single infusion of 1000 mg 305LO15 on day 1. The first SC injection (680 mg) was administered on day 8, followed by SC injections (680 mg) of 305LO15 every 4 weeks (Q4W) which continued for the entire treatment period of 5 months.

Preliminary pharmacodynamic performance results for patient Y are presented in Table 3. LDH levels, serving as a pharmacodynamic marker of hemolysis, were maintained at baseline from the first dose until day 43 (Table 3). As shown in Table 3, the results of the liposomal immunoassay (LIA) indicated that complement activity was completely inhibited at the end of the infusion on study start day, day 1. The 1000 mg dose of 305LO15 given on day 1 maintained complete complement inhibition up to until the SC dose of 680 mg 305LO15 on day 8. The SC dose on day 8 maintained complete complement inhibition until day 43.

Claims (14)

1. A method for treating or preventing a C5 related disease, comprising (a) administering intravenously to an individual a single intravenous dose of an anti-C5 antibody at a dose of 500 mg, 1000 mg or 1500 mg of the antibody; (b) subcutaneously administering two or more subcutaneous doses of the anti-C5 antibody to the subject (i) at a dose of 340 mg of the antibody once every week or once every two weeks, or (ii) 600 mg to 2000 mg of the antibody once every four weeks; wherein the anti-C5 antibody comprises a heavy chain variable domain (VH) sequence consisting of the amino acid sequence of SEQ ID NO: 2, a light chain variable domain (VL) sequence consisting of the amino acid sequence of SEQ ID NO: 6, a heavy chain constant domain sequence (CH ) consisting of the amino acid sequence of SEQ ID NO: 13, and a light chain constant domain (CL) sequence consisting of the amino acid sequence of SEQ ID NO: 18. 2. The method of claim 1, wherein the first subcutaneous dose of anti-C5 antibody is administered on the same day or 1-28 days after the intravenous dose of anti-C5 antibody. 3. The method of claim 1 or 2, wherein two or more subcutaneous doses of the anti-C5 antibody contain 680 mg of the antibody administered once every four weeks. 4. The method according to one of paragraphs. 1-3, where one intravenous dose is 1000 mg of antibody. 5. The method according to one of paragraphs. 1-4 wherein the subject has been treated with at least one pharmacological product intended for use in the treatment or prevention of the disease up to step (a), wherein one intravenous dose of anti-C5 antibody is administered after the final dose of the pharmacological product. 6. The method of claim 5, wherein a single intravenous dose of anti-C5 antibody is administered on the third day or 3 days after the final dose of the pharmacological product. 7. The method according to claim 5 or 6, wherein the pharmacological product contains an siRNA targeting C5 mRNA, or an anti-C5 antibody other than said anti-C5 antibody. 8. The method according to one of paragraphs. 5-7, where the pharmacological product contains eculizumab or its derivative. 9. The method according to one of paragraphs. 1-8, where intravenous and subcutaneous doses of antibody to C5 achieve a plasma concentration above 40 μg/ml. 10. The method according to one of paragraphs. 1-9 where complement activity is downregulated to less than 20% of baseline. 11. The method according to one of paragraphs. 1-10, where the C5-related disease is any disease selected from the group consisting of rheumatoid arthritis (RA); lupus nephritis; damage caused by ischemia-reperfusion; paroxysmal nocturnal hemoglobinuria (PNH); atypical hemolytic uremic syndrome (aHUS); dense deposit disease (DDD); macular degeneration; HELLP syndrome (hemolysis, increased activity of liver enzymes and thrombocytopenia); thrombotic thrombocytopenic purpura (TTP); spontaneous loss of the fetus; Pauci-immune vasculitis; bullous epidermolysis; recurring fetal loss; multiple sclerosis (MS); traumatic brain injury; and damage caused by myocardial infarction, cardiopulmonary bypass or hemodialysis; resistant generalized myasthenia gravis (gMG); and neuromyelitis of the optic nerve (NMO).

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